Document X7Vd5pOpX4OZ53qv9OeqZwxr4

T-7485: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE FINAL REPORT WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER 454-116 3M ENVIRONMENTAL LAB PROJECT NO. E03-0583 FIFRA Guideline 71-4 OECD Guideline 206 AUTHORS: Sean P. Gallagher Raymond L. Van Hoven Joann B. Beavers Mark Jaber STUDY INITIATION DATE: September 5, 2003 STUDY COMPLETION DATE: May 27, 2005 Submitted to 3M Corporation Environmental Laboratory 935 Bush Avenue St. Paul, Minnesota 55106 Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 Page 1 of 439 Contains No CBI Wildlife International, Ltd, Project Number 454-116 -2 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT SPONSOR: 3M Corporation TITLE: T-7485: A Reproduction Study with the Northern Bobwhite WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER 454-116 STUDY COMPLETION: May 27, 2005 The study was conducted in compliance with Good Laboratory Practice Standards as published by the U.S. Environmental Protection Agency, 40 CFR Part 792, 17 August 1989; OECD Principles of Good Laboratory Practice (ENV/MC/CHEM (98) 17); and Japan MAFF, 11 NohSan Notification No. 6283, Agricultural Production Bureau, 1 October 1999, with the following exceptions: Periodic analysis of avian feed and water for potential contaminants were performed using a certified laboratory and standard U.S. EPA analytical methods. STUDY DIRECTOR: Senior Biologist, Avian Toxicology SPONSOR'S REPRESENTATIVE. DATE Contains No CBI Wildlife International, Ltd. Project Number 454-116 -3 - QUALITY ASSURANCE STATEMENT This study was examined for compliance with Good Laboratory Practice Standards as published by the U.S. Environmental Protection Agency, 40 CFR Part 792, 17 August 1989; OECD Principles o f Good Laboratory Practice (ENV/MC/CHEM (98) 17); and Japan MAFF, 11 NohSan, Notification No. 6283, Agricultural Production Bureau, 1 October 1999. The dates o f all audits and inspections and the dates any findings were reported to the Study Director and Laboratory Management were as follows: ACTIVITY DATE CONDUCTED DATE REPORTED TO: STUDY DIRECTOR MANAGEMENT Test Substance Preparation Matrix Fortification Body Weight September 16, 2003 September 25, 2003 October 1, 2003 September 17, 2003 September 25, 2003 October 1, 2003 September 17,2003 September 30, 2003 October 2, 2003 Diet Preparation, Analytical Sampling Feed Consumption October 8, 2003 October 8, 2003 October 8, 2003 October 8, 2003 October 9, 2003 October 9, 2003 Observations January 6, 2004 January 6, 2004 January 6,2004 Candling Data Entry Data Entry January 29, 2004 March 10, 2004 March 15, 2004 January 29, 2004 March 10, 2004 March 15, 2004 January 29, 2004 March 10, 2004 March 15, 2004 Analytical Data and Draft Report Data Entry Protocol March 29-31 and April 1, 2004 April 12 and 13, 2004 April 26, 2004 April 1, 2004 April 13, 2004 April 26, 2004 April 2,2004 April 13,2004 April 27, 2004 Biological Data and Draft Report Final Report April 26-30, 2004 May 25-26, 2005 April 30, 2004 May 26, 2005 May 5, 2004 May 27, 2005 All inspections were study based unless otherwise noted. Marshall T lynson Quality As: rrance Program Supervisor DATE 57**t r Contains No CBI Wildlife International, Ltd, -4 REPORT APPROVAL Project Number 454-116 SPONSOR: 3M Corporation TITLE: T-7485: A Reproduction Study with the Northern Bobwhite WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER 454-116 STUDY DIRECTOR: Sean P Gallagher Senior Biologist, Avian Toxicology PRINCIPAL INVESTIGATOR: DATE MANAGEMENT: DATE Director o f Chemistry Contains No CBI Wildlife International, Ltd. Project Number 454-116 -5- TABLE OF CONTENTS Title Page..........................................................................................................................................1 Good Laboratory Practice Compliance Statement........................................................................... 2 Quality Assurance Statement........................................................................................................... 3 Report Approval............................................................................................................................... 4 Table of Contents............................................................................................................................. 5 Summary.......................................................................................................................................... 9 Introduction..................................................................................................................................... 10 O bjectives....................................................................................................................................... 10 Experimental Design.......................................................................................................................10 Materials and Methods....................................................................................................................11 Test Substance..........................................................................................................................11 Test Organisms.........................................................................................................................12 Identification............................................................................................................................ 12 Avian Feed and Water..............................................................................................................12 Diet Preparation........................................................................................................................13 Diet Sampling...........................................................................................................................13 Analytical Method for Diets.....................................................................................................14 Analytical Method for Serum, Liver and Eggs.........................................................................16 Study Phases.............................................................................................................................16 Housing and Environmental Conditions..................................................................................16 Observations.............................................................................................................................17 Necropsy and Tissue Collection...............................................................................................17 Adult Body Weight and Feed Consumption............................................................................18 Egg Collection and Storage......................................................................................................18 Candling and Incubation...........................................................................................................18 Hatching and Brooding............................................................................................................19 Egg Shell Thickness Measurements and Egg Content Collection.......................................... 20 Statistical Analyses.................................................................................................................. 20 Results and Discussion.................................................................................................................. 21 Analytical Results for Diets..................................................................................................... 21 Mortalities............................................................................................................................... 22 Clinical Observations.............................................................................................................. 23 Gross Necropsy ...................................................................................................................... 24 Adult Body W eight................................................................................................................. 24 Adult Feed Consumption......................................................................................................... 25 Reproductive Results............................................................................................................... 26 Egg Shell Thickness................................................................................................................ 27 Offspring Body Weights.......................................................................................................... 27 Liver Weights.......................................................................................................................... 27 Serum, Liver and Egg Analytical Results............................................................................... 27 Histopathology........................................................................................................................ 28 Conclusion..................................................................................................................................... 29 References...................................................................................................................................... 30 Contains No CBI Wildlife International, Ltd. Project Number 454-116 -6- TABLE OF CONTENTS PAGE 2 TABLES AND FIGURES Table 1. Mean Adult Body Weight (g) from a Northern Bobwhite Reproduction Study with T-7485.................................................................................31 Figure 1. Mean Adult Male Body Weight (g) from a Northern Bobwhite Reproduction Study with T-7485............................................................... 32 Figure 2. Mean Adult Female Body Weight (g) from a Northern Bobwhite Reproduction Study with T-7485............................................................... 33 Table 2. Mean Feed Consumption (g/bird/day) from a Northern Bobwhite Reproduction Study with T-7485 ............................................................... 34 Figure 3. Mean Feed Consumption (g/bird/day) from a Northern Bobwhite Reproduction Study with T-7485 ............................................................... 35 Table 3. Summary of Reproductive Performance from a Northern Bobwhite Reproduction Study with T-7485................................................................36 Figure 4. Mean Reproductive Performance from a Northern Bobwhite Reproduction Study with T-7485................................................................ 37 Table 4. Summary of Reproductive Performance, Normalized as Percentages (%), from a Northern Bobwhite Reproduction Study with T-7485........38 Figure 5. Mean Reproductive Performance, Normalized as Percentages (%), from a Northern Bobwhite Reproduction Study with T-7485.................................... 39 Table 5. Mean Egg Shell Thickness Measurements (mm) from a Northern Bobwhite Reproduction Study with T-7485................................................................................. 40 Table 6. Mean Body Weight (g) of Hatchlings and 14-Day Old Survivors from a Northern Bobwhite Reproduction Study with T-7485.................................... 41 Table 7. Summary of Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485................................................................ 42 Table 8. Mean Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 ....................................................................................................... 43 Contains No CBI Wildlife International, Ltd. Project Number 454-116 -7- TABLE OF CONTENTS PAGE 3 APPENDICES Appendix I. Diet and Supplement Formulations............................................................ .....44 Appendix II. Diet Preparation......................................................................................... .....46 Appendix III. Reproductive Parameters........................................................................... ......47 Appendix IV. Individual Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485............................... .....50 Appendix V. Adult Body Weight (g) from a Northern Bobwhite Reproduction Study with T-7485............................................................... .....54 Appendix VI. Feed Consumption (g/bird/day) from a Northern Bobwhite Reproduction Study with T-7485 .............................................. ......62 Appendix VII. Reproductive Performance by Pen from a Northern Bobwhite Reproduction Study with T-7485 .............................................. ......66 Appendix VIII. Reproductive Performance by Week and Pen from a Northern Bobwhite Reproduction Study with T-7485............................... ....100 Appendix IX. Egg Shell Thickness Measurements (mm) per Pen by Week from a Northern Bobwhite Reproduction Study with T-7485 ................... ....131 Appendix X. Mean Hatchling Body Weight (g) per Pen by Week from a Northern Bobwhite Reproduction Study with T-7485 ................... ....135 Appendix XI. Mean 14-Day Old Survivor Body Weight (g) per Pen by Week from a Northern Bobwhite Reproduction Study with T-7485......... ....139 Appendix XII. Adult Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485...................................................................................... .....143 Appendix XIII. Offspring Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485...................................................................................... .....147 Appendix XIV. Histopathology Report............................................................................... ....149 Appendix XV. Certificate of Analysis............................................................................... .....190 Appendix XVI. The Analysis of T-7485 in Avian Diet....................................................... ....193 Contains No CBI Wildlife International, Ltd. Project Number 454-116 -8- TABLE OF CONTENTS PAGE 3 APPENDICES (continued) Appendix XVII. Analytical Determination of T-7485 in Northern Bobwhite Tissues................209 Appendix XVIII. Diagram of Test Layout.................................................................................... 437 Appendix XIX. Changes to Study Protocol................................................................................ 438 Appendix XX. Personnel Involved in Study............................................................................. 439 Contains No CBI Wildlife International, Ltd. -9SUMMARY Project Number 454-116 STUDY: T-7485: A Reproduction Study with the Northern Bobwhite SPONSOR: 3M Corporation WILDLIFE INTERNATIONAL, LTD. PROJECT NUMBER 454-116 TEST DATES: Study Initiation - September 5, 2003 Experimental Start (OECD) - September 16, 2003 Experimental Start (EPA) - September 17, 2003 Photostimulation - November 5, 2003 First Eggs Set - December 11, 2003 Adult Termination - February 12, 2004 Biological Termination - April 6, 2004 Analytical Termination - December 15, 2004 Experimental Termination - December 15, 2004 TEST ANIMALS: Northern bobwhite (Colinus virginianus) AGE TEST ANIMALS: Approximately 18 weeks of age at the initiation of the test SOURCE TEST ANIMALS: K & L Quail 26 Thompson Flat Road Oroville, CA 95965 U.S.A. NOMINAL TEST CONCENTRATIONS: 0, 100, 300, and 900 ppm a.i. RESULTS: There were no treatment-related mortalities, overt signs of toxicity or treatment-related effects upon body weight or feed consumption at any of the concentrations tested. Additionally, there were no treatment-related effects upon any of the reproductive parameters measured at the 100, 300 or 900 ppm a.i. test concentrations. The no observed-effect concentration for northern bobwhite exposed to T-7485 in the diet during the study was 900 ppm a.i., the highest concentration tested. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 10 - INTRODUCTION This study was conducted by Wildlife International, Ltd. for 3M Corporation at the Wildlife Inter national, Ltd. avian toxicology facility in Easton, Maryland 21601. The biological portion of the test was conducted from September 17, 2003 until April 6, 2004. Raw data generated at Wildlife International, Ltd. and a copy of the final report are filed under Project Number 454-116 in archives located on the Wildlife International, Ltd. site. OBJECTIVES The objective of this study was to evaluate the effects upon the adult northern bobwhite (Colinus virginianus) of dietary exposure to T-7485 over a period of approximately 21 weeks. Effects on adult health, body weight, and feed consumption were evaluated. In addition, the effects of adult exposure to T-7485 on the number of eggs laid, fertility, embryo viability, hatchability, offspring survival, and egg shell thickness were evaluated. EXPERIMENTAL DESIGN Northern bobwhite (64 males and 64 females) were randomly distributed into one control group and three treatment groups. The test concentrations were selected in consultation with the Sponsor, based upon the results of a pilot reproduction study (Wildlife International, Ltd. Project Number 454-115. Group 1 2 3 4 T-7485 Treatment Groups Nominal Concentration (ppm a.i.) (Control) 0 100 300 900 Pens per Group 16 16 16 16 Birds per Pen Males Females 11 11 11 11 Each treatment and control group contained 16 pairs of birds with one male and one female per pen. Three treatment groups were fed diets containing either 100, 300 or 900 ppm a.i. of T-7485 for approximately 21 weeks. The control group was fed diet comparable to the treatment groups, but without the addition of the test substance. Contains No CBI Wildlife International, Ltd. - 11 - Project Number 454-116 All adult birds were observed daily throughout the test for signs of toxicity or abnormal behavior. Adult body weights were measured at test initiation, on Weeks 2, 4, 6, 8, and at adult termination and feed consumption was measured weekly throughout the test. At the beginning of Week 8, the photoperiod was increased to induce egg production. Following the start of egg production, eggs were set weekly for incubation. Weekly, eggs were selected by indiscriminate draw for egg shell thickness measurement and all remaining eggs were candled prior to incubation to detect egg shell cracks or abnormal eggs. Eggs were also candled twice during incubation to detect infertile eggs or embryo mortality. On Day 21 of incubation, the eggs were placed in a hatcher and allowed to hatch. Once hatching was completed, hatchlings were removed from the hatcher and the group body weight of the hatchlings by pen was determined. At 14 days of age, the average body weight by parental pen of all surviving offspring was determined. Upon completion of the test, statistical analyses were performed to determine statistically significant differences between groups. Egg contents from those eggs selected for eggshell thickness measurements were also collected for potential analysis. Samples of sera and liver were collected from all adults surviving until test termination and from selected offspring for potential analysis. Additionally, samples of liver, kidney and gonad were also collected from adults and selected offspring for histopathological examination. MATERIALS AND METHODS The study was conducted according to the procedures outlined in the protocol, "T-7485 A Reproduction Study with the Northern Bobwhite". The protocol was based on procedures outlined in the Environmental Protection Agency's Registration Guidelines Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation: Wildlife and Aquatic Organisms, Subsection 71-4; OECD Guideline 206; and the ASTM "Standard Practice for Conducting Reproductive Studies with Avian Species" (1,2,3). Test Substance The test substance, T-7485, was received from 3M Corporation on July 18, 2003 and was assigned Wildlife International, Ltd. identification number 6407 upon receipt. The test substance was a white crystalline solid and was identified as: PFBS; T-7485; L-7038; Lot # 5; CAS # 29420-49-3. The test material had a reported purity of 98.2% and an expiration date of August 31, 2008 (Appendix XV). The test substance was held under frozen conditions in locked storage at the Wildlife International, Ltd. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 12 - facilities in Easton, Maryland. Concentrations of the test substance in the diet were adjusted to 100% active ingredient. Therefore, dietary concentrations are expressed as parts per million active ingredient (ppm a.i.) in the diet. Test Organisms Seventy-eight pairs of pen-reared northern bobwhite were purchased from K & L Quail, 26 Thompson Flat Road, Oroville, CA 95965, U.S.A. At the start of acclimation, the bobwhite were apparently healthy and phenotypically indistinguishable from wild type. The birds were from the same hatch, approaching their first breeding season and had not been used in any previous testing. At the start of acclimation, a random number generating function in a spreadsheet program was used to randomize pen assignment for each bird. Immediately prior to test initiation, all potential study birds were examined for physical injuries and general health. Birds that did not appear healthy, either due to injury or inability to acclimate to laboratory conditions, or were outside the weight range for the test, were excluded from the study. All birds were approximately 18 weeks of age at test initiation (first day of exposure to test diet) and ranged in weight from 171 to 220 grams at test initiation. Sex of the birds was determined by a visual examination of the plumage. Identification Adult birds were identified by individual leg bands, each pen was identified with a unique number, and groups of pens were identified by project number and concentration. All eggs laid during the study were marked with the pen number using a permanent ink marking pen for identification. Hatchlings were identified by leg bands so that they could be traced to their parental pen of origin. Avian Feed and Water All adult birds and their offspring were given feed and water ad libitum during acclimation and testing. The basal diet fed to both adults and offspring was formulated to Wildlife International, Ltd. specifications by Agway Inc. (Appendix I, Table 1). The basal ration contained at least 27% protein and 2.5% fat, and no more than 3.8% fiber. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 13 - The basal diet contained approximately 1.1% calcium, derived from feedstuffs and the 0.9% limestone used in the formulation of the basal diet by Agway. While this level of calcium is sufficient for growth and maintenance rations, additional calcium is required in the ration of breeding birds for egg shell formation. Therefore, an additional 5% (w/w) of limestone (approximately 38.5% Ca) was added to the basal diet for the adults. This raised the calcium level in the diet for the breeding birds to approximately 3%, slightly above the minimum recommended for quail (2.3%) (4). Offspring received basal diet without test substance and without the addition of 5% supplemental limestone. Water was supplied by the town of Easton public water supply. All offspring received a watersoluble vitamin and electrolyte mix in their water (Appendix I, Table 2). Neither the adults nor offspring received any form of medication in their feed during the test. Feed and water were analyzed periodically in accordance with Wildlife International, Ltd. Standard Operating Procedures. Diet Preparation Test diets were prepared by mixing T-7485 into a premix that was used for weekly preparation of the final diet. Control diet and each of the three treated diets were prepared weekly beginning on September 17, 2003 and presented to the birds on Wednesday of each week. Dietary concentrations were adjusted for purity of the test substance and are presented as parts per million active ingredient (ppm a.i.). Details of the weekly preparation of test and control diets are shown in Appendix II. Diet Sampling Homogeneity of the test substance in the diet was evaluated by collecting six samples from each of the treated diets and one sample from the control diet on Day 0 of Week 1. Samples were collected from the top, middle and bottom of the left and right sections of the mixing vessel. Control and treatment group diet samples were also collected from the feed troughs on Day 7 of Week 1to assess stability of the test substance under actual test conditions. Additionally, a single sample was collected from the control and two samples were collected from each treatment group diet during Weeks 4, 8, 12, 16 and 20 of the test to measure/verify test concentrations. The diet samples were transferred immediately to the Wildlife International, Ltd. analytical chemistry facility, where they were stored frozen prior to analysis. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 14 - Analytical Method for Diets A liquid chromatography mass spectrometry (LC/MS/MS) based method was developed and verified at Wildlife International, Ltd. for the analysis of the T-7485 in avian diet (5). The verified analytical method consisted of sample weighing, extraction with methanol, vacuum filtration, micro filtration, and dilution into the calibration range of the LC/MS/MS methodology. The method was slightly modified in the present test to expedite sample processing and to reduce solvent volume. The change was centrifugation of an aliquot of each sample extract instead of vacuum filtration of the entire extract volume. Dilutions into the calibration range of the LC/MS/MS methodology were performed with methanol (intermediate dilutions, if necessary) and/or a solution of 50% methanol and 50% NANOpure or HPLC-grade reagent water (v/v). Samples were then analyzed by direct injection. A method flowchart is provided in Appendix XVI, Figure 1. Concentrations of T-7485 in extracts of the samples were determined by reverse-phase high performance liquid chromatography using a Hewlett-Packard Model 1100 High Performance Liquid Chromatograph (HPLC) with a Perkin-Elmer SCIEX API 3000LC Mass Spectrometer equipped with a Perkin-Elmer SCIEX TurboIonSpray ion source. Chromatographic separations were achieved using a Keystone PRISM RP column (50 mm x 2.0 mm, 3-pm particle size) fitted with a Keystone Javelin C18 guard column (20 mm x 2.0 mm I.D.). The instrument parameters are summarized in Appendix XVI, Table 1. All primary and secondary stock preparations were adjusted for the purity of the test substance (98.2%). A 1.00-mg a.i./mL primary stock solution of T-7485 in methanol was prepared by weighing 0.1018 g of the test substance and bringing to a final volume of 100 milliliters with methanol. Secondary stock solutions of T-7485 in methanol (100- and 10.0-mg a.i./L) were prepared by serial volumetric dilution of the primary stock. Calibration standards were prepared in a solution of 50% methanol and 50% HPLC-grade reagent water (v/v) by appropriate dilutions of the 100- or 10.0-mg a.i./L stock solutions of T-7485 in methanol. Two sets of calibration standards of T-7485 were prepared. The first set, ranging in concentration from 0.0500 to 0.500 mg a.i./L, was analyzed with the Week 1, Days 0 and 7 sample sets. The second set, ranging in concentration from 0.00500 to 0.0250 mg a.i./L, was analyzed with all subsequent sample sets. The second set of lower-level calibration standards was selected for use in the remainder of the Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 15 - study to allow for greater sample dilution. The larger dilution minimized matrix loading into the LC/MS/MS system. A calibration standard series was injected at the beginning and at the end of each analytical sequence, and one standard was injected, at a minimum, after every five samples. Linear regression equations were generated using peak area responses versus the respective concentrations of the calibration standards. A representative calibration curve is presented in Appendix XVI, Figure 2. The concentration of T-7485 in the samples was determined by substituting the peak area responses of the samples into the applicable linear regression equation. Representative ion chromatograms of low and high-level calibration standards from the 0.00500 to 0.0250 mg a.i./L calibration standard set are presented in Appendix XVI, Figures 3 and 4, respectively. Examples of equations used in calculations are presented in Appendix XVI, Table 2. The instrument limit of detection (LOD) was set based upon the injection volume (5.00 or 10.0 pL) and the lowest standard concentration (0.0500 or 0.00500 mg a.i./L) used in the study. The LOD was calculated as 0.250 ng on-column for Week 1, Days 0 and 7 sample analyses and was calculated as 0.0500 ng on-column for all subsequent sample sets. The method limit of quantitation (LOQ) for these analyses was set at 50 ppm a.i. based upon the lowest matrix fortification level analyzed concurrently with the samples. For Week 1, Days 0 and 7 analyses, the 0.0500 mg a.i./L low-level standard was equivalent to a calculated value of 20 ppm a.i. T-7485 in the matrix blank extract. For all subsequent sample intervals, the 0.00500 mg a.i./L low-level standard was equivalent to a calculated value of 25 ppm a.i. T-7485 in the matrix blank extract. Measured values greater than or equal to the ppm a.i. equivalent were reported. Along with the sample analyses, seven matrix blanks were analyzed to determine possible interferences. No interferences were observed at or above the ppm a.i. equivalent of the lowest standard during the sample analyses (Appendix XVI, Table 3). A representative ion chromatogram of a matrix blank sample is presented in Appendix XVI, Figure 5. Avian diet samples were fortified with T-7485 using the dry-mix technique at 50.0, 200, and 1000 ppm a.i. (seven fortifications at each level) and analyzed concurrently with the samples to determine mean procedural recoveries. The method yielded mean procedural recoveries of 95%, 91%, 112%, 109%, 105%, 106% and 103%. These values correspond to each sample set analyzed during the definitive study (Appendix XVI, Table 3). Sample measured concentrations were not corrected for the mean procedural Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 16 - recoveries. A representative ion chromatogram of a matrix fortification sample is presented in Appendix XVI, Figure 6. Analytical Method for Serum, Liver and Eggs Preparation and analysis of serum, liver and egg samples were conducted by the 3M Environmental Laboratory using methods validated by 3M Environmental Laboratory. In summary, the samples were weighed, homogenized in acetonitrile, extracted using a Zymark Rapid Trace Automated SPE System, and analyzed using liquid chromatography/mass spectrometry (LC/MS). The full description of the analytical method is presented in Appendix XVII. Study Phases The primary phases of the study and their approximate durations were: 1. Acclimation - Approximately 5 weeks. 2. Pre-photostimulation - 8 weeks. 3. Pre-egg laying (with photostimulation) - Approximately 3 weeks. 4. Egg laying - Approximately 10 weeks. 5. Post-adult termination (final incubation, hatching, and 14-day offspring rearing period) - 6 weeks. Housing and Environmental Conditions Housing and husbandry practices were conducted so as to adhere to the guidelines established by the National Research Council (6). The adult birds were housed indoors in batteries of pens manufac tured by Georgia Quail Farm Manufacturing (GQFM Model No. 0330), measuring approximately 27 X 51 cm. The pens had sloping floors that resulted in ceiling height ranging from 20 to 25 cm. The pens were constructed of galvanized wire mesh and galvanized sheeting. A diagram of the test layout is presented in Appendix XVIII. Each pen was equipped with feed and water troughs. Weekly, sufficient feed for the feeding period was placed in the trough for each pen and presented to the birds. During the feeding period additional feed was weighed and added to the troughs as needed. Water troughs were changed and water added as necessary to provide potable water (generally every 2-3 days). Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 17 - Only birds associated with this study were maintained in the study room in order to avoid excessive disturbances. The average temperature in the adult northern bobwhite study room during the course of the test was 23.2 1.0C (SD) with an average relative humidity of 34 17% (SD). The air handling system in the study room was designed to vent up to 15 room air volumes every hour and replace them with fresh air. The photoperiod in the adult northern bobwhite room was maintained by a time clock. The photoperiod during acclimation and the first seven weeks of the test was eight hours of light per day. The photoperiod was increased to 17 hours of light per day at the beginning of Week 8 to induce egg laying and was maintained at that length until the adult birds were euthanized. Throughout the test, the birds received a mean of approximately 203 lux (~ 19 ft. candles) of illumination provided by fluorescent lights that closely approximated noon-day sunlight. Observations The test birds were acclimated to the facilities and study pens for approximately five weeks prior to initiation of the test. During acclimation, all birds were observed daily. Birds exhibiting abnormal behavior or debilitating physical injuries were not used for the test. During the study, all adult birds were observed daily for signs of toxicity or abnormal behavior. Additionally, all offspring were observed daily from hatching until euthanasia. A record was maintained of all mortalities and clinical observations. Necropsy and Tissue Collection Adult birds that died or were euthanized during the course of the study were subjected to a gross necropsy. At the conclusion of the exposure period, all surviving adult birds were euthanized with carbon dioxide gas. Prior to euthanasia, blood samples were drawn from birds. The blood was separated and serum was collected for potential analysis. Livers were also weighed and collected. A portion of liver was collected for histopathological examination and the remaining liver was collected for potential analysis. Samples of kidney and gonad were also collected for histopathological examination. All histopathological samples were fixed in 10% buffered formalin and shipped to EPL in Sterling, Virginia for histopathology. Additionally, similar samples were collected from a single indiscriminately selected offspring from each pen (if available) at 16 days of age. Offspring were sampled from the last lot of offspring produced. Following sample collection, all carcasses were disposed of by incineration. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 18 - Adult Body Weight and Feed Consumption Adult body weights were measured at test initiation, on Weeks 2, 4, 6, 8, and at adult termination. Body weights were not measured during egg laying because of the possible adverse effects handling may have on egg production. Feed consumption for each pen was measured weekly throughout the test. Feed consumption was determined by weighing the freshly filled feeder on Day 0, recording the amount of any additional diet added during the week, and weighing the feeder and remaining feed at the end of the feeding period (Day 7). An attempt was made to minimize feed wastage by the birds by using externally mounted feeders designed with a "feed-saver" lip. The amount of feed wasted by the birds was not quantified, since the wasted feed was normally scattered and mixed with water and excreta. Therefore, feed consumption is presented as an estimate of total feed consumption. Egg Collection and Storage Eggs were collected daily from all pens, when available. The eggs were stored in a cold room until incubation. The cold room was maintained at a mean temperature of 13 0C (SD) with a mean relative humidity of approximately 84 6% (SD). Groups of eggs were identified by an alphabetic lot code. All eggs laid in a weekly interval were considered as one lot. Candling and Incubation At the end of the weekly interval, all eggs were removed from the cold room, counted and eggs selected by indiscriminate draw for egg shell thickness measurement. The remaining eggs were candled with a Speed King (Model No. 32) egg-candling lamp to detect egg shell cracks or abnormal eggs. Cracked or abnormal eggs were recorded and discarded. All eggs to be incubated were fumigated with formaldehyde gas in an airtight cabinet with a circulating fan for approximately two hours, to reduce the possibility of pathogen contamination prior to incubation. Formaldehyde gas was generated by combining 20 g of potassium permanganate and 19 ml of 37% commercial grade formalin in a porcelain bowl at the base of the airtight cabinet. All eggs not discarded or used for egg shell thickness measurements were placed in a NatureForm Incubator (Model No. NMC 4000). In the incubator the temperature was maintained at an average 37.4 0.0C (SD) with an average relative humidity of 54 0%. The incubator was equipped with a pulsator Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 19 - fan and blades that produced a mild breathing air movement designed to eliminate intracabinet temperature and humidity variation during incubation. In order to prevent adhesion of the embryo to the shell membrane, the incubator was also equipped with an automatic egg rotation device, designed to rotate the eggs from 50 off of vertical in one direction to 50 off of vertical in the opposite direction (total arc of rotation was 100) every two hours through Day 21 of incubation. Eggs were candled on Day 11 or 12 of incubation to determine embryo viability and on Day 21 to determine embryo survival. Hatching and Brooding On Day 21 of incubation, the eggs were placed in a Petersime Hatcher (Model No. S6H) and allowed to hatch. Pedigree baskets constructed of galvanized steel wire mesh were used to keep hatchlings separated by parental pen of origin. Eggs were not rotated in the hatcher. The average temperature in the hatching compartment was 37.2 0.0 C (SD), and the average wet bulb temperature was raised to 33.3 0.0 C (SD) (relative humidity of approximately 77%). All hatchlings, unhatched eggs, and egg shells were removed from the hatcher on Day 25 or 26 of incubation. The group body weight of the surviving hatchlings by pen was determined. Hatchlings were leg banded for identification by pen of origin and then routinely housed according to the appropriate parental concentration grouping in brooding pens until 14 days of age. The hatchlings were fed untreated diet without the addition of 5% supplemental limestone. At 14 days of age, the average body weight by parental pen of all surviving chicks was determined. The chicks were euthanized with carbon dioxide and disposed of by incineration. While the final lot of offspring (Lot J) was weighed at 14 days of age, chicks from this lot were maintained alive until 16 days of age for tissue sampling. Lot J offspring were disposed of by incineration following sampling. Hatchlings were housed in batteries of brooding pens manufactured by Beacon Steel Company (Model B735Q). Each pen measured approximately 72 X 90 X 23 cm high. The external walls and ceilings of each pen were constructed of galvanized wire mesh and galvanized sheeting. Floors were of galvanized wire mesh. Thermostats in the brooding compartment of each pen were set to maintain a temperature of approximately 38 C from the time of hatching until the birds were 14 days of age. The average ambient room temperature was 25.5 1.1C (SD) with an average relative humidity of 15 6% (SD). The photoperiod for the hatchlings was maintained by a time clock at 16 hours of light per day. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 20 - Egg Shell Thickness Measurements and Egg Content Collection Weekly throughout the egg laying period, one egg was collected, when available, from each of the odd numbered pens during odd numbered weeks (1,3,5, etc.) and from each of the even numbered pens during the even numbered weeks (2,4,6, etc.). Prior to being opened, eggs were rinsed with water to remove any excreta or feed adhering to the shell. The eggs were opened at the waist, the contents were removed and placed in a Whirl Pak and stored frozen for potential analysis. The empty shells were thoroughly rinsed with water, and allowed to air dry for at least one week at room temperature. The average thickness of the dried shell plus the membrane was determined by measuring five points around the waist of the egg using a micrometer. Measurements were made to the nearest 0.002 mm. Statistical Analyses Upon completion of the test, an analysis of variance (ANOVA) was performed to determine statistically significant differences between groups. Dunnett's multiple comparison procedure (7,8) was used to compare the three treatment means with the control group mean and assess the statistical significance of the observed differences. Sample units were the individual pens within each experimental group, except adult body weights where the sample unit was the individual bird. Percentage data were examined using Dunnett's method following arcsine square root transformation (see Appendix III for reproductive parameters). Each of the following parameters was analyzed statistically: 1. Adult Body Weight - Individual body weight was measured at test initiation, Weeks 2, 4, 6, 8 and at termination. Statistical comparisons were made between the control group and each treatment group at each weighing interval by sex. 2. Adult Feed Consumption - Feed consumption expressed as grams of feed per bird per day was examined by pen weekly during the test. Statistical comparisons were made between the control and each treatment group. 3. Eggs Laid of Maximum Laid - The number of eggs laid per female divided by the largest number of eggs laid by any one female. This transformation was used to convert the number of eggs laid to a percentile value less than or equal to 100. 4. Eggs Cracked of Eggs Laid - The number of eggs determined by candling to be cracked divided by the number of eggs laid, per pen. 5. Viable Embryos of Eggs Set - The number of viable embryos at the Day 10-12 candling was divided by the number of eggs set, per pen. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 21 - 6. Live 3-Week Embryos of Viable Embryos - The number of live embryos at the Day 21 candling was divided by the number of viable embryos, per pen. 7. Hatchlings of 3-Week Embryos - The number of hatchlings removed from the hatcher was divided by the number of live 3-week embryos, per pen. 8. 14-Day Old Survivors of Hatchlings - The number of 14-day old survivors was divided by the number of hatchlings, per pen. 9. Hatchlings of Eggs Set - The number of hatchlings was divided by the number of eggs set, per pen. 10. 14-Day Old Survivors of Eggs Set - The number of 14-day old survivors was divided by the number of eggs set, per pen. 11. Hatchlings of Maximum Set - The number of hatchlings per female divided by the largest number of eggs set from any one female. This transformation was used to convert the number of hatch lings to a percentile value equal to or less than 100. 12. 14-Day Old Survivors of Maximum Set - The number of 14-day old survivors per pen divided by the largest number of eggs set. 13. Egg Shell Thickness - The average egg shell thickness of indiscriminately selected eggs per pen was measured. 14. Offspring's Body Weight - The group body weights of surviving hatchlings and 14-day old survivors were measured by parental pen group. 15. Liver Weight - Liver weights were measured for adults and selected offspring. Statistical comparisons were made between the control group and each treatment group by age and sex. RESULTS AND DISCUSSION Mature northern bobwhite received T-7485 at nominal dietary concentrations of 100, 300 or 900 ppm a.i. for 21 weeks. A control group was maintained concurrently with the treatment groups. Analytical Results for Diets Diet samples were collected on Week 1, Day 0 of the test from the 100, 300 and 900 ppm a.i. test levels to evaluate the homogeneity of the test substance in the diet. The mean and standard deviation for the 100, 300 and 900 ppm a.i. test levels were 105 5.13 ppm a.i., 299 12.0 ppm a.i. and 880 16.0 ppm a.i., respectively. The corresponding coefficients of variation were 4.91%, 4.00% and 1.82% (Appendix XVI, Table 4). The mean measured T-7485 concentrations from the 100, 300 and 900 ppm a.i. test levels were 105%, 100%, and 98%, respectively, of the nominal concentrations. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 22 - Diet samples collected on Weeks 4, 8, 12, 16 and 20, Day 0 of the test from the 100, 300 and 900 ppm a.i. test levels were analyzed to verify the concentration of the test substance in the diet. The mean and standard deviation for the 100, 300 and 900 ppm a.i. test concentrations were 107 7.74 ppm a.i., 314 5.97 ppm a.i. and 961 36.8 ppm a.i., respectively. The corresponding coefficients of variation were 7.21%, 1.90% and 3.83% (Appendix XVI, Table 5). The mean measured T-7485 concentrations from the 100, 300 and 900 ppm a.i. test levels were 107%, 105% and 107%, respectively, of the nominal concentrations. A typical ion chromatogram of a test sample is shown in Appendix XVI, Figure 7. Analysis of diet samples collected from feeders after being held at ambient temperature for 7 days averaged 96%, 105%, and 112% of the Day 0 values for the 100, 300 and 900 ppm a.i. test concentrations, respectively (Appendix XVI, Table 6). Mortalities There were no treatment-related mortalities in the 100, 300 or 900 ppm a.i. treatment groups during the course of the test. There were three adult mortalities that were considered to be incidental to treatment that occurred during the test, one in the control group and one each in the 100 and 300 ppm a.i. treatment groups. The female in Pen 1608 of the control group was found dead during Week 12 of the test. Gross necropsy revealed feather loss and an open lesion on the head and neck. The breast muscle, spleen, liver and kidneys were noted to be pale and the ovary was developing. The male in Pen 1617 of the 100 ppm a.i. treatment group was found dead at the beginning of Week 6 of the test, having exhibited no prior signs of toxicity. Gross necropsy revealed bruising on the cranium and intracranial bleeding between the orbits. The final mortality was the female in Pen 1639 of the 300 ppm a.i. treatment group that was euthanized at the end of Week 14 due to its moribund condition. Four days prior to being euthanized, this female had been noted with a neck lesion. Gross necropsy revealed a large head and neck lesion, exposing the cranium and neck muscle, with necrotic margins. The bird was emaciated, with a loss of muscle mass and prominent keel. The spleen was noted to be large and pale, the kidneys were pale, the cecal contents were firm and the ovary was regressing. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 23 - No other mortalities occurred during the course of the study. Due to the nature of the lesions observed at necropsy, none of the mortalities that occurred were considered to be related to treatment. Clinical Observations No overt signs of toxicity were observed at any of the concentrations tested. Incidental clinical observations noted during the test included those that normally are associated with injuries and penwear, and were noted for the control group and all treatment groups. Such observations included foot, leg, and head lesions, feather loss and ventral head curl. In addition, lameness, loss of coordination, ruffled appearance, wing droop and lethargy also were observed and were generally associated with incidental injuries. While some incidental observations were persistent once noted (e.g. feather loss), most were transient in nature and subsided once an injury had healed. In the control group, incidental clinical observations were noted for eight individuals during the course of the test. Three birds were noted with feather loss, four birds were noted with foot lesions and associated lameness, two birds were noted with either bruising or a lesion on the head and one bird was noted with a lesion and bruising on the upper back. Additionally, one bird was also noted with right wing droop, one bird was noted with a growth above the left eye, and one bird was noted with a transient loss of coordination. Except for incidental findings, all birds in the control group were normal in appearance and behavior throughout the study. In the 100 ppm a.i. treatment group, incidental clinical observations were noted for five individuals during the course of the test. Two birds were noted with feather loss, one bird was noted with foot lesions and associated lameness and ruffled appearance, and two birds were noted with either bruising and/or a lesion on the head. Additionally, one bird was also noted with twisted primary feathers on the left wing, presumably due to a wing injury. Except for incidental findings, all birds in the 100 ppm a.i. treatment group were normal in appearance and behavior throughout the study. In the 300 ppm a.i. treatment group, incidental clinical observations were noted for seven individuals during the course of the test. One bird was noted with feather loss and four birds were noted with foot lesions, one of which was also noted with associated lameness. One bird was noted with swelling on the head, as well as a head lesion. Additionally, one bird was noted with a ventral head curl, a condition that is thought to be caused by a neck injury. One female in the 300 ppm a.i. treatment group Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 24 - was noted with head and neck lesions that resulted in her being euthanized. Prior to her being euthanized in a moribund condition, this bird exhibited depression, a reduced reaction to external stimuli (sound and movement), a ruffled appearance and lethargy. Except for incidental findings, all birds in the 300 ppm a.i. treatment group were normal in appearance and behavior throughout the study. In the 900 ppm a.i. treatment group, incidental clinical observations were noted for 11 individuals during the course of the test. Five birds were noted with feather loss and six birds were noted with foot lesions, three of which were also noted with associated lameness. One hen with foot lesions was also noted with a ruffled appearance and lethargy that were attributed to the injury. One bird was noted with bruising on the back and another bird was noted with bruising on the head and neck. Lesions were noted on the back of one bird and on the head of another bird. Two birds were noted with lesions around one eye, one of these birds was also noted with a ruffled appearance that was attributed to the eye lesion. Except for incidental findings, all birds in the 900 ppm a.i. treatment group were normal in appearance and behavior throughout the study. Gross Necropsy All surviving adults were subjected to gross necropsy following adult termination. All findings observed were considered unrelated to treatment. Necropsy findings are reported in Table 7 and Appendix IV. Adult Body Weight There were no apparent treatment-related effects upon adult body weight at any of the concentrations tested. No statistically significant differences between the control group and the 100, 300 or 900 ppm a.i. treatment groups were observed at any of the body weight intervals. Mean body weight measurements are presented in Table 1, and Figures 1 and 2. Individual body weight measurements are presented in Appendix V. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 25 - Adult Feed Consumption There were no apparent treatment-related effects upon feed consumption at the 100, 300, and 900 ppm a.i. test concentrations. No statistically significant differences between the control group and the 100 and 900 ppm a.i. treatment groups were observed at any of the feed consumption intervals. When compared to the control group, there was a slight, but statistically significant (p < 0.01), increase in feed consumption at the 300 ppm a.i. test concentration during Week 16 of the test. However, this difference was slight and was neither consistent nor concentration responsive. Since the difference represented an increase in the consumption of the treated diet, it was not considered to be treatment related. Mean feed consumption measurements are shown in Table 2 and Figure 3. Feed consumption measurements by pen are presented in Appendix VI. Estimated test substance intakes, daily dietary dose, for northern bobwhite were calculated by treatment group for the pre-egg production period, the egg production period and the overall adult period using the following formula: Daily Dietary Dose = Test Concentration (mg a.i./kg) x Daily Feed Consumption (g/bird/day) (mg a.i./kg body weight/day) Body Weight (g/bird) The mean body weight value is the mean of both male and female body weights. For the pre-egg production interval the body weights were averaged over Weeks 0, 2, 4, 6 and 8. For the egg-production interval body weights were averaged over Weeks 8 and 20 (adult termination). The accuracy of the estimated mean daily dietary dose may be impacted by differences in individual feed consumption, both within and between pens, and feed wastage. The estimated daily dietary doses are presented in the table below. Contains No CBI Wildlife International, Ltd. - 26 - Project Number 454-116 Estimated Maximum Mean Daily Dietary Dose of T-7485 (mg a.i./kg body weight/day) Test Interval (test weeks) Test Concentration (ppm a.i.) Mean Body Weight (g) Mean Feed Consumption (g/bird/day) Estimated Daily Dietary Dose (mg a.i./kg/day) Pre-Egg Production (Weeks 1 - 10) 0 100 300 900 196 196 196 197 16 0.0 17 8.5 17 25.2 17 76.4 Egg Production 0 212 22 0.0 (Weeks 11 - 21) 100 214 22 10.3 300 216 23 32.0 900 214 22 93.9 Over-all 0 200 19 0.0 (Weeks 1 - 21) 100 201 20 9.7 300 201 20 29.7 900 201 20 87.8 Reproductive Results There were no treatment-related effects upon reproductive performance at any of the concentrations tested. When compared to the control group, there were no statistically significant differences in any of the reproductive parameters measured in the 100, 300, or 900 ppm a.i. treatment groups. While not statistically significant, there was a slight reduction in the total number of eggs laid at the 900 ppm a.i. test concentration. However, reduction was primarily attributable to a single hen (Pen 1662), that exhibited foot lesions and did not lay any eggs during the last five weeks of the study. If data from that pen is excluded, the number of eggs laid per hen at the 900 ppm a.i. test concentration is comparable to the control value. Summaries of the reproductive data are presented in Tables 3 and 4, and in Figures 4 and 5. Reproductive parameters by pen are presented in Appendix VII and VIII. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 27 - Egg Shell Thickness There were no apparent treatment related effects upon egg shell thickness at any of the concentrations tested. When compared to the control group, there were no statistically significant differences in egg shell thickness in the 100, 300 or 900 ppm a.i. treatment groups. Egg shell thickness data are presented in Table 5 and Appendix IX. Offspring Body Weights There were no apparent treatment related effects upon offspring body weight at any of the concentrations tested. When compared to the control group, there were no statistically significant differences in the body weight of hatchlings or 14-day old survivors from the 100, 300 or 900 ppm a.i. treatment groups. Offspring body weight data are presented in Table 6, and Appendices X and XI. Liver Weights There were no apparent treatment related effects upon adult or offspring liver weights at any of the concentrations tested. When compared to the control group, there were no statistically significant differences in liver weights of adults or 14-day old offspring from the 100, 300 or 900 ppm a.i. treatment groups. Liver weight data are presented in Table 8 and Appendices XII and XIII. Serum, Liver and Egg Analytical Results At the conclusion of the exposure period, serum and liver samples were collected from all surviving adult birds. Serum and liver samples were also collected from indiscriminately selected offspring from each test pen from the last lot of offspring produced from the study (Lot J). Egg contents from those eggs selected for eggshell thickness measurements were collected for potential analysis. Following are summary tables of the results of serum, liver and egg content analyses for each test concentration. The full report for the analysis of serum, liver and egg contents is provided in Appendix XVII. Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 28 - Summary of Mean Concentrations of T-7485 in Serum and Liver Test Concentration 100 ppm a.i. 100 ppm a.i. 100 ppm a.i. 300 ppm a.i. 300 ppm a.i. 300 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. Group Adult Male Adult Female Offspring Adult Male Adult Female Offspring Adult Male Adult Female Offspring Mean (SD) Concentration Mean (SD) of T-7485 (ppm a.i.) in Concentration of T-7485 Serum (ppm a.i.) in Liver 16.5 (10.3) 14.6 (8.4) 0.0369 (0.0148) 3.25 (2.26) 3.52 (2.31) 0.0211 (0.0103) 27.9 (9.5) 37.8 (23.5) 0.0567 (0.0324) 7.78 (3.32) 11.1 (6.03) 0.0515 (0.0620) 68.2 (21.3) 104 (84.6) 0.133 (0.0743) 15.7 (4.78) 29.6 (19.7) 0.111(0.0792) Summary of Mean Concentrations of T-7485 in Egg Contents Treatment Group 100 ppm a.i., Lot B Eggs 100 ppm a.i., Lot G Eggs 300 ppm a.i., Lot B Eggs 300 ppm a.i., Lot G Eggs 900 ppm a.i., Lot B Eggs 900 ppm a.i., Lot G Eggs Lot B = 2ndweek of eggs set Lot G = 7thweek of eggs set Mean (SD) Concentration of T-7485 (ppm a.i.) in Egg Contents 7.65 (2.05) 14.0 (5.50) 23.6 (8.58) 31.4 (19.1) 50.5 (27.9) 92.6 (31.8) Histopathology Samples of liver, kidney and gonad were collected from all adult birds for histopathological examination. Similar samples were also collected from indiscriminately selected offspring from each test pen, from the last lot of offspring produced from the study (Lot J). The spleen from one adult female in the 900 ppm a.i. test concentration was noted as slightly enlarged during gross necropsy, and was also Contains No CBI Wildlife International, Ltd. Project Number 454-116 - 29 - collected for histopathological examination. All histopathological samples were fixed in 10% buffered formalin and shipped to EPL in Herndon, Virginia for histopathology. No lesions considered related to exposure to T-7485 were noted in the liver, kidney, ovary and testes of the adults or their offspring. The few findings in various tissues of the adults and offspring from the control and treatment groups were considered to be incidental and unrelated to treatment. Testes of adult males for all treatment groups exhibited spermatogenesis that was characterized by the presence of numerous mature spermatozoa in the seminiferous tubules. While a few males in the treatment groups exhibited decreased testicular diameter and/or seminiferous tubule degeneration, this result was not dose responsive and was most consistent with physicological regression. Additionally, similar findings were noted for several males in the control group. The pathology report provided by EPL is presented in Appendix XIV. CONCLUSION There were no treatment-related mortalities, overt signs of toxicity or treatment-related effects upon body weight or feed consumption at any of the concentrations tested. Additionally, there were no treatment-related effects upon any of the reproductive parameters measured at the 100, 300 or 900 ppm a.i. test concentrations. The no-observed-effect concentration for northern bobwhite exposed to T-7485 in the diet during the study was 900 ppm a.i., the highest concentration tested. Contains No CBI Wildlife International, Ltd. - 30 REFERENCES Project Number 454-116 1 U.S. Environmental Protection Agency. 1982. Pesticide Assessment Guidelines, FIFRA Subdivision E, Hazard Evaluation: Wildlife and Aquatic Organisms, subsection 71-4, Environmental Protection Agency, Office of Pesticide Programs. Washington, D.C. 2 Organization for Economic Cooperation and Development. 1984. Avian Reproduction Test. OECD Guideline for Testing of Chemicals. Guideline 206. Paris. 3 American Society for Testing and Materials. 1986. Standard Practice for Conducting Reproductive Studies with Avian Species. ASTM Standard E1062-86. Annual Book of ASTM Standards. Vol. 11.04. Philadelphia, PA. 15 pp. 4 Merck & Co., Inc. 1991. The Merck Veterinary Manual. Merck & Co. Rahway, NJ. 1832 pp. 5 Wildlife International, Ltd. 2003. Analytical Method Verificationfor the Determination o f T-7485 in Avian Diet. Wildlife International, Ltd. Project No. 454C-129. 6 National Research Council. 1996. Guide for the Care and Use o f Laboratory Animals. Washington, DC. National Academy Press. 125 pp. 7 Dunnett, C.W. 1955. A Multiple Comparison's Procedure for Comparing Several Treatments with a Control. Jour. Amer. Statis. Assoc. 50: 1096-1121. 8 Dunnett, C.W. 1964. New Tables for Multiple Comparisons with a Control. Biometrics 20: 482 491. Contains No CBI Table 1 Mean A d u lt Body W eight (g ) from a N o rth e rn B obw h ite R e p r o d u c tio n S tu d y w ith T -7 4 8 5 1 Experimental Group (ppm a.i.) Sex Control 100 300 900 Male Female Male Female Male Female Male Female Week 0 Change Week 0-2 Week 2 Change Week 2-4 Week 4 Change Week 4-6 Week 6 Change Week 6-8 Week 8 Change Week 8-Term Test Total Term Change 193 1 190 1 195 1 188 0 192 2 189 2 193 1 188 2 193 3 191 4 196 3 189 2 194 3 191 3 194 4 190 4 196 3 195 4 200 4 191 4 196 5 193 5 198 5 194 3 199 4 199 3 203 5 195 2 201 6 198 4 204 6 197 4 203 2 201 37 208 7 197 40 208 8 201 36 209 4 201 30 205 13 238 48 215 20 237 48 215 24 238 49 214 21 230 42 The means for body weights and body weight changes are calculated and rounded separately. Differences between control and each treatment group were not significant (p > 0 .0 5 ). 1 Only surviving birds were included in the calculations for each body weight interval. Project Number 454-116 Contains No CBI Figure 1 Mean Adult Male Body Height (g) from a N o r th e r n Bobuhite Reproduction Study uith T-7485 MALE BODV HEIGHT (g ) Project Number 454-116 to Week 0 Change Ueek 2 Change Ueek 4 Change Ueek 6 Change Ueek 8 Change Ueek T Total Uk 0 -2 Uk 2 -4 Uk 4 -6 Uk 6 -8 Uk 8 -T Change UE E KS o Concentrations, ppm a .i. 100 122300 139900 Contains No CBI Figure 2 Mean Adult Female Body Height (g) from a N o r th e rn Bobuhite Reproduction Study uith T-7485 Contains No CBI FEMALE BODV WEIGHT (g) tooo oo oo 3n id 3 a CO T3oJ a> cooo 2w0h1O3 s<CO33QDU* llllllllllllllllilllllllllllllllllllllllllll Wn to C33oU* IIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIIinTTT? ' O"yf . 3a jy00Iz0r\ <to-3IooDJtI2IDD= H0I0 I32D E ID ID 3a id 91 \-p9t jaqum^ p a ia ij - ee- Table 2 Mean Feed C onsum ption ( g / h i r d / d a y ) from a N o r th e r n B o b w h ite R e p r o d u c tio n S tu d y w ith T -7485 Experimental WEEKS Group (ppm a. i .) 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 Control 100 300 900 15 16 16 16 16 15 15 17 16 18 18 19 19 21 23 20 25 23 25 23 24 16 17 17 17 17 16 16 17 16 18 18 19 21 22 24 21 24 23 24 23 24 16 17 16 16 17 16 15 18 16 18 19 19 21 22 25 23** 26 25 26 23 24 16 17 17 17 17 16 16 18 16 17 18 19 20 22 24 22 25 24 25 22 24 OJ * * Significantly different from the control at p < 0.01 Project Number 454-116 Contains No CBI Mean Figure 3 Feed Consumption (g /b ir d /d a y ) from Reproduction Study uith T-7485 a Northern Bobuhite UC/)i Project Number 454-116 Concentrations, ppm a .i. 0 100 ........ 300 ...........900 Contains No CBI Tabi e 3 Summary o f R e p ro d u c tiv e Perform ance from a N o rth ern B obw h ite R ep ro d u c tio n S tu d y w ith T-7485 Reproductive Parameter Control Experimental Group (ppm a .i .) 100 300 Number of Replicates Total Eggs Laid Eggs Cracked Eggs Set Viable Embryos Live 3-Week Embryos Hatchlings 14-Day Old Survivors Eggs Laid/Hen Eggs Laid/Hen/Day1 14-Day Old Survivors/Hen 15 866 4 787 692 689 671 64 8 58 0.58 43 15 840 2 763 714 710 691 646 56 0.57 43 * Results for the 900 ppm a.i. group when Pen 1662 is excluded from calculations. Based on 99 days of eggs production. 15 897 6 814 753 746 703 674 60 0.60 45 900 900 * 16 815 7 736 697 693 674 633 51 0.51 40 15 801 6 725 688 684 665 625 53 0.54 42 to Os Project Number 454-116 Contains No CBI Figure 4 Mean R e p ro d u c tiv e P erfo rm an ce from a N o rth e rn Bobuhite Reproduction Study uith T -7 4 85 CC\L LJ OJ Project Number 454-116 Control Eggs Cracked lOOppm a .i. IZ2 Eggs U ia b le Set Embryos Live 3-uk Embryos Hatchlings M -D ay Old Survivors Contains No CBI Table 4 Summary o f R e p r o d u c t i v e P e r f o r m a n c e , N o r m a l i z e d a s P e r c e n t a g e s (%) from a Northern Bobwhite R eproduction Study w ith T-74851 Reproductive Parameter Control Number of Replicates 2 Total Eggs Laid Eggs Laid/Maximum Laid (%) Eggs Cracked/Eggs Laid (%) Viable Embryos/Eggs Set (%) Live 3-Week Embryos/Viable Embryos (%) Hatchlings/Live 3-Week Embryos (%) 14-Day Old Survivors/Hatchlings (%) Hatchlings/Eggs Set (%) 14-Day Old Survivors/Eggs Set (%) Hatchlings/Maximum Set (%) 14-Day Old Survivors/Maximum Set (%) 15 866 84 1 89 99 96 96 86 83 71 69 Experimental Group (ppm a.i.) 100 300 15 15 840 897 81 87 01 93 92 99 99 97 94 93 96 90 86 84 83 73 74 68 71 Differences between the control and each treatment groups were not significant (p > 0 .0 5 ). Values represent pen means for experimental group. Values for each pen are presented in Appendices VII and VIII. 2 Represents the total number of eggs laid in each group. *Results for the 900 ppm a.i. group when Pen 1662 is excluded from calculations. * 900 900 16 15 815 801 74 77 11 94 95 99 99 97 97 94 94 91 92 86 87 67 70 63 66 Contains No CBI Project Number 454-116 Figure 5 Mean R e p r o d u c tiv e P e rfo rm a n c e , N orm a lize d as P e rc e n ta g e s from a N orth ern Bobuhite Reproduction Study with T-7A85 Project Number 454-116 Maximum Laid Eggs Laid Eggs Set Uiable Embryos Live 3-Ueek Reproductive parameter Hatchlings E3o Concentrations, ppm a .i. 100 [771300 Egg900 Contains No CBI Table 5 Mean E g g s h e l l T h i c k n e s s M e a s u r e m e n t s (mm) from a Northern Bobwhite R eproduction Study w ith T-7485 Experimental Group (ppm a.i.) Control 100 300 900 N o . Eggs Measured 72 72 74 71 Shell Thickness Mean ( SD) 0.219 + 0.013 0.227 0.011 0.222 0.015 0.224 0.015 Differences between the control and each treatment groups were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Table 6 Mean B o d y W eig h t ( g ) o f H a t c h l i n g s and 1 4 -D ay O ld S u r v i v o r s from a Northern Bobwhite Reproduction Stu dy w ith T-7485 Experimental (PPM A . I .) Hatchlings Number Mean ( SD) 14-Day Old Survivors Number Mean ( SD) Control 100 300 900 671 6 0 690 6 + 1 703 6 0 672 6 0 648 26 2 646 26 3 674 27 2 633 27 2 The number of hatchlings weighed may differ from the total number of hatchlings since those hatchlings found dead were not weighed. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Table 7 Summary of Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485 Adult Birds Euthanized at Termination of the Test Males - Treatment Group (ppm a.i.) Control 100 300 900 Number ofbirds 16 15 16 16 External - feather loss 7 32 4 External - head lesion 0 00 1 External - foot/leg lesions 2012 External - wart-like growth below beak 0 0 1 0 Spleen - slightly enlarged 000 0 Liver - slightly mottled 0 110 Liver - friable 000 0 Abdominal cavity - extensive fat 00 10 Abdominal cavity - egg yolk peritonitis - - - - Ovary - hemorrhagic follicles --- - Testes - one testis small, ~1.0 - 1.25 cm 2 5 2 5 Testes - small, ~1.0 - 1.5 cm 10 12 Not remarkable 6 88 6 Females - Treatment Group (ppm a.i.) Control 100 300 900 15 16 15 16 14 11 12 13 120 0 3 0 14 0 00 0 0 00 1 0 00 0 0 00 1 0 00 0 0 012 100 0 - -- - -- 0 53 1 Project Number 454-116 Contains No CBI Table 8 Mean Liver Weights1(g) from a Northern Bobwhite Reproduction Study with T-7485 Experimental Group Adult Males Mean SD Adult Females Mean SD Offspring Males Offspring Females Mean SD Mean SD Control 0 ppm a.i. 4.134 + 1.446 7.228 0.644 1.013 + 0.111 1.082 + 0.118 100 ppm a.i. 4.068 + 0.901 7.576 1.575 1.113 0.183 1.030 + 0.129 300 ppm a.i. 4.187 1.692 7.202 1.112 1.207 + 0.122 1.292 0.287 900 ppm a.i. 3.926 1.068 6.523 1.403 1.107 0.228 1.071 + 0.169 ' Mean standard deviation. Differences between the control group and the treatment groups were not statistically significant (p>0.05, Dunnett's t-test). 1 U) Project Number 454-116 Contains No CBI W ildlife International, Ltd. Project Number 454-116 # # =B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B==B= INGREDIENTS Appendix I DIET AND SUPPLEMENT FORMULATIONS WILDLIFE INTERNATIONAL, LTD. GAME BIRD RATION1 PERCENT (%) Fine CornMeal Soy Bean Meal, 47.5%Protein Wheat Midds Agway Special, 60% Protein Alfalfa Meal, 20%Protein Dried Whey Ground Limestone Eastman CalPhos GL Ferm (Fermatco)2 Salt Iodized CM T-Premix 561 CM TM Premix 434 CHO CHL 60% 55# Liquimeth 40% Lysine 75% DL Methionine 055 Vit K 16 g/lb 50# Selenium 42.86 33.39 6.48 9.96 3.00 2.50 0.62 0.11 0.25 0.10 0.05 0.08 0.09 0.25 0.06 0.10 0.05 0.05 Total 100.00 VITAMIN AND MINERAL CONTENT AMOUNT ADDED PER POUND Vitamin D3 0.600 kiu/lb Vitamin A 2.000 kiu/lb Riboflavin 2.1992 mg/lb Niacin 21.6932 mg/lb Pantothenic Acid 5.2241 mg/lb Vitamin B12 6.0000 ug/lb Folic Acid 0.5169 mg/lb Biotin 65.9360 ug/lb Pyridoxine 1.4639 mg/lb Thiamine 0.9200 mg/lb Vitamin E 5.0000 iu/lb VitaminK (Menadione Dimethylpyrimidinol Bisulfite) 3.2400 mg/lb Manganese 115.0203 mg/kg Zinc 194.6601 mg/kg Copper 24.3007 mg/kg Iodine 2.4410 mg/kg Iron 150.4117 mg/kg Selenium 0.4749 mg/kg Beta-Carotene 1.1413 mg/lb Calcium 0.9750 % Chloride 0.2164 % Choline 1882.4116 mg/kg Cobalt 0.1139 mg/kg Magnesium 0.2503 % Phos 30 0.3908 % Phos 50 0.4719 % Potassium 1.1148 % Sodium 0.1335 % Sulfur 0.3478 % Phosphorus 0.6800 % Ca:PHOS 1.4338 ratio i The guaranteed analysis is a minimum of 27% protein, a minimum of 2.5% crude fat and a maximum of 3.8% crude fiber. 2 Fermentation By-Products (Source of Unidentified GrowthFactors) Contains No CBI Wildlife International, Ltd. -45 - Project Number 454-116 Appendix I Diet and Supplement Formulations Table 2 Vitamins and Electrolytes Concentrate GUARANTEED ANALYSIS Vitamin A Vitamin D Vitamin E Riboflavin d-Pantothenic Acid Niacin Vitamin B-12 MSBC Folic Acid Thiamine HC1 Pyridoxine Hydrochloride Ascorbic Acid Water Soluble Powder Per 4 oz. 2,500,000 1,000,000 1,000 750 1,250 2,500 2.5 1,000 65 250 250 3,750 Per lb. 10,000,000 IU 4,000,000 ICU 4.000 IU 3.000 Mg 5.000 Mg 10.000 Mg 10.0 Mg 4.000 Mg 260 Mg 1.000 Mg 1.000 Mg 15.000 Mg INGREDIENTS: Vitamin A Supplement, D-Activated Animal Sterol (source of Vitamin D3), Alpha Tocopheryl Acetate (source of Vitamin E). Riboflavin Supplement, d-Calcium Pantothenate, Niacin Supplement, Vitamin B-12 Supplement, Menadione Sodium Bisulfite (source of Vitamin K), Folic Acid, Thiamine HC1, Pyridoxine Hydrochloride, Ascorbic Acid, Sodium Chloride, Calcium Chloride, Magnesium Sulfate, Ferric Ammonium Citrate, Potassium Chloride and Dextrose. MIXING PROCEDURE: The vitamin and electrolyte mix was prepared as a ration of approximately 2 grams of Durvet vitamins and electrolytes to approximately 1 gallon of water. Contains No CBI W ildlife International, Ltd. -46- Project Number 454-116 Appendix II Diet Preparation Premixes for T-7485 were prepared on September 16, October 14, November 11 and December 9, 2003, and on January 5 and February 3, 2004. Nominal preparation was as follows: Control: 4018.0 g ration + 91 ml com oil 100 ppm a.i.: 3.7576 g T-7485 + 4014.2 g ration + 91 ml com oil 300 ppm a.i.: 11.2729 g T-7485 + 4006.7 g ration + 91 ml com oil 900 ppm a.i.: 33.8187 g T-7485 + 3984.2 g ration + 91 ml com oil For each o f the T-7485 premixes, the basal ration was weighed into a tared mixing bowl. The com oil was measured and added to the basal ration in the bowl. The bowl was placed on a Hobart mixer and the contents were mixed approximately 5 minutes. A portion o f the ration, sufficient to cover the blender mixing blades, was transferred to a Waring blender. The test substance was weighed into a tared plastic weigh boat and added to the ration in the Waring blender. The weigh boat was rinsed three times with ration from the mixing bowl, with the rinse being added to the blender contents. The ration and test substance in the Waring blender were blended approximately one minute. The blender contents were added to the ration in the Hobart mixing bowl. The blender was rinsed three times with uncontaminated ration from the mixing bowl, with the rinse being added to the contents o f the bowl. The Hobart mixing bowl was placed on a Hobart mixer and the contents were mixed approximately 10 minutes. If not used immediately after mixing, 1000.0 g aliquots o f the premix were weighed, placed in appropriately labeled plastic bags, reweighed, and stored frozen. As needed, the appropriate premix was incorporated into the final diet as follows: 0 ppm a.i.: 1000 g Premix + 7.55 kg ration + 450 g limestone 100 ppm a.i.: 1000 g Premix + 7.55 kg ration + 450 g limestone 300 ppm a.i.: 900 ppm a.i.: 1000 g Premix + 7.55 kg ration + 450 g limestone 1000 g Premix + 7.55 kg ration + 450 g limestone The diet was mixed for approximately 10 minutes in a Hobart mixer. Contains No CBI Wildlife International, Ltd. -47- Project Number 454-116 Appendix III Reproductive Parameters 1. Eggs Laid Definition - The number of eggs produced during the breeding season. Sensitivity - This is a parameter that is frequently affected by chemicals that cause reproductive impairment. It is also highly sensitive to the general conditions under which the test is conducted, and may be adversely affected by improper diet, crowding, excessive disturbances, etc. 2. Eggs Cracked Definition - Eggs determined to have cracked shells when inspected with a candling lamp. Fine cracks cannot be detected without utilizing a candling lamp, and if undetected, will bias the data by adversely affecting embryo development. Sensitivity - This parameter is not frequently affected as cracking is directly related to shell thickness, and few chemicals have caused egg shell thinning. Improper pen design, overcrowding, and pecking by the birds can also increase egg cracking. 3. Eggs Set Definition - All eggs placed under incubation, i.e., total eggs laid minus cracked eggs, mechanically damaged eggs, and those selected for egg shell thickness analysis. The reason for presenting this parameter is to establish a base o f reference for the following parameter - viable embryos. 4. Viable Embryos Definition - Eggs in which fertilization has occurred and embryonic development has begun. This is determined by candling the eggs 10-12 days after the initiation o f incubation. It is difficult to distinguish between infertility and an early embryonic death. Sensitivity - This is a frequently affected parameter with infertility or embryonic mortality caused by an unfavorable environment for fertilization in the oviduct, impotent males, or chemical residue in the egg. Contains No CBI Wildlife International, Ltd. -48 - Project Number 454-116 Appendix III Reproductive Parameters Page 2 5. Live Three-Week Embryos Definition - These are embryos that are developing normally after three weeks of incubation. This is determined by candling the eggs. Sensitivity - This parameter is seldom affected. 6. Hatchlings Definition - Embryos that mature, pip the shell, and liberate themselves from their eggs on Day 25 or 26 o f incubation. Sensitivity - This is a frequently affected parameter which is also highly sensitive to the conditions o f incubation, such as rate and angle o f rotation, humidity, and temperature. - Humidity: If too dry, chicks will stick to their shells. - Temperature: If too hot and humid, chicks will grow too fast and be too large for their shells; thereby not having the intra-egg mobility necessary to pip their shells. 7. Body Weight - Hatchlings Definition - The average weight o f hatchlings by parental pen o f origin, taken immediately upon removal from the hatcher. Hatchlings from each weekly lot of eggs are weighed. Mean weight for each week and mean weight by pen are reported. Sensitivity - This is an occasionally affected parameter and may reflect some residual or latent toxic effects from chemical residue in the egg. 8. 14-Day Old Survivors Definition - Birds that survive brooding for two weeks following hatch. Sensitivity - This is a seldom affected parameter and is probably more indicative of the conditions under which the birds were reared in battery brooders than the chemical to which the adults were exposed. Contains No CBI ,W ildlife International Ltd. -49- Project Number 454-116 Appendix III Reproductive Parameters Page 3 9. Body Weight - 14-Day Old Survivors Definition - The average body weight of the 14-day old survivors by parental pen o f origin, taken upon removal from the brooder unit at 14 days o f age. Fourteen-day old survivors from each weekly lot o f eggs are weighed. Mean weight for each week and mean weight by pen are reported. Sensitivity - This parameter is seldom affected, and more closely reflects the rearing practices utilized. 10. Egg Shell Thickness Definition - The thickness of the shell and the membrane o f the egg at its equator after having been opened, washed, and dried for at least one week at room temperature. Egg shell thickness measurements for each egg measured, and pen means are reported. Sensitivity - This is a seldom affected parameter; however, measurements must be taken very carefully to ensure accuracy. Possible mechanical errors include membrane curling at the measurement surface, and calcium deposits on the measurement surface. Contains No CBI Males External - feather loss External - foot lesions Testes - small, ~1.0 cm Testes - right testis small, -1.25 cm Testes - right testis small, -1.0 cm Not remarkable Appendix - IV - Table 1 Individual Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485 Birds Euthanized at Test Termination 0 ppm a.i. - Control Pens 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 __ _ X _ X _ XXXX - _X_ _ - - - - - - - - - - - - - - XX - - - - - - - - -X- - - - - - -X - - - - - - - - - - - - - - - - - - - - - - - - - - - -X- X- X - X - X - " - "XX " - - Project Number 454-116 Females External - feather loss External - head lesion External - old lesion on head External - foot lesions External - missing toe Ovary - hemorrhagic follicles Not remarkable a Bird found dead during test. Pens 1601 1602 1603 1604 1605 1606 1607 1608a 1609 1610 1611 1612 1613 1614 1615 1616 XXX X XX X _ XXX X XXX --- ---- - -------- - - - -X - - - - - - - - - - - - - - -X- - - - - - - - - - - -X-X - - - - - - - - - - - - - - - - - - - - - - -X - "--"-- --""--- Contains No CBI Appendix - IV - Table 2 Individual Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485 Birds Euthanized at Test Termination 100 ppm a.i. Males External - feather loss Liver - slightly mottled Testes - right testis small, -1.25 cm Testes - left testis small, ~1.0 cm Not remarkable Pens 1617a 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 - - - - -X. - _ - - - - - XX ------ ------- - X - - - - - - - -XX -X X " - - - - - -X - - - - - - - - - - - XX X - X X X " - X - - " X - Females External - feather loss External - head lesion External - bruising on head Not remarkable a Bird found dead during test. Pens 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 X . XX - XXX - XX - - XXX - - - - - - -X - - - - - - - - - - - - - -X - - - - - - - - - -X - -X " -X - -X X " - " Project Number 454-116 Contains No CBI Appendix - IV - Table 3 Individual Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485 Birds Euthanized at Test Termination 300 ppm a.i. Males External - feather loss External - wart-like growth (-0.25 cm dia.) under beak External - foot lesion Liver - slightly mottled Adominable cavity - extensive fat Testes - small, -1.25 cm Testes - right testis small, -1.25 cm Testes - left testis small, -1.25 cm Not remarkable Pens 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 _ _ _ _ _ X _ _X _ _ _ _ -X- - - - - - - - - - - - - - - - -X- - - - - - - - - - - - --- - -X- - --- - --- - - - - - -X- - - - - - - - - - - - - - - - - - X - - - - - - - - - - - - - - - - - - - -X - - - - - - - - - - - - - - - -X - X -X -X - -X -XX -X - -X to Project Number 454-116 Females External - feather loss External - foot lesion Abdominal cavity - egg yolk peritinitus Not remarkable Pens 1633 1634 1635 1636 1637 1638 1639a 1640 1641 1642 1643 1644 1645 1646 1647 1648 XXX X _ _ _ X XXX X X _ XX - - - - - - -X - - - - --- - - - - - - - - - - - - - - - -X - - - -XX - - - - - - -X - - a Bird euthanized during test. Contains No CBI Males External - feather loss External - foot lesions External - abcess (-0.05 cm dia.) above left eye Testes - small, ~1.5 and -1.25 cm Testes - small, -1.25 cm Testes - right testis small, -1.5 cm Testes - right testis small, -1.25 cm Testes - right testis small, -1.0 cm Not remarkable Appendix - IV - Table 4 Individual Gross Pathological Observations from a Northern Bobwhite Reproduction Study with T-7485 Birds Euthanized at Test Termination 900 ppm a.i. Pens 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 ___ X _ _ _ _X _X _X . - -X - -X - - - - -- - - - - - - -X - - - - - - - - - - - - X---- -------- --- - - - - - - - - - -X - - - - - -X - - - - - - - - - - - - - - - -X - -X - -X - - - - - - - - - - - - - -X - - - - - - - - - - - - X "X " "X -X X -X Project Number 454-116 Females Pens 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 External - feather loss XXX X X XXX _ XXXX _X _ External - foot/leg lesions X - - - - - - - - -- - -XXX Spleen - slightly enlarged (-1.5 x 1.0 cm) - - - - - - X - - - - - - - - - Liver - friable - - - - - - - - - - - - - -X - Abdominal cavity - egg yolk peritinitus - -X - - - - - - - - - - -X - Not remarkable - "- - " - -X "- - "- Contains No CBI Appendix V - Table 1 A du lt Body Weight (g) from a Northern Bobwhite Reproduction Study w ith T-7485 (Control, Males) Pen 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term Test Term 184 -4 199 0 182 5 201 -1 211 7 205 7 179 -3 203 -1 183 3 195 2 200 -1 179 2 179 1 214 -4 198 1 173 -5 193 1 13 4 180 -3 199 -2 187 5 200 3 218 6 212 5 176 1 202 5 186 0 197 3 199 0 181 0 180 3 210 4 199 7 168 4 193 3 14 3 177 2 197 5 192 3 203 2 224 4 217 8 177 5 207 3 186 3 200 1 199 1 181 -3 183 6 214 2 206 4 172 5 196 3 16 3 179 7 202 6 195 1 205 3 228 -2 225 -4 182 5 210 7 189 6 201 1 200 3 178 7 189 8 216 5 210 4 111 5 199 4 16 3 186 2 208 33 196 13 208 -2 226 -7 221 15 187 -1 217 -11 195 0 202 13 203 -4 185 13 197 -1 221 13 214 -28 182 -9 203 2 14 14 188 241 209 206 219 236 186 206 195 215 199 198 196 234 186 173 205 20 Total Change 4 42 27 5 8 31 7 3 12 20 -1 19 17 20 -12 0 13 14 The means for body weights and body weight changes are calculated and rounded separately. Project Number 454-116 Contains No CBI Appendix V - Table 2 A dult Body Weight (g) from a N orthern Bobwhite Reproduction Study w ith T-7485 (C on trol, Females) Pen 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term Test Term 196 1 197 4 191 0 191 0 174 3 177 2 179 -7 . 172 11 182 ii 193 8 211 5 216 3 202 -2 200 5 187 2 189 4 191 0 191 -2 197 -1 196 3 175 5 180 4 192 2 194 3 207 -4 203 7 178 -2 176 1 199 1 200 9 177 -2 175 1 201 191 179 183 201 219 205 193 189 199 184 197 210 177 209 176 4 205 -1 204 34 6 197 3 200 49 1 180 8 188 34 3 186 4 190 41 8 209 -2 207 25 7 226 9 235 22 0 205 1 206 22 2 195 6 201 - 8 197 -7 190 64 6 205 4 209 31 0 184 6 190 30 3 200 2 202 45 5 215 8 223 42 3 180 2 182 33 6 215 0 215 31 3 179 2 181 50 190 1 191 4 195 4 199 3 201 37 12 4 12 3 13 3 14 4 15 12 238 249 222 231 232 257 228 - 254 240 220 247 265 215 246 231 238 15 Total Change 42 58 48 52 50 46 26 _ 63 43 45 55 58 37 47 54 48 9 The means for body weights and body weight changes are calculated and rounded separately. - Data are not available due to adult mortality. Project Number 454-116 Contains No CBI Appendix V - Table 3 A dult Body Weight (g) from a Northern Bobwhite R eproduction Study w ith T-7485 ( 1 0 0 PPM A . I . , M a l e s ) Pen 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term 197 7 186 1 197 8 195 5 203 2 196 3 207 -2 197 -1 182 4 204 -5 210 1 189 6 182 -3 212 2 193 -12 174 3 204 4 187 -i 205 2 200 7 205 1 199 3 205 3 196 3 186 4 199 1 211 2 195 7 179 3 214 4 181 5 177 2 208 _ 186 4 207 10 207 8 206 4 202 4 208 4 199 1 190 4 200 3 213 7 202 8 182 4 218 5 186 -1 179 2 . 190 4 217 8 215 7 210 5 206 7 212 2 200 -1 194 7 203 2 220 -1 210 -1 186 9 223 11 185 5 181 6 194 2 225 -3 222 3 215 8 213 2 214 1 199 11 201 7 205 9 219 -6 209 1 195 12 234 4 190 28 187 19 Test Term 196 222 225 223 215 215 210 208 214 213 210 207 238 218 206 195 1 196 3 200 4 203 5 208 7 11 5 11 2 12 3 14 4 14 9 215 10 Total Change 10 25 30 20 19 8 13 26 10 3 21 25 26 25 32 20 9 The means for body weights and body weight changes are calculated and rounded separately. - Data are not available due to adult mortality. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix V - Table 4 Adult Body Weight (g) from a Northern Bobwhite Reproduction Study w ith T-7485 ( 1 0 0 PPM A . I . , F e m a l e s ) Pen 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Test Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Terra Term 203 2 196 -1 188 -3 176 -2 187 3 195 2 197 -5 182 2 180 2 196 1 189 -3 192 2 175 -2 184 2 181 -1 192 5 188 0 83 205 1 195 -18 185 4 174 3 190 4 197 2 192 3 184 0 182 4 197 6 186 5 194 1 173 7 186 0 180 3 197 11 189 2 96 206 2 177 22 189 2 177 1 194 3 199 2 195 5 184 -5 186 5 203 1 191 3 195 10 180 4 186 3 183 2 208 7 191 4 10 6 208 3 199 -7 191 8 178 5 197 2 201 4 200 3 179 -16 191 5 204 2 194 0 205 3 184 1 189 6 185 4 215 7 195 2 11 6 211 33 192 44 199 52 183 46 199 36 205 39 203 11 163 52 196 44 206 40 194 43 208 55 185 25 195 39 189 41 222 36 197 40 13 11 244 236 251 229 235 244 214 215 240 246 237 263 210 234 230 258 237 15 Total Change 41 40 63 53 48 49 17 33 60 50 48 71 35 50 49 66 48 13 The means for body weights and body weight changes are calculated and rounded separately. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix V - Table 5 A du lt Body Weight (g) from a Northern Bobwhite Reproduction Study w ith T-7485 ( 3 0 0 PPM A . I . , M a l e s ) Pen 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term Test Term 190 5 206 -1 193 2 200 2 197 5 201 -2 193 3 190 3 177 1 206 1 183 2 182 -1 184 2 186 2 194 6 182 2 192 2 92 195 1 205 -1 195 9 202 0 202 3 199 0 196 2 193 5 178 3 207 -2 185 8 181 3 186 5 188 1 200 3 184 1 194 3 93 196 5 204 2 204 5 202 4 205 7 199 11 198 5 198 8 181 6 205 3 193 7 184 5 191 7 189 3 203 7 185 0 196 5 83 201 6 206 -1 209 5 206 6 212 4 210 9 203 9 206 16 187 4 208 6 200 4 189 5 198 9 192 9 210 5 185 4 201 6 94 207 22 205 7 214 -4 212 -i 216 18 219 30 212 -7 222 -2 191 -7 214 20 204 4 194 4 207 8 201 9 215 14 189 8 208 8 10 11 229 212 210 211 234 249 205 220 184 234 208 198 215 210 229 197 215 17 Total Change 39 6 17 11 37 48 12 30 7 28 25 16 31 24 35 15 24 12 The means for body weights and body weight changes are calculated and rounded separately. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix V - Table 6 A dult Body Weight (g) from a Northern Bobwhite Reproduction Stu dy w ith T-7485 ( 3 0 0 PPM A . I . , F e m a l e s ) Pen 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term Test Term 205 -4 186 -1 197 2 184 8 197 -10 171 4 185 9 199 2 171 2 183 7 200 i 185 8 174 2 191 -3 185 2 205 1 189 2 11 5 201 '4 185 3 199 4 192 0 187 6 175 2 194 -1 201 4 173 0 190 4 201 i 193 3 176 -1 188 1 187 12 206 2 191 3 10 3 205 7 188 5 203 2 192 4 193 5 177 2 193 1 205 6 173 3 194 4 202 2 196 5 175 4 189 10 199 7 208 6 193 5 11 2 212 2 193 5 205 -8 196 6 198 7 179 2 194 1 211 -4 176 7 198 5 204 11 201 6 179 1 199 12 206 1 214 3 198 4 12 5 214 43 198 25 197 50 202 26 205 58 181 34 195 - 207 44 183 38 203 25 215 33 207 53 180 21 211 22 207 29 217 45 201 36 12 12 257 223 247 228 263 215 - 251 221 228 248 260 201 233 236 262 238 19 Total Change 52 37 50 44 66 44 _ 52 50 45 48 75 27 42 51 57 49 11 The means for body weights and body weight changes are calculated and rounded separately. - Data are not available due to adult mortality. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix V - Table 7 A dult Body Weight (g) from a Northern Bobwhite R eproduction Study w ith T-7485 ( 9 0 0 PPM A . I . , M a l e s ) Pen 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term Test Term 202 0 197 2 189 3 194 -3 204 -4 194 3 191 2 204 4 189 4 186 0 186 2 208 -8 188 2 193 6 179 3 184 2 193 1 84 202 10 199 2 192 6 191 8 200 2 197 -2 193 4 208 0 193 4 186 6 188 6 200 5 190 11 199 2 182 5 186 0 194 4 74 212 4 201 6 198 9 199 6 202 1 195 8 197 5 208 6 197 3 192 5 194 3 205 13 201 5 201 5 187 4 186 1 198 5 73 216 9 207 8 207 4 205 8 203 11 203 6 202 9 214 7 200 7 197 1 197 6 218 4 206 2 206 5 191 1 187 2 204 6 83 225 6 215 33 211 -5 213 8 214 22 209 -4 211 -ii 221 -6 207 9 198 -3 203 3 222 0 208 -7 211 15 192 -2 189 13 209 4 10 12 231 248 206 221 236 205 200 215 216 195 206 222 201 226 190 202 214 16 Total Change 29 51 17 27 32 11 9 11 27 9 20 14 13 33 11 18 21 12 The means for body weights and body weight changes are calculated and rounded separately. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix V - Table 8 A dult Body Weight (g) from a Northern Bobwhite Reproduction Study with T-7485 ( 9 0 0 PPM A . I . , F e m a l e s ) Pen 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 Mean SD Week 0 Change Week 2 Change Week 4 Change Week 6 Change Week 8 Change Week 0-2 Week 2-4 Week 4-6 Week 6-8 Week 8-Term Test Term 208 0 208 5 213 -1 212 4 216 27 179 1 180 3 183 2 185 4 189 23 180 0 180 4 184 0 184 3 187 40 171 2 173 2 175 3 178 -2 176 29 179 1 180 6 186 3 189 4 193 19 176 7 183 5 188 6 194 7 201 45 220 5 225 0 225 7 232 7 239 35 190 3 193 2 195 0 195 5 200 49 191 1 192 3 195 5 200 6 206 30 193 2 195 4 199 3 202 7 209 23 183 3 186 5 191 3 194 4 198 17 196 1 197 5 202 4 206 5 211 45 181 2 183 0 183 3 186 5 191 34 184 2 186 2 188 2 190 1 191 21 189 2 191 15 206 2 208 3 211 27 190 1 191 -1 190 1 191 5 196 9 188 2 190 4 194 3 197 4 201 30 12 2 13 4 13 2 13 2 15 11 243 212 227 205 212 246 274 249 236 232 215 256 225 212 238 205 230 20 Total Change 35 33 47 34 33 70 54 59 45 39 32 60 44 28 49 15 42 14 The means for body weights and body weight changes are calculated and rounded separately. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix VI - Table 1 Feed Consumption (g /b ird /d a y ) from a Northern Bobwhite Reproduction Stu dy w ith T-7485 (C o n tro l) WE E K S Pen 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 18 21 20 21 20 20 20 18 19 22 18 22 21 23 22 22 28 26 28 28 27 17 17 17 15 16 17 15 18 17 20 20 23 22 22 28 21 27 25 29 24 24 15 15 14 14 13 13 14 15 13 15 15 16 16 16 20 18 21 22 24 19 21 13 13 15 15 15 14 13 15 15 16 18 17 19 19 21 19 29 23 23 23 25 17 19 19 19 20 19 16 15 14 22 24 20 19 22 31 17 24 23 23 23 24 15 16 15 18 16 16 14 15 13 15 18 20 20 21 23 21 24 23 25 23 23 12 14 15 14 14 13 13 14 13 14 16 15 17 21 16 18 21 18 15 18 22 17 17 16 18 17 18 22 23 15 19 20 - - - - - - - - - - 15 16 15 15 16 15 15 15 16 16 20 26 26 25 28 24 28 26 26 25 25 16 16 16 15 16 15 16 20 18 19 20 21 20 23 23 21 # 23 27 23 # 16 16 15 15 16 15 16 19 19 20 17 18 20 22 24 20 24 26 26 25 25 14 14 14 13 14 12 13 15 14 16 15 14 13 17 20 17 23 21 22 21 23 15 15 17 17 17 16 17 18 16 18 18 17 19 22 24 19 25 23 25 24 26 17 17 18 25 20 17 17 20 21 22 22 24 26 25 27 23 27 25 29 25 25 16 17 17 17 17 16 16 16 16 18 18 20 19 23 26 22 26 24 23 21 24 13 13 13 14 13 13 12 15 13 16 16 17 16 14 19 20 22 22 23 20 23 Mean SD 15 16 16 16 16 15 15 17 16 18 18 19 19 21 23 20 25 23 25 23 24 222322333 3 333342 32 3 2 2 # Feed wet, data not used. - Data are not available due to adult mortality. Project Number 454-116 Contains No CBI Appendix VI - Table 2 Feed Consumption (g /b ird /d a y ) from a Northern Bobwhite R eproduction Study w ith T-7485 ( 1 0 0 PPM A . I . ) Pen 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 WE E K S 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 _19 17 16 17 13 - - - - - _ _ _ _ _ _ ___ . 15 16 17 13 14 17 16 16 18 18 15 18 20 22 22 19 20 20 20 20 22 15 18 19 16 23 27 20 20 18 17 18 24 23 25 29 21 25 23 24 23 23 17 18 19 21 17 15 16 17 16 17 16 16 17 19 21 21 23 22 23 22 22 16 16 16 15 17 15 16 16 13 17 18 18 22 22 25 24 25 26 26 24 26 18 18 17 18 17 16 15 19 17 18 21 20 20 25 25 24 27 26 27 26 26 15 15 16 17 17 14 14 16 15 15 15 16 18 16 19 19 22 23 26 23 22 17 20 18 17 17 12 15 16 15 16 19 20 21 24 26 22 27 24 25 27 30 15 16 17 16 17 16 16 16 17 18 18 22 23 24 25 21 24 22 25 22 23 18 17 18 18 17 16 17 19 18 18 18 21 23 20 22 18 19 21 21 20 20 16 15 17 16 16 15 15 14 14 16 18 18 19 24 26 22 25 24 26 24 26 17 17 17 18 19 18 19 18 15 19 21 18 22 25 27 21 25 24 24 24 24 15 15 16 18 17 17 15 16 14 20 20 19 25 22 22 18 22 21 21 20 23 15 18 16 17 16 16 18 18 18 18 18 19 20 23 24 20 28 25 29 23 24 14 15 16 15 14 13 14 15 16 19 20 25 21 23 24 21 26 24 24 21 23 15 17 16 15 16 15 15 17 17 17 18 18 19 25 26 23 26 25 26 24 24 Mean SD 16 17 17 17 17 16 16 17 16 18 18 19 21 22 24 21 24 23 24 23 24 121223 2 22 12 2 232 2 3232 2 - Data are not available due to adult mortality. Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix VI - Table 3 Feed Consumption (g /b ird /d a y ) from a Northern Bobwhite Reproduction Study w ith T-7485 ( 3 0 0 PPM A . I . ) Pen 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 WEEKS 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 15 16 15 15 16 19 16 20 18 21 21 23 20 22 29 24 28 27 29 26 26 19 19 15 16 18 17 16 18 17 18 18 20 21 25 27 24 25 24 24 23 23 18 18 17 18 18 16 15 19 18 25 24 23 29 25 30 24 30 30 28 25 27 16 18 18 16 17 16 15 18 15 17 14 16 21 23 23 22 25 26 26 23 24 17 14 19 16 21 15 15 18 22 17 19 20 23 27 28 27 28 24 27 24 25 14 14 14 12 14 14 14 16 13 13 14 14 18 21 21 20 22 20 23 19 20 18 20 18 18 20 19 18 22 18 20 22 22 22 _ - - - _ _ _ _ 17 18 17 18 17 16 15 18 17 18 20 19 23 9 28 25 28 28 28 25 25 16 15 15 15 16 15 16 18 18 16 18 17 23 22 25 20 30 29 31 29 28 16 16 16 16 15 15 16 17 15 16 18 17 18 20 22 22 23 21 24 23 23 15 16 16 16 16 16 16 18 18 20 20 24 20 24 26 26 27 25 28 25 24 16 15 16 16 15 14 14 15 17 17 16 18 21 23 23 24 25 24 24 22 23 15 15 14 13 14 13 12 18 14 19 19 19 20 20 22 18 21 25 21 20 20 15 16 14 14 15 14 15 17 16 18 18 18 18 19 21 22 23 20 21 21 21 17 18 18 17 20 17 16 16 16 18 17 17 20 25 28 22 26 24 27 24 25 17 19 17 18 16 15 14 18 15 18 20 24 25 24 26 24 26 24 26 22 25 Mean SD 16 17 16 16 17 16 15 18 16 18 19 19 21 22 25 23** 26 25 26 23 24 12 2 22212 23333 432 33322 - Data are not available due to adult mortality. * * Significantly different from the control at p < 0.01 Project Number 454-116 Contains No CBI Appendix VI - Table 4 Feed Consumption (g /b ird /d a y ) from a Northern Bobwhite R eproduction Study w ith T-7485 ( 9 0 0 PPM A . I . ) Pen 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 WEEKS 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 16 19 18 17 17 16 19 20 16 17 18 19 22 22 27 23 25 22 25 22 22 15 16 17 16 16 14 14 17 16 16 17 17 19 20 23 22 23 21 24 22 22 17 23 21 20 24 15 16 20 17 19 17 18 18 22 26 24 26 24 25 22 24 17 15 20 17 17 15 16 20 17 18 19 21 20 23 24 24 24 23 25 24 26 16 16 17 17 18 16 17 17 17 19 18 22 23 22 26 21 27 26 26 24 23 16 16 15 16 16 15 15 17 15 17 18 17 20 19 26 24 28 27 25 24 26 16 16 16 14 14 16 14 16 15 14 16 17 20 21 23 22 23 22 24 22 22 17 17 17 16 17 18 16 19 14 16 20 22 21 24 27 24 26 26 24 22 21 17 17 19 16 17 16 15 18 16 16 15 19 17 23 26 23 28 24 26 23 24 15 17 17 19 19 17 18 18 17 20 21 23 21 24 27 26 26 25 24 22 24 17 17 15 14 16 15 15 16 17 17 18 18 22 24 24 19 25 25 26 22 24 15 18 16 16 17 17 17 19 17 21 20 22 21 23 27 25 25 27 30 24 29 20 19 16 19 16 19 16 20 18 19 14 21 21 25 29 25 31 27 34 29 30 16 16 15 16 16 15 14 16 15 15 17 19 19 22 19 15 18 17 18 16 22 16 20 18 19 19 17 17 17 17 16 17 17 18 13 12 18 25 23 26 23 22 16 16 14 15 15 14 13 16 16 17 18 17 18 22 22 20 22 23 23 16 20 Mean SD 16 17 17 17 17 16 16 18 16 17 18 19 20 22 24 22 25 24 25 22 24 12222122 12 2 2234 332 3 3 3 Differences between control and this treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI R e p r o d u c tiv e Perform ance b y Pen Appendix VII - Table 1 E g g s L a i d / Maximum L a i d (%) from a Northern Bobwhite R eproduction Study w ith T-7485 Eggs Replicate Laid 0 PPM A. I . Max. Laid Arcsin % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 67 69 97 80.20 61 69 88 70.09 53 69 77 61.21 52 69 75 60.24 59 69 86 67.62 66 69 96 77.96 43 69 62 52.13 67 69 97 80.20 53 69 77 61.21 62 69 90 71.43 48 69 70 56.52 62 69 90 71.43 54 69 78 62.21 59 69 86 67.62 60 69 87 68.83 866 1035 58 69 70 84 67.26 10 8.40 Eggs Laid 100 PPM A.. 1 . Max. Laid Arcsin % Trans. - __ . 41 69 59 50.43 63 69 91 72.85 56 69 81 64.27 57 69 83 65.35 57 69 83 65.35 45 69 65 53.86 43 69 62 52.13 67 69 97 80.20 56 69 81 64.27 62 69 90 71.43 61 69 88 70.09 52 69 75 60.24 56 69 81 64.27 61 69 88 70.09 63 69 91 72.85 840 1035 56 69 80 81 65.18 11 8.34 Eggs Laid 300 PPM A.I. Max. Laid Arcsin % Trans. 61 69 88 70.09 46 69 67 54.74 69 69 100 90.00 61 69 88 70.09 63 69 91 72.85 56 69 81 64.27 - -- - 61 69 88 70.09 56 69 81 64.27 53 69 77 61.21 60 69 87 68.83 62 69 90 71.43 69 69 100 90.00 53 69 77 61.21 60 69 87 68.83 67 69 97 80.20 897 1035 60 69 60 87 70.54 9 9.88 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). Eggs Laid 900 PPM A. I . Max. Laid Arcsin % Trans. 62 69 90 71.43 52 69 75 60.24 44 69 64 52.99 53 69 77 61.21 43 69 62 52.13 64 69 93 74.38 58 69 84 66.47 46 69 67 54.74 41 69 59 50.43 63 69 91 72.85 59 69 86 67.62 58 69 84 66.47 57 69 83 65.35 14 69 20 26.77 40 69 58 49.59 61 69 88 70.09 815 1104 51 69 13 0 74 60.17 19 12.16 Project Number 454-116 Contains No CBI Eggs Appendix UII - F i g ur e l a Laid frorn a N o r t h e r n Bobuhite Reproduction Study uith T-7485 - Control aso NUMBER / PEN Project Number 454-116 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 mm Eggs Laid P en Number Data a re not a v a ila b le due to adult m o rta lity . 1613 1614 1615 1616 Contains No CBI Eggs Appendix UII - F i g u r e lb Leid from e Northern Bobuhite R e p r o d u c t i o n Study ui t h T - 7 4 8 5 - 100 PPM A.I. u Cl CL mL J O0N0 Project Number 454-116 1617 1<518 1619 1620 1621 1622 1623 1624 1625 1626 Pe n Nu mb e r TM Eggs Laid :$: D ata a r e not a v a i l a b l e d u e to a d u l t mor Contains No CBI NUMBER / PEN Eggs Appendix UII - F i g u r e l c Laid from a Northern Bobuhite R e pr o d u c t i o n Study ui t h T - 7 4 85 - 3 0 0 PPM A.I. 70 60 50 to 30 20 10 0 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 Pe n Nu mb e r EHH E g g s L a id 5ji D a t a a r e n o t a v a i l a b l e d u e t o a d u l t m o r t a l i t y . Osos Project Number 454-116 Contains No CBI NUMBER / PEN Eggs Appendix UII - F i g u r e Id Laid from a Northern Bobuhite R e p r o d u c t i o n Study ui t h T - 7 4 8 5 - 9 00 PPM A.I. 70 60 50 40 30 20 10 0 EHI Eggs Laid <i o Project Number 454-116 Contains No CBI R e p r o d u c tiv e Perform ance b y Pen Appendix VII - Table 2 E g g s C r a c k e d / E g g s L a i d (%) from a Northern Bobwhite Reproduction Study with T-7485 0 P P M A. I . Replicate Eggs Crack Eggs Laid Arcsin % Trans. 1 0 67 0 0.00 2 0 61 0 0.00 3 0 53 0 0.00 4 0 52 0 0.00 5 1 59 2 7.48 6 1 66 2 7.07 7 1 43 2 8.77 8 "- * 9 0 67 0 0.00 10 0 53 0 0.00 11 0 62 0 0.00 12 0 48 0 0.00 13 0 62 0 0.00 14 0 54 0 0.00 15 0 59 0 0.00 16 1 60 2 7.42 1 0 0 P P M A..1. Eggs Crack Eggs Laid Arcsin % Trans. _ _ _ 0 41 0 63 0 56 0 57 0 57 1 45 0 43 0 67 0 56 0 62 0 61 0 52 0 56 0 61 1 63 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 2 8.57 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 2 7.24 300 PPM A.I. Eggs Crack Eggs Laid Arcsin % Trans. 1 61 2 7.36 0 46 0 0.00 1 69 1 6.91 0 61 0 0.00 0 63 0 0.00 0 56 0 0.00 - -- - 0 61 0 0.00 1 56 2 7.68 0 53 0 0.00 0 60 0 0.00 1 62 2 7.30 0 69 0 0.00 0 53 0 0.0 0 0 60 0 0.00 2 67 3 9.95 Total Mean SD 4 866 0 58 17 1 2.05 1 3.54 2 840 0 56 08 0 1.05 1 2.79 6 897 0 60 16 1 2.61 1 3.88 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). 900 P P M A. I . Eggs Crack Eggs Laid Arcsin % Trans. 1 62 1 52 0 44 0 53 0 43 0 64 1 58 0 46 0 41 0 63 0 59 0 58 2 57 1 14 0 40 1 61 7 815 0 51 1 13 2 7.30 2 7.97 0 0.00 0 0.00 0 0.00 0 0.00 2 7.55 0 0.00 0 0.00 0 0.00 0 0.00 0 0.00 4 10.80 7 15.50 0 0.00 2 7.36 1 3.53 2 5.07 Project Number 454-116 Contains No CBI Eggs Appendix UII - F i g u r e 2a Cracked / Eggs Laid from a Northern Bobuhite Reproduction Study uith T-7485 - Control 70 60 50 LJ CL \ 40 C mL J 30 20 10 0 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 P en Number Eggs Laid ^ 3 Eggs Cracked Data a re not a v a ila b le due to adult m o rtality. ro Project Number 454-116 Contains No CBI 70 60 50 CL 40 \ aLtnJ 30 20 10 Eggs Appendix UII - F i g u r e 2b Cracked / Eggs Laid from a Northern Bobuhite R e pr o d u c t i o n Study with T - 7 4 8 5 - 100 PPM A.I. 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 Pen N u m b e r Eggs Laid ^ 3 Eggs Cracked Data a re not a v a ila b le due to adult m o rta lity . 1629 1630 1631 1632 Project Number 454-116 Contains No CBI Eggs Appendix UII - F i g ur e 2c Cracked / Eggs Laid from a Northern Bobuhite R e p r od uc t i on Study ui t h T - 7 48 5 - 3 00 PPM A.I. <4^1 Project Number 454-116 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 P en Number Eggs Laid Eggs Cracked ijc D ata a r e not a v a i l a b l e d ue to a d u lt m o r t a l i t y . 1645 1646 1647 1648 Contains No CBI Eggs Appendix UII - F i g u r e 2d Cracked / Eggs Laid from a Northern Bobuhite R e p r o d u c t i o n Study with T - 7 4 8 5 - 9 0 0 PPM A.I. -j NUMBED / PEN Project Number 454-116 Contains No CBI R e p r o d u c t i v e P erform an ce b y Pen Appendix VII - Table 3 V i a b l e E m b r y o s / E g g s S e t (%) from a Northern Bobwhite R eproduction Study w ith T-7485 0 PPM A. I . Viable Eggs Replicate Embr. Set Arcsin % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 58 62 94 75..29 44 56 79 62..42 49 49 100 90..00 45 47 96 78..10 46 53 87 68 .,69 59 60 98 82..58 36 38 95 76..74 - -- - 57 62 92 73..50 40 47 85 67..30 54 57 95 76..74 43 44 98 81..33 16 57 28 31..99 48 49 98 81..79 45 53 85 67..14 52 53 98 82..10 692 787 46 52 89 73 .05 11 7 18 13 .49 100 PPM A..1. Viable Eggs Embr. Set Arcsin % Trans. . 35 36 97 80..41 56 58 97 79..30 51 51 100 90..00 52 53 98 82..10 52 52 100 90.,00 37 39 95 76..91 25 38 66 54 .20 61 62 98 82 .70 45 50 90 71..57 57 57 100 90,.00 56 56 100 90..00 39 48 81 64..34 40 51 78 62..33 56 56 100 90 .00 52 56 93 74 .50 714 763 48 51 93 78 .56 10 8 10 11 .39 300 PPM t L.I . Viable Eggs Embr. Set Arcsin % Trans. 54 54 100 90..00 40 42 95 77 .40 60 63 95 77 .,40 39 56 70 56 .57 53 58 91 72 .93 49 51 96 78..58 - -- - 51 56 91 72 .61 39 50 78 62 .03 48 48 100 90..00 54 55 98 82 .25 55 56 98 82 .32 57 63 90 72 .02 42 48 88 69 .30 55 55 100 90 .00 57 59 97 79 .39 753 814 50 54 92 76 .85 7 6 9 9 .76 900 PPM A. I . Viable Eggs Embr. Set Arcsin % Trans. 54 56 96 79. 11 45 45 100 90..00 40 40 100 90..00 48 48 100 90..00 38 39 97 80..79 46 59 78 62 .00 47 53 89 70..34 39 41 95 77 .24 36 37 97 80,.54 55 58 95 76..85 53 54 98 82 .18 52 53 98 82 .10 51 51 100 90 .00 9 11 82 64 .76 33 36 92 73 .22 51 55 93 74 .35 697 736 44 46 94 78 .97 11 12 6 8 .71 - Data are not available due to adult mortality. Differences between the control and each treatment group were not siignificant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI NUMBER / PEN Uiable Appendix UII - F i g u r e 3a Embryos / Eggs Set from a Northern Bobuhite Reproduction Study uith T-7485 - Control 70 60 50 40 30 20 10 0 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 Pen Number ^=3 Eggs Set ^ 3 Uiable Embryos Data a r e not a v a ila b le due to adult mortality --a Project Number 454-116 Contains No CBI Uiable Appendix UII - F i g u r e 3b Embryos / Eggs Set from a Northern Bobuhite R e p r o d u c t i o n Study uith T - 7 4 8 5 - 100 PP(1 A.I. Project Number 454-116 CL -4 \ oo UCmLl 161? 1618 1613 1620 1621 1622 1623 1624 1625 1626 162? Pen Number Eggs Set Uiable Embryos ^ Data a re not a v a ila b le due to adult m o rtality 1628 1629 1630 1631 1632 Contains No CBI Uiable Appendix UII - F i g ur e 3c Embryos / Eggs Set from a Northern Bobuhite R e pr oduc t io n St udy uith T - 7 4 8 5 - 3 0 0 PPM A.I. 70 -VUO NUMBER / PEN Project Number 454-116 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 Pen Number TM Eggs Set ^ 3 Uiable Embryos Data a r e not a v a i l a b l e due to adult m o rtality. 1645 1646 1647 1648 Contains No CBI Uiable Appendi x UII - F i g u r e 3d Embryos / Eggs Set from a Northern Bobuhite R e p r o d u c t i o n Study ui t h T - 7 4 8 5 - SOO PPM A.I. ooo NUMBER / PEN Project Number 454-116 Contains No CBI R e p r o d u c tiv e Perform ance b y Pen Appendix VII - Table 4 L i v e 3 -Week E m b r y o s / V i a b l e E m b r y o s (%) from a Northern Bobwhite Reproduction S tu dy w ith T-7485 0 PPM A. I. Live Viable Arcsin Replicate 3-Week Embr. % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 58 58 100 90.00 44 44 100 90.00 49 49 100 90.00 45 45 100 90.00 46 46 100 90.00 59 59 100 90.00 36 36 100 90.00 _ -- - 57 57 100 90.00 40 40 100 90.00 54 54 100 90.00 43 43 100 90.00 15 16 94 75.52 46 48 96 78.22 45 45 100 90.00 52 52 100 90.00 689 692 46 46 99 88.25 11 11 2 4.65 100 PPM A..I . Live Viable Arcsin 3-Week Embr. % Trans. . 35 35 100 90.00 56 56 100 90.00 51 51 100 90.00 50 52 96 78.69 52 52 100 90.00 37 37 100 90.00 25 25 100 90.00 61 61 100 90.00 45 45 100 90.00 57 57 100 90.00 56 56 100 90.00 39 39 100 90.00 40 40 100 90.00 55 56 98 82.32 51 52 98 82.03 710 714 47 48 99 88.20 10 10 1 3.80 300 PPM A. I. Live Viable Arcsin 3-Week Embr. % Trans. 53 54 98 82.18 40 40 100 90.00 59 60 98 82.58 39 39 100 90.00 53 53 100 90.00 49 49 100 90.00 - -- - 50 51 98 81.95 38 39 97 80.79 47 48 98 81.70 54 54 100 90.00 54 55 98 82.25 57 57 100 90.00 41 42 98 81.12 55 55 100 90.00 57 57 100 90.00 746 753 50 50 99 86.17 7 7 1 4.26 900 PPM A. I . Live Viable Arcsin 3-Week Embr. % Trans. 54 54 100 90.00 45 45 100 90.00 40 40 100 90.00 48 48 100 90.00 38 38 100 90.00 46 46 100 90.00 46 47 98 81.61 37 39 95 76.91 36 36 100 90.00 54 55 98 82.25 53 53 100 90.00 52 52 100 90.00 51 51 100 90.00 9 9 100 90.00 33 33 100 90.00 51 51 100 90.00 693 697 43 44 99 88.17 11 11 1 4.07 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI NUMBER / PEN Live Appendix UII - F i g u r e 4a 3-Ueek Embryos / Uiable Embryos from a Northern Bobuhite Reproduction Study uith T-7485 - Control 70 60 50 40 30 20 10 0 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 16 H 1615 1616 Pen Number Uiable Embryos ^ 3 Live 3-Ueek Embryos sj; Data a r e not a v a i l a b l e due t o a d u l t m o r t a l i t y . oKo> Project Number 454-116 Contains No CBI Live Appendi x UII - F i g u r e 4b 3-Ueek Embryos / Uiable Embryos from a Northern Bobuhite R e p r o d u c t i o n Study ui t h T - 7 4 8 5 - 1GG PPM A.I. Project Number 454-116 LCJL \ ou>o CL LCJ :z: 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Pen Number Uiable Embryos ^ 3 Live 3-Ueek Embryos 2$c Data a r e not a v a i l a b l e due to a d u l t m o r t a l i t y . Contains No CBI Live Appendix UII - F i g u r e 4c 3-Ueeb Embryos / Uiable Embryos from a Northern Bobuhite R e p r od uc t i on Study ui t h T - 7 4 8 5 - 3 00 PPM A.I. 70 oo NUMBER / PEN Project Number 454-116 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 Pen Number Uiable Embryos 3 Live 3-Ueeb Embryos ^ Data a re not a v a ila b le due to adult mortality. 1645 1646 1647 1648 Contains No CBI Live Appendi x UII - F i g u r e 4d 3-Ueek Embryos / Uiable Embryos from a Northern Bobuhite R e p r o d u c t i o n Study ui t h 1 - 7 4 8 5 - 900 PPM A.I. 70 60 ' NUMBER / PEN Project Number 454-116 Contains No CBI R e p ro d u c tiv e Performance b y Pen Appendix VII - Table 5 H a t c h l i n g s / L i v e 3 -Week E m b ry o s (%) from a Northern Bobwhite R eproduction S tu dy w ith T-7485 0 PPM A. I . Number Live Arcsin Replicate Hatch. 3-Week % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 58 58 100 90.00 41 44 93 74.86 48 49 98 81.79 44 45 98 81.43 46 46 100 90.00 59 59 100 90.00 35 36 97 80.41 - -- - 57 57 100 90.00 39 40 97 80.90 53 54 98 82.18 43 43 100 90.00 10 15 67 54.74 44 46 96 77.96 43 45 96 77.83 51 52 98 82.03 671 689 45 46 96 81.61 12 11 8 9.06 100 PPM A..i . Number Live Arcsin Hatch. 3-Week % Trans. _ .. _ 35 35 100 90.00 55 56 98 82.32 51 51 100 90.00 48 50 96 78.46 52 52 100 90.00 37 37 100 90.00 22 25 88 69.73 61 61 100 90.00 44 45 98 81.43 56 57 98 82.39 54 56 96 79.11 39 39 100 90.00 39 40 97 80.90 50 55 91 72.45 48 51 94 75.96 691 710 46 47 97 82.85 10 10 4 6.94 300 PPM A. I . Number Live Arcsin Hatch. 3-Week % Trans. 46 53 87 68.69 40 40 100 90.00 57 59 97 79.39 37 39 95 76.91 49 53 92 74.05 47 49 96 78.34 - -- - 48 50 96 78.46 36 38 95 76.74 46 47 98 81.61 53 54 98 82.18 49 54 91 72.28 51 57 89 71.07 39 41 95 77.24 55 55 100 90.00 50 57 88 69.49 703 746 47 50 94 77.76 6 7 4 6.43 90Cl PPM A.I. Number Live Arcsin Hatch. 3-Week % Trans. 53 54 98 82.18 45 45 100 90.00 40 40 100 90.00 47 48 98 81.70 38 38 100 90.00 46 46 100 90.00 42 46 91 72.85 35 37 95 76.56 35 36 97 80.41 53 54 98 82.18 50 53 94 76.24 49 52 94 76.10 50 51 98 81.95 9 9 100 90.00 32 33 97 79.98 50 51 98 81.95 674 693 42 43 97 82.63 11 11 3 5.78 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). Project Number 454-116 Contains No CBI Appendix UII - F i g u r e 5a Hatchlings / Live 3-Ueek Embryos from a Northern Bobuhite Reproduction Study uith T-7485 - Control o-Jo NUMBER PEN Project Number 454-116 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 P en Number iasHl L i v e 3 - U e e k E mb r y o s l= = 1 H a t c h l i n g s ^ Data a r e not a v a ila b le due to adult mortality. 1613 1614 1615 1616 Contains No CBI Appendix UII - F i g u r e 5b Hatchlings / Live 3-Ueek Embrgos from a Northern Bobuhite R e pr o d u c t i o n St udy ui t h T - 7 4 8 5 - 100 PPM A.I. oocc Project Number 454-116 Pen Number H ill L i v e 3 - U e e k Embryos f----i H a t c h l i n g s ifi Data a r e not a v a il a b le due to adult m o rtality. Contains No CBI Appendix UII - F i g u r e 5c Hatchlings / Live 3-U eet Embryos from a Northern Bobuhite R e p r o d u c t i o n Study ui t h T - 7 4 8 5 - 3 0 0 PPM A.I. 00 NUMBER / PEN Project Number 454-116 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 P en Number Live 3-Ueek Embryos ^ 3 Hatchlings * Data a re not a v a ila b le due to adult mortality. 1645 1646 1647 1648 Contains No CBI Appendix UII - F i g u r e 5d Hatchlings / Live 3-Ueek Embryos from a Northern Bobuhite R e pr o d u c t i o n Study uith T - 7 4 8 5 - 900 PPM A.I. 70 60 ' NUMBER / PEN Project Number 454-116 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 ___ P en Number L i v e 3 - U e e k Embryos Hatchlings Contains No CBI Reproductive Performance by Pen Appendix VII - Table 6 1 4 -Day Old Survivors / Hatchlings (%) from a Northern Bobwhite Reproduction Study with T-7485 0 ]PPM A. I . 14-Day Number Arcsin Replicate Old Hatch. % Trans. 100 PPM A .I . 14-Day Number Arcsin Old Hatch. % Trans. 300 PPM A.I. 14-Day Number Arcsin Old Hatch. % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 54 58 93 74.77 41 41 100 90.00 47 48 98 81.70 42 44 95 77.69 44 46 96 77.96 56 59 95 76.97 35 35 100 90.00 - -" - 57 57 100 90.00 39 39 100 90.00 48 53 91 72.11 41 43 95 77.55 9 10 90 71.57 42 44 95 77.69 43 43 100 90.00 50 51 98 81.95 648 671 43 45 96 81.33 11 12 3 6.92 __ 34 35 97 80 .27 45 55 82 64 .76 47 51 92 73 .74 39 48 81 64 .34 49 52 94 76 .10 36 37 97 80 .54 15 22 68 55 .66 61 61 100 90..00 41 44 93 74 .86 55 56 98 82 .32 52 54 96 78..90 39 39 100 90.,00 38 39 97 80..79 48 50 96 78..46 47 48 98 81. 70 646 691 43 46 93 76. 83 11 10 9 9. 31 43 46 93 75 .20 40 40 100 90 .00 56 57 98 82 .39 35 37 95 76 .56 48 49 98 81 .79 43 47 91 73 .04 - -- - 46 48 96 78..22 35 36 97 80..41 46 46 100 90..00 53 53 100 90..00 46 49 94 75..67 49 51 96 78 .58 33 39 85 66 .,91 54 55 98 82 ..25 47 50 94 75. 82 674 703 45 47 96 79. 79 7 6 4 6 .60 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). 900i PPM A. I 14-Day Number Arcsin Old Hatch. % Trans. 53 53 100 90 .00 43 45 96 77 .83 40 40 100 90 .00 45 47 96 78 .10 35 38 92 73 .68 43 46 93 75..20 35 42 83 65..91 32 35 91 72 .98 33 35 94 76..17 52 53 98 82 .,10 44 50 88 69..73 48 49 98 81..79 47 50 94 75..82 8 9 89 70..53 31 32 97 79..82 44 50 88 69 .73 633 674 40 42 94 76 .84 11 11 5 6. 83 Project Number 454-116 Contains No CBI 14-Dag Appendi x UII - F i g u r e 6 a Old S u r v i v o r s / H a t ch l i ng s from a No r t he r n Bobuhite Reproduction Study uith T-7485 - Control 70 K> NUMBER / PEN Project Number 454-116 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 P en Number W Hatchlings i i 1.4-Day Old S u r v i v o r s jji Data are not a v a il a b le due to adult m o rtality. 1613 1614 1615 1616 Contains No CBI 14-Dag Ap pe ndi x UII - F i g u r e 6b Old S u r v i v o r s / H a t c h l i n g s from a No r the rn Bobuhite Re p r o du c t i on Study ui t h T - 7 48 5 - 100 PPM A.I. Project Number 454-116 LCdl VO \ U) au. 0C5ZD3 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 Pen Number Hatchlings ^ 14-Da g Old S u r v i v o r s Data a r e not a v a ila b le due to adult mortality. 1629 1630 1631 1632 Contains No CBI NUMBER / PEN M-Day Appendi x UII - F i g u r e 6c Old S u r v i v o r s / H a t c h l i n g s from a No r the rn B obuhi t e R e p r o d u c t i o n Study ui t h T - 7 4 8 5 - 300 PPM A.I. 70 60 ' 50 40 30 20 10 0 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 Pen Numb e r Hatchlings EE3 M - D a y Old S u r v i v o r s * Data are not a v a ila b le due to adult mortality. Project Number 454-116 Contains No CBI 14-Day Appendix UII - F i g u r e 6 d Old S u r v i v o r s / H a t c h l i n g s from a N o r t he r n Bobuhi t e R e pr o d u c t i o n St udy ui t h T - 7 4 8 5 - 3 0 0 PPM A.I. NUMBER / PEN Project Number 454-116 1619 1650 1651 1652 ta rn H a t c h l i n g s 1653 1651 1655 1656 1657 1658 1659 1660 P en Number t----< 1.4-Day Old S u r v i v o r s 1661 1662 1663 1661 Contains No CBI R e p r o d u c tiv e P erform ance b y Pen Appendix VII - Table 7 H a t c h l i n g s / E g g s S e t (%) from a Northern Bobwhite Reproduction Study w ith T-7485 0 p p m a .:r. Number Eggs Replicate Hatch. Set Arcsin % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 58 62 94 75.29 41 56 73 58.83 48 49 98 81.79 44 47 94 75.37 46 53 87 68.69 59 60 98 82.58 35 38 92 73.68 " -- - 57 62 92 73.50 39 47 83 65.63 53 57 93 74.64 43 44 98 81.33 10 57 18 24.76 44 49 90 71.37 43 53 81 64.25 51 53 96 78.80 671 787 45 52 86 70.03 12 7 20 14.23 100 PPM A . 1 . Number Eggs Hatch. Set Arcsin % Trans. __ _ 35 36 97 80 .41 55 58 95 76 .85 51 51 100 90 .00 48 53 91 72 .11 52 52 100 90 .00 37 39 95 76 .91 22 38 58 49..54 61 62 98 82 .70 44 50 88 69..73 56 57 98 82..39 54 56 96 79..11 39 48 81 64..34 39 51 76 60..98 50 56 89 70..89 48 56 86 67. 79 691 763 46 51 90 74 .25 10 8 11 10. 93 300 PPM A.I. Number Eggs Hatch. Set Arcsin % Trans. 46 54 85 67 .36 40 42 95 77 .40 57 63 90 72 .02 37 56 66 54 .37 49 58 84 66 .80 47 51 92 73 .74 - -- - 48 56 86 67..79 36 50 72 58 .05 46 48 96 78 ,.22 53 55 96 79 ,.01 49 56 88 69..30 51 63 81 64 .12 39 48 81 64 .34 55 55 100 90..00 50 59 85 67.,01 703 814 47 54 86 69.,97 6 6 9 8 ..89 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). 900 PPM A. I Number Eggs Hatch. Set Arcsin % Trans. 53 56 95 76..62 45 45 100 90..00 40 40 100 90 .00 47 48 98 81..70 38 39 97 80..79 46 59 78 62..00 42 53 79 62 .90 35 41 85 67..51 35 37 95 76 .56 53 58 91 72..93 50 54 93 74 .,21 49 53 92 74 .05 50 51 98 81..95 9 11 82 64 ..76 32 36 89 70..53 50 55 91 72..45 674 736 42 46 91 74. 93 11 12 7 8. 53 Project Number 454-116 Contains No CBI R e p r o d u c tiv e Perform ance b y Pen Appendix VII - Table 8 1 4 - D a y O l d S u r v i v o r s / E g g s S e t (%) from a Northern Bobwhite R eproduction S tu d y with T-7485 0 PPM A. I . 14-Day Eggs Replicate Old Set Arcsin % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 54 62 87 68 .95 41 56 73 58 .83 47 49 96 78 .34 42 47 89 70 .96 44 53 83 65 .66 56 60 93 75 .04 35 38 92 73 .68 " -- - 57 62 92 73 .50 39 47 83 65..63 48 57 84 66 .59 41 44 93 74 .86 9 57 16 23 .41 42 49 86 67 ..79 43 53 81 64 .25 50 53 94 76. 24 648 787 43 52 83 66 .92 11 7 19 13 .16 100 PPM A..I . 14-Day Eggs Old Set Arcsin % Trans. _ __ . 34 36 94 76 .37 45 58 78 61 .74 47 51 92 73 .74 39 53 74 59 .07 49 52 94 76 .10 36 39 92 73 .90 15 38 39 38 .92 61 62 98 82..70 41 50 82 64 .90 55 57 96 79..20 52 56 93 74 .50 39 48 81 64 .34 38 51 75 59..68 48 56 86 67. 79 47 56 84 66. 37 646 763 43 51 84 67. 96 11 8 15 10. 85 300 PPM i.I. 14-Day Eggs Old Set Arcsin % Trans. 43 54 80 63 .17 40 42 95 77 .40 56 63 89 70 .53 35 56 63 52 .24 48 58 83 65 .47 43 51 84 66..67 - -- - 46 56 82 65..00 35 50 70 56..79 46 48 96 78.,22 53 ' 55 96 79..01 46 56 82 65..00 49 63 78 61..87 33 48 69 56. 01 54 55 98 82. 25 47 59 80 63. 19 674 814 45 54 83 66. 86 7 6 11 8. 99 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). 900 PPM A. I 14-Day Eggs Old Set Arcsin % Trans. 53 56 95 76 .62 43 45 96 77 .83 40 40 100 90 .00 45 48 94 75 .52 35 39 90 71 .32 43 59 73 58 .62 35 53 66 54 .35 32 41 78 62 .06 33 37 89 70 .80 52 58 90 71..24 44 54 81 64 .51 48 53 91 72 .11 47 51 92 73 .74 8 11 73 58 ..52 31 36 86 68 .12 44 55 80 63 .43 633 736 40 46 86 69. 30 11 12 10 8 .93 Project Number 454-116 Contains No CBI R e p r o d u c tiv e Perform ance b y Pen Appendix VII - Table 9 H a t c h l i n g s / Maximum S e t (%) from a Northern Bobwhite R eproduction Study w ith T-7485 0 PPM A.I. Number Max. Replicate Hatch. Set Arcsin % Trans. 100 PPM A.I. Number Max. Hatch. Set Arcsin % Trans. 300 PPM A.I. Number Max. Hatch. Set Arcsin Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 58 63 92 73.64 41 63 65 53.78 48 63 76 60.79 44 63 70 56.69 46 63 73 58.70 59 63 94 75.41 35 63 56 48.19 - -- - 57 63 90 72.02 39 63 62 51.89 53 63 84 66.52 43 63 68 55.71 10 63 16 23.48 44 63 70 56.69 43 63 68 55.71 51 63 81 64.12 671 945 45 63 71 58.22 12 0 19 12.59 - -- 35 63 56 48.19 55 63 87 69.12 51 63 81 64.12 48 63 76 60.79 52 63 83 65.30 37 63 59 50.03 22 63 35 36.22 61 63 97 79.74 44 63 70 56.69 56 63 89 70.53 54 63 86 67.79 39 63 62 51.89 39 63 62 51.89 50 63 79 62.98 48 63 76 60.79 691 945 46 63 73 59.74 10 0 16 10.84 46 63 73 58.70 40 63 63 52.83 57 63 90 72.02 37 63 59 50.03 49 63 78 61.87 47 63 75 59.74 - -- - 48 63 76 60.79 36 63 57 49.11 46 63 73 58.70 53 63 84 66.52 49 63 78 61.87 51 63 81 64.12 39 63 62 51.89 55 63 87 69.12 50 63 79 62.98 703 945 47 63 74 60.02 6 0 10 6.78 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). 900 PPM A.I. Number Max. Hatch. Set Arcsin % Trans. 53 63 84 66.52 45 63 71 57.69 40 63 63 52.83 47 63 75 59.74 38 63 60 50.95 46 63 73 58.70 42 63 67 54.74 35 63 56 48.19 35 63 56 48.19 53 63 84 66.52 50 63 79 62.98 49 63 78 61.87 50 63 79 62.98 9 63 14 22.21 32 63 51 45.45 50 63 79 62.98 674 1008 42 63 11 0 67 55.16 18 11.04 Project Number 454-116 Contains No CBI R e p ro d u c tiv e Performance b y Pen Appendix VII - Table 10 1 4 - D a y O l d S u r v i v o r s / Maximum S e t (%) from a Northern Bobwhite R eproduction S tu dy with T-7485 0 PPM A .:I . 14-Day Max. Replicate Old Set Arcsin % Trans. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 Total Mean SD 54 63 86 67 .79 41 63 65 53 .78 47 63 75 59 .74 42 63 67 54 .74 44 63 70 56 .69 56 63 89 70 .53 35 63 56 48 .19 - -- - 57 63 90 72 .02 39 63 62 51..89 48 63 76 60..79 41 63 65 53..78 9 63 14 22..21 42 63 67 54 .74 43 63 68 55. 71 50 63 79 62. 98 648 945 43 63 69 56. 37 11 0 18 11. 72 100 PPM A .1. 14-Day Max. Old Set Arcsin % Trans. _ __ _ 34 63 54 47 .28 45 63 71 57 .69 47 63 75 59 .74 39 63 62 51 .89 49 63 78 61 .87 36 63 57 49..11 15 63 24 29..21 61 63 97 79..74 41 63 65 53 .78 55 63 87 69..12 52 63 83 65..30 39 63 62 51..89 38 63 60 50. 95 48 63 76 60. 79 47 63 75 59. 74 646 945 43 63 68 56. 54 11 0 17 11. 38 30CI PPM I . 14-Day Max. Old Set Arcsin % Trans. 43 63 68 55 .71 40 63 63 52 .83 56 63 89 70 .53 35 63 56 48 .19 48 63 76 60 .79 43 63 68 55..71 - -- - 46 63 73 58..70 35 63 56 48..19 46 63 73 58 .70 53 63 84 66..52 46 63 73 58..70 49 63 78 61.,87 33 63 52 46..36 54 63 86 67..79 47 63 75 59. 74 674 945 45 63 71 58. 02 7 0 11 7 .16 - Data are not available due to adult mortality. Differences between the control and each treatment group were not significant (p > 0 .0 5 ). 900 PPM A. I 14-Day Max. Old Set Arcsin % Trans. 53 63 84 66 .52 43 63 68 55 .71 40 63 63 52 .83 45 63 71 57 .69 35 63 56 48 .19 43 63 68 55 .71 35 63 56 48 .19 32 63 51 45 .45 33 63 52 46..36 52 63 83 65 ,.30 44 63 70 56..69 48 63 76 60..79 47 63 75 59 .74 8 63 13 20..88 31 63 49 44 .55 44 63 70 56. 69 633 1008 40 63 11 0 63 52 .58 17 10. 81 Project Number 454-116 Contains No CBI R e p r o d u c t i v e P erfo rm a n ce b y Week and Pen Appendix VIII - Table la Eggs L a id b y Week and Pen from a Northern Bobwhite R eproduction Study w ith T-7485 Experimental Group (ppm a .i .) 1 LOT A B C D Control E F G H I J 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 5 3 025 63 -4 4 3 02 3 13 7 3445 65-7 3 606444 66555 66-7 4 6264 5 5 7 7 65 6 66-7 5 74 6 67 6 7 7655 74 -7 67 7 7 77 7 7 75667 6-7 667 7 677 7 7767 7 7 -7 5 77 7 67 7 7 7 666 7 1- 7 667 7 67 7 7 7767 7 0-7 7 67 7 67 7 7 7777 7 5 -7 7 877 677 Totals 44 68 78 91 96 97 101 93 95 103 Totals 67 61 53 52 59 66 43 _ 67 53 62 48 62 54 59 60 - Data are not available due to adult mortality 1LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 866 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen A ppendix V III - Table lb E ggs L a id b y Week and Pen from a N orthern B obw hite R eproduction S tu d y w ith T-7485 Experimental Group (ppm a .i .) PENS ----------------------------- ----------------------------------1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 1 LOT A B C D 100 E F G H I J -0110 0 1 061 3 202 1 4 -2 6 31 4 3 0 6 8 5 44 4 5 4 -5 7 57 5 2 0 7 7 6 655 6 6 -5767 6 4 3 767 766 7 4 -5 7 77 6 4 7 7 5 6 756 7 8 -5777 7 4 6 767 767 7 7 -4 777 7 6 7 7 6 7 766 7 8 -5767 7 7 6 657 776 7 7 -5777 7 7 7 76 7 777 7 7 -5 7 77 8 7 7 7 6 7 767 7 8 Totals 22 59 79 88 94 97 99 97 102 103 Totals _ 41 63 56 57 57 45 43 67 56 62 61 52 56 61 63 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 840 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen Appendix V III - Table lc Eggs L a id b y Week and Pen from a N orthern B obwhite R eprodu ction S tu d y w ith T-7485 Experimental Group (ppm a.i.) PENS ------------------------ -- ---------------------------------------------------------------------------------- 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 Totals 1 LOT A B C D E F G H I J 3 072 3 3 -2 2 01 1 8 217 4 076 6 4 -4 3 35 6 6 44 5 5176 6 5-76476 6 366 74777 7-65667 7 677 7766 7 6 -7 7 76 7 7 677 7777 7 6 -7 6 77 7 6 777 77777 6-76 77 78 577 7576 6 6 -7 7 57 6 7 676 77777 7-77 777 6 777 78777 6-77 77 8 8 778 42 67 81 96 100 102 102 95 104 108 Totals 61 46 69 61 63 56 . 61 56 53 60 62 69 53 60 67 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 897 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform ance b y Week and Pen Appendix V III - Table Id E ggs L a id b y Week and Pen from a N orthern B obwhite R eproduction S tu dy w ith T-7485 Experimental Group (ppm a .i .) 1 LOT A B C D 900 E F G H I J 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 2 1663 1664 50 0102 0 006 3 3002 3 54 132 7 7305 6 553 5 7 54 444 7 442 5 644 54 7 65 663 6 7 54 5 6 664 0 7 765567 7657 577207 77 5657 5 557 6 7702 7 66 5757 7 657 7 6706 7 766767 7667 76707 7 77 6767 7 587 7 77064 77 6767 7 667 6 7708 5 Totals 25 68 73 82 89 88 94 99 98 99 Totals 62 52 44 53 43 64 58 46 41 63 59 58 57 14 40 61 815 LOT A - Eggs Set During W eek 13; LOT B - Eggs Set During W eek 14; etc. 2Female noted with foot lesions beginning during W eek 16 of the test, and did not lay eggs after Week 16. Project Number 454-116 Contains No CBI Appendix UIII - F ig ur e 1 Eggs Laid per Hen from a Nort hern B obuhi t e R e pr oduc t io n Study u i t h T- 74 8 5 Project Number 454-116 Concentrations, ppm a .i. 0 ----100 300 .... 900 Contains No CBI Appendix UIII - F i g u r e 2 Hens Laying per Pen from a Northern Bobuhite Reproduction Study uith T-7485 o NUMBER Project Number 454-116 C oncentrations, ppm a .i. 0 ----1 0 0 ........ 300 ......... 900 Contains No CBI Appendix UIII - F i g u r e 3 Eggs per Layi ng Hen from a Nor the rn B obuhi t e R epr oduc t io n Study ui t h T- 7 48 5 Project Number 454-116 Concentrations, ppm a .i. 0 " " 100 ........... 300 ........... 900 Contains No CBI R e p r o d u c tiv e P erform ance b y Week and Pen A ppendix V III - T able 2a Eggs C racked b y Week and Pen from a N orthern B obw hite R eproduction S tu dy w ith T-7485 Experimental Group (ppm a. i .) PENS -------------- -- ----------------------------------------------- 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 1 LOT A B C D Control E F G H I J 00 0000 0-0 0 0000 00 00 000 0 0 -0 0 0 00 0 0 0 00 000 0 1 -0 0 000 0 00 00 0000 0-0 0 000000 00 000 0 0 -0 0 000 0 00 00 000 0 0 -0 0 000 0 0 0 00 001 0 0 -0 0 0 00 0 0 0 00 000 0 0 -0 0 000 0 0 0 00 0000 0-0 0 0000 01 00 000 1 0 -0 0 000 0 0 0 Totals 0 0 1 0 0 0 1 0 1 1 Totals 0 0 0 0 1 1 1 _ 0 0 0 0 0 0 0 1 4 - Data are not available due to adult mortality ' LOT A - Eggs Set During W eek 12; LOT B - Eggs Set During W eek 13; etc. Project Number 454-116 Contains No CBI R e p r o d u c tiv e P e rfo rm a n c e b y Week and Pen A p p en d ix VIII - T a b le 2b Eggs C racked b y Week and Pen from a N orthern Bobwhite R eproduction S tu dy w ith T-7485 Experimental Group (ppm a.i.) PENS --------------------- ------------------ ------------------------------ ---------------------------- 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 1 LOT A B C D 100 E F G H I J . 000000 0 0000 00 0 0 -0 0 0 0 0 0 0 00 0 0 00 0 0 -0 0 0 0 0 0 0 0 00 0 00 0 0 -0 0 0 0 0 0 0 0 00 0 00 0 0 -00 0 000 0 0000 00 0 1 -00 0 0 0 0 0 0 00 0 00 0 0 -0 0 0 0 01 0 0 00 0 00 0 0 -00 0 000 0 000 0 00 0 0 -000 00 0 0 000 0 00 0 0 -0 0 0 0 0 0 0 0 00 0 00 0 0 Totals - 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 - Data are not available due to adult mortality 1LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 0 0 0 0 1 0 1 0 0 0 2 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen Appendix V III - Table 2c Eggs C ra ck ed b y Week and Pen from a N orthern Bobwhite R eproduction S tu d y w ith T-7485 Experimental Group (ppm a .i .) PENS ----------------------------- ---------------------------------- 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 1 LOT A B C D 300 E F G H I J 00 0 000 - 01 0 0 100 0 0 00 1 00 0 - 0 00 0 000 0 1 00 000 0 - 00 0 0 00 0 0 0 100000 -000 0 00000 00 0 000 - 0 00 0 00 0 0 0 00 000 0 - 00 0 0 000 0 0 00 000 0 - 00 0 0 00 0 01 000 00 0 -0 0 0 0 00 0 0 0 000000 -000 0 00000 00 0 000 - 0 0 0 0 000 0 0 Totals 2 2 0 1 0 0 1 0 0 0 Totals 1 0 1 0 0 0 _ 01 0 0 10002 - Data are not available due to adult mortality 1 LOT A - Eggs Set During W eek 12; LOT B - Eggs Set During W eek 13; etc. 6 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P e rfo rm a n c e b y Week and Pen Appendix V III - Table 2d Egg-s C ra c k e d b y Week and Pen from a N orthern B obw hite R eprodu ction S tu dy w ith T-7485 Experimental Group (ppm a. i .) 1649 1650 1651 1652 1653 1654 1655 1656 1657 PEN S 1658 1659 1660 1661 1662 1663 1664 1 LOT A B C D 900 E F G H I J 0000 0 0 00 0 000 0 00 0 0000 0 0 0 0 0 000 000 0 0000 0 0000 000 0100 1000 0 0 00 0 000 0 00 1 0000 0 0 00 0 000 1 0 0 0 0000 0 0 00 0 000 0 00 0 0100 0 0 00 0 000 0 00 0 0000 0 0 00 0 000 0 00 0 0000 0 0 00 0 000 0 0 0 0 0000 0 010 0 0001000 Totals 0 0 1 2 1 0 1 0 0 2 Totals 1 1 0 0 0 0 1 0 0 0 0 0 2 1 0 1 7 ^LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13;: etc, Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform ance b y Week and Pen A ppendix V III - Table 3a Eggs S e t b y Week and Pen from a N orthern B obwhite R eproduction S tu d y w ith T-7485 Experimental Group (ppm a.i .) 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 1 LOT A B C D Control E F G H I J 4 3 0246 2 -33 2 0130 3 72 4 35 5 5 -72 6 0 634 3 564 54 6 4 -6 4 5 2 544 5 76 6465 6 -74 7 3 656 5 67 5 54 7 3 -6 6 6 7 676 7 76 5566 6 -75 6 6 757 6 67 6657 6 -65 6 7 666 7 76 6 566 1 -75 6 6 757 6 67 6667 0 -67 5 7666 5 76 7 6 7 5 5 -7 6 8 6 7 57 6 Totals 36 62 69 83 88 90 92 86 86 95 Totals 62 56 49 47 53 60 38 _ 62 47 57 44 57 49 53 53 - Data are not available due to adult mortality LOT A - Eggs Set During W eek 12; LOT B - Eggs Set During W eek 13; etc. 787 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen A ppendix V III - Table 3b Eggs S e t b y Week and Pen from a N orthern B obw hite R ep ro d u ctio n S tu d y w ith T-7485 Experimental Group (ppm a .i .) PENS 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Totals LOT A B C D 100 E F G H I J 0010 000 5 122 0204 1621 33 0 6 7534 35 3 5656 510 6 6564 556 4757 541 7 5766 573 5676 637 6 5574 666 4767 645 7 5766 676 4676747 6 6675668 4757 675 6 4 767 576 5676 767 6 6676 767 4767 776 7 5766 67 7 17 52 71 79 85 89 91 89 95 95 Totals 36 58 51 53 52 39 38 62 50 57 56 48 51 56 56 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 763 1 1-- > NJ Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen Appendix V III - Table 3c E ggs S e t b y Week and Pen from a N orthern Bobwhite R eproduction S tu d y w ith T-7485 Experimental Group (ppm a .i .) PENS -- -- -------------------- -- -- ------------------------------------ 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 1 LOT A B C D 300 E F G H I J 20622 3 -2 000 072 0 7 4 06 56 3 - 3 32 5 5634 3 416655 -754 6 6535 6 637676-5556 6757 6 675666 -7675 7666 7 76767 5-6667 6667 6 576766-7 576 7756 6 74 7 56 5 - 6 74 7 575 7 5 676767 -7676 7576 7 77767 5-6767 7767 6 Totals 33 58 74 87 93 94 93 87 97 98 Totals 54 42 63 56 58 51 _ 56 50 48 55 56 63 48 55 59 - Data are not available due to adult mortality LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 814 OJ Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen A ppendix V III - T able 3d Eggs S e t b y Week and Pen from a N orthern B obwhite R eprodu ction Stu dy w ith T-7485 Experimental Group (ppm a.i. ) PENS 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 1 LOT A B C D E F G H I J 4001 0 2 0 0 0 623 0 013 53122 672 0 4 64 5 256 4 334 3 7 3 4 1 5 54 3 4 3 7 54653 5 74 4 4 656 305 6645 5 7 664 7475 2 07 76555 654 5 6 667 026 55474 7 6 6 4 7 66 6 057 7566 6 6 7 5 6 6 757 076 6757 5 76 5 7 7 676 054 76666 66 56 6 666 084 Totals 22 60 63 72 81 81 85 92 90 90 Totals 56 45 40 48 39 59 53 41 37 58 54 53 51 11 36 55 736 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen A ppendix V III - Table 4a V ia b le E m bryos b y Week and Pen from a N orthern B obwhite R eproduction S tu dy w ith T-7485 Experimental Group (ppm a .i .) PENS ---------------------------------- -- __________________________ 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 1 LOT A B C D Control E F G H I J 2 0 0 12 5 0 -00 1 0 13 0 2 704 355 5 -506 04 34 3 5 04 546 4 -64 5 124 4 5 756465 6 -74 7 3356 5 6 7 5 54 7 3 -6 6 6 7 3 7 6 7 76 5566 6 -756 62 57 6 67 6 657 6 -654 7166 7 666 566 1 -756 6056 6 67 6 657 0 -65 5 7 06 1 5 6 6 7 535 5 -76 8 6 04 5 6 Totals 17 54 59 79 85 85 85 77 72 79 Totals 58 44 49 45 46 59 36 _ 57 40 54 43 16 48 45 52 - Data are not available due to adult mortality LOT A - Eggs Set During W eek 12; LOT B - Eggs Set During W eek 13; etc. 692 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P e rfo rm a n c e b y Week and Pen Appendix V III - Table 4b V ia b le Embryos b y Week and Pen from a N orthern Bobwhite R ep ro d u ctio n S tu d y w ith T-7485 Experimental Group (ppm a .i .) PENS --------------------------------------------------------------1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 1 LOT A B C D 100 E F G H I J -001 0 00 0 4 022 0103 -162 1 33 0 6 75 3 0352 -5556 51 0 6 6 56 035 5 -4 75 7 54 0 7 4 7 6 6 57 3 -5676 63 1 6 55 74465 -4 76 7 64 4 7 5 7 6 6 67 6 -4 67 6 74 7 6 6 6 7 5 56 8 -3657 66 3 6 2 7 66 576 -567 6 76 5 6 66 7 646 7 -4 76 6 7 6 5 7 4 7 6 6 4 7 7 Totals 13 47 63 77 76 88 90 81 90 89 Totals _ 35 56 51 52 52 37 25 61 45 57 56 39 40 56 52 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 714 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen A ppendix V III - Table 4c V ia b le E m bryos b y Week and Pen from a N orthern B obwhite R eproduction S tu dy w ith T-7485 Experimental Group (ppm a .i .) PENS ------------------------------------------ -- _____________ _____ 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 1 LOT A B C D 300 E F G H I J 204202 -00000200 6 4 0634 3 - 30244634 2 406055 -604 6653 5 6 627 076 -5456675 7 6 675366 -76757666 7 767665 -65676667 6 576766 -65767756 6 74 7564 -564 7563 7 5 676767 -76767576 7 77667 5 -6767774 7 6 Totals 18 48 61 79 90 92 92 81 97 95 Totals 54 40 60 39 53 49 51 39 48 54 55 57 42 55 57 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 753 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen A ppendix V III - T able 4d V ia b le Embryos b y Week and Pen from a N orthern Bobwhite R eprodu ction S tu dy w ith T-7485 Experimental Group (ppm a.i.) PENS 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 1 LOT A B C D E F G H I J 40010 0000 3230 013 53 122 050 0 4 64 5 043 43 343 4 34 1 54 33 4 27 54653 5544 4 656 305 664556564 7475207 66554 544 5 6 667 026 45474 7664 7 666 056 75666 6756 6 757 076 67575 7657 7 676 044 76666 665 5 6 666 084 Totals 17 44 57 70 79 77 83 92 89 89 Totals 54 45 40 48 38 46 47 39 36 55 53 52 51 9 33 51 697 LOT A - Eggs Set During W eek 12; LOT B - Eggs Set During Week 13; etc. Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen Appendix V III - Table 5a L iv e T h ree-W eek Embryos b y Week and Pen from a N orthern B obw hite R eproduction S tu d y w ith T-7485 Experimental Group (ppm a.i.) 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 1 LOT A B C D Control E F G H I J 2 0 012 5 0 -00 1 00 2 0 2 7 0 4 3 5 5 5 -5 0 6 04 3 4 3 5 0 4 54 6 4 -64 5 12 4 4 5 75 64 65 6 -74 7 33 5 6 5 6 7 554 7 3 -66 6 73 7 6 7 76 5 56 6 6 -75 6 62 4 7 6 6 7 6 65 7 6 -6 5 4 71 6 6 7 66 656 6 1 -75 6 60 5 6 6 67 66 5 7 0 -6 5 5 70 6 1 5 66 7535 5 -76 8 604 5 6 Totals 15 54 59 79 85 84 85 77 72 79 Totals 58 44 49 45 46 59 36 - 57 40 54 43 15 46 45 52 - Data are not available due to adult mortality 1LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 689 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen Appendix V III - T able 5b L iv e T hree-W eek Embryos b y Week and Pen from a N orthern Bobwhite R eprodu ction S tu dy w ith T-7485 Experimental Group (ppm a. i .) 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 1 LOT A B C D 100 E F G H I J -001 0 0004 022 0 103 -162 1 3306 753 0 352 -5555 5106 6 56 0 355 -4756 5407 4 76 6 573 -5676 6316 5 574 465 -476 7 644 7 5 76 6 676 -467 6 74 76 6 67 5 567 -3657 6636 2 766 566 -567 6 7 656 6 67 6 467 -4766 7657 4 766 477 Totals 13 47 62 76 76 88 89 80 90 89 Totals - 35 56 51 50 52 37 25 61 45 57 56 39 40 55 51 - Data are not available due to adult mortality 1LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 710 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen Appendix V III - Table 5c L iv e T h ree-W eek Em bryos b y Week and Pen from a N orthern B obw hite R eprodu ction S tu d y w ith T-7485 Experimental Group (ppm a .i .) 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 1 LOT A B C D 300 E F G H I J 204202 - 0000 0200 6 4 0 6 34 3 - 3 0 2 4 4 63 4 2 405 05 5 - 6 04 6 653 5 6 62 7 07 6 - 54 56 675 7 6 675366 -76757666 7 767665 -6 567 6667 6 576766 - 6576 6756 6 74 7564 - 4 64 7 5637 5 576767 -7 566 7576 7 776 67 5 - 6 7 67 773 7 6 Totals 18 48 60 79 90 92 91 80 94 94 Totals 53 40 59 39 53 49 _ 50 38 47 54 54 57 41 55 57 - Data are not available due to adult mortality 1LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 746 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erfo rm a n ce b y Week and Pen Appendix V III - Table 5d L i v e T h ree-W eek Em bryos b y Week and Pen from a N orthern B obwhite R eprodu ction S tu d y w ith T-7485 Experimental (ppm a .i .) PENS 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 1 LOT A B C D 900 E F G H I J 4 001 0 0 000 32 3 0 013 5 312 2 04 0 0 4 64 5 04 3 4 3 34 3 4 34 1 54 3 3 4 2 7 54653 5544 4656 305 6 645 5 6564 74 7 5 2 07 6 6554 5435 6667 026 4 5474 7 664 6 66 6 056 7 566 6 6 756 67 57 076 67575 7657 7676 044 76666 664 5 6666 084 Totals 17 43 57 70 79 76 82 92 89 88 Totals 54 45 40 48 38 46 46 37 36 54 53 52 51 9 33 51 693 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 1 bO K) Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen A p p en d ix V III - T a b le 6a H a tc h lin g s b y Week and Pen from a N orthern B obw hite R eproduction S tu d y w ith T-7485 Experimental (ppm a. i .) 1601 1602 1603 1604 1605 1606 1607 1608 1609 PENS 1610 1611 1612 1613 1614 1615 1616 1 LOT A B C D Control E F G H I J 20 012 5 0 001 0020 2 704355 5 -506 0134 3 504 54 6 4 - 64 4 1144 5 74 64 6 5 6 -7 4 7 3 2 56 5 67 554 7 3 -6 6 6 7376 7 765566 6 -756 6246 6 666657 6 -654 7165 7 656466 1 -756 6046 5 676657 0 -64 5 7051 5 6665354 -768 6045 6 Totals 15 51 57 77 85 83 83 73 70 77 Totals 58 41 48 44 46 59 35 - 57 39 53 43 10 44 43 51 - Data are not available due to adult mortality 1LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 671 , 123 Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen A ppen dix V III - T able 6b H a tc h lin g s b y Week and Pen from a N orthern Bobwhite R eprodu ction S tu d y w ith T-7485 Experimental Group (ppm a.i.) PENS ----------------------------- --------------------- ------ _----------------------------------------------- 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Totals LOT A B C D 100 E F G H I J -0010 0004 022 0 103 -1621 3306 753 0 342 -5555 5106 6550 354 -4756 54 07 4766 573 -5676 6316 5564 455 -4767 642 74 766 654 -4675 7466 657 5 557 -3656 6636 2766 466 -56767656 6676 467 "4 666 7657 4 7 6 6 4 77 13 46 60 76 74 81 85 78 90 88 Totals - 35 55 51 48 52 37 22 61 44 56 54 39 39 50 48 - Data are not available due to adult mortality LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 691 1 NJ Project Number 454-116 Contains No CBI R e p r o d u c tiv e P erform an ce b y Week and Pen A ppendix V III - Table 6c H a tc h lin g s b y Week and Pen from a N orthern B obw hite R eproduction S tu dy w ith T-7485 Experimental (ppm a .i .) PENS 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 1 LOT A B C D 300 E F G H I J 20310 2 - 00 0 0 02 0 0 5 40624 3 -3 02 4 44 34 1 2 0 5 04 4 - 5 0 3 5 54 3 5 6 52 704 5 - 54 5 6 6 64 7 6 575366 -757 57566 5 76 766 5 -6 5 6 7 666 7 5 476766 -54 7 65756 5 54 756 4 -4 6 4 7 562 7 5 575767 -756 64476 6 77 667 5 - 6 7 6 7 773 7 6 Totals 15 44 51 72 85 91 86 77 88 94 Totals 46 40 57 37 49 47 - 48 36 46 53 49 51 39 55 50 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 703 Project Number 454-116 Contains No CBI R e p r o d u c t i v e P e r f o r m a n c e b y Week and Pen A p p en d ix V II I - T able 6d H a t c h l i n g s b y Week and Pen from a Northern Bobwhite R eproduction Stu dy w ith T-7485 Experimental Group (ppm a.i.) PENS 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 Totals LOT A B C D 900 E F G H I J 4 000 0 000 0 313 0 003 5312 2 030 0 364 5 043 4 3 34 3 4 33 1 54 3 3 4 2 7 44653 5444 4656 305 6645 5 646 4 74 6 5 2 07 66554 5434 666 7 025 4 5474 765 4 64 6 6 056 7566 6 6756 674 6 076 6757 5 765 7 766 6 044 76666 654 5 666 6 084 14 41 56 68 77 74 79 90 88 87 Totals 53 45 40 47 38 46 42 35 35 53 50 49 50 9 32 50 674 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. 1 K) Os Project Number 454-116 Contains No CBI R e p r o d u c tiv e Perform ance b y Week and Pen A p p e n d ix V I I I - T a b le 7a 1 4 -Day Old S u r v i v o r s b y Week and Pen from a Northern Bobwhite Reproduction S tu dy w ith T-7485 Experimental Group (ppm a. i .) PENS --------------------------------- ------------------------------ 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 1 LOT A B C D Control E F G H I J 1 0 0 12 4 0 - 0 0 0 0 0 1 0 2 6032545 -505 01343 4 04 54 64 -64 4 1 144 5 746464 6 -74 72 2565 675 54 73 -66 5 72 76 7 7655666 -756 62466 6 66 63 7 6 -653 61 55 7 5563661 -756 6 0464 6766570 -64 570515 666 5354 -767 6 045 6 Totals 11 46 56 75 83 83 78 70 70 76 Totals 54 41 47 42 44 56 35 _ 57 39 48 41 9 42 43 50 648 - Data are not available due to adult mortality 1 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. Project Number 454-116 Contains No CBI R e p r o d u c t i v e Perform ance b y Week and Pen A p p en d ix V II I - T a b le 7b 1 4 -Day O ld S u r v i v o r s b y Week and Pen from a Northern Bobwhite R eproduction Study w ith T-7485 Experimental Group (ppm a.i.) PENS -------------------------------- -- ---------------------------------------------------------- --------------- 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Totals 1 LOT A B C D E F G H I J 0000 0004 012 0 103 -0320 32 06 7 52 0242 -5354 5106 5550 354 -4443 4407 4 756 553 -5564 6306 5564 4 55 -4756 64 07 3766 654 -4674 644 6 5575 557 -3556 56362 766466 -5676 76366676466 -4666 7657 4 766 4 77 11 38 56 65 69 76 80 76 87 88 Totals - 34 45 47 39 49 36 15 61 41 55 52 39 38 48 47 - Data are not available due to adult mortality LOT A - Eggs Set During W eek 12; LOT B - Eggs Set During Week 13; etc. 646 Project Number 454-116 Contains No CBI R e p r o d u c t i v e P erfo rm a n ce b y Week and Pen A p p e n d ix VIII - T a b le 7c 1 4 -Day Old S u r v i v o r s b y Week and Pen from a Northern Bobwhite Reproduction S tu dy w ith T-7485 Experimental (ppm a. i .) PENS 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 1 LOT A B C D 300 E F G H I J 203000-0000 02004 406142 -30244424 i 105044 -503 5 5425 6 52 604 5 - 54 5 6 5547 6 575366 - 74 7 5 6 546 5 667664 -5567 6657 5 376766 -5476 56554 547564 -4 64 7 4 617 4 575767 -65664476 6 776665-6767 7737 6 Totals il 41 49 69 81 87 82 74 87 93 Totals 43 40 56 35 48 43 - 46 35 46 53 46 49 33 54 47 Data are not available due to adult mortality 1. ' LOT a - Eggs Set During Week 12; LOT B - Eggs Set During W eek 13; etc. 674 tVoO Project Number 454-116 Contains No CBI R e p r o d u c t i v e P e r fo r m a n c e b y Week and Pen A p p e n d ix V I I I - T a b le 7d 1 4 -D ay O ld S u r v i v o r s b y Week and Pen from a N orthern Bobwhite R eproduction Study w ith T-7485 Experimental Group (ppm a.i. ) PENS ---------------------------- -- ---------------------------------1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 1 LOT A B C D 900 E F G H I J 40000 0000 3130002 531 12 020 0 364 4 04 3 43342 433 1 543 3 32 6 44652 5344 435 6 304 664 55 54 54 74 5 5 2 0 5 66554 5424 666 7 02 5 4 54 73 644 4 536 6 04 5 75656 5455 664 5 076 65575 7657 756 6 044 766 66 6544 666 5 084 Totals 13 38 53 62 72 73 70 82 85 85 Totals 53 43 40 45 35 43 35 32 33 52 44 48 47 8 31 44 633 LOT A - Eggs Set During Week 12; LOT B - Eggs Set During Week 13; etc. Project Number 454-116 Contains No CBI Appendix IX - Table 1 E g g s h e l l T h i c k n e s s (mm) p e r Pen b y Week from a Northern Bobwhite Reproduction S tu dy w ith T-7485 (C o n tro l) Pen 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 IblZ 1613 1614 1615 1616 A 0.211 0.218 0.211 0.221 0.251 0.227 0.2 02 B 0.200 0.227 0.216 " 0.204 0.188 0.222 - No eggs available. C 0.216 0.221 0.200 0.236 0.192 0.231 0.227 0.214 D 0.213 0.222 0.235 - 0.209 0.218 0.209 0.233 E 0.227 0.241 0.187 0.250 0.199 0.231 0.236 0.224 Week s FG H I J Mean SD [n] 0.221 0.227 0.226 _ 0.204 0.213 0.203 0.226 0.204 0.226 0.201 0.242 0.204 0.246 0.226 0.227 0.210 0.230 0.226 0.206 0.226 0.193 0.212 0.208 0.232 0.206 - 0.209 0.241 0.231 0.221 0.208 0.214 0.226 0.209 0.217 0.192 0.219 Mean and standard deviation of pen means : 0.213 0.211 0.230 0.224 0.202 0.226 0.235 0.009 0.007 0.009 0.006 0.011 0.007 0.017 5 5 4 5 5 5 4 0.205 0.206 0.240 0.219 0.230 0.197 0.218 0.222 0.011 0.003 0.009 0.006 0.004 0.009 0.010 0.008 5 5 5 4 5 5 5 5 Mean 0.219 SD 0.013 72 Project Number 454-116 Contains No CBI Appendix IX - Table 2 E g g s h e l l T h i c k n e s s (mm) p e r Pen b y Week from a Northern Bobwhite R eproduction Study w ith T-7485 ( 1 0 0 PPM A . I . ) Pen 1617 1618 1619 lb2U lbZ J.bZ 1623 lb^4 162 b lb Z b 1627 1628 1629 Ibb U lb j 1 lb6Z A _ 0.265 " 0.203 0.219 0.227 ~ 0.192 B 0.242 0.189 0.226 " 0.221 0.229 0.226 0.224 C 0.246 0.211 0.217 0.236 0.221 0.200 0.219 D 0.229 0.230 0.230 0.211 0.229 0.232 0.226 0.247 E 0.241 0.225 0.222 0.240 0.229 0.211 0.205 Weeks FG 0.220 0.230 0.226 0.226 0.231 0.228 0.225 0.250 0.246 0.222 0.226 0.243 0.230 0.209 0.213 H 0.226 0.250 0.237 0.227 0.227 0.237 0.227 0.228 I 0.248 0.221 0.227 0.246 0.232 0.213 0.213 J 0.231 0.244 0.242 0.221 0.237 0.230 0.222 0.244 Mean and standard deviation of pen means : - No eggs available. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). Mean SD [n] 0.230 0.249 0.229 0.220 0.232 0.219 0.221 0.237 0.229 0.228 0.231 0.208 0.225 0.208 0.239 0.008 0.009 0.024 0.006 0.007 0.010 0.007 0 .Oil 0.006 0.004 0.004 0.006 0.002 0.010 0.012 5 5 5 4 5 5 4 5 5 5 5 4 5 5 5 Mean SD 0.227 0.011 72 1 u> NJ i Project Number 454-116 Contains No CBI Appendix IX - Table 3 E g g s h e l l T h i c k n e s s (non) p e r P e n b y Week from a Northern Bobwhite Reproduction S tu d y w ith T-7485 ( 3 0 0 PPM A. I . ) Pen 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 A 0.231 0.214 0.191 0.247 0.215 0.180 0.180 B 0.221 0.199 0.269 0.191 0.229 0.188 0.213 C 0.227 0.226 0.182 0.213 0.216 0.221 0.206 D 0.236 0.226 0.209 0.249 0.226 0.241 0.182 0.221 E 0.226 0.236 0.204 - 0.220 0.231 0.219 0.219 Weeks FG 0.236 0.231 0.216 0.241 0.227 0.247 0.200 0.226 0.227 0.226 0.2 04 _ 0.214 0.228 0.226 0.221 H 0.258 0.241 0.222 0.246 0.226 0.255 0.192 0.226 I 0.228 0.229 0.203 0.221 0.228 0.226 0.220 J 0.246 0.238 0.203 0.212 0.222 0.241 0.209 0.226 Mean and standard deviation of pen means : - No eggs available. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). Mean SD [n] 0.228 0.244 0.226 0.231 0.197 0.210 0.002 0.010 0.008 0.008 0.010 0.009 5 4 5 5 5 5 0.243 0.223 0.218 0.224 0.243 0.214 0.194 0.209 0.222 0.020 0.014 0.016 0.008 0.009 0.020 0.010 0.018 0.006 5 5 5 5 5 5 5 5 5 Mean 0.222 SD 0.015 74 1 U) OJ Project Number 454-116 Contains No CBI Appendix IX - Table 4 E g g s h e l l T h i c k n e s s (mm) p e r Pen b y Week from a N orthern Bobwhite R eproduction Study w ith T-7485 ( 9 0 0 PPM A . I . ) Weeks 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 JLbbl) 1661 1662 1663 1664 0.220 " " " 0.230 0.213 0.175 0.241 0.237 0.211 0.222 0.231 0.206 0.219 0.242 0.237 0.191 0.236 0.207 0.219 0.208 0.208 0.184 0.247 0.257 0.226 0.222 0.229 0.239 0.210 0.226 0.231 0.227 0.246 0.211 0.221 0.220 - 0.199 0.258 0.239 0.211 0.229 0.226 0.231 0.229 0.246 0.222 0.226 0.215 0.221 0.232 0.219 0.184 0.256 0.246 0.213 0.232 0.229 " 0.229 0.229 0.222 0.222 0.231 0.211 0.222 0.212 0.210 0.204 0.266 0.242 0.227 0.227 0.227 0.226 Mean and standard deviation of pen means : - No eggs available. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). 0.229 0.189 0.234 0.253 0.215 0.244 0.235 0.218 0.211 0.226 0.223 0.229 0.218 0.222 0.213 0.223 0.008 0.012 0.010 0.010 0.016 0.008 0.008 0.008 0.003 0.004 0.004 0.002 0.010 0.024 0.005 0.009 5 5 4 5 4 5 4 5 4 5 5 5 4 2 4 5 Mean 0.224 SD 0.015 71 u> Project Number 454-116 Contains No CBI Appendix X - Table 1 Mean H a t c h l i n g B o d y W e i g h t ( g ) p e r Pe n b y IVeeJfc from a Northern Bobwhite Reproduction Study w ith T-7485 (C o n tro l) Weeks [No. of Hatchlings] Pen A [ n] B [ n] C[ n] D[ n] E[ n] F [ n] G[ n] H [ n] I[ n] J[ n] Mean Total SD Hatch 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 lb 1J 1614 1615 1616 5[ 2] [ 0] [ 0] 5[ 1] 5 [ 2] 5[ 5] [ 0] [ 0] [ 0] 5[ 1] [ 0] [ 0] 5[ 2] [ 0] 5[ 2] 5[ 7] [ 0] 5[ 4] 5 [ 3] 6[ 5] 5[ 5] 5[ 5] 6[ 5] [ 0] 6[ 6] [ 0] 5 [ 1] 6[ 3] 6[ 4] 5[ 3] 5[ 5] [ 0] 6[ 4] 5 [ 5] 6[ 4] 6[ 6] 6[ 4] - 6[ 6] 5[ 4] 5 [ 4] 5[ 1] 5[ 1] 6[ 4] 6[ 4] 5[ 5] 5[ 7] 6[ 4] 5[ 6] 6[ 4] 6 [ 6] 6[ 5] 5[ 6] - 6[ 7] 6 [ 4] 5[ 7] 6[ 3] 6 [ 2] 6[ 5] 6[ 6] 5 [ 5] 6[ 61 6[ 7] 5 [ 5] 5 [ 5] 6[ 4] 6[ 7] 6[ 3] _ 6[ 6] 6 [ 6] 5[ 6] 6[ 7] 5[ 3] 5[ 7] 61 6] 5 [ 7] 61 7] 6 [ 6] 6 [ 5] 5 [ 5] 6 [ 61 6[ 61 6 [ 61 _ 6 [ 7] 61 5] 5[ 6] 61 61 5[ 2] 5 [ 4] 6[ 6] 5[ 6] 61 6] 6 [ 61 5 [ 61 6 [ 6] 6 [ 5] 6 [ 7] 6 [ 6] 7 [ 61 6 [ 5] 6 [ 4] 7 [ 7] 6 [ 1] 5[ 6] 6 [ 5] 6 [ 7] 6 [ 6] 6 [ 5] 6 1 6] 6 [ 4] 6 [ 6] 6 [ 6] 6 [ 1] 6 [ 7] 6 [ 5] 6 [ 6] 7 [ 6] [ 0] 61 4] 7 [ 6] 6 [ 5] 6 [ 6] 7 [ 7] 5 [ 6] 6[ 6] 6 [ 5] 6 [ 7] [ 0] 7 [ 61 6 [ 4] 6[ 5] 6 [ 7] [ 0] 6[ 5] 7 [ 1] 6 [ 5] 6 [ 6] 6 [ 6] 5 [ 6] 6 [ 5] 6 [ 3] 6 [ 5] 6 [ 4] 7 [ 7] 6[ 61 6[ 8] 7 [ 6] [ 0] 6 [ 4] 7 [ 5] 61 6] 6 0 58 6 0 41 5 0 48 6 0 44 6 0 46 6 0 59 6 0 35 6 0 57 6 0 39 6 0 53 6 0 43 5 0 10 5 0 44 6 0 43 6 0 51 Mean 6 SD 0 671 The number of hatchlings weighed may differ from the total number of hatchlings since those hatchlings found dead were not weighed. - Data are not available due to adult mortality. SD = Standard deviation of mean body weight, by parental pen, by week. 1 u> L/i Project Number 454-116 Contains No CBI Appendix X - Table 2 Mean H a t c h l i n g B o d y W e ig h t ( g ) p e r Pen b y Week from a Northern Bobwhite R eproduction Study w ith T-7485 ( 1 0 0 PPM A . I . ) Pen 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 A [ n] _ [ 0] [ 0] 4[ 1] [ 0] [ 0] [ 0] [ 0] 6[ 4] [ 0] 6[ 2] 6[ 2] [ 0] 5[ 1] [ 0] 5[ 3] B [ n] . 5[ i] 5[ 6] 5 [ 2] 4 [ 1] 6[ 3] 6[ 3] [ 0] 6[ 6] 5 [ 7] 6 [ 5] 6[ 3] [ 0] 6[ 3] 5[ 4] 6 [ 2] C[ n] 6[ 5] 6 [ 5] 5[ 5] 5[ 5] 6 [ 5] 6[ 1] [ 0] 61 6] 6 [ 6] 6 [ 5] 6 [ 5] [ 0] 61 3] 5[ 5] 6 [ 4] D[ n] 6 [ 4] 5 [ 7] 5[ 5] 5 [ 6] 6 [ 5] 6 [ 4] [ 0] 6 [ 7] 5[ 4] 6 [ 7] 61 6] 5 [ 6] 6 [ 5] 5 [ 7] 6 [ 3] Weeks [No. of Hatchlings] E[ n] F [ n] G[ n] H [ n] 7 [ 5] 61 6] 5 [ 7] 51 6] 6 [ 6] 6 [ 3] 5 [ 1] 6[ 6] 5[ 5] 6 t 5] 6 [ 6] 6 [ 4] 6 [ 4] 6 [ 5] 6 [ 5] 7 [ 4] 6 [ 7] 5[ 6] 61 7] 6 [ 6] 7 [ 4] 4 [ 2] 6 [ 7] 5[ 3] 7 t 7] 6 C 6] 5[ 6] 6[ 6] 6 [ 5] 6 [ 4] 7[ 4] 6 [ 6] 5 [ 7] 6 [ 5] 6 [ 7] 7 [ 4] 5[ 6] 7 [ 6] 5[ 6] 7 [ 5] 6 [ 7] 5[ 5] 6 [ 5] 6[ 5] 6 [ 7] 7 [ 3] 6 [ 6] 5[ 5] 6 C 6] 6 [ 6] 6 [ 6] 5[ 3] 7 [ 6] 5[ 2] 6 [ 7] 6 [ 6] 6 [ 6] 6[ 4] 6 [ 6] 6[ 6] I [ n] 7 [ 5] 6 t 6] 61 7] 6 [ 6] 6 [ 7] 6[ 6] 5[ 5] 7 [ 6] 5[ 6] 7 [ 6] 6 [ 7] 5 [ 6] 7 [ 4] 6[ 6] 6 [ 7] J[ n] Mean Total SD Hatch 7 [ 4] 7 [ 6] 6[ 6] 6[ 6] 6 [ 7] 7 [ 6] 5[ 5] 7 [ 7] 5 [ 4] 7 [ 7] 7 [ 6] 6[ 6] 6[ 4] 7 [ 7] 7 [ 7] 6 0 35 6 0 55 5 0 51 5 0 48 6 0 52 6 0 37 5 0 22 7 0 61 5 0 43 7 0 56 6 0 54 5 0 39 6 0 39 6 1 50 6 0 48 Mean 6 SD 1 690 The number of hatchlings weighed may differ from the total number of hatchlings since those hatchlings found dead were not weighed. - Data are not available due to adult mortality. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). SD = Standard deviation of mean body weight, by parental pen, by week. 1 U) Os i Project Number 454-116 Contains No CBI Appendix X - Table 3 Mean H a t c h l i n g B o d y W eigh t (g ) p e r Pen b y Week from a Northern Bobwhite Reproduction Study w ith T-7485 ( 3 0 0 PPM A . I . ) Weeks [No. of Hatchlings] Pen A [ n] B [ n] C[ n] D[ n] E[ n] F [ n] G[ n] H [ n] I[ n] J[ n] Mean SD Hatch 1633 lb J4 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 5[ 2] [ 0] 5 [ 3] 5[ 1] [ 0] 5 [ 2] [ 0] [ 0] [ 0] [ 0] [ 0] 5[ 2] [ 0] [ 0] 5 [ 5] 5[ 4] [ 0] 5[ 6] 5[ 2] 6[ 4] 5[ 3] 5[ 3] [ 0] 6[ 2] 5[ 4] 5 [ 4] 5 [ 4] 5[ 3] 5 [ 4] 6 [ 1] 5 [ 2] [ 0] 5[ 5] [ 0] 6[ 4] 5 [ 4] - 6 [ 5] [ 0] 6 [ 3] 5 [ 5] 5[ 5] 5[ 4] 5[ 3] 5[ 5] 5[ 6] 5[ 5] 5 [ 2] 6[ 7] [ 0] 6[ 4] 5 [ 5] - 6[ 5] 6 [ 4] 6[ 5] 6 [ 6] 6[ 6] 5[ 6] 5[ 4] 5 [ 7] 5[ 6] S [ 5] 6 [ 7] 6 [ 5] 6[ 3] 6 [ 6] 5[ 6] - 6 [ 7] 6[ 5] 6 [ 7] 6[ 5] 6 [ 7] 6[ 5] 6[ 6] 5 [ 6] 6[ 5] 6[ 7] 6[ 6] 6 [ 7] 6[ 6] 6[ 6] 5 [ 5] _ 6[ 6] . 6 [ 5] 6[ 6] 6[ 7] 6 [ 6] 5[ 6] 6 [ 6] 5[ 7] 6[ 5] 5[ 4] 6[ 7] 6 [ 6] 5[ 7] 6 [ 6] 5[ 6] _ 6 [ 5] 6 [ 4] 5[ 7] 6 [ 6] 6[ 5] 5 [ 7] 6[ 5] 5 [ 6] 5[ 5] 6[ 5] 6[ 4] 6[ 7] 6[ 5] 7 [ 6] 6[ 4] . 6[ 4] 6[ 6) 6 [ 4] 6[ 7] 6[ 5] 5[ 6] 6[ 2] 5[ 7] 6[ 5] 6[ 5] 6[ 7] 6 [ 5] 6 [ 7] 7 [ 6] 6 [ 7] 6[ 7] 6 [ 5] 6[ 6] 6[ 6] 6 [ 4] 6[ 4] 6[ 7] 5[ 6] 6[ 6] 6[ 7] 6 [ 7] 7 [ 6] 7 [ 6] 6 [ 7] 6[ 5] 6 [ 6] 6[ 7] 6[ 6) 7 [ 7] 7 [ 7] 6[ 7] 5[ 3] 6[ 7] 7[ 6] 6 0 46 6 0 40 6 0 57 6 1 37 6 0 49 5 0 47 6 0 48 6 0 36 6 0 46 6 0 53 6 0 49 5 0 51 5 0 39 5 0 55 6 1 50 Mean 6 SD 0 703 The number of hatchlings weighed may differ from the total number of hatchlings since those hatchlings found dead were not weighed. - Data are not available due to adult mortality. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). SD = Standard deviation of mean body weight, by parental pen, by week. 1 1 Project Number 454-116 Contains No CBI Appendix X - Table 4 Mean H a t c h l i n g B o d y W eig h t (g ) p e r Pen b y Week from a Northern Bobwhite R eproduction Study w ith T-7485 ( 9 0 0 PPM A . I . ) Pen 1649 lb5U 1651 lb b2 1653 1654 1655 1656 1657 1658 1659 lb6U 1661 bb2 Ibbb lb b4 A[ n] 6 [ 4] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] 6[ 3] 5[ 1] 6[ 3] [ 0] [ 0] [ 0] 5[ 3] B [ n] 6 [ 5] 6[ 3] 5[ 1] 5[ 2] 6[ 2] [ 0] S[ 3] [ 0] [ 0] 6[ 3] 5[ 6] 6[ 4] 5[ 5] [ 0] 6 [ 4] 5[ 3] C[ n] 6[ 4] 6[ 3] 5[ 3] 6[ 4] 5[ 3] 5 [ 4] 5[ 3] 5t 3] 5[ 1] 6 [ 5] 5[ 4] 6 [ 3] 5[ 3] S[ 4] 6[ 2] 5[ 7] D[ n] 6[ 4] 6[ 4] 5 [ 6] 5 [ 5] 5[ 3] 5[ 5] 5[ 4] 6[ 4] 5 [ 4] 6[ 4] 5[ 6] 6[ 5] 5[ 6] 6[ 3] [ 0] 5[ 5] Weeks [No. of Hatchlings] E[ n] F [ n] G[ n] H [ n] 7 [ 6] 6[ 6] 6[ 4] 6[ 5] 6[ 5] 5[ 6] 5[ 4] 6 [ 6] 5[ 4] 6[ 7] 5[ 4] 6 [ 6] 5[ 5] 7 [ 2] [ 0] 6[ 7] 7 [ 6] 6 1 6] 6[ 5] 6[ 5] 6[ 4] 6 [ 5] 6[ 4] 6[ 2] 6[ 4] 6[ 6] 5 [ 6] 6 [ 6] 5 [ 7] [ 0] 61 2] 5[ 5] 7 [ 4] 6[ 5] 6 [ 4] 6 1 7] 6 [ 4] 6 [ 7] 6[ 6] 6 1 5] 6[ 4] 6 [ 6] 5[ 4] 6 [ 6] 5[ 6] [ 0] 6 [ 5] 6 [ 6] 7 [ 7] 6 1 5] 6 [ 6] 6 [ 6] 6 1 6] 6[ 6] 5 [ 7] 6 [ 5] 5[ 6] 6 [ 6] 5[ 7] 6[ 4] 5[ 6] [ 0] 6 [ 7] 6 [ 6] I [ n] 7 [ 6] 6 [ 6] 6 [ 5] 6 [ 1] 6 [ 5] 6[ 7] 6[ 6] 6[ 5] 6 [ 7] 7 [ 7] 5[ 6] 7[ 6] 6 [ 6] [ 0] 7 [ 4] 6[ 4] J[ n] Mean Total SD Hatch 7 [ 7] 6 0 53 6 [ 6] 6 0 44 6 [ 6] 6 0 40 6 [ 6] 6 0 47 7 [ 6] 6 0 38 6 [ 6] 5 0 46 6[ 5] 6 0 42 6 [ 4] 6 0 34 6 [ 5] 6 1 35 7[ 6] 6 0 53 5 [ 6] 5 0 50 6 [ 6] 6 0 49 6 [ 6] 5 0 50 [ 0] 6 1 9 7 [ 8] 6 1 32 6[ 4] 5 0 50 Mean 6 The number of hatchlings weighed may differ from the total number of hatchlings since those hatchlings found dead were not weiaheH Differences between the control and this treatment group were not significant (p > 0 .0 5 ). SD = Standard deviation of mean body weight, by parental pen, by week. SD 0 672 Project Number 454-116 Contains No CBI Appendix XI - Table 1 Mean 1 4 -Day O ld S u r v i v o r s Body W eigh t ( g ) p e r Pen b y Week from a Northern Bobwhite Reproduction Study w ith T-7485 (C o n tro l) Pen 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 A[ n] B [ n] C[ n] D[ n] Weeks [No. of 14-Day Old Survivors] E[ n] F [ n] G[ n] H [ n] I[ n] 22 [ l] [ 0] [ 0] 20 [ 1] 26 [ 2] 26 [ 4] [ 0] - [ 0] [ 0] [ 0] [ 0] [ 0] 21 [ 1] [ 0] 23 [ 2] 26 [ 6] [ 0] 26 [ 3] 25 [ 2] 28 [ 5] 27 [ 4] 29 [ 5] 27 [ 5] [ 0] 24 [ 5] [ 0] 24 [ 1] 22 [ 3] 26 [ 4] 21 [ 3] 28 [ 4] [ 0] 26 [ 4] 25 [ 5] 27 [ 4] 28 [ 6] 29 [ 4] - 25 [ 6] 22 [ 4] 24 [ 4] 23 [ 1] 19 [ 1] 26 [ 4] 23 [ 4] 26 [ 5] 23 [ 7] 23 [ 4] 27 [ 6] 25 [ 4] 26 [ 6] 28 [ 4] 23 [ 6] - 30 [ 7] 27 [ 4] 21 [ 7] 26 [ 2] 20 [ 2] 24 [ 5] 26 [ 6] 25 [ 5] 22 [ 6] 22 [ 7] 28 [ 5] 23 [ 5] 24 [ 4] 28 [ 7] 26 [ 3] - 26 [ 6] 26 [ 6] 25 [ 5] 28 [ 7] 22 [ 2] 22 [ 7] 24 [ 6] 25 [ 7] 22 [ 7] 22 [ 6] 30 [ 53 24 [ 5] 22 [ 6] 26 [ 6] 25 [ 6] _ 28 [ 7] 26 [ 5] 21 [ 6] 28 [ 6] 23 [ 2] 26 [ 4] 24 [ 6] 23 [ 6] 26 [ 6] 23 [ 6] 29 [ 6] 26 [ 6] 24 [ 3] 27 [ 7] 25 [ 61 _ 31 [ 6] 23 [ 5] 19 [ 3] 30 [ 6] 21 [ 1] 22 [ 5] 26 [ 5] 25 [ 7] 27 [ 5] 26 [ 5] 27 [ 6] 30 [ 3] 29 [ 6] 27 [ 6] 28 [ 1] 29 [ 7] 26 [ 5] 23 [ 6] 28 [ 6] [ 0] 26 [ 4] 27 [ 6] 27 [ 4] 29 [ 6] 29 [ 7] 26 [ 6] 30 [ 6] 26 [ 5] 30 [ 7] [ 0] 31 [ 6] 25 [ 4] 25 [ 5] 30 [ 7] [ 0] 25 [ 5] 23 [ 1] 26 [ 5] - Data are not available due to adult mortality. SD = Standard deviation of mean body weight, by parental pen, by week. u> o J[ n] Mean 14-Day SD Total 32 [ 6] 31 [ 6] 27 [ 6] 31 [ 5] 28 [ 3] 31 [ 5] 29 [ 4] 26 25 27 26 26 28 26 3 54 4 41 i 47 3 42 2 44 2 56 2 35 35 [ 7] 6] 27 [ 7] 33 [ 6] [ 0] 24 [ 4] 34 [ 5] 30 [ 6] 29 26 24 29 21 24 26 25 3 57 2 39 2 48 2 41 29 2 42 3 43 2 50 Mean 26 SD 2 648 , LO Project Number 454-116 Contains No CBI Appendix XI - Table 2 Mean 1 4 -D ay O ld S u r v i v o r s B o d y W e ig h t ( g ) p e r Pen b y Week from a Northern Bobwhite R eproduction Stu dy w ith T-7485 ( 1 0 0 PPM A . I . ) Weeks [No. of 14-Day Old Survivors] 14-Day Pen A[ n] B [ n] C[ n] D[ n] E[ n] F[ n] G[ n] H [ n] I[ n] J[ n] Mean SD Total 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 _ [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] 26 [ 4] [ 0] 23 [ 1] 23 [ 2] [ 0] 29 [ 1] [ 0] 26 [ 3] _ [ 0] 24 [ 3] 26 [ 2] [ 0] 29 [ 3] 25 [ 2] [ 0] 27 [ 6] 22 [ 7] 26 [ 5] 30 [ 2] [ 0] 25 [ 2] 23 [ 4] 31 [ 2] 26 [ 5] 25 [ 3] 24 [ 5] 26 [ 4] 28 [ 5] 27 [ 1] [ 0] 28 [ 6] 24 [ 5] 26 [ 5] 29 [ 5] [ 0] 29 [ 3] 26 [ 5] 25 [ 4] 24 [ 4] 23 [ 4] 18 [ 4] 20 [ 3] 21 [ 4] 23 [ 4] [ 0] 29 [ 7] 23 [ 4] 25 [ 7] 22 [ 5] 22 [ 6] 25 [ 5] 21 [ 5] 29 [ 3] 28 [ 5] 26 [ 5] 22 [ 6] 25 [ 4] 25 [ 6] 23 [ 3] [ 0] 27 [ 6] 22 [ 5] 27 [ 5] 27 [ 6] 23 [ 4] 27 [ 4] 25 [ 5] 29 [ 5] 25 [ 4] 25 [ 7] 22 [ 5] 27 [ 6] 30 [ 6] 27 [ 4] [ 0] 29 [ 7] 22 [ 3] 28 [ 7] 28 [ 6] 20 [ 6] 28 [ 6] 25 [ 5] 24 [ 4] 29 [ 4] 25 [ 6] 21 [ 7] 24 [ 4] 28 [ 6] 27 [ 4] 22 [ 4] 26 [ 6] 19 [ 5] 26 [ 5] 26 [ 7] 23 [ 5] 31 [ 5] 24 [ 5] 30 [ 7] - Data are not available due to adult mortality. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). SD = Standard deviation of mean body weight, by parental pen, by week. 29 [ 3] 25 [ 5] 25 [ 5] 25 [ 6] 31 [ 5] 26 [ 6] 22 [ 3] 30 [ 6] 14 [ 2] 25 [ 7] 24 [ 6] 24 [ 6] 32 [ 4] 27 [ 6] 32 [ 6] 29 [ 5] 25 [ 6] 28 [ 7] 25 [ 6] 27 [ 7] 25 [ 6] 23 [ 3] 30 [ 6] 22 [ 6] 29 [ 6] 25 [ 7] 22 [ 6] 32 [ 4] 31 [ 6] 32 [ 6] 27 [ 4] 24 [ 6] 27 [ 6] 26 [ 6] 30 [ 7] 28 [ 6] 21 [ 5] 30 [ 7] 26 [ 4] 31 [ 7] 30 [ 6] 22 [ 6] 35 [ 4] 30 E 7] 37 [ 7] 27 25 24 25 28 26 22 28 22 27 26 22 29 26 30 2 34 1 45 3 47 2 39 3 49 2 36 1 15 1 61 3 41 2 55 2 52 1 39 3 38 3 48 4 47 Mean 26 SD 3 646 Project Number 454-116 Contains No CBI Appendix XI - Table 3 Mean 1 4 -Day O ld S u r v i v o r s Body W e ig h t (g) p e r Pen b y Week from a N orthern Bobwhite R eproduction Study w ith T-7485 ( 3 0 0 PPM A . I . ) Weeks [No. of 14-Day Old Survivors] 14-Day Pen A [ n] B [ n] C[ n] D[ n] E[ n] F [ n] G[ n] H [ n] I [ n] J[ n] Mean SD Total 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 23 [ 2] [ 0] 23 [ 3] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] 21 [ 2] [ 0] [ 0] 22 [ 4] 27 [ 4] [ 0] 29 [ 6] 26 [ 1] 27 [ 4] 27 [ 23 23 [ 3] [ 0] 25 [ 2] 27 [ 4] 24 [ 4] 24 [ 4] 24 [ 2] 23 [ 4] 27 [ 1] 24 [ i] [ 0] 29 [ 5] [ 0] 28 [ 4] 25 [ 4] - 27 [ 5] [ 0] 25 [ 3] 26 [ 5] 25 [ 5] 25 [ 4] 22 [ 2] 24 [ 5] 24 [ 6] 20 [ 5] 25 [ 2] 28 [ 6] [ 0] 28 [ 4] 22 [ 5] - 26 [ 5] 25 [ 4] 26 [ 5] 27 [ 6] 24 [ 5] 22 [ 5] 25 [ 4] 27 [ 7] 21 [ 6] 23 [ 5] 26 [ 7] 31 [ 5] 30 [ 3] 28 [ 6] 27 [ 6] - 28 [ 7] 28 [ 4] 24 [ 7] 28 [ 5] 29 [ 6] 21 [ 5] 27 [ 4] 26 [ 6] 28 [ 5] 24 [ 6] 26 [ 6] 30 [ 7] 27 [ 6] 29 [ 6] 27 [ 4] - 30 [ 5] 24 [ 5] 26 [ 6] 27 [ 7] 27 [ 6] 24 [ 6] 30 [ 5] 27 [ 7] 28 [ 5] 29 [ 3] 26 [ 7] 28 [ 6] 26 [ 7] 30 [ 6] 25 [ 6] - 27 [ 5] 28 [ 4] 26 [ 7] 28 [ 6] 23 [ 5] 23 [ 6] 29 [ 5] 26 [ 5] 25 [ 4] - Data are not available due to adult mortality. Differences between the control and this treatment group were not significant (p > 0 .0 5 ). SD = Standard deviation of mean body weight, by parental pen, by week. 27 [ 5] 27 [ 4] 29 [ 7] 24 [ 5] 31 [ 6] 28 [ 4] - 29 [ 4] 29 [ 6] 26 [ 4] 31 [ 7] 26 [ 4] 23 [ 6] 28 [ 1] 31 [ 7] 29 [ 4] 28 [ 5] 29 [ 7] 29 [ 5] 25 [ 7] 31 [ 6] 27 [ 7] - 33 [ 6] 25 [ 5] 27 [ 6] 28 [ 6] 27 [ 4] 25 [ 43 29 [ 7] 27 [ 6] 28 [ 6] 30 [ 7] 27 [ 7] 38 [ 6] 33 [ 6] 38 [ 6] 33 [ 53 - 33 [ 6] 27 [ 7] 30 [ 6] 36 [ 7] 32 [ 7] 24 [ 7] 31 [ 3] 34 [ 73 30 [ 6] 26 27 30 27 30 27 29 27 26 29 27 23 28 28 26 3 43 1 40 3 56 3 35 3 48 3 43 3 46 2 35 2 46 3 53 3 46 1 49 3 33 3 54 3 47 Mean 27 SD 2 674 Project Number 454-116 Contains No CBI Appendix XI - Table 4 Mean 1 4 -Day O ld S u r v i v o r s B ody W eig h t ( g ) p e r Pen b y Week from a Northern Bobwhite R eproduction S tu dy w ith T-7485 ( 9 0 0 PPM A . I . ) Weeks [No. of 14-Day Old Survivors] ------------------------------------------------- 14-Day Pen A [ n] B [ n] C[ n] D [ n] E[ n] F[ n] G[ n] H [ n] I[ n] J[ n] Mean SD Total 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 24 [ 4] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] [ 0] 29 [ 3] 26 [ 1] 26 [ 3] [ 0] [ 0] [ 0] 22 [ 2] 32 [ 5] 26 [ 3] 19 [ 1] 19 [ 1] 32 [ 2] [ 0] 28 [ 2] [ 0] [ 0] 27 [ 3] 27 [ 6] 31 [ 4] 25 [ 4] [ 0] 25 [ 4] 27 [ 3] 24 [ 4] 27 [ 3] 25 [ 3] 20 [ 4] 29 [ 2] 24 [ 4] 25 [ 3] 22 [ 3] 18 [ 1] 27 [ 5] 22 [ 4] 30 [ 3] 22 [ 3] 24 [ 3] 28 [ 2] 26 [ 6] 26 [ 4] 24 [ 4] 22 [ 6] 23 [ 5] 30 [ 2] 28 [ 5] 26 [ 3] 21 [ 4] 23 [ 4] 28 [ 4] 27 [ 3] 27 [ 5] 25 [ 6] 27 [ 3] [ 0] 23 [ 4] 27 [ 6] 25 [ 6] 27 [ 4] 24 [ 5] 29 [ 5] 24 [ 5] 27 [ 4] 22 [ 5] 21 [ 4] 27 [ 7] 25 [ 4] 26 [ 5] 25 [ 5] 26 [ 2] [ 0] 26 [ 5] 27 [ 6] 26 [ 6] 26 [ 5] 22 [ 5] 28 [ 4] 25 [ 5] 30 [ 4] 25 [ 2] 28 [ 4] 27 [ 6] 26 [ 6] 27 [ 6] 26 [ 7] [ 0] 20 [ 2] 24 [ 5] 25 [ 4] 25 [ 5] 29 [ 4] 22 [ 7] 27 [ 3] 25 [ 6] 29 [ 4] 23 [ 4] 24 [ 4] 25 [ 5] 25 [ 3] 26 [ 6] 24 [ 6] [ 0] 26 [ 4] 25 [ 5] 28 [ 7] 27 [ 5] 26 [ 6] 26 [ 5] 32 [ 6] 24 [ 5] 31 [ 4] 27 [ 5] 23 [ 5] 28 [ 6] 27 [ 6] 31 [ 4] 24 [ 5] [ 0] 27 [ 7] 29 [ 6] 33 [ 6] 28 [ 5] 30 [ 5] 24 [ 7] 30 [ 5] 26 [ 7] 30 [ 6] 23 [ 5] 28 [ 7] 29 [ 7] 27 [ 5] 30 [ 6] 27 [ 6] [ 0] 27 [ 4] 29 [ 4] 31 [ 7] 26 [ 6] 29 [ 6] 24 [ 6] 32 [ 6] 26 [ 6] 32 [ 5] 27 [ 4] 31 [ 4] 27 [ 6] 25 [ 6] 34 [ 6] 30 [ 5] [ 0] 31 [ 8] 30 [ 4] 28 26 26 23 30 25 29 24 25 28 26 29 26 26 27 26 3 53 1 43 3 40 2 45 2 35 1 43 2 35 2 32 4 33 1 52 1 44 3 48 2 47 18 3 31 2 44 Mean 27 SD 2 633 Differences between the control and this treatment group were not significant (p > 0 .0 5 ). SD = Standard deviation of mean body weight, by parental pen, by week. Project Number 454-116 Contains No CBI - 143 - Project Number 454-116 Appendix XII - Table 1 Adult Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 Control (0 ppm a.i.) Male Pen Liver Female Liver 1601 1602 1603 1604 1605 1606 1607 1608 1609 1610 1611 1612 1613 1614 1615 1616 Mean SD 3.893 8.937 5.079 4.930 3.184 3.622 2.296 3.808 4.253 3.709 4.208 3.357 3.087 4.031 3.825 3.922 4.134 1.446 7.981 7.912 7.231 6.756 8.459 7.057 6.240 7.411 6.329 6.879 6.556 7.750 7.572 7.464 6.818 7.228 0.644 - Data not reported due to mortality. Contains No CBI - 144 - Project Number 454-116 Appendix XII - Table 2 Adult Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 100 ppm a.i. Pen 1617 1618 1619 1620 1621 1622 1623 1624 1625 1626 1627 1628 1629 1630 1631 1632 Mean SD Male Liver 3.064 4.088 4.624 5.039 4.796 3.486 3.819 3.014 3.553 3.673 3.906 3.401 3.882 4.134 6.545 4.068 0.901 Female Liver 8.091 7.866 6.812 7.065 7.595 7.099 5.060 8.996 6.967 9.093 7.284 6.855 12.255 6.930 6.715 6.529 7.576 1.575 - Data not reported due to mortality. Differences from the control group were not statistically significant (p>0.05, Dunnett's t-test). Contains No CBI - 145 - Project Number 454-116 Appendix XII - Table 3 Adult Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 300 ppm a.i. Male Pen Liver Female Liver 1633 1634 1635 1636 1637 1638 1639 1640 1641 1642 1643 1644 1645 1646 1647 1648 Mean SD 3.381 3.577 3.525 3.870 3.697 9.874 3.756 4.226 2.901 4.451 4.008 2.960 6.127 3.484 4.006 3.147 4.187 1.692 6.926 6.082 8.607 6.401 8.139 5.154 6.792 8.035 7.000 9.105 8.546 7.576 6.446 6.085 7.141 7.202 1.112 - Data not reported due to mortality. Differences from the control group were not statistically significant (p>0.05, Dunnett's t-test). Contains No CBI - 146- Project Number 454-116 Appendix XII - Table 4 Adult Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 900 ppm a.i. Male Pen Liver Female Liver 1649 1650 1651 1652 1653 1654 1655 1656 1657 1658 1659 1660 1661 1662 1663 1664 Mean SD 5.790 6.179 3.183 4.908 4.679 # 3.334 3.464 3.112 3.022 3.598 3.906 3.024 4.708 3.050 2.928 3.926 1.068 5.963 5.975 7.119 6.007 6.000 7.811 6.457 6.807 9.105 7.627 8.379 6.894 6.782 3.270 5.238 4.938 6.523 1.403 # Data not available due to recording error. Differences from the control group were not statistically significant (p>0.05, Dunnett's t-test). Contains No CBI - 147 - Project Number 454-116 Appendix XIII Offspring Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 Control (0 ppm a.i.) Parental Pen # 1601 1602 1603 1605 1611 1612 1614 Mean SD Male Liver 1.247 1.016 1.024 0.957 0.948 0.991 0.908 1.013 0.111 Parental Pen # 1604 1606 1607 1609 1610 1615 1616 Mean SD Female Liver 1.076 1.209 0.989 1.183 0.870 1.125 1.122 1.082 0.118 Mean 1.048 SD 0.116 100 ppm a.i. Parental Pen # Male Liver Parental Pen # Female Liver 1621 1.163 1625 0.916 1627 1.113 1630 1.019 1631 1.025 1632 1.443 Mean SD 1.113 0.183 1618 1619 1620 1622 1623 1624 1626 1628 1629 Mean SD 1.225 1.087 0.876 0.837 0.971 1.023 1.196 1.024 1.034 1.030 0.129 Group Mean 1.063 SD 0.152 Differences fromthe control group were not statistically significant (p>0.05, Dunnett's t-test). Contains No CBI - 148 - Project Number 454-116 Appendix XIII Offspring Liver Weights (g) from a Northern Bobwhite Reproduction Study with T-7485 300 ppm a.i. Parental Pen # 1633 1635 1640 1641 1642 1643 1646 Mean SD Male Liver 1.004 1.356 1.171 1.158 1.199 1.355 1.208 1.207 0.122 Parental Pen # 1634 1636 1637 1638 1644 1645 1647 1648 Mean SD Female Liver 1.079 0.914 1.664 1.163 1.550 0.993 1.513 1.461 1.292 0.287 Group Mean 1.253 SD 0.222 900 ppm a.i. Parental Pen # Male Liver Parental Pen # Female Liver 1652 1654 1655 1656 1657 1660 1661 Mean SD 0.838 0.933 1.494 0.988 1.309 1.042 1.147 1.107 0.228 1649 1650 1651 1653 1658 1659 1663 1664 Mean SD 1.294 0.926 0.777 1.002 1.096 1.248 1.147 1.080 1.071 0.169 Group Mean 1.088 SD 0.192 Differences fromthe control group were not statistically significant (p>0.05, Dunnett's t-test). Contains No CBI ,W ildlife International Ltd. - 149- Appendix XIV Histology Report Project Number 454-116 Contains No CBI - 150 - EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 W ILDLIFE INTERNATIONAL, LTD. STUDY NUMBER 454-116 EPL PROJECT NUMBER 212-030 T-7485: A REPRO DUCTIO N STUDY W ITH THE NO RTHERN BOBW HITE PATHOLOGY REPORT Submitted by: Experimental Pathology Laboratories, Inc. P.O. Box 474 Herndon, VA 20172-0474 (703) 471-7060 Submitted to: Wildlife International, Ltd. Easton, MD 21601 May 14, 2004 Contains No CBI  EPL -151 EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 TABLE OF CONTENTS PATHOLOGY SUM M ARY....................................................................................... 1 COMPLIANCE STATEM ENT.................................................................................. 5 QUALITY ASSURANCE FINAL CERTIFICATION............................................... 6 ADULT EUTHANASIA SUMMARY INCIDENCE TABLES M ales.................................................................................................................... 1-1 Females............................................................................................................... I-2 HISTOPATHOLOGY INCIDENCE TABLES Males.................................................................................................................... 11-1 Females............................................................................................................... II-5 CORRELATION OF GROSS AND MICROSCOPIC FINDINGS M ales.................................................................................................................... 111-1 Females...................................................... III-6 OFFSPRING EUTHANASIA SUMMARY INCIDENCE TABLES Males.................................................................................................................... IV-1 Females............................................................................................................... IV-2 HISTOPATHOLOGY INCIDENCE TABLES M ales................................................................................................................... V-1 Females............................................................................................................... V-3 Contains No CBI - 152 - Project Number 454-116 PATHOLOGY SUMMARY Contains No CBI EPL - 153 - EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 W ILDLIFE INTERNATIONAL, LTD. STUDY NUMBER 454-116 EPL PROJECT NUMBER 212-030 T-7485: A REPRO DUCTIO N STUDY W ITH THE NO RTHERN BOBW HITE PATHOLOGY SUMMARY Light microscopic examination was performed on sections of selected tissues from adult male and fem ale northern bobwhite ( Colinus virginianus) which w ere untreated or received various concentrations of the test article (T-7485) in the feed for an approximate five-month period. Selected tissues from 16-day-old offspring of the adult bobwhite were also exam ined by light microscopy. The objective of this study was to evaluate the effects of dietary exposure to T -7 4 8 5 upon adult bobwhite ( Colinus virginianus) over an approximate five-month period. T h e experim ental design for selection of adults and offspring for histopathological evaluation was as follows: Experimental Group Adults Males Females Offspring3 Male Fem ale Control (0 ppm a.i.) 100 ppm a.i. 300 ppm a.i. 900 ppm a.i. 16 16 16 16 16 16 16 16 6 6 7 7 8 9 8 8 a Offspring did not receive the test substance in the diet. All offspring were euthanized on Day 16 post-hatching. MATERIALS AND METHODS At the completion of the approxim ate five-month study interval, all surviving adult bobwhite were euthanized. Necropsies were performed by Wildlife International, Ltd., and tissue samples were collected for histopathological evaluation. Selected offspring were euthanized at Day 16 post-hatching, and samples were collected for histopathological evaluation by Wildlife International, Contains No CBI  EPL - 154- EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 Wildlife International, Ltd. Study Number 454-116 Ltd. Selected tissues from adults and offspring were fixed in 10% neutral buffered formalin and sent to Experimental Pathology Laboratories, Inc. (EPL, Inc.), where they were em bedded in paraffin and m ade into hematoxylin and eosin-stained sections on glass microscope slides. The following tissues from adults and offspring w ere exam ined by light microscopy as available: liver, kidney, and gonad (ovary or testis). Because of a gross observation, the spleen from 900 ppm a.i. adult male 462 was also submitted, processed, and evaluated. All tissues required by protocol are presented in the Histopathology Incidence Tables. Microscopic findings for each tissue exam ined from each bobwhite are listed in the Histopathology Incidence Tables. Microscopic changes w ere graded one to five depending on severity. Nongradable findings are listed as present (P). All findings for all animals are summarized by sex, age group, and treatm ent group in the Sum m ary Incidence Tables, together with the total num ber of anim als in each group for which the tissues w ere exam ined. During histopathological evaluation, one offspring (0 ppm a.i. B829) which had been submitted as a male was discovered to be a fem ale, and was therefore included in the fem ale tables. A tabulation of gross observations noted at necropsy with the corresponding microscopic change, if appropriate, is presented in the Correlation of Gross and Microscopic Findings tables. The entries in these tables were transcribed from records provided by the sponsor. Additional gross observations noted at trimming are indicated as gross lesions found at trimming in the Correlation of Gross and Microscopic Findings tables. RESULTS All adults and offspring survived to the end of their respective study intervals, with the exception of the following adults: 100 ppm a.i. m ale 3 85 found dead approximately 16 weeks before scheduled euthanasia, 0 ppm a.i. female 368 found dead approximately 10 weeks before scheduled euthanasia, and 300 ppm a.i. fem ale 430 euthanized approximately seven weeks before scheduled 2- - Contains No CBI - 155 - EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 Wildlife International, Ltd. Study Number 454-116 euthanasia due to her moribund condition. This low, sporadic, early mortality was considered incidental and unrelated to treatment. Although the exact cause(s) of moribundity and/or mortality in these birds was not determined, they did exhibit gross necropsy observations of "intracranial bleeding," head and neck lesions with "necrotic margins," and/or emaciation which m ay have contributed to the birds' early demise. No effects considered related to test article administration w ere noted in liver, kidney, testis, and ovary of male and fem ale adult bobwhite or their offspring. Testes of most adult males exhibited spermatogenesis characterized by the presence of numerous mature sperm atozoa in the seminiferous tubules. A few males exhibited decreased testicular diameter and/or seminiferous tubule degeneration most consistent with physiological regression. In addition, testes of 100 ppm a.i. m ale 385 (which died about 16 weeks before scheduled euthanasia) had morphology consistent with physiological sexual immaturity. Offspring testes w ere listed as immature because they lacked all evidence of sperm atogenesis, had small seminiferous tubules with absent or narrow lumens, and had germinal epithelium with decreased cell layers (compared to adult testes). This morphology was consistent with normal physiological immaturity in young (16-day old) birds rather than a pathological effect. Ovaries of several control and treated adult fem ales exhibited serosal eosinophilic material (amorphous sheets and globules of deep m agenta material adhered to the serosal surfaces and consistent with extra follicular egg yolk). In one adult bird (0 ppm a.i. fem ale 376), the material was associated with serosal infiltrates of macrophages and heterophils (chronic active inflammation). Ovarian serosal eosinophilic material corresponded to grossly observed "egg yolk peritonitis" in one case (300 ppm a.i. fem ale 448). Similar eosinophilic material was adhered to the kidney serosal surfaces of a few adult females. Ovaries of all offspring fem ales were characterized by numerous oocytes and very small, nonpedunculated follicles completely em bedded in a prominent -3 - Contains No CBI  EPL -156- EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 Wildlife International, Ltd. Study Number 454-116 ovarian cortex. This morphology is consistent with normal physiological immaturity in young (16-day old) birds rather than a pathological effect. A few other findings w ere noted in various tissues of adult and offspring bobwhite. T h ese generally minimal to mild changes had similar incidences and m ean severity in control and treated groups. Som e findings had low, sporadic incidences or even occurred only in a single individual bird. All w ere considered incidental and unrelated to treatment. CONCLUSIONS AND SUMMARY All adult and offspring bobwhite survived to their respective scheduled euthanasias except for three adults (one 100 ppm a.i. male, one 0 ppm a.i. fem ale, and one 300 ppm a.i. fem ale) which were found dead or euthanized due to moribundity approximately 7-16 weeks before scheduled euthanasia. This low, sporadic early mortality was considered incidental and unrelated to treatm ent. No lesions considered related to test article administration were noted in liver, kidney, ovary, and testis of adult male and fem ale bobwhite or their offspring. T h e few findings in various tissues of adult and offspring bobwhite from control and treated groups were considered incidental and unrelated to test article administration. V 1ARGAARRKHT7A M. G R U E B B E L, D VM , PhD, Diplomate, A C V P Veterinary Pathologist M M G /cb T 6 6 '-A Date -4Contains No CBI  EPL - 157- EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 COM PLIANCE STATEM ENT Client Name Wildlife International, Ltd. EPL Project Coordinator Dr. Margarita M. Gruebbel Client Study 454-116__________________ EPL Pathologist Dr. Margarita M. Gruebbel Species Northern Bobwhite__________ NEPumL bPerorject 212-030 Study Title T-7485: A Reproduction Study with the Northern Bobwhite Test Article T-7485 T h e Histopathology portions of the above-referenced study w ere conducted in com pliance with the Good Laboratory Practice regulations of the Environmental Protection Agency as stipulated by 40 C FR 160; O EC D Principles of Good Laboratory Practice [E N V /M C /C H EM (98)17]; and Japan M AFF (11 NohSan, Notification No. 6283, Agricultural Production Bureau, 1 October 1999). EPL Project Coordinator _____ f l W \M, 2004___________ Date ' \ -5 Contains No CBI - 158- EPL EXPERIMENTAL PATHOLOGY LABORATORIES, INC. Project Number 454-116 QUALITY ASSURANCE FINAL CERTIFICATION Study Title: T-7485: A Reproduction Study with the Northern Bobwhite Client Study: 454-116 EPL Project Coordinator: Dr. Margarita M. Gruebbel EPL Project Number: 212-030 EPL Pathologist: Dr. Margarita M. Gruebbel TEhxepeforilmloewnitnagl Pasaptheocltosgoyf Lthaibsosrtautdoyriewse, rIencin. sDpeactetesdinbsyptehcetioQnusawliteyreAspseurrfaonrmceedUannitdof findings reported to the EPL Project Coordinator and Management are indicated below. Area Inspected _In_s_p_e_c_ti_o_n_______________Date_s_______________R_e_p_o_r_ti_n_g EPL Project Sheets Project Setup Histology Setup Data Review Draft Report Final Report 3/2/04; 4/5/04; 4/26/04 3/4/04; 4/5/04 3/8/04; 4/5/04 4/12/04; 4/13/04 4/29,30/04 5/14/04 3/2/04; 4/5/04; 4/26/04 3/4/04; 4/5/04 3/8/04; 4/5/04 4/12/04; 4/13/04 4/30/04 5/14/04 Date reported to Study Director/Management_____________5/14/04 Form No. 6-2 (7/2/99) -6- Contains No CBI - 159 - Project Number 454-116 SUM M ARY INCIDENCE TABLES ADULT EUTHANASIA Contains No CBI - 160 - Project Number 454-116 SUMMARY INCIDENCE TABLE 454-116 Adult Euthanasia Male Bobwhite KIDNEY (NO. EXAMINED) Cortex, Interstitium, Mineralization Cortex, Tubules, Dilatation Cortex, Tubules, Regeneration Glomerulus, Mineralization Infiltrate, Mononuclear Cell Serosa, Eosinophilic Material LIVER (NO. EXAMINED) Bile Ducts, Hyperplasia Hepatocytes, Atrophy, Diffuse Hepatocytes, Dissociation, Diffuse Hepatocytes, Fatty Chanqe, Diffuse Hepatocytes, Necrosis, Centrilobular Hepatocytes, Necrosis, Focal Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell Periportal Inflammation, Granulomatous TESTIS (NO. EXAMINED) Decreased Diameter Immature Infiltrate, Mononuclear Cell Seminiferous Tubules, Degeneration GROUP GROUP CONTROL 100PPM (16) (16) GROUP 300PPM (16) GROUP 900PPM (16) 4 748 1 4 373 7 486 (16) (16) (16) (16) 11 11 12 13 11 12 11 9 10 21 2 (16) (16) (16) (16) 21 1 1 112 2 11 EPL Experimental Pathology Laboratories, Inc. Contains No CBI - 161 - Project Number 454-116 SUMMARY INCIDENCE TABLE 454-116 Adult Euthanasia Female Bobwhite KIDNEY (NO. EXAMINED) Cortex, Interstitium, Mineralization Cortex, Tubules, Dilatation Cortex, Tubules, Regeneration Glomerulus, Mineralization Infiltrate, Mononuclear Cell Serosa, Eosinophilic Material LIVER (NO. EXAMINED) Bile Ducts, Hyperplasia Hepatocytes, Atrophy, Diffuse Hepatocytes, Dissociation, Diffuse Hepatocytes, Fatty Change, Diffuse Hepatocytes, Necrosis, Centrilobular Hepatocytes, Necrosis, Focal Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell Periportal Inflammation, Granulomatous OVARY (NO. EXAMINED) Hemorrhage Infiltrate, Mononuclear Cell Serosa, Eosinophilic Material Serosa, Inflammation, Chronic Active SPLEEN (NO. EXAMINED) Congestion GROUP GROUP CONTROL 100PPM 0 6 ) (16) GROUP 300PPM (16) GROUP 900PPM (16) 7 342 1 1 4 344 6 284 4 13 (16) (16) (16) (16) 21 1 1 12 13 14 9 1 1 7 996 3222 1 (16) 0 6 ) (16) (16) 1 11 1 4 442 1 (1) 1 EPL Experimental Pathology Laboratories, Inc. Contains No CBI - 162 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLES ADULT EUTHANASIA Contains No CBI - 163 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE GROUP CONTROL 454-116 Adult Euthanasia A Male Bobwhite N I M A L 3333333333333333 55 5 56 66 6 6 7 7 7 7 7 8 8 3579 13579 135 79 13 KIDNEY XX XXXX Cortex, Interstitium. Mineralization 1 11 1 Cortex, Tubules, Dilatation Cortex. Tubules. Reaeneration Glomerulus. Mineralization Infiltrate, Mononuclear Cell 1 1 1 1 122 1 1 111 Serosa, Eosinophilic Material LIVER Bile Ducts, Hyperplasia Hepatocvtes, Atrophy, Diffuse Hepatocvtes. Dissociation, Diffuse Hepatocvtes, Fatty Chanqe, Diffuse Hepatocvtes, Necrosis, Centrilobular Hepatocvtes. Necrosis. Focal Inflammation, Chronic PeriDortal infiltrate. Mononuclear Cell Periportal Inflammation, Granulomatous TESTIS Decreased Diameter Immature Infiltrate, Mononuclear Cell Seminiferous Tubules, Degeneration 4 3 12 3 1 2 11 21 1 11 11 1 111111 11 X XXXXXXXX XX 2 XX 2 1 13 EPL E x p e rim e n ta l P a th o lo g y Labora to rie s, Inc. 11-1 Key: X=Not Remarkable N=No Section l=lncomp[ete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 164- Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Male Bobwhlte KIDNEY Cortex, Interstltlum, Mineralization Cortex, Tubules, Dilatation Cortex, Tubules. Reaeneratlon Glomerulus, Mineralization Infiltrate, Mononuclear Cell Serosa, Eosinoohilic Material LIVER Bile Ducts. Hvoerolasla Heoatocvtes. Atroohv. Diffuse Hepatocvtes, Dissociation, Diffuse Heoatocvtes, Fattv Chanae, Diffuse Heoatocvtes, Necrosis, Centrllobular Heoatocvtes, Necrosis, Focal Inflammation, Chronic PerlDortal Infiltrate, Mononuclear Cell Periportal Inflammation. Granulomatous TESTIS Decreased Diameter Immature Infiltrate. Mononuclear Cell Seminiferous Tubules. Deqeneration GROUP 100PPM A N I M A L 3u 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 8 889999900000 111 5 79 13 579 135 79 135 XXX X X 1 11 1111 11 11 1 2 1 X 2 1 2 3 111222 5 11 11 111 1 11 1 1 XX XX XXXX XXX XXX P 1 EPL E x p e rim e n ta l P a th o lo g y Labora to rie s, Inc. II-2 Key: X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P^Present B^Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 165 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Male Bobwhite KIDNEY Cortex, Interstitium, Mineralization Cortex. Tubules, Dilatation Cortex, Tubules. Reaeneration Glomerulus. Mineralization Infiltrate, Mononuclear Cell Serosa, Eosinophilic Material LIVER Bile Ducts, Hyperplasia Hepatocvtes. Atrophy, Diffuse Hepatocvtes, Dissociation, Diffuse Hepatocvtes, Fatty Chanae, Diffuse Hepatocvtes, Necrosis. Centrilobular Hepatocvtes. Necrosis, Focal Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell Periportal Inflammation, Granulomatous TESTIS Decreased Diameter Immature Infiltrate, Mononuclear Cell Seminiferous Tubules. Deaeneration GROUP 300PPM A N I M A L 4 4444 4444 44 44444 1122222333334444 7 9 13579 135 7 9 1357 XX 11 11 2 112 1 12 11 11 1 11 1 XX 21 1 12 5 1 1 111 32 4 133 1 1111 X X X X X X Xm XXXXXXXX 1 1 _L_ I EPL E x p e rim e n ta l P a th o lo g y L a b ora to rie s, Inc. 11-3 Key: X=Not Remarkable N=No Section l=lncomptete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 166 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Male Bobwhlte KIDNEY Cortex. Interstitium. Mineralization Cortex. Tubules. Dilatation Cortex, Tubules. Reaeneration Glomerulus. Mineralization Infiltrate. Mononuclear Cell Serosa. Eosinophilic Material LIVER Bile Ducts, Hyperplasia Hepatocvtes. Atroohv. Diffuse Hepatocvtes. Dissociation, Diffuse Hepatocvtes. Fattv Chanae. Diffuse Hepatocvtes. Necrosis. Centrilobular Hepatocvtes. Necrosis. Focal Inflammation. Chronic Periportal Infiltrate. Mononuclear Cell Perioortal Inflammation. Granulomatous TESTIS Decreased Diameter Immature Infiltrate. Mononuclear Cell Seminiferous Tubules. Deaeneration GROUP 900PPM A N I M A L 44444 44444444444 4555556666677777 9 1357 9 13 5 79 135 79 X XX 1 111 1 1 11 2 21 111 111 4 5 113 13 1 11 232 1 11111 1 1111 1 1 XXX X XXXXXXX 2 XX 21 1 EPL E xperim ental Pathology Laboratories, Inc. 11-4 Key; X=Not Remarkable N=No Section ^Incomplete A=Autolysis 1=mimmal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m^missing one paired organ u=unscheduled sac./death Contains No CBI - 167 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Female Bobwhlte KIDNEY Cortex, Interstitium, Mineralization Cortex, Tubules, Dilatation Cortex, Tubules, Reaeneration Glomerulus, Mineralization Infiltrate, Mononuclear Cell Serosa, Eosinophilic Material LIVER Bile Ducts. Hyperplasia Hepatocvtes. Atrophy, Diffuse Heoatocvtes, Dissociation, Diffuse Hepatocvtes. Fatty Chanae. Diffuse Hepatocvtes. Necrosis, Centrilobular Hepatocvtes, Necrosis, Focal Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell Periportal Inflammation, Granulomatous OVARY Hemorrhaae Infiltrate, Mononuclear Cell Serosa. Eosinophilic Material Serosa. Inflammation. Chronic Active GROUP CONTROL A N I M A L 3 3 3 3 3 3 3 3u 3 3 3 3 3 3 3 3 5 5 5 6 6 6 66 7 7 7 7 7 8 8 8 4 6 8 0 2 4 68 0 2 4 6 8 0 2 4 X XX X 1 11 1 11 1 2 1 X 2 1 1 X 11 1 11 X 11 21 2 122 1 112 11 2 111 11 11 XX XXXXX X XXX 2 1 PP PP 2 EPL Experim ental Pathology Laboratories, Inc. 11-5 Key: X=Not Remarkable N=No Section ^Incomplete A=Autolysis 1=minimal 2--slight/mild 3smoderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 168 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Female Bobwhite KIDNEY Cortex, Interstltium. Mineralization Cortex. Tubules. Dilatation Cortex, Tubules, Reaeneration Glomerulus. Mineralization Infiltrate. Mononuclear Cell Serosa. EosinoDhilic Material LIVER Bile Ducts. HvDerolasia Heoatocvtes. Atroohv. Diffuse Heoatocvtes. Dissociation. Diffuse Heoatocvtes. Fattv Chanae. Diffuse Heoatocvtes. Necrosis. Centrilobular Heoatocvtes. Necrosis. Focal Inflammation. Chronic Perioortal Infiltrate. Mononuclear Cell Perioortal Inflammation. Granulomatous OVARY Hemorrhaae Infiltrate. Mononuclear Cell Serosa, EosinoDhilic Material Serosa. Inflammation. Chronic Active GROUP 100PPM A N I M A L 3333333444444444 8899999000001111 68024 6802468n?4R XX X XXX X ?1 1 1 21 11 PP pP 22 2 1 123 11 23 113 1 1 1 11 2 11 11 1 11 XX X X XXX XX XX 2 PP Pp I EPL E x p e rim e n ta l P a th o lo g y Labora to rie s, Inc. H-6 Key; X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimai 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 169 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Female Bobwhite KIDNEY Cortex, Interstitium, Mineralization Cortex, Tubules. Dilatation Cortex. Tubules. Reaeneration Glomerulus, Mineralization Infiltrate. Mononuclear Cell Serosa. Eosinoohilic Material LIVER Bile Ducts, Flvoerplasia HeDatocvtes. Atrophy, Diffuse Hepatocvtes, Dissociation, Diffuse Flepatocvtes, Fatty Chanqe, Diffuse Flepatocvtes. Necrosis, Centrilobular Flepatocvtes. Necrosis, Focal Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell Periportal Inflammation. Granulomatous OVARY Flemorrhaae Infiltrate, Mononuclear Cell Serosa. Eosinoohilic Material Serosa. Inflammation, Chronic Active GROUP 300PPM A N I M A L 4 4 4 4 4 4 4u 4 4 4 4 4 4 4 4 4 1 2 2 2 2 23 3 3 3 3 4 4 4 4 4 8 0 2 4 6 80 2 4 6 8 0 2 4 6 8 X XX 1 1 11 2 2 1 11 1 1 111 1 11 1 P X 2 1 12 12 1 222 232 11 11 1111111 11 X XXX XXXXXX XX P PPP EPL E xperim ental Pathology Laboratories, Inc. 11-7 Key: X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 170 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Adult Euthanasia Female Bobwhite KIDNEY Cortex. Interstitium, Mineralization Cortex. Tubules. Dilatation Cortex. Tubules. Reaeneration Glomerulus. Mineralization Infiltrate, Mononuclear Cell Serosa, EosinoDhilic Material LIVER Bile Ducts. HvDerolasia Heoatocvtes. Atroohv. Diffuse Heoatocvtes. Dissociation. Diffuse HeDatocvtes, Fattv Chanae. Diffuse Heoatocvtes. Necrosis. Centrilobular Heoatocvtes. Necrosis, Focal Inflammation, Chronic Perioortal Infiltrate. Mononuclear Cell Perioortal Inflammation. Granulomatous OVARY Hemorrhaae Infiltrate. Mononuclear Cell Serosa. EosinoDhilic Material Serosa. Inflammation. Chronic Active SPLEEN Conaestion GROUP 900PPM A N I M A L 44444 44444444444 55 555 66 666 7777 78 02468 02468024680 XX XXX X X 11 11 11 PP X 11 11 P X ? ? 1 21 1?3 111 1 11 11 1 1 11 ? XX XXX XX X XX XX X 1 P P 2 - _ ziz: EPL Experim ental P athology Laboratories, Inc. 11-8 Key: X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 171 - Project Number 454-116 CORRELATION OF GROSS AND M ICRO SCO PIC FINDING S ADULT EUTHANASIA Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Animal Number 355 TESTIS Sex: Client Topography / Site 371 TESTES 381 TESTIS Males Group Identification: Client Gross Observations Right -1 .25 cm -1 .0 cm Right -1 .0 cm CONTROL - Sacrificed Microscopic Observations No Correlating Lesion Decreased Diameter (TESTIS) Decreased Diameter --K>a Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. 111-1 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 4 5 4 -1 1 6 A d u lt E uthanasia Species: Bobwhite Animal Number 393 TESTIS 401 TESTIS 403 TESTIS 407 TESTIS 409 TESTIS 415 LIVER Sex: Client Topography / Site Males Group Identification: Left ~1 cm Client Gross Observations Right ~1.2 cm Right -1 .2 5 cm Right -1.2 5 cm Right -1 .2 5 cm Slightly mottled 100PPM - Sacrificed No Correlating Lesion No Correlating Lesion No Correlating Lesion No Correlating Lesion No Correlating Lesion Hepatocytes, Fatty Change, Diffuse -o Project Number 454-116 EPL Experim ental Pathology Laboratories, Inc. III-2 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 A d u lt E uthanasia Species: Bobwhite Animal Number 385 TESTIS Sex: Client Topography / Site Males Group Identification: Client Gross Observations Bilateral - 0.2 x 0.4 - 0.5 cm, black (noted at gross trimming) 100PPM - Unscheduled Sac./Death Microscopic Observations Immature --i Project Number 454-116 EPL Experim ental Pathology Laboratories, Inc. lil-3 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 A dult E uthanasia Species: Bobwhite Sex: Animal Number 427 LIVER Client Topography / Site 433 TESTES 443 TESTIS 445 TESTIS Males Group Identification: Client Gross Observations Slightly mottled -1.25 Right -1.25 cm Left -1 .2 5 cm 300PPM - Sacrificed Microscopic Observations Hepatocytes, Fatty Change, Diffuse No Correlating Lesion (TESTIS) No Correlating Lesion No Correlating Lesion -j Project Number 454-116 EPL Experim ental P athology Laboratories, Inc. II-4 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Animal Number 449 TESTIS Sex: Client Topography / Site TESTIS 451 TESTIS 453 TESTIS 459 TESTIS 463 TESTIS 465 TESTIS 469 TESTES Males Group Identification: Client Gross Observations Right -1.25 cm Left -1.5 cm Right -1 .5 cm Right is -1.25 cm Right -1.25 cm Right -1.0 cm Right -1.25 cm -1.25 cm 900PPM - Sacrificed Microscooic Observations No Correlating Lesion No Correlating Lesion No Correlating Lesion No Correlating Lesion Decreased Diameter No Correlating Lesion No Correlating Lesion No Correlating Lesion (TESTIS) O-js Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. III-5 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Sex: Animai Number 384 Client Topography / Site REPRODUCTIVE Females Group Identification: Client Gross Observations Hemorrhagic follicles CONTROL - Sacrificed Microscopic Observations Hemorrhage (OVARY) Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. III-6 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Animal Number 368 LIVER Sex: Client Topography / Site KIDNEYS Females Pale Pale Group Identification: Client Gross Observations CONTROL - Unscheduled Sac./Death Hepatocytes, Fatty Change, Diffuse No Correlating Lesion (KIDNEY) 00 Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. III-7 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Sex: Animal Number Client Topography / Site KtPKODUCTIVE Females Group Identification: Client Gross Observations Egg yolk peritonitis 300PPM - Sacrificed Serosa, Eosinophilic Material (OVARY) Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Sex: Animal Number 430 Client Topography / Site KIDNEYS OVARY Females Pale Regressing Group Identification: Client Gross Observations 300PPM - Unscheduled Sac./Death Microscopic Observations Cortex, Tubules, Dilatation (KIDNEY) No Correlating Lesion oo o Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. III-9 Contains No CBI CORRELATION OF GROSS AND MICROSCOPIC FINDINGS 454-116 Adult Euthanasia Species: Bobwhite Sex: Animal Number 454 Client Topography / Site REPRODUCTIVE 462 SPLEEN 478 LIVER REPRODUCTIVE Females Group Identification: Client Gross Observations Egg yolk peritonitis Slightly enlarged spleen ~1.5 cm x 1.0 cm Friable Egg yolk peritonitis 900PPM - Sacrificed Microscopic Observations No Correlating Lesion (OVARY) Congestion Flepatocytes, Dissociation, Diffuse No Correlating Lesion (OVARY) Project Number 454-116 EPL Experimental Pathology Laboratories, Inc. 111-10 Contains No CBI - 182 - Project Number 454-116 SUM M ARY INCIDENCE TABLES O FFSPRING EUTHANASIA Contains No CBI - 183 - Project Number 454-116 SUMMARY INCIDENCE TABLE 454-116 Offspring Euthanasia Male Bobwhite KIDNEY (NO. EXAMINED) Infiltrate, Mononuclear Cell GROUP GROUP CONTRO _ 100PPM (6) (6) 1 GROUP 300PPM (7) 1 GROUP 900PPM (7) LIVER (NO. EXAMINED) Hepatocytes, Fatty Chanqe, (6) (6) (7) (7) Diffuse Hepatocytes, Vacuolization, Diffuse 3 375 Inflammation, Chronic 2 12 Periportal Infiltrate,___________________ Mononuclear Cell 13 TESTIS (NO. EXAMINED)______________ (6) (6) (6) (7) Immature 6 667 EPL Experimental Pathology Laboratories, Inc. Contains No CBI - 184 - Project Number 454-116 SUMMARY INCIDENCE TABLE 454-116 Offspring Euthanasia Female Bobwhite KIDNEY (NO. EXAMINED) Infiltrate, Mononuclear Cell LIVER (NO. EXAMINED) Hepatocytes, Fatty Change, Diffuse Hepatocytes, Vacuolization, Diffuse Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell OVARY (NO. EXAMINED) Immature |G R O U P HONTRO . (8) 1 GROUP 100PPM (9) 1 GROUP 300PPM (8) GROUP 900PPM (8) 1 (8) (9) (8) (8) 1 4 244 2 23 1 11 (7) (8) (6) (7) 7 867 EPL Experimental Pathology Laboratories, Inc. Contains No CBI - 185 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLES OFFSPRING EUTHANASIA Contains No CBI - 186- Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Offspring Euthanasia Male Bobwhite KIDNEY Infiltrate, Mononuclear Cell GROUP CONTROL A N I M A BBBBB B L 888888 12367 7 2 3530 6 X XXX X 1 LIVER XX Hepatocvtes, Fattv Chanae, Diffuse Hepatocvtes, Vacuolization,_________ Diffuse Inflammation. Chronic 111 11 Periportal Infiltrate, Mononuclear Cell TESTIS Immature mm p p PPPP GROUP ____________100PPM GGGGGG 222333 478001 54 1293 XXXXXX XXX ? ?1 mmmmm Pp p p p p EPL Experimental Pathology Laboratories, Inc. V-1 Key: X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 187 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Offspring Euthanasia Male Bobwhite . KIDNEY Infiltrate, Mononuclear Cell LIVER Hepatocvtes, Fattv Chanae. Diffuse Hepatocvtes. Vacuolization. Diffuse Inflammation. Chronic Periportal Infiltrate. Mononuclear Cell TESTIS Immature GROUP GROUP 300PPM_______________________ 900PPM A N I M A PPPPp p P L 111 11 12 34 7788 1 28273 80 X X X XX X 1 Ww Ww w w w 6666666 12 34 4 56 7 85159 9 XXXXXXX X 1132 1 12 1 1 mN PPPPp P 1 2 211 11 111 mmm p Pp p p PP _I___ z e z : |. EPL Experimental Pathology Laboratories, Inc. V-2 Key: X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 188 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Offspring Euthanasia Female Bobwhite KIDNEY Infiltrate, Mononuclear Cell LIVER Hepatocvtes. Fattv Chanae. Diffuse Hepatocvtes, Vacuolization, Diffuse Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell OVARY Immature GROUP GROUP CONTROL_________________________100PPM A N I M A BBBBB BBB L 88888 888 2 3444 588 9 0 147 928 XXX XXXX 1 XXX 12 2 11 2 1 N PPP PPPP GGGGGGGGG 222222222 234566789 932305694 XXXXXXXX 1 XXX X 1 21 11 N PPPPPPPP EPL Experimental Pathology Laboratories, Inc. V-3 Key: X=Not Remarkable N=No Section !=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI - 189 - Project Number 454-116 HISTOPATHOLOGY INCIDENCE TABLE 454-116 Offspring Euthanasia Female Bobwhite KIDNEY Infiltrate, Mononuclear Cell LIVER Heoatocvtes, Fatty Chanqe. Diffuse Heoatocvtes. Vacuolization, Diffuse Inflammation, Chronic Periportal Infiltrate, Mononuclear Cell OVARY Immature GROUP 300PPM__________________ GROUP 900PPM A N I M A PPPPPPPP L 11111222 356690 12 9 5 159 550 XXXXX XXX X XX WWWWWWWW 56666666 90 124578 7 30 1925 1 XXXXX XX 1 XX X 1 2 22 11 1 1 NN PPPP PP 11 32 1 N PPPPPPp | EPL Experimental Pathology Laboratories, Inc. V-4 Key: X=Not Remarkable N=No Section l=lncomplete A=Autolysis 1=minimal 2=slight/mild 3=moderate 4=moderately severe 5=severe/high P=Present B=Benign M=Malignant m=missing one paired organ u=unscheduled sac./death Contains No CBI ,W ildlife International Ltd - 1 9 0 - Appendix XV Certificate of Analysis Project Number 454-116 Contains No CBI ,W ildlife International Ltd - 191 - Project Number 454-116 c: i-i Test Name Purity^ C E R T IF IC A T E O F A N A LY S IS Revision #1 (15-September-03) Exygen Research COA Reference #: 023-067 3M Product: PFBS,I.ot#5 Purity: 98.2% Specifications Appearance ' Identification NMR A M etals (IC P/M S)/ 1. C alcium 2. M agnesium 3. Sodium 2 4. Potassium 2 . 5. N ickel : : . 6. iron 7. M anganese Total % Im purity (NMR,) Total % Impurity (LC/M S) ' ' : Total % Impurity : (GC/M S) 7t Residual Solvents TGA ; Purity by DSC inorganic A nions (IC) 1. C hloride 2. Fluoride : 3. B rom ide s : 4. Nitrate I 5. N itrite i 6. Phosphate | 7. Sulfate ! O rganic A cids ` (IC ) 1. T FA 2. l 'F P A 3. HFBA 4. NFPA Elem ental Analysis": 1. C arbon 2. H ydrogen 3. N itrogen 4. Sulfur 5. Fluorine White, crystalline solid Theoretical Value = 14.2% Theoretical Value = 0.0% Theoretical Value = 0% Theoretical Value = 0.5% Theoretical Value = 50.6% Result 98.2 % C onform s Positive : ! 1. 0.002 w t./w t.% 2. <0.001 w t/w t.% 3. 1.142 w t/w t.% ' 4. 8.442 wt.Avt.% ' 5. <0.001 wt./wt.% 6. 0.002 wt./wt.% 7. <0.001 wt./wt.% , 0.9 wt./wt.% 0.774 w t/w t.% None Quantified 0.02 w t/w t.% 99.69% .. 1. <0.015 wl.Avt.% 2. 0.101 w t/w t.% ; 3. <0.040 wt.Avt.% ! 4, <0.009 wt./wt.% 5. <0.006 wt.Avt.% 6. <0.006 w t/w t.% 7. <0.07S wt.Avt.% 1. <0.1 wl.Avt.% 2. <0.1 w t/w t.% 3. <0.1 wt./wl.% 4. <0.25 w t/w t.% 1. 2. 3. 4. ,,5 10.4 wt.Avt.% 0.311 w t/w t.% 3.94 wt.Avt.% 9.46 wt.Avt.% 38.1 wt.Avt.% COA023-067 Page 1 o f 2 30.58 Research Drive Suite College, PA 6801. USA T: 800.281.3219 r- h i .1 t ?? i n n Contains No CBI ,Wildlife International Ltd - 192- Project Number 454-116 Precise Research. Proven Results. C E R T IF IC A T E O F A N A L Y S IS Revision #1 (lS-September-03) Exygcn Research COA Reference #: 023-067 Date of Last Analysis: 08/31/01 Expiration Date: 08/31/08 ............. < v: .. Storage Conditions: Frozen Rc-assessment Date: 08/31/08 1 Purity - 100% - (Total NMR impurities, 0.90% + Total LC/MS impurities, 0.774% + Total Metal impurities, 0.004% + Total Anion Impurities, 0.101 + Residual Solvents TfiA, 0.02%) " TotaLuqpurity fifoni'alf tests = 1.790% Purity = 100% - .799% = 98.2% 'Potassium and Sodium arepresent-as counterions and thiis are excluded from the purity calculation. !"" '"T";:1'-; : ! ;.+ : : VI'FA HFBA NFPA PFPA Trifhtoroacetic acid Heptafiuorolmtyfic aeid Nonafkioropentenptc-acid ' Pentafluotoppanoic acid! 'Theoretical value calculations based on the etnpnica] formula, GtFijSCbTC (MW=338.2) Scientist, iygen Research Vice-President - Operation?, Exygen Research Date Contains No CBI W ildlife International, Ltd. - 193 - Project Number 454-116 Appendix XVI The Analysis of T-7485 in Avian Diet Contains No CBI ,W ildlife International Ltd. - 194- Project Number 454-116 Appendix XVI Table 1 Typical LC/MS/MS Operational Parameters INSTRUMENT: ANALYTICAL COLUMN: Hewlett-Packard Model 1100 High Performance Liquid Chromatograph with a Perkin-Elmer SCIEX API 3000LC Mass Spectrometer equipped with a Perkin-Elmer SCIEX TurboIonSpray ion source. Operated in multiple ion reaction monitoring (MRM) mode. Keystone PRISM RP column (50 mm x 2.0 mm I.D., 3-pm particle size) GUARD COLUMN: Keystone Javelin Cm column (20 mm x 2.0 mm I.D.) OVEN TEMPERATURE: 40C STOP TIME: 3.00 minutes FLOW RATE: 200 pL/minute MOBILE PHASE: 10% HPLC-Grade Reagent Water with 0.02M Ammonium Formate: 90% Methanol INJECTION VOLUME: 5.0 pL T-7485 RETENTION TIME: Approximately 2.1 minutes T-7485 MONITORED MASS: 299.0 amu -> 98.9 amu Contains No CBI W ildlife International, Ltd. - 195 - Project Number 454-116 Appendix XVI Table 2 Example Calculations Sample number 454-116-28, nominal concentration o f 100 ppm a.i. in avian feed Peak Area = 285711 Y-intercept = 45978.2266 Slope = 22645852 Primary Dilution: Initial Weight (Wi) = 10.0 g Original Final Volume (Vj)= 100 mL Secondary Dilution: Initial Volume (V;) = 0.0500 mL Final Volume (Vf) = 50.0 mL Overall Dilution Factor (Vi/W; x V /V ) = 10000 mL/g = 10000 L/Kg (peak area - y-intercept) T-7485 at instrument (mg a.i./L) = slope (285711 -45978.2266) 22645852 = 0.01059 mg a.i./L T-7485 in sample (mg a.i./Kg) = T-7485 at instrument (mg a.i./L) x overall dilution factor (L/Kg) T-7485 in sample (mg a.i./Kg) = 0.01059 mg L a.i, 10000 L x Kg = 105.9 mg a.i./Kg 105.9 ppm a.i. Percent o f Nominal Concentration = T-7485 (ppm a.i.) in sample T-7485 (ppm a.i.) nominal x 105.9 ~TnrT X 100= 106% Instrument Software: MacQuan, version 1.6. Contains No CBI W ildlife International,Ltd, Project Number 454-116 Number (454-116-) MAB-1 MAS-1 MAS-2 MAS-3 MAB-2 MAS-4 MAS-5 MAS-6 MAB-3 MAS-7 MAS-8 MAS-9 MAB-4 MAS-10 MAS-11 MAS-12 Appendix XVI Table 3 Matrix Blank and Fortifications Analyzed Concurrently with the Samples Sample Type Interval Concentration of T-7485 (ppm a.i.) Fortified Measured1,2 Percent Recovery2 Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Week 1, Day 0 Week 1, Day 0 Week 1, Day 0 Week 1, Day 0 Week 1, Day 7 Week 1, Day 7 Week 1, Day 7 Week 1, Day 7 Week 4, Day 0 Week 4, Day 0 Week 4, Day 0 Week 4, Day 0 Week 8, Day 0 Week 8, Day 0 Week 8, Day 0 Week 8, Day 0 0.00 50.0 200 1000 0.00 50.0 200 1000 0.00 50.0 200 1000 0.00 50.0 200 1000 <20.0 43.3 198 993 <20.0 41.0 182 992 <25.0 57.1 234 1050 <25.0 55.9 222 1042 86.7 99.1 99.3 _ 81.9 91.1 99.2 _ 114 117 105 - 112 111 104 Mean Recovery (?) 95 91 112 109 1Less than values correspond to the ppm a.i. equivalent concentration of the low-level calibration standard in the matrix blank extract. 2Results were generated using MacQuan version 1.6 software. Manual calculations may differ slightly. - 196- Contains No CBI W ildlife International, Ltd, Project Number 454-116 Number (454-116-) MAB-5 MAS-13 MAS-14 MAS-15 MAB-6 MAS-16 MAS-17 MAS-18 Appendix XVI Table 3 (Continued) Matrix Blanks and Fortifications Analyzed Concurrently with the Samples Sample Type Interval Concentration of T-7485 (ppm a.i.) Fortified1 Measured1,2 Percent Recovery Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Week 12, Day 0 Week 12, Day 0 Week 12, Day 0 Week 12, Day 0 Week 16, Day 0 Week 16, Day 0 Week 16, Day 0 Week 16, Day 0 0.00 50.0 200 1000 0.00 50.0 200 1000 <25.0 52.3 209 1055 <25.0 54.1 210 1042 _ 105 105 106 - 108 105 104 Mean Recovery G) 105 106 MAB-7 MAS-19 MAS-20 MAS-21 Matrix Blank Matrix Fortification Matrix Fortification Matrix Fortification Week 20, Day 0 Week 20, Day 0 Week 20, Day 0 Week 20, Day 0 0.00 50.0 200 1000 <25.0 51.3 204 1045 103 102 105 1Less than values correspond to the ppm a.i. equivalent concentration of the low-level calibration standard in the matrix blank extract. 2Results were generated using MacQuan version 1.6 software. Manual calculations may differ slightly. 103 -197- Contains No CBI Wildlife International, Ltd. Project Number 454-116 Nominal Concentration (ppm a.i.) 100 300 900 Sample I.D. Number (454-116-) 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 Appendix XVI Table 4 Homogeneity of T-7485 in Avian Diet Location Sampled in Mixing Vessel Measured T-7485 Concentration (ppm a.i.) Top Left Top Right Middle Left Middle Right Bottom Left Bottom Right Top Left Top Right Middle Left Middle Right Bottom Left Bottom Right Top Left Top Right Middle Left Middle Right Bottom Left Bottom Right 104 104 102 97.0 112 108 299 312 303 282 311 289 870 880 876 859 903 894 Mean g) Standard Deviation (SD) Coefficient of Variation (CV) %= 105 ppm a.i. SD = 5.13 ppm a.i. CV = 4.91% %= 299 ppm a.i. SD = 12.0 ppm a.i. CV = 4.00% = 880 ppm a.i. SD = 16.0 ppm a.i. CV= 1.82% Mean Percent of Nominal 105 100 98 198 - Contains No CBI W ildlife International, Ltd. Project Number 454-116 Nominal Concentration (ppm a.i.) Sample I.D. Number (454-116-) 27 34 41 48 55 Appendix XVI Table 5 Verification o f T-7485 Concentrations in Avian Diet Interval (Day 0 of W eek-) Measured T-7485 Concentration1 (ppma.i.) Mean () Standard Deviation (SD) Coefficient of Variation (CV) 4 <25.0 \ = <25.0 ppm a.i. 8 <25.0 12 <25.0 16 <25.0 20 <25.0 Percent of Nominal -- - -- Mean Percent of Nominal 28 4 106 106 29 4 119 119 35 8 103 = 107 ppm a.i. 103 36 8 107 SD = 7.74 ppm a.i. 107 42 12 102 CV = 7.21% 102 43 12 121 121 49 16 110 110 50 16 109 109 56 20 99.8 99.8 57 20 97.3 97.3 1Less than values correspondto the ppm a.i. equivalent concentration of the low-level calibration standard in the matrix blank extract. 107 -199- Contains No CBI Wildlife International, Ltd, Project Number 454-116 -200 - Appendix XVI Table 5 (Continued) Verification o f T-7485 Concentrations in Avian Diet Nominal Concentration (ppm a.i.) Sample I.D. Number (454-116-) Interval (Day 0 of Week -) Measured T-7485 Concentration1 (ppm a.i.) Mean () Standard Deviation (SD) Coefficient of Variation (CV) 300 30 4 323 31 4 312 37 8 310 = 314 ppm a.i. 38 8 320 SD= 5.97 ppm a.i. 44 12 311 CV= 1.90% 45 12 323 51 16 315 52 16 309 58 20 312 59 20 306 900 32 33 39 4 983 4 949 8 938 40 8 929 46 12 1013 47 12 990 53 16 9911 54 16 989 60 20 900 61 20 928 1 Result corresponds to the mean of duplicate re-injections of the original sample extract. %= 961 ppm a.i. SD = 36.8 ppm a.i. CV = 3.83% Percent of Nominal 108 104 103 107 104 108 105 103 104 102 109 105 104 103 113 110 1101 110 100 103 Mean Percent of Nominal 105 107 Contains No CBI W ildlife International,Ltd Project Number 454-116 Appendix XVI Table 6 Ambient Stability of T-7485 in Avian Diet During the Northern Bobwhite Reproduction Study Nominal Concentration (ppm a.i.) 0 100 300 900 Sample Number (454-116-) 1 2-7 8-13 14-19 Week 1, Day 0` Mean Measured (ppm a.i.) -7485 Concentration Mean Percent of Nominal Sample Number (454-116-) Week 1, Day 7 Measured2 (ppm a.i.) Mean Measured (ppm a.i.) <20.0 105 299 880 105 100 98 20 <20.0 <20.0 21 95.2 101 22 106 23 300 313 24 326 25 989 982 26 975 1Day 0 values are from homogeneity samples presented in Table 4. 2Less than values correspond to the ppm a.i. equivalent concentration of the low-level calibration standard in the matrix blank extract. Mean Percent of Day 0 96 105 112 -201 - Contains No CBI ,W ildlife International, Ltd -202- Project Number 454-116 Appendix XVI Analytical Method Flowchart for the Analysis of T-7485 in Avian Diet Prepare matrix fortification samples in the desired avian feed stock using the dry mix technique. Weigh 10-g samples of the matrix blank, matrix fortification and test samples into weigh boats and transfer to 8-oz. French square glass bottles. Record weights. i For each sample, measure 100 mLs of methanol with a graduated cylinder and transfer into the French square bottle. i Cap bottles and place on shaker table. Allow the samples to shake for a minimum of 30 minutes at approximately 250 rpm. I Transfer an aliquot of each sample into a 20-mL scintillation vial and centrifuge at a setting of 1500 rpm for approximately 10 minutes. I Transfer an aliquot of each sample from the scintillation vial into a 3-mL B-D syringe fitted with a 0.45 gm Acrodisc and filter into a new scintillation vial. 4 Prepare appropriate dilution(s) to bring final concentration into the calibration range of the LC/MS/MS methodology. Use methanol for intermediate dilutions, if required. For all final dilutions, use 50% methanol:50% HPLC-grade reagent water (v/v) dilution solvent. 4 Ampulate and submit samples for LC/MS/MS analysis. Figure 1. Analytical method flowchart for the analysis of T-7485 in avian diet. Contains No CBI W ildlife International, Ltd, -203 - Appendix XVI T-7485 299.0->98.9 No Internal Standard Weighted (1/x) Intercept = 45978.2266 Slope = 22645852.0000 Correlation Coeff. = 0.99756 Area Project Number 454-116 Figure 2. Representative calibration curve for T-7485. Contains No CBI ,W ildlife International Ltd -204- Project Number 454-116 Appendix XVI T7485_1 LCMS-6 STD 0.00500 mg a.i./L 2.9S In 1 period T-7485 No Internal Standard Use Area 1: 2.98 MRM, 179 scans 299.0->98.9 Noise Thres. 10.0 Quant Thres. 1.0 Min. Width 12 Mult. Width 10 Base. Width 50 RT Win. (secs) 20 Smooth Expected RT 1 2.23 Area 148860 Height 21966 Start Time End Time Integration Width Retention Time Integration Type 1.94 2.78 0.84 2.13 A BB Tue. Nov 11, 2003 13:06 intensity: 100000 cps Figure 3. Representative ion chromatogram of a low-level T-7485 standard, 0.00500 mg a.i./L. Contains No CBI W ildlife International, Ltd -205 - Project Number 454-116 Appendix XVI T7485_5 LCMS-10 STD 0.0250 mg a.i./L 2.98 in 1 period T-7485 No Internal Standard Use Area 1: 2.98 MRM, 179 scans 299.0->98.9 Noise Thres 10.0 Quant Thres. 1.0 Min. Width 12 Mult. Width 10 Base. Width 50 RT Win. (secs) 20 Smooth 1 Expected RT 2.23 Area 585024 Height 75727 Start Time 1.94 End Time 2.81 Integration Width 0.87 Retention Time 2.13 Integration Type A BB Tue, Nov 11, 2003 13:22 intensity: 100000 cps Figure 4. Representative ion chromatogram of a high-level T-7485 standard, 0.0250 mg a.i./L. Contains No CBI ,W ildlife International Ltd -206- Project Number 454-116 Appendix XVI T7485_7 454-116- MAB-3 Tue, Nov 11, 2003 13:30 2.98 in 1 period T-7485 No internal Standard Use Area 1: 2.98 MRM, 179 scans 299.0->98.9 Noise Thres. 10.0 Quant Thres. 1.0 Min. Width 12 Mult. Width 10 Base. Width 50 RT Win. (secs) 20 Smooth 1 Expected RT 2.23 Area 0 Height 0 Start Time End Time Integration Width Retention Time Integration Type 0.00 0.00 0.00 0.00 100-1 90807060504030 20100-* 3140 31 0.52 69 61 1.02 97 91 1.52 intensity: 100000 cps 'T 127 121 2.03 147 165 151 2.53 Scan Time Figure 5. Representative ion chromatogram of a matrix blank sample, (454-116-MAB-3). The arrow indicates the approximate retention time of T-7485. 10-g sample, Overall dilution = 5000X. Contains No CBI W ildlife International, Ltd -207- Project Number 454-116 Appendix XVI T7485_9 454-116- MAS-8 Tue, Nov 11, 2003 13:38 2.98 in 1 period T-7485 No Internal Standard Use Area 1: 2.98 MRM, 179 scans 299.0->98.9 Noise Thres. 10.0 Quant Thres. 1.0 Min. Width 12 Mult. Width 10 Base. Width 50 RT Win. (secs) 20 Smooth 1 Expected RT 2.23 Area 311191 Height 45599 Stari Time End Time Integration Width Retention Time 1.94 2.78 0.84 2.13 Integration Type A-BB intensity: 100000 cps Figure 6. Representative ion chromatogram o f a matrix fortification (454-116-MAS-8). Nominal concentration 200 ppm a.i., 10-g sample, Overall dilution = 20000X. Contains No CBI ,W ildlife International, Ltd -208 - Project Number 454-116 Appendix XVI T7485_13 28 Tue, Nov 11, 2003 13:54 454-116- 2.98 in 1 period T-7485 No Internal Standard Use Area 1: 2.98 MRM, 179 scans 299.0->98.9 Noise Thres. 10.0 Quant Thres. 1.0 Min. Width Mult. Width 12 10 Base. Width 50 RT Win. (secs) 20 Smooth Expected RT 1 2.23 Area 285711 Height 40320 Start Time 1.94 End Time Integration Width 2.78 0.84 Retention Time 2.13 Integration Type A - BB intensity: 1 0 0 0 0 0 cps Figure 7. Representative ion chromatogram of an avian diet sample from Week 4, Day 0, (454-115-28). Nominal concentration 100 ppm a.i., 10-g sample, Overall dilution = 10000X. Contains No CBI ,W ildlife International, Ltd -209 - Project Number 454-116 Appendix XVII Analytical Determination o f T-7485 in Northern Bobwhite Tissues Contains No CBI -210- Project Number 454-116 Study Title T-7485: A Reproduction Study with the Northern Bobwhlte Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Requirement 40 PR Part 792 A uthor Mark E. Ellefson Phase Initiation Date August 20, 2004 Phase Completion Date Upon Signing Performing Laboratory 3M Environmental Laboratory Building2-3E-09 935 BushAvenue St. Paul, MN 55106 Project Identification 3M Environmental Laboratory Study # E03-0583 Wildlife International Ltd: 454-116 Total Number of Pages 227 Contains No CBI -211 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues This page has been reserved for specific country requirements. 3M Environmental Laboratory Page 2 of 227 Contains No CBI Report E03-0583 -212- Project Number 454-116 ! T-7485: A Reproduction Study with the Northern Bobwhita Phase: Analytical Determination of PFBS In Northern Bobwhita Tissues GLP Compliance Statement Study Title: T-7485: A Reproduction Study with The Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Study Identification Number: E03-0583 This analytical phase was conducted in compliance with Toxic Substances Control Act (TSCA) Good Laboratory Practice (GLP) Standards with the exceptions listed below: Exceptions to GLP compliance: 40 CFR 792.105 (b): The stability of the test substance in the test system (quail tissues), in the sample homogenate, and in the sample extract is not known. 40 CFR 792.105 (e): The stability of the test (and reference) substance under the storage conditions at the test site is not known. 3M Environmental Laboratory Page 3 of 227 Contains No CBI Report E03-0583 -213 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Quality Assurance Statement Study Title: T-7485: A Reproduction Study with The Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Study Identification Number: E03-0583 The analytical phase was audited by the 3M Environmental Laboratory Quality Assurance Unit (QAU), as indicated in the following table. The findings were reported to the study director and laboratory management. In s p e c t io n D a t e s 08/05/2004 - 08/06/2004 09/03/2004, 09/16/2004, 09/21/2004, 09/27/2004 10/14/2004, 10/18/2004 -10/21/2004 11/01/2004 - 11/05/2004, 11/10/2004 11/12/2004,11/15/2004 -11/16/2004 12/20/2004-12/22/2004 1/5/2005 - 1/7/2005, 1/10/2005 Phase Protocol Amendment In-phase (Liver) Serum Data Liver Data Egg Data Final Report Date Reported t M anagement S t u d y D ir e c t o r 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 2/23/2005 03/<xhs` Date 3M Environmental Laboratory Page 4 of 227 Contains No CBI Report E03-0583 -214- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table of Contents GLP Compliance Statement.... Quality Assurance S tatem ent. Table of C ontents..................... List of T a b le s .............................. Study Inform ation...................... S um m a ry..................................... P urp ose........................................ Reference Substance................. Specimen Receipt and Storage Method S um m ary........................ Preparatory M e th o d ................... Analytical M ethod........................ Method V a lid a tio n ....................... Analytical R esults......................... Data S u m m a ry............................. Statement of C onclusion............ Statistical M etho ds....................... R eferences..................................... List of Attachm ents........................ Signature P a g e ............................. Attachm ent A: Extraction and Analytical Method, ETS-8-231.2, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices" ................................... Attachm ent B: Data Summary Tables............................................................ Attachm ent C: Sample Chrom atogram s......................................... Attachm ent D: Study Protocol and Protocol Am endm ents........................................ ..... 3 ..... 4 ..... 5 .... 6 .... 8 .....9 .... 11 ....11 ....11 ....12 ....12 ...12 ... 13 ...18 ...20 ...30 ...32 .,32 ..32 .33 3M Environmental Laboratory Page 5 of 227 Contains No CBI Report E03-0583 -215 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues List of Tables Table 1. Summary Table, C4F9S 0 3' Concentration in Serum ........................................................ g Table 2, Summary Table, C4F9S 0 3` Concentration in Liver.................................................. -)0 Table 3. Summary Table, C4F9S 0 3' Concentration in E g g .................................................. Table 4, Reference Substance....................................................................... -10 . Table 5. Specimen Receipt and S to ra g e .............................................................. 1, Table 6. Extracted Serum C alibration........................................................... 13 Table 7. Extracted Liver C alibration..................................................................... 14 Table 8. Extracted Egg C alibration..................................................................... 14 Table 9. Limits of Detection and Q uantitation..................................................................... 15 Table 10. Absolute Recovery o f Sample E xtraction........................................................ Table 11. Dilution V erification......................................................... j5 Table 12. Extract S ta b ility................................................................. Table 13. Uncertainty Calculation................................................................ ^ 17 Table 14. Matrix Blanks: PFBS in Control Serum ....................................................... Table 15. Matrix Blanks: PFBS In Control Liver...................................... Table 16. Serum QC Sample Recovery.............................................. 18 Table 17. Liver QC Sample R e co ve ry............................................................................. Table 18. Egg QC Sample Recovery.............................................................................. Table 19. C4F9S 0 3- Data Summary, 100 ppm Dose Group, Adult Female Quail S eru m ....20 Table 20. C4F9S 0 3' Data Summary, 100 ppm Dose Group, Adult Male Quail S e ru m .........21 Table 2 1 . C4F9S 0 3'D a ta Summary, 100 ppm Dose Group, Offspring Quail Serum .............21 Table 22. C4F9S 0 3' Data Summary, 300 ppm Dose Group, Adult Female Quail S eru m ................... 22 Table 23. C4F9S 0 3 Data Summary, 300 ppm Dose Group, Adult Male Quail S e ru m ........................22 Table 24. C4F9S 0 3` Data Summary, 300 ppm Dose Group, Offspring Quail Serum .......................... 23 Table 25. C4F9S 0 3` Data Summary, 900 ppm Dose Group, Adult Female Quail S eru m ....................23 Table 26. C4F9S 0 3 Data Summary, 900 ppm Dose Group, Adult Male Quail S e ru m .........................24 Table 27. C4F9S 0 3` Data Summary, 900 ppm Dose Group, Offspring Quail S erum ..........................24 Table 28. C4F9S 0 3' Data Summary, 100 ppm Dose Group, Adult Female Quail Liver...................... 25 Table 29. C4F9S 0 3' Data Summary, 100 ppm Dose Group, Adult Male Quail L iv e r........................... 25 Table 30. C4F9S 0 3` Data Summary, 100 ppm Dose Group, Offspring Quail Liver............................. 26 Table 31. C4F9S 0 3` Data Summary, 300 ppm Dose Group, Adult Female Quail Liver....................... 26 Table 32. C4F9S 0 3` Data Summary, 300 ppm Dose Group, Adult Male Quail L iv e r............................27 Table 33. C4F9S 0 3' Data Summary, 300 ppm Dose Group, Offspring Quail Liver............................. 27 Table 34. C4F9SQ3' Data Summary, 900 ppm Dose Group, Adult Female Quail Liver..................... 28 3M Environmental Laboratory Page 6 of 227 20 20 Contains No CBI -2 1 6 - P ro ject N u m b er 4 5 4 -1 1 6 R eport E 03-0583 T -7 4 85 : A Reproduction Study w ith the Northern Bobwhite Phase: A nalytical D eterm ination o f PFBS in Northern Bobw hite Tissues Table 35. C4FbS 03 DataSummary, 900 ppm Dose Group, Adult Male Quail Liver..................... 28 Table 36. C4F9SO3 DataSummary, 900 ppm Dose Group, Offspring QuailLiver......................... 29 Table 37. C^eSOs Data Summary, Quail Egg Lot B .................................................... 29 Table 38. C4FS03 Data Summary, Quail Egg Lot G .................................................... 30 Table 39. Summary of Results: C4F9S O / Concentration in Serum..................................................31 Table 40. Summary of Results: C4FbS0 3' Concentration in Liver....................................................31 Table 41. Summary Table, C4FS03 Concentration in Egg............................................................ 32 3M Environm ental Laboratory Page 7 o f 227 Contains No CBI -217- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Study Information Sponsor 3M Environmental Technology and Safety Services 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 Sponsor Representative Dr. John Newsted ENTRIX East Lansing Office 4295 Okemos Road, Suite 101 Okemos, Ml 48864 Study Director Sean P. Gallagher, Senior Avian Biologist Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 Principal Analytical Investigator, Analytical Phase Mark E. Ellefson 3M Environmental Technology and Safety Services 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 Study Location Testing Facility, Analytical Phase 3M Environmental Laboratory 3M Environmental Technology and Safety Services 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 Study Personnel, Analytical Phase William K. Reagen, Ph.D., Testing Facility Management, Analytical Phase Mark E. Ellefson, Principal Analytical Investigator, Analytical Phase Cindy M. Carlson, Analytical Chemist Mark L. Anderson, Analytical Chemist Study Dates Study Initiation: 10 September 2003 Experimental Initiation: 30 January 2003 Experimental Completion: 01 April 2003 Analytical Phase Completion: Date of signing Location of Archives All original raw data, protocol, and analytical report have been archived at the 3M Environmental Laboratory according to 40 CFR Part 792. The test substance and analytical reference standard reserve samples are archived at the 3M Environmental Laboratory according to 40 CFR Part 792. 3M Environmental Laboratory Page 8 of 227 Contains No CBI Report E03-0583 -218- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Summary The concentration of the C4F9S 0 3` anion of perfluorobutane sulfonate, potassium salt (PFBS, C4F9SO3 K+) was determined in the liver, serum, and eggs of adult bobwhite quail and their offspring in a control group and three dose groups consisting of 100, 300 and 900 pg PFBS/gram feed. Extracted matrix standards were used to establish a calibration curve. Concentrations of C4F9SO3 are reported after correction for the purity and the potassium contribution to the mass of the PFBS reference substance used. Table 1 summarizes data from the serum samples. Table 2 summarizes data from the liver samples. Table 3 summarizes data from the egg samples. Table 1. Summary Table, C4F9SO3" Concentration in Serum* Docn Group and G .iidcr , Number of > i im i Samples ' S'mm ^ ug'pFS^g0 R 1111 1p`-<iults 100 ppm, Male 13 16.5 12.3 5.81-37.7 100 ppm, Female 100 ppm Offspring 300 ppm, Male 300 ppm, Female 15 14.6 13.1 15 0.0369* 0.0334* 16 27.9 26.3 15 37.8 27.8 2.49-33.1 < 0.0278 - 0.0552 14.2-42.9 12.2-102 300 ppm Offspring 15 0.0567 0.0433 0.0245-0.119 900 ppm, Male 16 68.2 63.4 34.3-98.8 900 ppm, Female 16 104 82.2 30.5-370 900 ppm Offspring 15 0.133 0.117 0.0413-0.320 'The uncertainty for the serum sample values is +/-15% as calculated in the validation. Mean and Median reported for samples in which analyte was detected. For more specific information see tables in Attachment A The control serum samples that showed measurable levels of target analyte are listed individually in Table 14 3 For individual sample values see Attachment A 3M Environmental Laboratory Page 9 of 227 Contains No CBI Report E03-0583 -219- Project Number 454-116 T-/485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 2. Summary Table, C4F9S 03' Concentration in Liver* Dose Group and Gender Number of Liver Samples Mean Concentration ig PFBS g Li,er Median Concentration, ug PFBS/g Liver Range of Results 100 ppm, Male 17 3.25 2.71 0.808-8.89 100 ppm, Female 16 3.52 3.91 0.396-8.19 100 ppm Offspring 15 0.0211* 0.0233* <0.0137-0.0301 300 ppm, Male 15 7.78 7.52 1.42-14.2 300 ppm, Female 16 11.1 9.08 3.57-23.5 300 ppm Offspring 15 0.0515* 0.0288* <0.0138 - 0.200 900 ppm, Male 16 15.7 15.1 6.73 - 23.7 900 ppm, Female 16 29.6 23.5 9.73-77.4 900 ppm Offspring 15 0.111 0.0935 0.0340 - 0.313 TThe uncertainty for the liver sample values is +/- 46% as calculated in the validation. *Mean and Median reported for samples in which analyte was detected. For more specific information see tables in Attachment A The control liver samples showing measurable levels of target analyte are listed individually in Table 15. Table 3. Summary Table, C4F9S 03' Concentration in Egg* Do,p Group Number of Egg Samples IV in Com. miration u j PFBS g Egg Mi chan Concentration ug PFBS'g Egg 100 ppm, LotB 7 7.65 7.14 300 ppm, Lot B 7 23.6 25.2 900 ppm, Lot B 8 50.5 39.7 100 ppm, Lot G 8 14.0 12.4 300 ppm, Lot G 7 31.4 27.6 900 ppm, Lot G 8 92.6 92.4 TThe uncertainty of the egg sample values is +/-15% ascaiculatedlnTheTaiidation" Rang 'fP olii1 4.92-10.8 13.3-36.5 33.8-118 8.44-26.2 16.1-73.1 52.6 -137 None of the egg samples from the control group demonstrated measurable levels of target analyte at a level greater than 50% of the LLOQ. 3M Environmental Laboratory Page 10 of 227 Contains No CBI Report E03-0583 -220- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Purpose The purpose of this analytical phase of the study was to quantify C4F9S 0 3' in the liver, serum, and eggs of both control and dosed Northern Bobwhite Quail and their offspring. Reference Substance Table 4. Reference Substance Reference Substance Acren, m and Structure Identification Number Source Solubility in Water Purity Stou jiConditions Expu lti ->n Date Potassium Perfluorobutane sulfonate** PFBS C4F9S03'K+ TCR-282 3M Specialty Chemicals 56.6 mg/mL 97.3% Frozen * According to E00-1877, "Determination of the Solubility of Potassium Perfluorobutanesulfonate (PFBS) in Water" **The vapor pressure and octanol/water partition coefficient are not available. 1/17/2007 Specimen Receipt and Storage During the "In-Life " phase of the study, Northern Bobwhite serum, liver, and egg samples were collected by W ildlife International, Ltd. and sent to the 3M Environmental Laboratory for analysis. The specimens were received at the 3M Environmental Laboratory, assigned a laboratory tracking (LIMS) number (E03-0583) and were stored at -20C +/- 10C until extraction and analysis. Table 5. Specimen Receipt and Storage Sample Giutip Adult Serum Offspring Serum Adult Liver Offspring Liver Eggs Receipt Di te 2/19/2004 4/01/2004 2/19/2004 4/01/2004 2/24/2004 Receipt Ci ndition Frozen Frozen Frozen Frozen Frozen 3M Environmental Laboratory Page 11 of 227 Contains No CBI Report E03-0583 -221 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Method Summary Preparatory Method Preparation and analysis was conducted using the 3M Environmental Laboratory analytical method ETS-8-231, "Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry" with several modifications (noted specifically in the protocol amendment). In summary, the samples were weighed, homogenized in acetonitrile, extracted using a Zymark Rapid Trace automated SPE system, and analyzed using liquid chromatography/mass spectrometry (LC/MS). All sample calculations were based on sample weight rather than volume. Analytical Method Samples were analyzed via ETS-8-231, "Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry." Specific parameters used are listed below. Analytical Equipment Liquid Chromatograph: Hewlett-Packard Series 1100 Liquid Chromatograph system Analytical column: Keystone BetasilTM C 18 2x50mm, 5pm particle size Column temperature: 30 C Stop Time: 9.00 minutes Flow rate: 300 pL/min Injection volume: Variable between 2 and 20 pL Mobile phase components: Solvent A: 2.0 mM ammonium acetate in ASTM Type I water Solvent B: HPLC Grade Methanol Solvent Gradient: Time %B 0.00 10% 0.50 10% 0.80 60% 3.50 100% 5.50 100% 6.00 10% 9.00 10% Mass Spectrometer: Hewlett-Packard Series 1100 API/Mass Spectrometer Detector Software: Agilent ChemStation'" A.08.03 Capillary Voltage: 3000 V Gain = 1.0 EMV Mode: Electrospray Negative Gas Temperature: 350 C Drying Gas: 10 L /min. Nebulizer Pressure: 30 psig Analysis Type: Selected Ion Monitoring (SIM) Compound PFBS Ion fm/z) 299 Fraamentor (volts) 100 3M Environmental Laboratory Page 12 of 227 Contains No CBI Report E03-0583 -222- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Method Validation The analytical method used In this study has been validated for PFOS (Perfluorooctane sulfonate, C8F17S 0 3-) In rat liver and serum and is described In the 3M analytical report E02-1042, "Method and Validation Report for `Solid Phase Extraction and Analysis o f Fluorochemical Compounds from Biological Matrices Method Number: ETS-8-231.1.'" It has also been cross-validated for PFOS in quail and mallard liver and serum, as reported in the 3M analytical report E01-1245, "Analytical Phase Report for PFOS: A Reproduction Study with the Northern Bobwhite." Additional validation elements were performed during the course of these analyses, as detailed below, to extend the validation to PFBS in quail serum, liver and eggs. Specificity Specificity was achieved via chromatographic resolution o f the analyte in conjunction with mass spectral detection of an ion with a mass to charge ratio (m/z) which Is characteristic and specific to the target analyte. Matrix spike recoveries in each matrix (see tables 16 through 18) and example chromatograms of matrix spike samples are included in attachment C of this report to demonstrate method specificity. No other peaks were present with the mass and retention time of the analyte Selectivity The presence of interfering substances was investigated in each matrix by analysis of the control group samples. Instances where the amount of target analyte present was greater than 50% of the LLOQ are documented in tables 14 and 15 Calibration Extracted calibration curves were generated using control matrix during the course of the study. Curve points that did not meet the acceptance requirements of the analytical method were deactivated and the curve was recalculated. Calibration points were also evaluated for stability using successive injections spanning several days. The following acceptance criteria were applied (accuracy within 15% or +20% for the LLOQ, quadratic curve, not weighted, zero not included or forced). The relative standard deviation of the recovery at each concentration was determined and is presented below: Table 6. Extracted Serum Calibration Serum Curve EC-040901-5ng/mL EC-040901 -1 Ong/mL EC-040901-25ng/mL EC-040901-50ng/mL E C -0 4 0 9 0 1-7 5 n g /m L EC-040901 -1 OOng/mL EC-040901-250ng/mL EC-040901 -500ng/mL E C -0 4 0 9 0 1- 7 50 ng /m L EC040901 -1000ng/mL 'Relative Percent Difference (RPD) Number of Observations 4 4 8 9 9 9 9 9 5 2 Recovery '*, RSD 9.0% 3.1% 8.8% 4.7% 1.4% 1.4% 2.7% 1.1% 2.1% 0.0%* 3M Environmental Laboratory Page 13 of 227 Contains No CBI Report E03-0583 -223 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 7. Extracted Liver Calibration Liver Curve EC-041004-5ppb EC-041004-1 Oppb EC-041004-25ppb EC-041004-50ppb EC-041004-75ppb EC-041004-1 OOppb EC-041012d-25 EC-041012d-50 EC-041012d-75 EC-041012d-100 EC-041012d-175 EC-041012d-250 EC-041012d-375 EC-041012d-500 EC-041012d-750 EC041012d-1000 Relative Percent Difference (RPD) Number of Observations 3 3 4 4 4 4 3 4 4 4 4 4 4 4 2 2 Recovery, % RSD 1.5% 0.8% 2.7% 1.5% 4.0% 3.7% 8.9% 6.1% 2.2% 0.5% 1.5% 0.9% 0.9% 1.2% 0.2%* 0.3%* Table 8. Extracted Egg Calibration Egg Curve EC-041202-50 EC-041202-75 EC-041202-100 EC-041202-175 EC-041202-250 EC-041202-375 EC-041202-500 EC-041202-750 EC041202-1000 Number of Observations 6 6 6 6 6 6 6 6 5 Recovery ` RSD 7.9% 1.8% 1.0% 1.9% 1.1% 0.5% 0.5% 1.4% 0.7% 3M Environmental Laboratory Page 14 of 227 Contains No CBI Report E03-0583 -224 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Lower Limit of Detection (LLOD) and Lower Limit of Quantitation (LLOQ) For each matrix, six matrix spikes were prepared ranging in concentration from 0.25 ng/mL to 5.0 ng/mL. The instrument response was evaluated against the theoretical concentration using a linear regression tool in Excel. The LLOD and LLOQ were calculated using the resulting standard error and slope. There are no acceptance limits defined for this phase of the validation. The results are listed below: Table 9. Lower Limits of Detection and Quantitation ? Matrix Serum Liver Egg Method Precision and Accuracy LLOD, nq/mL LLOQ nqmiL 0.732 2.20 0.620 1.86 1.92 5.76 Quality control samples were prepared according to the requirements of the method and the results are presented in tables 16 through 18. The calculated %RSD values represent data collected and averaged over more than one day for serum and liver. The egg data was all collected on the same day. The data passed within acceptance limits. Absolute Recovery and M atrix Effect Blank matrix was spiked at three levels and evaluated against a solvent curve. There are no acceptance limits for this phase of the validation. The results are presented below. Table 10. Absolute Recovery of Sample Extraction Serum Spike-1 Serum Spike-2 Serum Spike-3 Liver Spike-1 Liver Spike-2 Liver Spike-3 Egg Spike-1 Egg Spike-2 Egg Spike-3 Theoretical ( onccnlration (ng niL) 25.11 176.7 401.8 25.11 176.7 401.8 25.11 176.7 401.8 Measured l l p p ^ ^ p s i S Concentration ' (ng/nLj Recovery. 29.43 117% 203.4 115% > ULOQ (255.8) NA 16.72 67% 103.4 59% 233.7 58% 13.23 53% 83.06 47% 161.2 40% The absolute recovery data demonstrates the need to use an extracted calibration curve when quantifying results. Tables 16, 17, and 18 display matrix spike (QC sample) recoveries ranging from 88.6 - 109% in Northern Bobwhite serum, liver, and eggs. The QC samples were quantified using the appropriate extracted calibration curves. Ruggedness The sample data presented in this study represents the work of four analysts and at least two different sets o f equipment (the exception being the Rapid Trace solid phase extraction apparatus). The intra-day accuracy and precision data passed within acceptance limits. 3M Environmental Laboratory Page 15 of 227 Contains No CBI Report E03-0583 -225 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Carry-over Instrument carryover was evaluated throughout the entire course of this study. There were no instances of instrument carry-over following a high concentration standard or sample (area counts after a high standard were less than 50% of the LLOQ area counts). The data passed within acceptable limits. Range of Quantitation This method may be used for any samples whose values fall within the range of the calibration curve (between the LLOQ and approximately 1 ppm). The LOQ and LOD were discussed above. System Suitability The system suitability parameters discussed in ETS-8-231.2 (triplicate injections of one standard with retention time %RSD less than 2% and area count %RSD less than 5% at the opening and close o f each analytical sequence) were applied during the course of this study and are sufficient for the additional techniques and matrices used in this study. The data passed within acceptable limits. Dilution Scheme The dilution scheme used in this study was limited to the variation of injection volumes on the analytical instrumentation. No samples were actually physically diluted with solvent or blank matrix. Quality control samples were evaluated using this technique and passed within the acceptable limits. It is expected that the serum and egg extracts will be representative of the liver dilution recovery. T---a--b--l-e---1--1--.--D---i-l-u--t--i-o--n---V---e--r-i-f-i-c--a--t-i-o---n1---------------------------- 1-------------------------- 1----------------------- Curve I T lie o n tu ai On-Column Sample Injection Concentration. Concentration. Injection Volume Recovery Sample Info ng/inL 1 ng.'mL Volume. uL uL 1 % Extracted Serum Curve Point 102 85.3 4 5 104% Extracted Serum Curve Point 102 38.9 2 5 95.1% Egg Matrix Spike Egg Matrix Spike 25.6 25.6 84.4 83.9 6 2 110% 6 2 109% Egg Matrix Spike 25.6 82.6 6 2 108% 3M Environmental Laboratory Page 16 of 227 Contains No CBI Report E03-0583 -226- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Stability The stability of the test substance in the test system (tissues) and in the homogenate is not known and was not investigated during the course o f this study. Target analyte stability in the serum extracts stored in autovials was investigated by analyzing the following matrix spikes (prepared on 12/22/2004) against a newly extracted curve: Table 12. Extract Stability Spike ID L S -0 4 1222-1 L S -0 4 1222-2 L S -0 4 1222-3 M S -0 4 1 222-1 MS-041222-2 MS-041222-3 H S -0 4 1222-1 H S -0 4 1222-2 HS-041222-3 Recovery on 01 20/2005 u 83% 96% 85% 105% 103% 105% 100% 107% 102% 1 Recovery on 12/23/2004 103% 110% 102% 109% 104% 107% 104% 109% 104% It is expected that the serum extract will be representative of the liver and egg extract stability. The data passed within acceptable limits. Uncertainty The expanded uncertainty was calculated using the model provided in Attachment A of ETS-8 231.2. The results are as follows: Table 13. Uncertainty Calculation Standard Deviation of all QC Samples Square o f the Standard Deviation Homogeneity Standard Deviation Square o f the Homogeneity Standard Deviation Uncertainty Expanded Uncertainty Q uail Scrum 0.0751 0.00565 7.5% 15% Q uail Liver 0.110 0.0121 0.200 0.0400 23% 46% Q uail Egg 0.0750 0.00563 7.5% 15% 3M Environmental Laboratory Page 17 of 227 Contains No CBI Report E03-0583 -227- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues A nalytical Results For a detailed listing of analytical results, refer to Attachment A: Data Tables. Regressions. Quadratic curve fits were applied to calibration standards and sample data to facilitate quantitation over the concentration range appropriate to the data. All calibration curves had least-square fits of 0.990 or greater. Calibration Standards. Extracted matrix standards ranging In concentration from approxim ately 2.5 (or 5.0) to 1000 ng analyte per mL MeOH were used for the calibration curves. High/low calibration standards that were not within the 15% (20% for the lower limit o f quantitation) criteria were deactivated. W here appropriate, high calibration standards were deactivated to improve quantitation over the range appropriate to the data. System Suitability Samples. Three system suitability standards were analyzed before the initial calibration curve and after the last CCV. These samples had area counts with an RSD o f <5% and a retention time RSD of <2% when evaluated independently. Continuing Calibration Verification (CCV). A minimum of one calibration check was analyzed at least every ten samples to monitor instrumental drift. All continuing calibration verification checks were within 15%. Lim it of Quantitation (LOQ). The lower limit of quantitation (LLOQ) was equal to the lowest standard in the calibration curve, with a level o f accuracy within 20%. The upper limit of quantitation (ULOQ) was equal to the concentration of the highest curve point with an accuracy o f 15%. Solvent Blanks. All of the solvent blanks in this study had area counts < 50% o f the LLOQ. Matrix Blanks. The control group of samples was used to evaluate endogenous levels of analyte in the samples. The following samples had analyte present in quantities above the LLOQ: Table 14. Matrix Blanks: PFBS In Control Serum Sample Number Sani| h- Description E03-0583-72849 E03-0583-72850 E03-0583-72854 E03-0583-72858 E03-0583-72859 E03-0583-72861 E03-0583-72862 E03-0583-72866 E03-0583-72869 E03-0583-72870 E03-0583-72871 E03-0583-72872 E03-0583-72876 E03-0583-72877 E03-0583-72878 E03-0583-73110 E03-0583-73116 454-116-63 Band #354 454-116-64 Band #355 454-116 - 68 Band #359 454-116 - 72 Band #363 454-116 - 73 Band #364 454-116-75 Band #366 454-116 - 76 Band #367 454-116- 79 Band #371 454-116-82 Band #374 454-116-83 Band #375 454-116-84 Band #376 454-116-85 Band #377 454-116- 89 Band #381 454-116-90 Band #382 454-116 - 91 Band #383 454-116-449 B844 Lot J offsprinq 454-116-455 B882 Lot J offsprinq Concentration o f PFBS in Scrum Sample ug/g 2.80** 0.0440* 0.124* 0.0653* 0.0280* 0.441* 0.0334* 0.0234* 0.0238* 0.0209* 0.0272* 0.0427* 0.0288* 0.0379 0.0751 0.145* 0.0906* 3M Environmental Laboratory Page 18 of 227 Contains No CBI Report E03-0583 -228 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues * At least one replicate was < LLOQ; a value equal to 1/2 LLOQ was assigned for calculation purposes. *One replicate was > ULOQ; a value equal to the ULOQ was assigned for calculation purposes Table 15. Matrix Blanks: PFBS in Control Liver Sample Number Sample Description Concentration of PFBS in Liver Sample, tig g E03-0583-72976 454-116 - 187 Band # 353 0.0207* E03-0583-72977 454-116 - 188 Band #354 0.0382 E03-0583-72978 454-116- 189 Band #355 0.0128* E03-0583-72979 454-116- 190 Band #356 0.00958* E03-0583-72980 454-116- 191 Band #357 0.0136 E03-0583-72981 454-116- 192 Band #358 0.0123* E03-0583-72983 454-116 - 194 Band #360 0.0123* E03-0583-72984 454-116- 195 Band #361 0.0145* E03-0583-72986 454-116- 197 Band #363 0.0118* E03-0583-72987 454-116- 198 Band #364 0.00945* E03-0583-73164 454-116-503 B823 0.0629* E03-0583-73164** 454-116-503 B823 0.0843* E03-0583-73165 454-116-504 B829 0.129 E03-0583-73165** 454-116-504 B829 0.0843* E03-0583-73169 454-116-508 B844 0.0131* * At least one replicate was < LLOQ; a value equal to 1/2 LLOQ was assigned for calculation purposes. " The sample was analyzed twice. The LLOQ for each serum and liver replicate is presented in Appendix B. Method Blanks. All of the method blanks in this study had area counts < 50% of the LLOQ. Quality Control (QC) Sam ples. Blank matrix was spiked and extracted with the samples according to the SOP. Results are as follows: Table 16. Serum QC Sample Recovery Theon-ticdl Concentration ni Extnrt 1 i1 r of Extracts -nalyzcd Mom Recovery of Repjicates Relative Stun Lini De>nti ii 25.58 ng/mL 11* 100% 5.29% 102.3 ng/mL 13 100% 6.73% 255.8 ng/mL 11 93.6% 9.18% 'Two of the 25.58 ng/g QC samples were spiked at the wrong concentration. Those samples were excluded from the above calculations (the average represents nine values). 3M Environmental Laboratory Page 19 of 227 Contains No CBI Report E03-0583 -229- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS In Northern Bobwhite Tissues Table 17. Liver QC Sample Recovery Theoictlcal Concentration in Extract Number of Extracts Analyzed 25.58 ng/mL 6 255.8 ng/mL 6 767.3 ng/mL 6 Mean Recovery of Replicates 90.2% 95.7% 8 8 .6 % Relative Slaruhid Deviation 3.44% 8.08% 7.63% Table 18. Egg QC Sample Recovery Theoretical Concentration in Extract Num i >>f Extracts Analyzed 25.58 ng/mL 6 102.3 ng/mL 6 511.5 ng/mL 6 Mean Ret overy of Replicates 109% 103% 95% Relative Standard Deviation 1.13% 4.89% 7.60% Data Summary Tables 19 through 38 summarize individual sample data and represent the average of triplicate analysis except where noted. Representative chromatograms are presented in Attachment F. The data has been corrected for purity of the PFBS reference substance used for the calibration curve. Table 19. C4F9S 0 3` Data Summary, 100 ppm Dose Group, Adult Female Quail Serum* Wildlife Sample lilcntifu i i LIMb Identification Average Concenti dticn >t PFBS in Scrum ut| ij 45 4 -1 1 6 - 105 Bhnu fr ^98 E03-0583-72893 454-116-101 Band #394 E03-0583-72889 454-116- 115 Band #408 E03-0583-72903 454-116- 103 Band #3 96 E03-0583-72891 454-116- 117 Band #410 E03-0583-72905 454-116- 107 Band #4 00 E03-0583-72895 454-116- 109 Band #4 02 E03-0583-72897 454-116- 113 Band #4 06 E03-0583-72901 4 5 4-1 16- 111 Band # 4 0 4 E03-0583-72899 45 4 -1 1 6 - 121 Band # 4 1 4 E03-0583-72909 454-116-93 Band #386 E03-0583-72881 4 5 4-1 16-9 9 Band #392 E03-0583-72887 45 4 -1 1 6 -9 7 Band #390 E03-0583-72885 454-116- 123 Band #416 E03-0583-72911 454-116-95 Band #388 E03-0583-72883 * The uncertaintyforthe serum samples is +/-15% as calculated In the validation. 2.49 4.85 6.43 8.53 8.96 12.5 12.6 13.1 14.0 14.3 17.2 22.4 23.1 25.8 33.1 3M Environmental Laboratory Page 20 of 227 Contains No CBI Report E03-0583 -230- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 20. C4F9S03' Data Summary, 100 ppm Dose Group, Adult Male Quail Serum1 Wildlife Sample Identification LIMS Identification Averaqc Concenti ation nf PFBS in Serum ug'g 454-116- 104 Band #397 E03-0583-72892 5.81 454-116 - 106 Band #3 99 E03-0583-72894 6.77 454-116 - 120 Band #413 E03-0583-72908 9.84* 454-116 -9 8 Band #391 E03-0583-72886 10.6 454-116- 108 Band #401 E03-0583-72896 11.1 454-116- 100 Band #393 E03-0583-72888 11.4 454-116 - 118 Band #411 E03-0583-72906 12.3 454-116 - 112 Band #4 05 E03-0583-72900 13.2 454-116-96 Band #389 E03-0583-72884 13.9 454-116- 122 Band #4 1 5 E03-0583-72910 20.3 454-1 16- 110 Band # 4 0 3 E03-0583-72898 30.4 454-1 16- 102 Band # 3 9 5 E03-0583-72890 31.7 454-116-94 Band #387 E03-0583-72882 37.7 TThe uncertainty for the serum samples is +/-15% as calculated In the validation. ` This value represents the average of duplicates as there was insufficient sample for triplicate analysis. Table 21. C^FoSOT Data Summary. 100 ppm Dose Group, Offspring Quail Serum* Wildlife Sample Identification 1i\1t> Identification Average Conrrnti itimi of PrBS in Scrum urg g 454-116 -4 5 7 G229 Lot J E03-0583-73118 0.0552 4 5 4 -1 1 6 -4 5 8 G233 Lot J E 0 3 -0 5 8 3 -7 3 1 19 0.0334 454-1 16-4 59 G242 Lot J E03-0583-73120 0.0450 454-1 16-4 60 G245 Lot J E03-0583-73121 0.0241 454-116-461 G253 Lot J E03-0583-73122 0.0246 454-116 -4 6 2 G260 Lot J E03-0583-73123 <0.0701* 45 4-116-463 G265 Lot J E03-0583-73124 < 0.0293 454-1 16-4 64 G274 Lot J E03-0583-73125 0.0209 454-1 16-4 65 G276 Lot J E03-0583-73126 <0.0311 454-1 16-4 66 G281 Lot J E03-0583-73127 < 0.0278 454-116-467 G289 Lot J E03-0583-73128 <0.0318 454-1 16-4 68 G294 Lot J E03-0583-73129 < 0.0286* 4 5 4 -1 16-4 69 G302 Lot J E03-0583-73130 0.0550 454-1 16-4 70 G309 Lot J E03-0583-73131 < 0.0301 454-116-471 G313 Lot J E03-0583-73132 < 0.0389 r The uncertainty for the serum samples is +/-15% as calculated in the validation. ` This value represents the average of duplicates as there was insufficient sample for triplicate analysis. 3M Environmental Laboratory Page 21 of 227 Contains No CBI Report E03-0583 -231 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 22. C4F9S 0 3' Data Summary, 300 ppm Dose Group, Adult Female Quail Serum1 W ildlife Sample Identification LIMS Identification Average Concenti ation of PFBS in Serum, ug/g 454-116- 154 Band #4 48 E03-0583-72943 12.2 454-116 - 1 4 6 1Band # 4 4 0 45 4-1 16- 127 Band #4 2 0 E03-0583-72935 E03-0583-72915 17.7 20.7 454-116- 150 Band # 4 4 4 E03-0583-72939 21.8 4 5 4 -1 1 6 - 152 Band # 4 4 6 E03-0583-72941 23.8 454-116- 133 Band #4 2 6 E03-0583-72921 23.8* 454-116- 144 Band #4 38 E03-0583-72933 24.7 454-116- 125 Band #4 18 E03-0583-72913 27.8 45 4-116- 140 Band #4 3 4 E03-0583-72929 38.5 454-116- 129 Band #4 22 E03-0583-72917 40.5 4 5 4 -1 1 6 - 131 Band # 4 2 4 E03-0583-72919 42.9 454-116- 142 Band #4 3 6 E03-0583-72931 45.7 454-116- 135 Band #4 28 E03-0583-72923 57.7 4 5 4-1 16-1 48 Band #4 42 E03-0583-72937 67.3 454-116- 138 Band #4 32 E03-0583-72927 102 TThe uncertainty for the serum samples is +/-15% as calculated in the validation. *This value represents the average of duplicates as one of the samples was lost during extraction. Table 23. C4F9S 0 3` Data Summary, 300 ppm Dose Group, Adult Male Quail Serum*' Vlildlitc Sample Idi utili i ion LIMS Identification Average Concentration of PFBS in Slrum ug j 45 4-1 16- 134 Band # 4 2 7 E03-0583-72922 454-116 - 147 Band #441 E03-0583-72936 454-116- 126 Band #4 19 E03-0583-72914 454-116 - 143 Band #4 3 7 E03-0583-72932 45 4-116- 132 Band # 4 2 5 E03-0583-72920 45 4-1 16- 124 Band #4 1 7 E03-0583-72912 454-116- 130 Band #4 23 E03-0583-72918 4 5 4 -1 1 6 - 151 Band # 4 4 5 E03-0583-72940 454-116-145 Band #439 E03-0583-72934 454-116- 153 Band # 4 4 7 E03-0583-72942 454-116- 139 Band # 4 3 3 E03-0583-72928 454-116- 137 Band #431 E03-0583-72926 45 4-1 16- 128 Band #421 E03-0583-72916 454-1 16-1 49 Band #4 43 E03-0583-72938 4 5 4 -1 1 6 - 141 Band # 4 3 5 E03-0583-72930 454-116- 136 Band # 4 2 9 E03-0583-72924 r The uncertainty for the serum samples is +/-15% as calculated in the validation. 14.2 14.4 16.7 19.4 20.9 24.0 25.8 25.9 26.6 31.1 31.7 36.9 36.9 37.1 42.7 42.9 3M Environmental Laboratory Page 22 of 227 Contains No CBI Report E03-0583 -232 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 24. C4F9S 03' Data Summary, 300 ppm Dose Group, Offspring Quail Serum* Wildlife Sample Identification LIMS Identification Average Concentration <f PFBS in Soium u j | 454-116 - 481 P188 Lot J E03-0583-73142 0.0245 454-116 - 478 P 172LotJ E03-0583-73139 0.0252 454-116 -4 8 6 P220 Lot J E03-0583-73147 0.0269 454-116 -4 7 6 P161 Lot J E03-0583-73137 0.0315 454-116-484 P210LotJ E03-0583-73145 0.0329 45 4-116-483 P205 Lot J E03-0583-73144 0.0395* 454-116-477 P165LotJ E03-0583-73138 0.0421 454-116-482 P199LotJ E03-0583-73143 0.0433 454-116 - 485 P 215LotJ E03-0583-73146 0.0472 454-116-479 P177LotJ E03-0583-73140 0.0489 454-116 - 475 P 155LotJ E03-0583-73136 0.0690 454-116-472 P132 Lot J E03-0583-73133 0.0956 454-116-474 P148LotJ E03-0583-73135 0.0996 454-116-480 P183 Lot J E03-0583-73141 0.105* 454-116-473 P139 Lot J E03-0583-73134 0.119 TThe uncertainty for the serum samples is +/-15% as calculated in the validation. This value represents the average of duplicates as there was insufficient sample for triplicate analysis. Table 25. C4F9S 03' Data Summary, 900 ppm Dose Group, Adult Female Quail Serum* Wildlife Smipl Id n itir.itmn 1----------------------Liv a 11 indication. - Average Conce ntr iti n f ' PFBS in Serum uu fj 454-116- 184 Band #478 E03-0583-72973 454-116- 186 Band # 4 8 0 E03-0583-72975 454-116- 174 Band #468 E03-0583-72963 454-116- 178 Band #472 E03-0583-72967 454-116- 180 Band #474 E03-0583-72969 454-116- 170 Band #464 E03-0583-72959 454-116- 182 Band # 4 7 6 E03-0583-72971 454-116- 156 Band #4 50 E03-0583-72945 454-116- 176 Band #470 E03-0583-72965 454-116- 166 Band #460 E03-0583-72955 454-116- 168 Band # 4 6 2 E03-0583-72957 454-116- 160 Band #4 54 E03-0583-72949 454-116- 158 Band #452 E03-0583-72947 454-116- 172 Band #466 E03-0583-72961 454-116-162 Band #456 E03-0583-72951 454-116- 164 Band #458 E03-0583-72953 TThe uncertainty for the serum samples is +/-15% as calculated in the validation. 30.5 39.1 43.8 48.6 49.7 57.3 70.8 77.4 87.0 90.4 93.1 97.7 162 167 178 370 3M Environmental Laboratory Page 23 of 227 Contains No CBI Report E03-0583 -233 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 26. C4F9S0 3 ' Data Summary, 900 ppm Dose Group, Adult Male Quail Serumt Wildlife Sample Id- iitification LIMS Identification Average Concentration <f PFS in Scrum ug q 454-116- 155 Band #449 E03-0583-72944 4 5 4 -1 1 6 - 161 Band # 4 5 5 E03-0583-72950 45 4-116- 183 Band #477 E03-0583-72972 4 5 4 -1 1 6 - 171 Band # 4 6 5 E03-0583-72960 454-116- 177 Band #471 E03-0583-72966 454-116 - 185 Band #479 E03-0583-72974 454-116- 175 Band #469 E03-0583-72964 454-116- 165 Band #459 E03-0583-72954 454-116- 167 Band #461 E03-0583-72956 454-116- 169 Band #463 E03-0583-72958 454-116- 157 Band #451 E03-0583-72946 454-116- 173 Band #467 E03-0583-72962 4 5 4 -1 1 6 - 181 Band # 4 7 5 E03-0583-72970 454-116- 159 Band #453 E03-0583-72948 454-116- 163 Band #457 E03-0583-72952 45 4-1 16- 179 Band #473 E03-0583-72968 TThe uncertainty for the serum samples is +/-15% as calculated in the validation. 34.3 43.1 44.3 47.1 58.8 59.3 60.3 61.3 65.6 67.0 74.9 86.5 94.9 95.7 98.8 98.8 Table 27. C4F9SO3" Data Summary, 900 ppm Dose Group, Offspring Quail Serum1 Wildlife Siimpli. Id n` 1 L MS 11. iitification Average ConcuiitratiLii >f PFBS in Serum, ug/g 4 5 4 -1 1 6 -4 9 6 W649 Lot J E 0 3 -0 5 8 3 -7 3 1 57 0.0413* 454-1 16-5 01 W681 Lot J E03-0583-73162 0.0604 454-116 - 491 W621 Lot J E03-0583-73152 0.0686 4 5 4 -1 1 6 -4 9 5 W 645 Lot J E03-0583-73156 0.0754 4 5 4 -1 1 6 -4 9 4 W641 Lot J E03-0583-73155 0.0833 4 5 4 -1 1 6 -4 9 7 W 652 Lot J E03-0583-73158 0.0836 4 5 4 -1 1 6 -4 9 8 W659 Lot J E03-0583-73159 0.105* 4 5 4 -1 1 6 -4 9 9 W 669 Lot J E03-0583-73160 0.117* 4 5 4 -1 1 6 -4 9 0 W 617 Lot J E03-0583-73151 0.138 454-116 - 500 W 675 Lot J E03-0583-73161 0.147** 454-116 - 488 W 6 O3 L0 U E03-0583-73149 0.168** 4 5 4-1 16-4 93 W635 Lot J E03-0583-73154 0.170 454-116-487 W597 LotJ E03-0583-73148 0.187 454-116 - 492 W 628 Lot J E03-0583-73153 0.230 4 5 4 -1 1 6 -4 8 9 W 610 Lot J E03-0583-73150 0.320 'T h e uncertainty for the serum samples is +/-15% as calculated in the validation. *This value represents the average of duplicates as there was insufficient sample for triplicate analysis. **This value represents only one replicate as there was insufficient sample for duplicate analysis. 3M Environmental Laboratory Page 24 of 227 Contains No CBI Report E03-0583 -234- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 28. C4F9SQ3' Data Summary, 100 ppm Dose Group, Adult Female Quail Liverf Wildlife Sample Identification t\..~ ... ,, _ i_ M3 Id-unification Average Concentration of PFBS in Liver mj y 454-116 - 228 Band # 394 E03-0583-73017 0.396 454-116 -23 2 Band # 398 E03-0583-73021 0.622 454-116-242 Band #408 E03-0583-73031 0.920 454-116-244 Band #410 E03-0583-73033 1.28 45 4-116-230 Band #396 E03-0583-73019 1.95 454-116 - 248 Band #4 14 E03-0583-73037 2.19 45 4-116-238 Band # 404 E03-0583-73027 2.29 454-116-240 Band #406 E03-0583-73029 3.83 454-116-250 Band #416 E03-0583-73039 3.99 4 5 4 -1 16-2 36 Band # 402 E03-0583-73025 4.09 45 4-116-246 Band #412 E03-0583-73035 4.29 454-116-234 Band # 400 E03-0583-73023 4.51 454-116-220 Band #386 E03-0583-73009 5.06 4 5 4 -1 16-2 24 Band # 390 E03-0583-73013 5.30 454-1 16-2 26 Band # 392 E03-0583-73015 7.37 454-1 16-2 22 Band # 388 E03-0583-73011 8.19 ' The uncertainty for the liver samples is +/- 46% as calculated in the validation. Table 29. C4F9S 0 3' Data Summary, 100 ppm Dose Group, Adult Male Quail Liver1 Wildlife Sample Identifkuli n LIMS Identification Average Concenti alien t PFBS in Liver, ug/g 45 4 -1 1 6 -2 4 1 Band # 4 0 7 E03-0583-73030 454-116 -231 Band # 3 9 7 E03-0583-73020 454-116 -2 3 5 Band #401 E03-0583-73024 4 5 4-1 16-2 25 Band #391 E03-0583-73014 454-116 -2 4 9 Band #4 1 5 E03-0583-73038 454-116 -2 3 3 Band # 399 E03-0583-73022 454-1 16-2 47 Band #4 13 03-0583-73036 454-116 -2 2 7 Band # 393 E03-0583-73016 454-116 -2 4 5 Band #411 E03-0583-73034 454-1 16-2 39 Band # 405 E03-0 583-73028 454-116-243 Band # 409 E03-0583-73032 454-116 -251 Band #417 E03-0583-73040 454-1 16-2 23 Band # 389 E03-0583-73012 454-116 -2 1 9 Band #3 85 E03-0583-73008 454-116 -2 3 7 Band # 403 E03-0583-73026 454-116-221 Band #3 87 E03-0583-73010 454-116 -2 2 9 Band # 395 E03-0583-73018 TThe uncertainty for the liver samples is +/- 46% as calculated in the validation. 0.808 0.878 1.24 1.41 1.73 1.76 1.77 2.09 2.71 2.76 3.71 3.95 4.09 5.22 5.74 6.53 8.89 3M Environmental Laboratory Page 25 of 227 Contains No CBI Report E03-0583 -235 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 30. C4F8S 0 3' Data Summary, 100 ppm Dose Group, Offspring Quail Liver* Wildlife Sample LL i tification Llf.lS Identification Average Concentration of PFBS in Livci ug j 454-116-523 G274 E03-0583-73184 454-116-519 G245 E03-0583-73180 454-116-530 G313 E03-0583-73191 454-116 -52 2 G265 E03-0583-73183 454-116 -5 2 5 G281 E03-0583-73186 454-116-516 G229 E03-0583-73177 454-116-517 G233 E03-0583-73178 454-116-518 G242 E03-0583-73179 45 4-116-529 G309 E03-0583-73190 454-116-521 G260 E03-0583-73182 454-116-524 G276 E03-0583-73185 454-116-527 G294 E03-0583-73188 454-116-526 G289 E03-0583-73187 45 4-116-528 G302 E03-0583-73189 454-116 - 520 G253 E03-0583-73181 TThe uncertainty for the liver samples is +/- 46% as calculated in the validation. *Used 1/2 LLOQ for at least one of the replicate calculations 0.00985* 0.0233* 0.0301 <0.0137 <0.0138 <0.0139 <0.0139 <0.0139 <0.0139 <0.0140 <0.0140 <0.0141 <0.0143 <0.0148 <0.0152 Table 31. C4F9S 0 3` Data Summary, 300 ppm Dose Group, Adult Female Quail Liver* Wildlife Sanipl * 1i nil tu n LIMS Identification Average Conce iltraim i >t PFBS in Li.cr ug g 4 5 4-1 16-2 64 Band # 430 E03-0583-73053 3.57 45 4 -1 1 6 -2 8 2 Band # 448 E03-0583-73071 4.32 4 5 4-1 16-2 74 Band # 440 E03-0583-73063 5.94 45 4-1 16-2 54 Band # 420 E03-0583-73043 6.18 45 4-1 16-2 78 Band # 444 E03-0583-73067 6.92 454-116-280 Band #446 E03-0583-73069 7.85* 454-116-260 Band #426 E03-0583-73049 8.47 454-116-272 Band #438 E03-0583-73061 8.54 454-116-252 Band #418 E03-0583-73041 9.63 454-116-258 Band #424 E03-0583-73047 12.4 4 5 4-1 16-2 68 Band # 434 E03-0583-73057 12.7 454-116-270 Band #436 E03-0583-73059 12.7 4 5 4 -1 1 6 -2 5 6 Band # 422 E03-0583-73045 15.4 4 5 4 -1 1 6 -2 7 6 Band # 442 E03-0583-73065 17.7 4 5 4-1 16-2 62 Band # 428 E03-0583-73051 22.3 4 5 4 -1 1 6 -2 6 6 Band # 432 E03-0583-73055 23.5 TThe uncertainty for the liver samples is +/- 46% as calculated in the validation. 'Value represents the average of duplicate samples as the triplicate was lost during extraction. 3M Environmental Laboratory Page 26 of 227 Contains No CBI Report E03-0583 -236- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 32. C4F9S 03' Data Summary, 300 ppm Dose Group, Adult Male Quail Liver1 Wildlife Sample Identification t MS irli ntification Average Concentration of PFBS in Livci mi ij 454-116-261 Band #427 E03-0583-73050 454-116-275 Band #441 E03-0583-73064 454-116 - 271 Band # 4 3 7 E03-0583-73060 454-116 -253 Band #419 E03-0583-73042 454-116 -25 9 Band # 425 E03-0583-73048 454-116-279 Band #445 E03-0583-73068 454-116 -26 7 Band #433 E03-0583-73056 454-116 -277 Band #443 E03-0583-73066 454-116 -273 Band #439 E03-0583-73062 454-116 -25 7 Band #423 E03-0583-73046 454-116 -281 Band #447 E03-0583-73070 454-116 -265 Band #431 E03-0583-73054 454-116-255 Band #421 E03-0583-73044 454-116-269 Band #435 E03-0583-73058 454-116-263 Band #429 E03-0583-73052 * The uncertainty for the liver samples is +/- 46% as calculated in the validation. 1.42 3.77 3.80 5.51 6.99 7.31 7.48 7.52 8.18 8.42 9.43 9.67 11.3 11.7 14.2 Table 33. C4F9S 03' Data Summary, 300 ppm Dose Group, Offspring Quail Liver1 Wildlife S inipl I i i ificati i 1IV S Idi ntification Average.' Ci nci'iiliati ai f PFBS in Liver urj g 4 5 4 -1 1 6 -5 3 8 P177 E03-0583-73199 4 5 4 -1 1 6 -5 3 9 P183 E03-0583-73200 454-116-536 P165 E03-0583-73197 454-1 16-5 44 P215 E03-0583-73205 4 5 4 -1 1 6 -5 3 2 P139 E03-0583-73193 45 4-116-534 P155 E03-0583-73195 4 5 4 -1 1 6 -5 4 3 P210 E03-0583-73204 4 5 4 -1 1 6 -5 3 3 P148 E03-0583-73194 454-116-531 P132 E03-0583-73192 454-116-542 P205 E03-0583-73203 454-1 16-5 45 P220 E03-0583-73206 454-116-541 P199 E03-0583-73202 4 5 4 -1 1 6 -5 3 5 P161 E03-0583-73196 45 4-116-540 P188 E03-0583-73201 4 5 4 -1 1 6 -5 3 7 P172 E03-0583-73198 * The uncertainty for the liver samples is +/- 46% as calculated in the validation. ` Used 1/2 LLOQ for at least one of the replicate calculations 0.00965* 0.0115* 0.0161 0.0240 0.0272 0.0288 0.0305 0.0346 0.0369 0.146 0.200* <0.0138 <0.0141 <0.0141 <0.0144 3M Environmental Laboratory Page 27 of 227 Contains No CBI Report E03-0583 -237 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 34. C4F9SQ3' Data Summary, 900 ppm Dose Group, Adult Female Quail Liver1 Wildlife Sample Identification LII7IS Identification Average Concentration of PFBS in Liver ug g 4 5 4-1 16-3 10 Band #476 E03-0583-73099 9.73 4 5 4-1 16-3 14 Band #480 E03-0583-73103 12.1 454-116-306 Band #472 E03-0583-73095 14.1* 454-116 -3 0 2 Band #468 E03-0583-73091 14.7* 454-116 -30 8 Band #474 E03-0583-73097 14.7 454-116-298 Band #464 E03-0583-73087 17.2 454-1 16-3 12 Band #478 E03-0583-73101 17.5** 454-116 -304 Band #470 E03-0583-73093 22.8 454-1 16-2 94 Band # 460 E03-0583-73083 24.3 45 4-1 16-2 84 Band # 450 E03-0583-73073 25.3 4 5 4 -1 16-2 88 Band # 454 E03-0583-73077 28.9 454-116-296 Band #462 E03-0583-73085 35.2 454-116-290 Band #456 E03-0583-73079 46.1 454-116-300 Band #466 E03-0583-73089 53.4 45 4-116-286 Band # 452 E03-0583-73075 59.6 4 5 4 -1 16-2 92 Band # 458 E03-0583-73081 77.4* TThe uncertainty for the liver samples is +/- 46% as calculated in the validation. *This value represents the average of duplicates, as one replicate was lost during extraction. " This value represents only one replicate, as two replicates were lost during extraction. Table 35. C4F9S 0 3' Data Summary, 900 ppm Dose Group, Adult Male Quail Liver* Wildlifo Sampli l i n ificalion LIMS Identification Average Concentration of PFBS in Liu r ug j 454-116-311 Band #477 E03-0583-73100 6.73 454-116-283 Band #449 E03-0583-73072 10.4 454-116-285 Band #451 E03-0583-73074 10.9 454-1 16-2 89 Band #455 E03-0583-73078 12.0 4 5 4 -1 1 6 -2 9 9 Band # 465 E03-0583-73088 12.7* 454-116-313 Band #479 E03-0583-73102 13.5 454-116-301 Band #467 E03-0583-73090 13.9 454-116-305 Band #471 E03-0583-73094 14.7* 45 4 -1 1 6 -2 9 7 Band # 463 E03-0583-73086 15.5 454-116-303 Band #469 E03-0583-73092 16.8 454-116 - 291 Band # 4 5 7 E03-0583-73080 17.3 45 4-116-293 Band # 459 E03-0583-73082 18.7 454-116-287 Band #453 E03-0583-73076 19.2 454-116-295 Band #461 E03-0583-73084 21.1 454-116-307 Band #473 E03-0583-73096 23.5 454-116-309 Band #475 E03-0583-73098 23.7** TThe uncertainty for the liver samples is +/- 46% as calculated in the validation. *This value represents the average of duplicates, as one replicate was lost during extraction. " This value represents only one replicate, as two replicates were lost during extraction. 3M Environmental Laboratory Page 28 of 227 Contains No CBI Report E03-0583 -238 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 36. C4F9S 0 3' Data Summary, 900 ppm Dose Group, Offspring Quail Liver1 Wildlife Sample Identification LIMB Id ntitication Average Cencintration i f PFBS m Liver, ug/g 454-116 -549 W617 E03-0583-73210 0.0340 454-116 - 552 W635 E03-0583-73213 0.0484 454-116 -555 W649 E03-0583-73216 0.0492 454-116 - 556 W652 E03-0583-73217 0.0499 454-116 - 559 W675 E03-0583-73220 0.0509 454-116 -5 5 0 W621 E03-0583-73211 0.0548 454-116 - 553 W641 E03-0583-73214 0.0617 4 5 4 -1 1 6 -5 6 0 W681 E03-0583-73221 0.0935 454-116 - 557 W659 E03-0583-73218 0.107 454-116 -548 W610 E03-0583-73209 0.109 454-116 - 547 W603 E03-0583-73208 0.134* 454-116-554 W645 E03-0583-73215 0.162 454-116- 546 W597 E03-0583-73207 0.199* 454-116-558 W669 E03-0583-73219 0.207 454-116 -551 W628 E03-0583-73212 0.313 TThe uncertainty for the liver samples is +/- 46% as calculated in the validation. ` This value represents the average of duplicates, as one replicate was lost during extraction. Table 37. C4F9S 0 3 Data Summary, Quail Egg Lot BT Wildlife Sample Idontificatirn I D 'm ' Group LIMS Identification ', I Band #388 Eqq Contents Lot B Band #392 Eqq Contents Lot B Band #396 Egg Contents Lot B Band #404 Eqq Contents Lot B Band #408 Eqq Contents Lot B Band #412 Eqq Contents Lot B Band #416 Eqq Contents Lot B Band #424 Eqq Contents Lot B Band #428 Eqq Contents Lot B Band #432 Eqq Contents Lot B Band #436 Eqq Contents Lot B Band #440 Eqq Contents Lot B Band #444 Eqq Contents Lot B Band #448 Eqq Contents Lot B Band #452 Eqg Contents Lot B Band #456 Egq Contents Lot B Band #460 Eqq Contents Lot B Band #464 Eqq Contents Lot B Band #468 Egg Contents Lot B Band #472 Egg Contents Lot B Band #476 Egg Contents Lot B Band #480 Egg Contents Lot B 100 ppm a.i. 100 ppm a.i. 100 ppm a.i. 100 ppm a.i. 100 ppm a.i. 100 ppm a.i. 100 ppm a.i. 300 ppm a.i. 300 ppm a.i. 300 ppm a.i. 300 ppm a.i. 300 ppm a.i. 300 ppm a.i. 300 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. 900 ppm a.i. ` E03-0583-72583 E03-0583-72584 E03-0583-72585 E03-0583-72586 E03-0583-72587 E03-0583-72588 E03-0583-72589 E03-0583-72590 E03-0583-72591 E03-0583-72592 E03-0583-72593 E03-0583-72594 E03-0583-72595 E03-0583-72596 E03-0583-72597 E03-0583-72598 E03-0583-72599 E03-0583-72600 E03-0583-72601 E03-0583-72602 E03-0583-72603 E03-0583-72604 TThe uncertainty for the egg samples is +/-15% as calculated in the validation. Average Con ulti il n i t PFBS in Egg, ug/g 8.17 7.14 10.8 7.04 4.92 5.89 9.59 16.9 36.5 15.6 28.6 13.3 25.2 29.3 47.2 40.6 51.6 36.5 37.2 118 38.9 33.8 3M Environmental Laboratory Page 29 of 227 Contains No CBI Report E03-0583 -239- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 38. C4F9S 0 3' Data Summary, Quail Egg Lot G* Wildlife Sample Identir ition Dose Group LIMS Identification Band #386 Eqg Contents Lot G 100 ppm a.i. E03-0583-72735 Band #390 Egg Contents Lot G 100 ppm a.i. E03-0583-72736 Band #394 Egg Contents Lot G 100 ppm a.i. E03-0583-72737 Band #398 Egg Contents Lot G 100 ppm a.i. E03-0583-72738 Band #402 Egg Contents Lot G 100 ppm a.i. E03-0583-72739 Band #406 Egg Contents Lot G 100 ppm a.i. E03-0583-72740 Band #410 Egg Contents Lot G 100 ppm a.i. E03-0583-72741 Band #414 Egg Contents Lot G 100 ppm a.i. E03-0583-72742 Band #418 Egg Contents Lot G 300 ppm a.i. E03-0583-72743 Band #422 Egg Contents Lot G 300 ppm a.i. E03-0583-72744 Band #426 Egg Contents Lot G 300 ppm a.i. E03-0583-72745 Band #434 Egg Contents Lot G 300 ppm a.i. E03-0583-72746 Band #438 Egg Contents Lot G 300 ppm a.i. E03-0583-72747 Band #442 Egg Contents Lot G 300 ppm a.i. E03-0583-72748 Band #446 Egg Contents Lot G 300 ppm a.i. E03-0583-72749 Band #450 Egg Contents Lot G 900 ppm a.i. E03-0583-72750 Band #454 Egg Contents Lot G 900 ppm a.i. E03-0583-72751 Band #458 Egg Contents Lot G 900 ppm a.i. E03-0583-72752 Band #462 Egg Contents Lot G 900 ppm a.i. E03-0583-72753 Band #466 Egg Contents Lot G 900 ppm a.i. E03-0583-72754 Band #470 Egg Contents Lot G 900 ppm a.i. E03-0583-72755 Band #474 Egg Contents Lot G 900 ppm a.i. E03-0583-72756 Band #478 Egg Contents Lot G 900 ppm a.i. E03-0583-72757 The uncertainty for the egg samples is +/- 15% as calculated in the validation. Average Coik cntration of PFBS in Egg u j j 12.3 12.5 13.1 12.1 26.2 10.2 8.44 16.8 21.9 73.1 16.1 27.6 29.0 31.1 21.1 62.7 137 123 119 52.6 89.6 61.4 95.1 Attachm ent A contains data tables detailing individual sample data, and presents the LLOQ for each replicate analyzed. Statement of Conclusion In both serum and liver the adult females generally showed higher levels of analyte than the adult males within the same dose group. Concentration of PFBS in serum, liver, and eggs increased with dose level. Low levels of target analyte measured in the control group samples are not expected to significantly affect the Interpretation of results for the other dose groups. The LLOQs for individual replicates are presented In Appendix B for reference. The method used for extraction and analysis of the samples in this study has been validated for PFBS in quail serum, liver and eggs. See tables 39, 40, and 41 for additional summary information. 3M Environmental Laboratory Page 30 of 227 Contains No CBI R eport E 03-0583 -240- Project Number 454-116 T -7485: A Reproduction Study with the Northern Bobwhite Phase: A nalytical Determ ination of PFBS in Northern Bobwhite Tissues Table 39. Summary ofResults: C4F9SO3'Concentration in Serumt DoseGroup and Gender Numberof Serum Samples M ew Concentration, ug PFBS/g Serum ravom--- Concentration,ug PFBS/g Serum Range o f ResuSs 100 ppm, Male 13 16.5 12.3 5.81-37.7 100 ppm, Female 15 14.6 13.1 2.49-33.1 100 ppm Offspring 15 0.0369* 0.0334* <0.0278-0.0552 300 ppm, Male 16 27.9 26.3 14.2-42.9 300 ppm, Female 15 37.8 27.8 12.2-102 300 ppm Offspring 15 0.0567 0.0433 0.0245-0.119 900 ppm, Male 16 68.2 63.4 34.3 - 98.8 900 ppm, Female 16 104 82.2 3 0 .5 - 370 900 ppm Offspring 15 0.133 0.117 0.0413 - 0.320 TThe uncertainty for the serum samples is 15% as calculated in the validation. Mean and Median reported for samples inwhich analyte was detected. For more specific information see tables inAttachment A Table 40. SummaryofResults: C<FS03"Concentration in L)vert Dose Group and Gender Numberof Liver Samples Mam PFBS/g Liver co n ctw um if ug PFBSfeUvar Range o fResults 100 ppm, Male 17 3.25 2.71 0.808 - 8.89 100 ppm, Female 16 3.52 3.91 0.396-8.19 100 ppm Offspring 15 0.0211* 0.0233* <0.0137-0.0301 300 ppm, Male 15 7.78 7.52 1.42-14.2 300 ppm, Female 16 11.1 9.08 3.57 - 23.5 300 ppm Offspring 15 0.0515* 0.0288* <0.0138-0.200 900 ppm, Male 16 15.7 15.1 6.73 - 23.7 900 ppm, Female 16 29.6 23.5 9.73 - 77.4 900 ppm Offspring 15 0.111 0.0935 0.0340-0.313 TThe uncertainty for the liver samples is 46% as calculated inthe validation. Mean and Median reported for samples inwhich analyte was detected. For more specific information see tables in Attachment A 3M Environm ental Laboratory Page 31 o f 227 Contains No CBI Report E03-0583 -241 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Table 41. Sum m ary Table, C4F9S 0 3' C oncentration in Eggt Dose Group A. , 100 ppm, Lot B Number of e<)3 i Samples Mean Concentration, ug PFBS/g Egg Median Concentration, ug PFBS/g Egg 7 7.65 7.14 300 ppm, Lot B 7 23.6 900 ppm, Lot B 8 50.5 100 ppm, Lot G 8 14.0 300 ppm, Lot G 7 31.4 25.2 39.7 12.4 27.6 900 ppm, Lot G 8 92.6 92.4 f The uncertainty for the egg samples is +/-15% as calculated in the validation. Range of Results 4.92-10.8 13.3-36.5 33.8-118 8.44 - 26.2 16.1-73.1 52.6-137 Statistical M ethods Statistical methods were limited to calculating means, standard deviations, and % coefficient of variation. References 1. W ildlife International, Ltd. Protocol No. 454-116 "T-7485: A Reproduction Study with the Northern Bobwhite". 2. 3M Environmental Laboratory Standard Operating Procedure ETS-8-231.2, "Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry" . List of Attachments Attachment A: Method ETS-8-231.2 Attachment B: Data Tables Attachment C: Example Chromatograms Attachment D: Protocol and Amendments 3M Environmental Laboratory Page 32 of 227 Contains No CBI -242 - Project Number 454-116 Report E03-0583 T-748S: A Reproduction S tu d / with the Northern Bobwhfte Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Signature Page We certify that this report is a true and complete representation o f the data for this study: tA ju^ f i d i l a S ea n P . G allagher. StudvDirector JohWMerwsted, Sponsor Representative 3w 5 Date ---------------------T J ----------------------Date M a rk fc. E llefson, Principal AnalyffeaTEnvestigator ^ Date W illiam K. R eagen, 3M Environmental Laboratory Management Date 3M Environmental Laboratory Page 33 of 227 Contains No CBI Report E03-0583 -243 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Attachment A: Extraction and Analytical Method, ETS-8-231.2, "Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices" 3M Environmental Laboratory Page 34 of 227 Contains No CBI Report E03-0583 :, , -244- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite . Phase: Apal:y|ical Determination of PFBS in Northern Bobwlpite;Ts$ijps 3M Environm ental Laboratory Method Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry Method Number: ETS-8-231.2 Adoption Date: 13 November 2001 Revision Date: Upon Signing Effective Date: Upon Signing Approved By: William K. Reagen Laboratory Manager O/ Date ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Various Matrices 3M Environmental Laboratory Page 1 of 23 Page 35 of 227 Contains No CBI Report E03-0583 -245 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite of 9 F ^,ip .N p ^h e fo ^b.w hitp 1 Scope and Application This method describes the extraction of target analytes from various biological matrices using solid phase extraction (SPE) followed by separation, identification, and quantitation using high-performance liquid chromatography mass spectrometry (HPLC/MS) or tandem mass spectrometry (HPLC/MS/MS). This method has been validated for perfluorooctane sulfonate (PFOS) in mouse and rat liver and sera using HPLC/MS/MS based on the FDA guidance document, ``Guidance for Industry - Bioanalytical Method Validation, May 2001". Additionally, the performancebased validations in quail and mallard matrices confirmed that the method can be used in matrices other than the rat and mice matrices already validated, provided the concurrent quality control samples document the acceptability of each data set as shown in 3M Studies E01-1256 titled "Analytical Phase Report for PFOS: A Reproduction Study with the Mallard" and E01-I245 titled "Analytical Phase Report for PFOS: A Reproduction Study with the Northern Bobwhite." 3M Report E02-1042 titled "Method and Validation Report for `Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices Method Number: ETS-8-231.1 is a compilation of the results of these studies. These procedures are appropriate for perfluorinated acids, alcohols, amides, sulfonates and other fluorinated compounds in any biological matrix provided that quality control sample results meet the precision and accuracy requirements of the study. 2 Method Summary This method easily accommodates a significant range of initial sample weights and volumes, which is essential when preparing samples of limited quantities or when large sample sizes are required to provide low detection limits. When using a 1 gram SPE cartridge, sample amounts typically ranging from 0.02 to 1.0 mL or g are prepared (fluids diluted, tissues homogenized, etc.) at a 1/6, 1/10, or 1/20 dilution using reagent water. An aliquot of the dilution/homogenate is spiked with the appropriate surrogate or analyte mixture. Acetonitrile (ACN) is added as an extraction solvent and also serves to precipitate the proteins. The sample is capped, mixed, and put on the centrifuge to clarify the supernatant. The supernatant is transferred to a clean tube, diluted with water, and passed through a pre-conditioned C|8 SPE cartridge. Finally, the analytes of interest are eluted from the SPE cartridge and analyzed by HPLC/MS or HPLC/MS/MS. ' Depending on sample availability and detection limits required, the use of SPE cartridges of different sizes provides flexibility for varying initial sample sizes and final elution volumes. Sufficient quality control samples are incorporated into each study to ensure that precision and accuracy of the study are understood. The required quality control elements of this performance based method meet the cross validation requirements based on the FDA guidance documents. 3 Definitions 3.1 Dilution A dilution expressed as 1:5 or 1/6 is defined as: 1 mL of sample + 5 mLs of diluent for a total of 6 mLs combined, unless otherwise noted. 3.2 SPE cartridge A column containing an open solvent reservoir, retaining frit, sorbent bed, retaining frit, and luer tip. The sorbent bed is bonded silica which is designed to selectively retain or elute the compounds of interest depending on the solvent conditions. The compounds of interest can then be separated from difficult matrices and introduced into an appropriate solvent for analysis. SPE cartridges are available with a wide range of sorbent beds in a variety of sizes, allowing the analyst to process a wide array of sample types and volumes. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 2 of 23 Page 36 of 227 Contains No CBI -246 - Project Number 454-116 Report E03-0583 VPVetH !h a V T-7485: A Reproduction Study with the Northern Bobwhite 3.3 Reagent grade water Water with no detectable concentration(s) of the target analyte(s) or less than 1/2 the limit of quantitation (LOQ). 3.4 Quality control sample Sample used to determine extraction precision and accuracy and to estimate the uncertainty of results generated. 4 Warnings and Cautions 4.1 Health and Safety Warnings Always wear appropriate personal protective equipments such as protective gloves, eye protection, and appropriate clothing when working with biological matrices, solvents, chemicals and instrumentation. For potential hazard information refer to material safety data sheets, packing material, the 3M Environmental Laboratory's Chemical Hazard Review, the 3M Guide to Laboratory Practices or other information as appropriate. 4.2 Cautions The analyst must be familiar with the laboratory equipment and potential hazards including, but not limited to, the use of biological materials, solvents, high temperatures, pressurized gas and solvent lines, high voltage, and vacuum systems. Refer to the appropriate equipment procedure or operator manual for additional information and cautions. 5 Interferences To minimize interferences, Teflon should not be used for sample storage or any part of instrumentation that comes in contact with the sample or extract. Co-extracted matrix components may enhance or suppress the measured analyte signal in the mass spectrometer. These effects are minimized using this SPE technique, but the precision and accuracy of spike results must be evaluated for possible effects of co-extracted matrix interferences that may be present. 6 Instrumentation, Supplies, and Materials The following instrumentation, supplies, and materials are used while performing this method. Equivalent instrumentation, supplies, and materials may be used in place of those listed. 6.1 Instrumentation and Equipment Vortex mixer, VWR, Vortex Genie 2, or equivalent Ultra-Turrax T25 tissue homogenizer with various size blades, or equivalent Vacuum Pump SPE Extraction Manifold Centrifuge, Mistral 1000, IEC, or equivalent Shaker, Eberbach, VWR, or equivalent Balance (+/- 0.1000 g) Micromass, Quattro II or Ultima triple quadrupole Mass Spectrometer equipped with an electrospray ionization source, or equivalent HP 1100 or Agilent low pulse solvent pumping system, solvent degasser, column compartment, and autosampler, or equivalent ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 3 o f 23 Page 37 of 227 Contains No CBI -247- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite of PFBS m Npfftff ^bjiytytp /Ijifsgp? 6.2 Supplies and Materials Eppendorf or disposable pipettes, plastic or glass Dissecting scalpels Polypropylene bottles, (Nalgene or equivalent), various sizes Volumetric flasks, glass, type A, various sizes 40 mL glass vials (ICHEM or equivalent) Plastic sampule vials, (Wheaton or equivalent), 6 mL (or other appropriate size) Centrifuge tubes, polypropylene, various sizes Labels, various sizes Graduated pipettes, glass Syringes, capable of measuring 5 pL to 1000 pL Bottle-Top Dispenser (capable of dispensing 5mL of solvent) SPE extraction cartridges, for example 1 g, Sep-Pak 6 cc tri-functional C,*(Waters), or other sizes and adsorbents 75 mL sample reservoir (or other appropriate size) Crimp cap glass autovials and caps, crimpers HPLC analytical column: Keystone Betasil Cl 8, 2 X 50 mm, 5 pm, or equivalent 7 Reagents and Standards Reagent grade water, ASTM Type I Milli-QTM, Kandiyohi purchased drinking water, or equivalent Acetonitrile, HPLC grade or equivalent Methanol, HPLC grade or equivalent Ammonium acetate, reagent grade or equivalent Various biological fluids or tissues from supplier or study sponsor for use as control matrix 7.1 Control Matrix Tissue/Fluid Preparation This method requires that a control matrix be used for the preparation of matrix-extracted calibration standards, control spikes, and method blanks. Therefore, an amount of control matrix should be prepared to provide a sufficient amount throughout a study. Typically an aliquot of 40mL is mixed with 200 mL of reagent water or 40g of matrix is homogenized with 200 g reagent water and used as control matrix. Other amounts may be used to maintain the same dilution ratio as the samples or additional control matrix may be prepared during a study as needed. Control matrix is prepared from one of three sources: 1) a study control matrix from a study control received with a sample set; 2) a commercially obtained sample of the same matrix as the study matrix; or 3) a surrogate matrix, also obtained commercially, but of a different type than the study matrix, (eg. if rat is used to generate calibration curves and QCs for a mouse study). 7.2 Reagents Preparation Prepare a 2.0 mM ammonium acetate solution for use as a mobile phase for the liquid chromatograph: Weigh approximately 0.300 g ammonium acetate. Pour into a 2000 mL volumetric container containing reagent grade water, mix until all solids are dissolved, and bring to volume using reagent grade water. Store at room temperature. Note: When preparing different volumes or concentrations than those listed for reagents and target analyte standard preparation adjust accordingly. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 4 o f 23 Page 38 of 227 Contains No CBI -248- Project Number 454-116 Report E03-0583 ; n, j T-7485: A Reproduction Study with the Northern Bobwhite PS^e.:; 6faly(i<:^ | J ] ^ | 1| ] i n ^ p Qf l?FBjp;Npjr|ttP|'n B^bwfjite Tissue? 7.3 Target analyte standard preparation Prepare target analyte standard(s) for the matrix-extracted calibration curve. Multi-component analyte standards are acceptable. The following is an example only and may or may not be appropriate for all standard preparations. Weigh approximately 100 mg of target analyte into a 100 mL volumetric flask and record the actual weight in the standard logbook or other appropriate location. Bring to volume with methanol, or other appropriate solvent, for a stock standard of approximately 1000 ppm ((Xg/mL). The calculated concentration of this stock standard should be corrected for purity and should reflect the fluorochemical anion concentration that is measured. Dilute the stock solution with methanol for a working standard solution of approximately 50 ppm. Example calculation: lOOOpg ml. x 5 mL/100 mL = 50|Xg/mL. Additional dilutions or serial dilutions are made as appropriate. If surrogate standards or internal standards are part of the study, these stock solutions are prepared using the same procedures outlined above. Additional information regarding the appropriate use of surrogates or internal standards is found in section 9 of this method. Surrogates and internal standards are not required by this method, but may be used for individual studies. 8 Sample Handling It is expected that samples are received at the appropriate conditions and must be kept at the appropriate conditions until the extraction is performed, (i.e. frozen for biological samples). The condition of samples upon receipt and storage conditions throughout the study should be appropriately documented. Allow liquid samples and homogenized tissue samples to thaw/warm to room temperature prior to extraction. Matrix-extracted standards used to calibrate the instrument response for each target analyte are prepared at least once during a study. With each extraction batch, matrix extracted quality control standards must be prepared to verify the extraction efficiency. Alternately, an entire matrix-extracted curve may be prepared with each extraction set. Extracted standards and samples are stored in capped autovials until analysis. Extracted standards and samples may be stored at room temperature or refrigerated at approximately 4C until analysis can be performed. If required by the study protocol, 3 post-extraction matrix spikes (i.e. a known amount of the target analyte spiked into an extracted matrix blank) may be stored with the extracted standards and samples for evaluation to demonstrate extract stability. Alternately the precision and accuracy of matrix-extracted quality control samples may be used to document analyte stability during the tme frame of analysis. Results will be documented in the report. 9 Quality Control 9.1 Blanks 9.1.1 Solvent Blank An aliquot of methanol or other appropriate solvent is analyzed periodically throughout each analytical sequence as a solvent blank to serve as a check for background concentrations of analyte or cross over contamination between injections. Solvent blanks are not extracted. 9.1.2 Method Blank An aliquot of 1.0 mL of reagent water, or other amount as appropriate, is used as a method blank. Method blanks are extracted and analyzed with each extraction set following this procedure and may or may not be spiked with surrogate depending on study requirements. Method blanks may serve as a measure of analyte background concentrations not attributable to endogenous levels in the control matrix. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 5 o f 23 Page 39 of 227 Contains No CBI -249 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite gvApaltficM f^ipination qf l?F^Sjt|,Np^tt||i'p qqbwljitj 9.1.3 Matrix Blank An aliquot of 1.0 mL or 1.0 g of matrix (diluted or homogenized) is used as a matrix blank. Other amounts are used, as appropriate, in keeping with the amount of sample available for extraction. Matrix blanks are prepared from one of three sources: 1) a study control matrix from a study control received with a sample set; 2) a commercially obtained sample of the same matrix as the study matrix; or 3) a surrogate matrix, also obtained commercially, but of a different type than the study matrix, (eg. if rat is used to generate calibration curves and QCs for a mouse study). The matrix to use is the same as the matrix used for the curve. If limited matrix is available, the number of method and matrix blanks may be adjusted, as well as the initial volumes/weights, and will be noted in the study protocol or in the raw data. Study control matrix curve - if the study control matrix is used for the curve, prepare four (4) matrix blanks using the study control matrix (two spiked with surrogate and two not spiked). If surrogates are not part of a study, duplicates are acceptable. Commercially obtained matrix curve - if a commercially obtained matrix is used for the curve, prepare four (4) matrix blanks using the same commercially available matrix (two spiked with surrogate and two not spiked). If surrogates are not part of a study, duplicates are acceptable. 1 Surrogate matrix curve - if a surrogate matrix is used for the curve, prepare four (4) matrix blanks using the same surrogate matrix and prepare four (4) matrix blanks using the study control matrix, if available. If study control matrix is not available, prepare four (4) matrix blanks using a commercially available matrix of the same type as the study matrix (two spiked with surrogate and two not spiked). If surrogates are not part of a study, duplicates are acceptable. 9.2 Sample Replicate Samples replicates are prepared according to each study protocol, project outline, or as detennined by the analyst. 9.3 Surrogate standard If surrogate standard is a component of the study, all samples are spiked with surrogate standard prior to extraction to obtain an appropriate concentration within the target calibration curve, with the exception of blank samples as described in section 9.1. Typically surrogate standard is spiked into the 1.0 mL or 1.0 g, or other appropriate volume, diluted/homogenized sample removed for extraction. 9.4 Initial Calibration A minimum of one set of matrix-extracted standards is prepared and analyzed to establish the relationship of analyte concentration to instrument response for the target iluorochemical analytes. Details are provided in Section 10 of this method. 9.5 Quality Control (QC) Sample Quality control (QC) samples are used to determine extraction precision and accuracy and to estimate the uncertainty o f results generated. Typically 1.0 mL of the same matrix used to prepare the initial calibration curve is used for each QC sample, and the target analyte standard is spiked into the 1.0 mL or 1.0 g diluted/homogenized sample removed for extraction. All spiking should be performed at the same step in the extraction, (i.e. If the curve is spiked into the 1.0 mL or 1.0 g diluted/homogenized sample removed for extraction then QC samples, surrogate spike, etc. should also be spiked at that time.) A minimum of nine (9) quality control samples (QC) will be prepared for each matrix during the course of a study to allow the needed statistical calculations to be made. When study samples are extracted in three or fewer extraction batches, a minimum of three QC samples must be prepared (one at each level) on each day of sample extraction, and should total nine or more. When 3 or more extraction batches are prepared, fewer than three QC samples may be prepared with each batch provided the method or protocol QC frequency requirements are met. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 6 o f 23 Page 40 of 227 Contains No CBI -250- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite vPh:auses:eAdn,alyticcaluDrerteernmti,naltoionr off-P+FaBaSvinsNiorrothnersn Bhobwh>ite Tissues The total number of QC samples within a study should be at least 5% of the number of unknown samples or nine total QCs, whichever is greater. The levels listed below are target guidelines to be used when spiking QC samples and may represent sample concentrations diluted into the range of the calibration curve: Low level: 3X to 5X the lower limit of quantitation (LLOQ), Mid-level: equivalent to a point near the middle of the calibration curve, High level: 80% of the upper limit of quantitation (ULOQ) QC samples extracted with a particular sample are initially analyzed in the same analytical run to demonstrate acceptable quality control for the batch. After the QC samples have been successfully analyzed, the QC samples do not need to be re-injected when a subset of samples from a batch are analyzed since the acceptance criteria for the batch have already been met. The extracted calibration curve and continuing calibration verification samples will document the acceptable perfonnance of the instrumentation. Each QC is expected to show an accuracy 85-115% of expected and a precision of 15%. A minimum of 2/3 of all QC samples analyzed within an analytical run must meet these criteria. If not, the set must either be re-analyzed or re extracted. 9.6 Continuing Calibration Verification Sample (CCV) One or more initial calibration curve points are re-injected after not more than 10 sample injections to verify the continued accuracy of the initial calibration curve. Typically, two CCV sample levels are analyzed after every tenth sample injection. Solvent blanks are not considered samples and are not included as such for determining when QC samples will be analyzed. Each CCV is expected to show an accuracy of 85-115% of expected. If not, the set should either be re-analyzed or re-extracted. 9.7 Matrix Spike (MS) Sample Matrix spike samples are not a required component of this method, but may be required by protocol or project plan. If the matrices for the curve are not the same as the sample matrix to be analyzed, or the protocol/project outline requires matrix spike samples; the specific criteria regarding matrix spikes will be documented in the protocol/project outline and/or in a note to file. 9.8 Control Charts The percent recovery for each QC sample for each analyte and matrix will be control charted as per SOP ETS-4-26, "Control Charts for Laboratory Analyses". 9.9 Sample Dilution Any sample with an analyte area greater than that of the highest acceptable standard will need to be diluted into the range of the calibration curve. If samples are diluted into the range of the curve during analyses and enough sample remains, a post-run dilution validation may be performed to verify sample values. To perform the dilution validation, one sample will be separated into two representative samples (i.e. two 1.0 mL aliquots for fluid samples or two 1.0 gram amounts for tissue samples, or other amount as determined by the analyst and documented in a note to file) then diluted using two procedures. The first procedure consists of diluting the sample with additional blank matrix prior to extraction (fluid adding fluid), while the second procedure consists of diluting the extract with solvent post-extraction (methanol extract adding additional methanol solvent). If the relative percent difference is not within 15% for these two samples, additional testing may be performed to determine which value is a correct representation of the sample concentration as determined by the analyst and documented in a note to file. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 7 of 23 Page 41 of 227 Contains No CBI -251 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite 10 Calibration and Standardization 10.1 Instrument Calibration At least one calibration curve will be prepared from extracted matrix standards, in the same matrix as the samples if available, per study. It will consist of a minimum of nine concentration levels. Additional calibration curves may be extracted on separate sample extraction dates, as detennined by the analyst. Transfer 1.0 mL of diluted control fluid or 1.0 g of homogenized control tissue to a 15 mL centrifuge tube using a disposable plastic pipette. This will be repeated while preparing aliquots for the calibration curve. Be sure to vortex mix or shake by hand the control matrix container for approximately 5 seconds between aliquots to ensure a homogenous sample is removed. Record each standard weight or volume on the weight/volumes sheet or extraction worksheet, as appropriate. Four 1.0 mL or 1.0 g aliquots, or other appropriate amount as described in 9.1.2, of control matrix serve as matrix blanks. A total of 9 standards, four matrix blanks, and four method blanks are prepared in addition to the QC samples and test samples. The number of standards and blanks may be adjusted as per section 9.1, or as determined by the analyst. Use Attachment C, or other appropriate form, as an aid in calculating the concentrations of the working standards. Refer to section 12 to calculate the actual concentration of analyte in each calibration standard and QC sample. Typically the target analyte standard is spiked into the 1.0 mL diluted fluid or 1.0 g homogenized tissue sample removed for extraction. All spiking should be preformed at the same step in the extraction, (i.e. If the curve is spiked into the 1.0 mL or 1.0 g diluted/homogenized sample removed for extraction then QC samples, surrogate spike, etc. should also be spiked at that time.) Analyze the extracted matrix calibration curve prior to each set of extracts. The curve equation may be determined by quadratic regression analysis, weighted 1/x, and external standard using the peak areas of the target analyte(s) using MassLynx or other suitable software. Other suitable regressions may be used as appropriate. Any level outside 85% - 115% of nominal must be deactivated, and regression re-calculated, except the LLOQ which must be within 20% of nominal. All levels must show a response greater than twice that of the blank. A minimum of six (6) levels may be used to plot the curve in any one set, or the set will be re-analyzed. 11 Procedures 11-1 Tissue/Solid Sample Preparation Obtain tissue/solid samples. Cut approximately 1.000 g of tissue (+/- 0.100 g), or other appropriate amount, using a dissecting scalpel. This part of the procedure is best performed quickly, not allowing the tissue to thaw. Weigh approximately 1.000 g of solid sample (+/-0.100 g). Weigh the sample directly into a tared plastic sample vial. Record the weight on the weight/volume sheet, extraction worksheet, or other appropriate location. Return unused tissue/solid to the appropriate storage conditions after extraction amounts have been removed. Weigh 5 mL (or 10 mL) of reagent water into the tared sampule vial containing the tissue/solid sample then record the weight on the weight/volume sheet. (For solid samples that don't require homogenization, skip the remaining steps in this section.) Homogenize the sample. Put the Ultra-Turrax grinder probe in the sample and grind for approximately 2 minutes, or until the sample is homogeneous. ' ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 8 of 23 Page 42 of 227 Contains No CBI -252 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Take the grinder apart and clean it with methanol after each sample. Refer to ETS-9-52 for more infonnation. If an amount other than 1.000 g (not within +/- 0.100 g) is removed for an initial weight, adjust the water volume accordingly to maintain a 1/6 dilution, (e.g. if 0.5 g is removed for extraction, add a total of 2.5 mL of water.), or other ratio as documented in the protocol, the project outline, or a note to file. 11.2 Fluid/Liquid Sample Preparation Obtain fluid/Iiquid sample and allow it to thaw/warm at room temperature or in lukewarm water. Label a 15 mL polypropylene centriiuge tube with the study number, sample ID, extraction date and analyst initials. See attached worksheet (Attachment A or similar worksheet) for documenting the remaining steps. Vortex mix the sample for approximately 15 seconds, then transfer 1.0 mL of fluid/Iiquid, or other available amount, to a plastic sampule vial, or other appropriate plastic container. Return unused fluid/Iiquid to the appropriate storage conditions after extraction amounts have been removed. Add 5.0 mL of reagent water to the 1.0 mL of fluid/Iiquid for a 1/6 dilution, or 19 mL of reagent water to the 1.0 mL for a 1/20 dilution. If a volume other than 1.0 mL is removed for an initial volume, adjust the water volume accordingly to maintain the same dilutions as above. 11.3 Tissue and Fluid Sample Extraction After tissue/solid or fluid/Iiquid samples have been prepared according to sections 11.1 and 11.2, vortex mix or shake by hand the diluted/homogenized sample for approximately 15 seconds then transfer 1.0 mL, or other appropriate volume, to a clean 15 mL polypropylene centrifuge tube. Return unused diluted/homogenized portions to the appropriate storage condition after extraction amounts have been removed. Record the volume of diluted fluid/Iiquid removed on the extraction worksheet, (Attachment A or similar worksheet). Record the weight of the homogenized tissue/solid (removed as a 1.0 mL volume or other volume) on the weight/volume sheet, the extraction worksheet, or other appropriate location. Spike blanks, samples, and standards, ready for extraction with surrogate standard as described in this method. Spike each calibration standard matrix with the appropriate amount of standard as described in this method for the calibration curve standards and each QC sample. Vortex mix the calibration curve samples and QC samples for approximately 5 seconds. To each sample and standard, add 5.0 mL of acetonitrile, cap, and vortex mix or shake by hand approximately 15 seconds. Place all samples on the shaker at an appropriate speed for 20 minutes to adequately mix (a setting of approximately 300 rpm on the models listed in section 6.1). Remove from the shaker and centrifuge at an appropriate speed for 10 minutes to adequately pellet the precipitate (a setting of approximately 2000 rpm on the models listed in section 6.1). Add 40.0 mL of reagent grade water to a clean 50 mL centrifuge tube. Remove samples from the centrifuge and decant the supernatant into the water in the 50 mL tube, taking care not to introduce any of the matrix solids into the solution. Cap and mix by inverting several times. In this step the order of addition may be changed (i.e. the sample may be put into the centrifuge tube and then the water added). ' Attach the reservoir to the SPE cartridge and attach this reservoir/cartridge unit to a vacuum manifold. NOTE: When running the vacuum, set the vacuum chamber at approximately 20 kPA - to give an approximate elution flow of 5-7 mL/min. Flows may vary through cartridges and the kPA may be raised for slow tubes and drying after most have been drawn down. Prepare the SPE cartridge by washing twice with approximately 5.0 mL of methanol, followed by approximately two 5.0 mL aliquots of water, taking care not to allow the column to run to dryness after each wash. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 9 of 23 Page 43 of 227 Contains No CBI -253 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite After washing is complete, pour the sample into the reservoir/cartridge unit and allow all of the liquid to pass through the column to dryness. Run the vacuum on high for approximately 5 minutes to adequately dry each SPE cartridge. Place a collection 15 mL polypropylene centrifuge tube under each cartridge and elute with 2.0 mL of methanol. Spike extracted blanks, samples, and standards with internal standard as described in this method. Label each glass autovial, as appropriate, with the study number, vial file archive number, identification number/gender/timepoint (if applicable) or LIMS number, matrix, final solvent, analyte components (if needed), extraction type, extraction date, and analyses) performing the extraction. Transfer each eluant to a glass autovial and cap. 11.4 Extract Analysis 11.4.1 Software set-up Prior to analysis, a sample list must be created using the following parameters and naming conventions: instrument designator letter, last 2 digits of test year-month-day, and a letter that will increase through the alphabet with each additional list for that day. Other unique sample list names may be used. Example Sample List: IYYMMDDa or A020204a I = Initial of the instrument name (A = "Amelia") YY = Test year (02) MM = Test month (02) DD = Test day (04) a = First sample list (run) of the day (the next sample list will end with V, the next 'c', and so on.) Filenames are created by using the instrument designator letter, the last 2 digits of the test year-month-day, and a 3 digit sequential file number that starts with 1 and increases by one for each filename. Other unique filenames may be used. Example filename: IYYMMDD### or A020204001 I = Initial of instrument name YY = Test year MM = Test month DD = Test day ### = 3-digit sequential file number starting with 1 through 999 (001) Also, as part of the sample list, assign a mass spectrometer acquisition method for acquiring data, an inlet file to designate HPLC analytical parameters, a bottle number, an injection volume, and individual sample descriptions. Typically, analytical batch run sequences begin with a minimum of three system suitability injections, several solvent blanks, and a calibration curve. The end of the analytical sequence should be marked by a closing CCV. Typically, two CCV samples should be injected after every tenth sample, and all sample extracts must be bracketed by the opening calibration curve and a closing CCV. Solvent blanks should be analyzed periodically to monitor possible analyte carryover and are not considered sample extracts but may be included as such. 11.4.2 HPLC set-up Load the autosampler tray according to the sample list prepared above. ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 10 of 23 Page 44 of 227 Contains No CBI -254 - Project Number 454-116 Report E03-0583 D o C ir n ti Mv; m a v T-7485: A Reproduction Study with the Northern Bobwhite n Northern E ^ b w tjitj/F i^ i^ Download the HPLC inlet file created in section 12.4.1. Record actual conditions in the instrument logbook, or other appropriate location. The following instrument conditions are provided as an example: Sample injection volume = 10 pL Injections/sample = 1 Cycle time = 10.0 minutes Flow rate = 300 pL/min. Column: Mobile phase: Solvent gradient: Keystone Betasil C|8, 2 x 50mm, 5 um particle size, or equivalent Solvent A = 2 mM Ammonium Acetate in ASTM Type I water Solvent B - Methanol (HPLC grade) Time %B 0.00 10% 1.00 10% 5.50 95% 7.50 95% 8.00 10% 11.4.3 Instrument set-up The following instrument conditions are provided as an example: Drying gas 250-400 liters/hour ES nebulizing gas 10-15 liters/hour Source block temperature = 150C Desolvation temperature = 250C These settings may change in order to optimize the response. Please refer to the instrument tune page, and ETS-9-24, "Operation and Maintenance of Micromass Triple Quadrupole Mass Spectrometers Fitted With Atmospheric Pressure Ionization Sources," for additional details. 12 Data Analysis and Calculations 12.1 Calculations If alternative calculations are used, they will be documented in the raw data. Calculate the matrix amount contained in the initial dilution using the following equations: Weight of Tissue Extracted (g ) Initial Sample Weight (g) Homogenate Sample Weight (g) x Weight of Homogenate Aliquot (g) Volume of Fluid Extracted (mL) Initial Sample Volume (mL) Diluted Sample Volume (mL) x Volume of Fluid Aliquot (mL) ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 11 o f 23 Page 45 of 227 Contains No CBI -255 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite o /fF ^ S jn Nprthem E^byypitp Xipppps Calculate actual concentrations of analyte in calibration standards using the following equation: Concentration (ng/mL)in FinalSPE Eluent = SPik-e-Concentratlon (ug/mL>X SPlked Amount (mL)x IQOOng (mL) Final SPE Eluent lug Calculate actual concentration of analyte in fluid (jig/mL): (AR (n^mE eluent) * DF) x Final Eluent Volume (mL) x Volume of Fluid Extracted (mL) 1.0 = 1000 ng (pg/mL) Calculate actual concentration of analyte in tissue (pg/g): (AR (ng/mL eluent) * DF) x Final Eluent Volume (mL) x 1,0 qg - (jig/g) Weight of tissue Extracted (g) 1000 ng AR = Analytical result from MassLynx summary DF = Dilution factor introduced when sample extract is diluted to fall within the calibration curve range. Calculate spike percent recoveries using the following equation: n%/ tR,ecovery = -O--b--s-e-r-v--e-d--R--e--s-u-l-t------U--n-s-p-i-k-e--S--a-m--ip--l-e--R--e-s-u--l-t x 100 Spiking Level Calculate relative standard deviation using the following equation: Relative Standard Deviation Standard Deviation Mean 100 Calculate percent deviation using the following equation: .. __ . . Calculated Cone. - Expected Cone. % Deviation = --------------------------- --------------- x 100 Expected Cone. 13 Method Performance 13.1 System Suitability System suitability will be determined prior to the start and at the completion of each analytical run. Prior to the calibration curve and after the last sample of the run three (3) mid-level unextracted calibration standards will be analyzed. The peak area precision and retention time precision will be monitored at the beginning and the end of the run separately. The peak area precision must be equal to or less than 5.0% RSD, the precision of the retention time must be equal to or less than 2.5% RSD. If any item of the system suitability fails, system maintenance must be completed prior to running a second set of system suitability samples. The system suitability must pass before starting the calibration. If system suitability fails at the completion of a run, the sample set must be reanalyzed. 13.2 Uncertainty Estimation The combined uncertainty associated with this method, as determined by the validation results, is as follows: 29% for tissue analyses and 9% for fluid analyses. Refer to SOP ETS-12-12, "Estimation of Uncertainty Measurements" and ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices P a g e l2 o f2 3 page 0f 2 2 7 Contains No CBI -256- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite P ^ je r ^ a l| '| ic ^ | g^tgr^nEjtgp of fl'FBS in Nprthefn Bobw^itp Ttei||ip? the attached uncertainty estimation report (Attachment E) for additional information on calculating uncertainty for methods. The uncertainty estimation may change per matrix as data are collected and added to control charts, or as control charts are updated according to ETS-8-26, "Control Charts for Laboratory Analyses". 13.3 Quantitation The coefficient of determination value for the calibration curve, plotted by regression using the peak areas of the analyte(s), must be 0.990 or better. During the validation it was determined that four curve models adequately describe the concentration-response relationship for this method: linear-null weighted; linear- 1/x weighted; quadratic-null weighted, and quadratic-1/x weighted. However, it has been determined that quadratic-l/x weighted, external standard will most often be used for sample quantitation. All active calibration curve points must be within 15% of the theoretical value with the exception of the LOQ point, which may deviate up to 20%. Calibration standards with peak areas less than two times the curve matrix blank will be deactivated to disqualify a data range that may be affected by background levels ofthe analyte. A valid calibration curve must contain at least 6 active points above and including the LOQ. If the curve cannot meet these criteria, the sample set must be reanalyzed or reextracted. 13.4 Accuracy and Precision Two thirds of all quality control and continuing calibration verification samples are expected to show an accuracy of 85-115%. Quality control samples are expected to show a precision of 15%. 14 Pollution Prevention and Waste Management Sample waste is disposed of in noninfectious biohazard waste containers. Flammable solvent waste is disposed of in high BTU containers. Glass pipette waste is disposed of in broken glass containers located in the laboratory. 15 Records Complete the extraction worksheet attached to this method, or other applicable worksheet, and store with the study raw data. Each page generated for a study must contain the following information (if applicable): study/project or instrument number, acquisition method, integration method, sample name, extraction date, dilution factor (if applicable), and analyst. Other information may be added if applicable to the study. Print the tune page, sample list, and acquisition method from MassLynx to include with the study raw data. Copy these pages and tape into the instrument runlog, if applicable. Plot the calibration curve by the appropriate regression. Print these graphs and store with the study raw data. Print data integration summary, integration method, and chromatograms from MassLynx, and store with the study raw data. Summarize data using suitable software (Excel 7.0 or LIMS) and store in the study folder. Back up electronic data to appropriate medium. Record in study notebook the file name and location of backup electronic data. Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 47 of 227 Contains No CBI -257- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite K - alicrl*i2 f s 16 Attachments Attachment A: Extraction Worksheet Examples Attachment B: Sample Weight/Volume Worksheet Attachment C: Calibration Standard Concentration Worksheet Attachment D: Dilutions Summary Worksheet Attechmetn E: Uncertainty Estimation Report 17 References 3M report - E01-1277, "Method Validation for ETS-8-231" 3M Method - ETS-4-26, "Control Charts for Laboratory Analyses" 3M SOP - ETS-12-12, "Estimation of Uncertainty Measurements" 3M Equipment Procedure - ETS-9-24, "Operation and Maintenance of Micromass Triple Quadrupole Mass Spectrometers Fitted With Atmospheric Pressure Ionization Sources" 3M Equipment Procedure - ETS-9-52, "Operation and Maintenance of a Tissue Grinder" FDA guidance document, "Guidance for Industry - Bioanalytical Method Validation, May 2001 " 3M report - E01-1256, "Analytical Phase Report for PFOS: A Reproduction Study with the Mallard" 3M report - E01-1245, "Analytical Phase Report for PFOS: A Reproduction Study with the Northern Bobwhite" 3M report - E02-1042, `Method and Validation Report for `Solid Phase Extraction and Analysis of Fluorochemical Compounds from Biological Matrices Method Number: ETS-8-231.1"' 3M Standard Operating Procedure - ETS-12-1, "Analytical Definitions, Abbreviations, Acronyms, and Symbols" 18 Affected Documents None. 19 Revisions Revision Number Revision Description 1 Minor formatting changes. Added detailed information to all sections concerning the extraction procedure, analytical procedure, and calculations. Added attachments and references. 2 The minor changes in this revision will not affect the method validation results: Section 1 added reference to validated method, added clarification to the scope of the method, and removed fish matrix. Section 9.6, 10.1 added text for consistency in spiking calibration curves and quality control samples. Section 9.8 added control chart section. Section 12 updated calculations and corrected serum final concentration units. Section 13.2 added uncertainty estimation. Section 13.3 added text describing which curve model to use during sample analysis. Section 16 added additional extraction sheets and uncertainty estimation report. Section 17 added additional references. Added clarification to text throughout method. Revision Date 02/18/02 10/04/02 ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 14 of 23 Page 48 of 227 Contains No CBI -258 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite imeni may ?Pt*haIsjeS: fAint,alyt'ical "Determination 0/ P,FB'S In N1orthern B>oIb'.whIite' i Iss`ues Study Number: Prep Date: Analysts initials: Box#: Attachment A - Extraction Worksheet Example 1 Method Revision: ETS-8-231.2 Matrix: Sample Tlmepoint: Sample Number Sample Number or description Volume/Weigh of diluted sample removed Amount and Amount and spike mix surrogate spike Type of column Elution solven used mix used used and lot and volume Comments Blank matrix TN-A- : Amount weiqhed/allquoted: a/mL 1. Homogenize sample 2. Aliquot 1 mL of diluted matrix into 15 mL polypropylene tube (record volume or weiqht accordinqly) 3. Spike samples accordingly 4. Add mL of ACN (TN-A- U o each diluted sample and shake or vortex mix 5. Shake sample for 20 min at a settina of rom (Shaker ) 6. Centrifuqe sample for 10 min at a settina of rpm (Centrifuae ) 7. Add 40 mL of water to 50 mL polypropelene centrifuqe tube. 8. Decant extract into centrifuge tubes with water 9. Shake sample slightly to ensure proper mixing 10. Attach 6 mL C18 SPE cartridges and 75 mL reservoirs to vacuum manifold 11. Condition column with two washes of ~5 mL MeOH (TN-A- ) - do not allow column to qo to dryness 12. Wash column with two washes of -5 mL water - do not allow column to qo to dryness 13. Filter sample through conditioned column, discarding filtrate 14. Allow column to qo to dryness. After dripplnq stops, draw a hiqh vacuum throuqh column for at least 5 minutes. 15. Elute column with solvent ( TN-A- ) into appropriate 15 mL centrifuqe tube 16. Spike samples with uL of internal standard # . cone. 17. Transfer sample into appropriately labeled autovial and cap Signature/Date Note: In vacuum steps above set the vacuum chamber at approximately 20 kPA - this should give approximately 5-7 mL/min elution flow Flows may vary through cartridges - kPA may be raised for slow tubes and drying after most have been drawn down and shut off. E TS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 15 o f 23 Page 4g Qf 22/ Contains No CBI -259- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite o). ,Phase: Aijaly|ca[ Deterrnyn^Upn i?Ft|SJn Nprthern E^jbyytjitp Attachment A - Extraction Worksheet Example 2 % #%_ t< , xlaamt, ; tp*... M #*_ j' ___ '' ' ^ - - ' ^ t *^ v **<,#vy%, ^%a,-/ * j alit $**.a w ,* t __ ,,___ __ J...J. . ....... H . .....,....... . 31, ,, ...,# ^ # `^m4 &n m^0m,tm.: ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 16 of 23 Rage 5Q of 227 Contains No CBI -260 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite .^ijial|'.tic^l! fjeterrnin3ti.on of PFBS.in Northern Bqbwhite Ttssups Attachment A - Extraction Worksheet Example 3 ___ i^s&SCLvv. i ..,.... - mmmi%#||I ... . : ' ,... m$mmm .... mmfA_ . ......mtnwrnJ&m* 'mirn&m,..i j #U6____ _,, mt^m**M it .43K#Mr * G<*H,nd*H ukHj* iw#i>........ ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratory Compounds from Various Matrices Page 17 of 23 Page 51 of 227 Contains No CBI -261 - Project Number 454-116 Report E03-0583 it : T-7485: A Reproduction Study with the Northern Bobwhite ... :pba^e;;Afal^icatDaterrnin^p t^ |'F^jn,,Np|;|h^ ^bvyhitp,Tis|fi^ Attachment B - Sample Weight/Volume Worksheet Prep Date(s): Analyses): Sample Matrix: Method/Revision: Study Number: Equipment Number: Final Solvent & TN Number: Sample ID Initial Wt./Vol. g/mL/L Water added Volume/Weigh Wt./Vol. Removed g/mL mL/g Date Comments Removed/ Analyst 1 Form Completion Verified By:. - .....*......- ... ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical 3M Environmental Laboratiory Compounds from Various Matrices Page 18 o f 23 Page 52 of 227 Contains No CBI -262- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite 11 1r : i r i , : , , ! {.gha,fe;^falyficalDaterniinat!on o,f P F fS ip Northern Bobwhite Tissues Prep date(s): Analyte(s): Sample matrix: Method/revision Attachment C: Calibration Standard Concentration Worksheet Standard number: Equipment number: Final solvent and TN#: Blank Tissue or Fluid/identifler: Analyte mix std approx. 0.500 ug/mL: Analyte mix std approx. 5.00 ug/mL: Analyte mix std approx. 50.0 ug/mL: Surrogate std approx. 100 ug/mL: Actual concentrations of standards in the analyte mix Analyte Std cone ug/mL 0.500 0.500 0.500 0.500 0.500 5.00 5.00 5.00 5.00 5.00 5.00 50.0 50.0 All Amount spiked mL 0.0015 0.0030 0.0080 0.0160 0.0320 0.0056 0.0080 0.0160 0.0240 0.0320 0.0400 0.005 0.006 All Final Volume: mL 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 2.00 Calculated concentrations of standards in relation to die final 2.0 niL solvent 2.0 mL Final Volume Analyte Final cone. ng/mL 0.375 0.750 2.00 4.00 8.00 14.0 20.0 40.0 60.0 80.0 100 125 150 Surrogate Std cone ng/mL 100 Surrogate ' Final cone ng/mL 0.500 ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Various Matrices Page 19 of 23 3M Environmental Laboratory Page 53 of 227 Contains No CBI -263 - Project Number 454-116 Report E03-0583 ; T-7485: A Reproduction Study with the Northern Bobwhite j Phaser AhalyficaJ fjp tg r^in a ^p cfl?Fj|^,,iq Npfth^fi E^bwhitp fTi^sups Study: Dilution Date/Analyst: Box Number: Sam ple N um ber or D escription 1/ Attachment D: Dilutions Summary Worksheet Solvent/TN Number: Extraction Date/Anaiyst: M atrix/Tim epoint: D ilu tio n s 1/ 11 1/ 1/ 1/ 1/ C o m m e n ts V erified By: 1 N otes: 1/10 dilution = 1 mL of sample + 9 mL o f solvent Form Completion Verified By: ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Various Matrices Page 20 of 23 3M Environmental Laboratory Page 54 of 227 Contains No CBI -264- Project Number 454-116 Report E03-0583 i V.. T-7485: A Reproduction Study with the Northern Bobwhite ; P ^ e ;A|ialy)jc g i jcjp ofRFflS.in.Northpn ^Qbwhite ETS-8-231.1 Attachment E: Uncertainty Estimation Report U ncertainty Estim ation A full validation for the use of this method for the determination of PFOS has been performed on rat liver and serum; a partial validation has been performed with PFOS on mouse liver and serum. A method validation report has been issued for these validations, as well as a performance-based validation for the analysis of PFOS in mallard and quail liver and serum. This report presents precision and bias evaluations through this method; the results of these evaluations will help in estimating the uncertainty for the use of this method. 1. Method summary The method includes the following procedures: 2. Major sources of uncertainty The following describes each of several major sources ofuncertainty in laboratory determinations, the nature of the uncertainty associated with each, and an estimation ofthe extent of the error. Mass - Balances are used for measuring the weight of the sample aliquot for analysis (using an analytical balance), only for solid matrices (e.g., liver.) measuring the weight of analytical standard used for the preparation of a stock standard solution(using an analytical balance.) As discussed in SOP ETS-12-012, the uncertainty associated with the standard weights can be ignored. Random uncertainties associated with balance measurements were estimated using calibration data from one analytical balance: ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Various Matrices Page 21 of 23 3M Environmental Laboratory Page 55 of 227 Contains No CBI Report E03-0583 -265 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Aijialy|ical pet^n|n^i(3n of jn Nprthf Balance ID Logbook pages Standard deviation Square of standard deviation n 118453734 00062-7 - 00062-9 9.58E-04 9.17E-07 118 However, the uncertainty associated by the sample and neat standard weighing process is included in the validation studies using QC samples. No additional factor is needed. Volume - Volumetric flasks are used for: measuring the final volume of spiking solutions. measuring the final volume of calibrating solutions. Pipettors are used for: measuring the volume of the sample aliquot, only for liquid matrices (e.g., serum.) measuring the volume of water added before homogenizing. measuring the aliquot taken for clean-up. measuring the final volume of extract. measuring volume aliquots of standard solutions in the preparation of spiking solutions. measuring volume aliquots of standard solutions in the preparation of calibrating solutions. As discussed in SOP ETS-12-012, random uncertainties associated with volume determinations were estimated using calibration data from several pipettors: Equipment type Logbook Dates Standard deviation Square of standard deviation n Pipettors 01054 4/5/02 - 7/28/02 1.33E-02 1.76E-04 69 However, the uncertainty associated by the volume measurements of aliquots and extracts as well as of standard solutions is included in the validation studies using QC samples. No additional factor is needed. Calibration Standards - Uncertainties associated with the neat standards are not supplied with fluorochemical compounds. Since they are solid compounds, the random uncertainties associated with their purity is considered negligible. No additional factor is needed. Standardization Process - The standardization (or calibration) process for most instrumental analyses introduces uncertainty because of the inexact nature of the process. With a correlation coefficient of 0.990, the minimum required by the method, the standard error of estimate may be 10 20%. However, the uncertainty associated by the standardization process is included in the validation studies using QC samples. No additional factor is needed. Extraction and Analysis Process - With recoveries typically being acceptable with an accuracy of 70% to 130%, and with a precision of 15%, it is expected that the greatest contribution to the uncertainties will be associated with the extraction and analysis process. The full validation data generated with PFOS in rat liver and serum were used to estimate this uncertainty, using all the % recoveries obtained during validation, including all sets, whether they met the target recoveries or not. It is assumed to follow a normal distribution. ETS-8-231,2 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Various Matrices Page 22 of 23 3M Environmental Laboratory Page 56 of 227 Contains No CBI -266- Project Number 454-116 Report E03-0583 ; ,45X.. T-7485: A Reproduction Study with the Northern Bobwhite c|_fj'F^Sjn.Nprthern Bobwhite Tissues Method validation of Matrix Standard deviation Square of standard deviation n ETS-8-231 Rat liver Rat serum 2.06E-01 8.75E-02 4.24E-02 7.66E-03 116 52 Homogeneity - Not a parameter in the method diagram above, sample homogeneity appears to be a significant effect in the analytical procedure for solid samples (e.g., liver). For liquid samples, it is expected that this parameter will not be significant. The homogeneity of the liver sub-sample is not taken into account by the recoveries of the QC samples during validation, since spiking occurs after the 1-g subsample has been taken. However, as discussed in the EURACHEM/CITAC Guide QUAM:2000.P1, it is not practical to measure homogeneity directly. Following the same rationale presented in that document, an uncertainty factor of 0.20 is selected for this parameter. It is assumed to follow a normal distribution. 3. An analysis of the effect of contributing sources on combined uncertainty A summary table of the combined uncertainties and expanded uncertainties (k = 2) in typical analyses at the 3M Laboratory are therefore: SUMMARY OF SOURCES OF UNCERTAINTY Neat standard Homogeneity standard deviation Validation data standard deviation Homogeneity square of standard deviation Validation data square of standard deviation Combined uncertainty Expanded uncertainty rat liver negligible 2.00E-01 2.06E-01 4.00E-02 4.24E-02 29% 57% rat serum negligible - 8.75E-02 - 7.66E-03 9% 18% With combined uncertainty, a function of the individual uncertainties Uj : And the expanded uncertainty, U : with k = 2 for 95% confidence interval. As an example, for rat liver: U =kxuc uc = V (2.00--01)2 + (2 .0 6 -0 1 )2 = V (4.0 0 -0 2 + 4 .2 4 -0 2 ) = 0.287 = 29% 77 = 2x0.287 = 0.574 = 57% ETS-8-231.2 Solid Phase Extraction and Analysis of Fluorochemical Compounds from Various Matrices Page 23 of 23 3M Environmental Laboratory Page 57 of 227 Contains No CBI -267 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Attachment B: Data Summary Tables 3M Environmental Laboratory Page 58 of 227 Contains No CBI 3M Environmental Laboratory Quail Serum Matrix Spikes File Sam ple Nam e ___________________ Sam ple Info BQ902CM0.D 846-'! Control Serum Low Spike. 8/30/04 C M C /M LA /M E C 0 0 902044.0 7 2848-5 Control Serum' Low Spike , 8/30/04 C M C /M LA /M CC D0917152.D LS-040907-1 Low Matrix Spike; Serum; 07 Sept 04; M LA/CM C/M EE D0917155.D LS-040907-2 D0917156.D LS-040907-3 D0921120.D LS-040909-1 D0921121.D LS-040909-2 D0921122.D LS-040909-3 D1223045.D LS-041222-1 Low Matrix Spike; Serum; 07 Sept 04; M LA/CM C/M EE Low Matrix Spike; Serum; 07 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE Low Spike; Ext 12/22/04 CM C /M LA/VJT D1223046.D LS-041222-2 D1223047.D US-041222-3 Low Spike; Ext 12/22/04 CM C /M LA/VJT Low Spike; Ext 12/22/04 CM C/M LA/VJT D0902076.D 72857-4 D0902077.D 72857-5 Control Serum-Med Spike, 8/30/04 C M C /M LA/M EE Control Serum-Med Spike, 8/30/04 C M C /M LA/M EE D0907032.D 72873-4m ed spike Control Serum, 8/30/2004 M LA/CM C/M EE D0907033.D 72873-5m ed spike Control Serum, 8/30/2004 M LA/CM C/M EE D0917157.D MS-040907-1 Med Matrix Spike; Serum; 07 Sept 04; M LA/CM C/M EE D0917158.D MS-040907-2 Med Matrix Spike; Serum; 07 Sept 04; M LA /C M C /M EE D0917159.D MS-040907-3 Med Matrix Spike; Serum; 07 Sept 04; M LA /C M C /M EE D0921123.D MS-040909-1 DQ921127.D M S-040909-2 D0921128.D MS-040909-3 D1222044.D MS-041222-1 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE Med Spike; Ext 12/22/04 C M C /M LA/VJT D1222045.D MS-041222-2 D1222046.D MS-041222-3 Med Spike; Ext 12/22/04 C M C /M LA/VJT Med Spike; Ext 12/22/04 C M C /M LA/VJT_________ D 0902112.D 72865-5 D0917163.D HS-040907-1 D0917164.D HS-040907-2 D0917165.D HS-040907-3 D0921129.D HS-040909-1 D0921130.D HS-040909-2 D0921131.D HS-040909-3 D1222047. D HS -041222-1 D1222051.D HS-041222-2 D1222052.D HS-041222-3 Control Serum-High Spike, 8/30/04 C M C /M LA /M EE Control Serum-High Spike, 8/30/04 CM C /M LA /M EE High Matrix Spike; Serum; 07 Sept 04; M LA /C M C /M EE High Matrix Spike; Serum; 07 Sept 04; M LA /C M C /M EE High Matrix Spike; Serum; 07 Sept 04; M LA /C M C /M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE 900 ppm Dose Group; Serum; 13 Sept 04; M LA/CM C/M EE High Spike; Ext 12/22/04 CM C /M LA/VJT High Spike; Ext 12/22/04 CM C /M LA/VJT High Spike; Ext 12/22/04 CM C /M LA/VJT RT 11QQ1".922 Area 30-16284 3109410 3.265 303978 3.265 305582 3.265 320574 3.258 220379 3.258 229699 3.258 211781 3.468 437884 3.467 465674 3.468 433404 1.908 2524596 1.901 2526086 3.28 4136012 3.28 3823096 3.265 1100348 3.265 1094469 3.265 1100342 3.259 790455 3.258 774172 3.258 777920 3.475 4835384 3.475 4686940 3.474 4774809 1.866 5350750 1.845 5422888 3.265 2318987 3.265 2377903 3.265 2410907 3.258 1659451 3.258 1662421 3.258 1707702 3.475 9066961 3.475 9380430 3.482 9026117 Concentration of PFB S in Extract, O n- colum n, ng/mL 447t4 4240 24.74 24.87 26.13 24.62 25.88 23.46 26.42 28.16 26.13 96.72 96.78 114.2 104.8 94.15 93.62 94.15 104.0 101.7 102.2 111.4 106.3 109.3 222.5 226.0 213.4 219.7 223.3 236.6 237.1 244.5 266.9 279.2 265.3 Recovery, % Average Recovery, % ^53>/u Spiking Error Spiking Error 96.7% 97.2% 102% 96.2% 101% 91.7% 103% 110% 102% 94.5% 94.6% 112% 102% 92.0% 91.5% 92.0% 102% 99.4% 99.9% 109% 104% 107% 100% 87% 88% 83% 86% 87% 92% 93% 96% 104% 109% 104% %RSD zr Cfl ZJ H o Tj n P1 MO =o3: OcQ . O 5' "-n0=c3o CD c CO Q __ < CD Iof^ s 3CD Page 1 of 1 i to 00 1 I,-ot o' a ze o 4^ 4^ Page 59 of 227 Contains No CBI Q uail S eru m . C o ntrol Pirnnp 3M Environmental Laboratory S am ple File Name D102S032.D 72848-1 D1025033.D 72848-2 D1025036.D 72848-3 D1025039.D 72849-1 D1025040.D 72849-2 D1025044.D 72849-3 D1025045.D 72850-1 D1025046.D 72850-2 D1025047.D 72850-3 D1025048.D 72851-1 D1025051.D 72851-2 D1025052.D 72851-3 D1025053.D 72853-1 D1025054.D 72853-2 D1025055.D 72853-3 D1025059.D 72854-1 D1025060.D 72854-2 D1025061 ,D 72854-3 D1025062.D 72855-1 D1025063.D 72855-2 D1025066.D 72855-3 D1025067.D 72857-1 D1025068.D 72857-2 D1025069.D 72857-3 D1025075.D 72858-1 D1025076.D 72858-2 D1025077.D 72858-3 D1025078.D 72859-1 D1025081 .D 72859-2 D1025082.D 72859-3 D1025083.D 72860-1 D1025084.D 72860-2 D1025085.D 72860-3 D1025089.D 72861-1 D1025090.D 72861-2 D1025091 ,D 72861-3 D1025092.D 72862-1 D1025093.D 72862-2 D1Q25096.D 72862-3 D1025097.D 72864-1 D1025098.D 72864-2 D1025099.D 72864-3 D1025100.D 72865-1 D1025104.D 72865-2 D1025105.D 72865-3 Concentration o f PFBS in Extract, On-Colum n Concentration of w . .. , ^C N (in P F B S ^ n r - 0 -1 i Area ----------- ngm iL------ < LLOQ 3.482 3.482 5879 13771 < LLOQ < LLOQ 3.475 6607 2.558 3.475 1648417 662.0 3.475 22122177 767.3 3.482 3.482 46551 33837 12.31 8.369 3.482 12819 2.558 3.482 3.482 3.482 8838 5617 12652 < LLOQ < LLOQ < LLOQ 3.482 3.482 3.482 14625 14193 21237 < LLOQ < LLOQ < LLOQ 3.482 3.482 3.481 207984 10094 12157 63.32 2.558 2.558 3.489 3.489 3.482 14643 13986 16135 < LLOQ < LLOQ < LLOQ 3.482 3.482 3.482 14274 14017 15460 < LLOQ < LLOQ < LLOQ 3.489 3.489 3.489 21603 23246 107301 2.558 2.558 31.28 3.489 38604 9.844 3.488 13222 2.558 13443 2.558 3.489 3.489 3.489 8270 8276 14749 < LLOQ < LLOQ < LLOQ 3.489 8694 2.558 3.489 3710 2.558 709330 236.0 3.489 3.489 3.489 45405 13277 16030 11.95 2.558 2.558 3.489 3.489 9199 9376 < LLOQ < LLOQ 3.489 9905 < LLOQ 3.489 3.489 3.489 14512 14109 13446 < LLOQ < LLOQ < LLOQ p lcto r" Factor 0.1 00..11 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 PFBSna/mLtraCt' Wehi9 h t fS e r7 ' in h o m o ^ n at e, W eight o f _ng/m L _hom ogenate), g , g Aliquot, g < LLOQ < LLOQ < LLQQ 0.2558 66.20 76.73 1.231 0.8369 0.2558 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLQQ 6.332 0.2558 0.2558 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.2558 0.2558 3.128 0.9844 0.2558 0,2558 < LLOQ < LLOQ < LLOQ 0.2558 0.2558 23.60 1.195 0.2558 0,2558 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0 .3 3 6 3 0.3363 0,3363 0.1189 0.1189 0.1189 0.2353 0.2353 0.2353 0.3921 0.3921 0.3921 0.3886 0.3886 0.3886 0.1413 0.1413 0.1413 0.3968 0.3968 0,3968 0.3778 0.3778 0.3778 0.3795 0.3795 0,3795 0.3909 0.3909 0.3909 0.3985 0.3985 0.3985 0.3834 0.3834 0.3834 0.3973 0.3973 0,3973 0.1356 0.1356 0.1356 0.2845 0.2845 0.2845 6.744 6.744 6.744 2.532 2.532 2.532 4.810 4.810 4.810 7.780 7.780 7.780 7.779 7.779 7.779 2.732 2.732 2.732 8.056 8.056 8.056 7.620 7.620 7.620 7.609 7.609 7.609 7.752 7.752 7.752 7.985 7.985 7.985 7.710 7.710 7.710 8.097 8.097 8.097 2.830 2.830 2.830 5.655 5.655 5.655 0.7338 0.7887 0.7996 0.7518 0.7104 0,8092 0.7610 0.7409 0.7661 0.7602 0.7459 0,7822 0.7567 0.7599 0.7453 0.7439 0.7869 0.7644 0.7490 0.7655 0.7702 0.7601 0.7596 0.7697 0.7542 0.7847 0.7848 0.7335 0.7582 0.7668 0.7842 0.7610 0.7852 0.7419 0.7723 0.7690 0.7199 0.7472 0.7433 0.7690 0.7610 0.7447 0.7670 0.7698 0.7223 Serum SampTM ug/q < LLOQ < LLOQ < LLOQ 0.0 4.16 4.23 0.0694 0.0484 0.0143 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.346 0.0132 0.0136 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0143 0.0137 0.168 0.0559 0.0141 0.0139 < LLOQ < LLOQ < LLOQ 0.0146 0.0140 1.30 0.0710 0.0146 0.0147 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ Average, ug/g < LLOQ 0.0440 < LLOQ < LLOQ < LLOQ < LLOQ 0 ,0 6 5 3 < LLOQ 0.0334 < LLOQ < LLOQ LLOQ On- Colum n, LLOQ on Dilution Corrected fo r Colum n, Factor On D ilution ng/m L Column Factor 5.115 5.115 5.115 0.1 0.1 0.5115 0.5115 0.5115 5.115 5.115 5.115 0.1 0.1 0.5115 0.5115 0.5115 5.115 5.115 5.115 0.1 0.1 0.1 0.5115 0.5115 0.5115 5.115 5.115 5.115 0.5115 0.5115 5.115 5.115 5.115 0.5115 0.5115 0.5115 5.115 5.115 5.115 0.5115 0.5115 0.5115 5.115 5.115 5.115 0 .5 1 1 5 0 .5 1 1 5 0.5115 5.115 5.115 5.115 0 .5 1 1 5 0 .5 1 1 5 0.5115 5.115 5.115 5.115 0 .5 1 1 5 0.5115 0.5115 5.115 5.115 5.115 0.5115 0.5115 0.5115 5.115 5.115 5.115 0.5115 0.5115 0.5115 5.115 5 .1 1 5 5.115 0.5115 0 .5 1 1 5 0.5115 5.115 5.115 5.115 0 .5 1 1 5 0 .5 1 1 5 0.5115 5 .1 1 5 5 .1 1 5 5.115 0 .5 1 1 5 0 .5 1 1 5 0.5115 5 .1 1 5 5 .1 1 5 5.115 0 .5 1 1 5 0 .5 1 1 5 0.5115 LLOQ in M atrix, Average ug/ g ug/g 0 .0 2 9 4 0.0273 0,0269 0 .0 3 0 3 0.0321 0 ,0 2 8 2 0 .0 2 8 8 0 .0 2 9 6 0 .0 2 8 6 0 .0 2 7 9 0 .0 3 0 2 0 .0 2 9 0 0 .0 2 8 6 0 .0 2 7 3 0 .0 2 8 4 0 .0 2 8 3 0 .0 2 8 8 0.0280 0.0264 0.0272 0.0291 0.0285 0.0283 0.0285 0.0285 0.0281 0.0286 0.0274 0.0274 0.0291 0.0281 0-027B 0.0274 0.0283 0.0274 0.0291 0.0280 0.0281 0.0304 0.0293 0.0294 0.0291 0.0294 0,0300 0.0278 0.0277 0.0296 0 .0 2 8 0 0 .0 2 8 5 0.G272 0 .0 2 8 6 0.0284 0 .0 2 8 3 0.0297 0.0295 "zOUr CD *>3do"<) H O 9? %> I3 CD 5O)* "oS OC 3o 2, o tj 3 T03I cCo CO Q . __ CD II co 3- 3 =w! 5<S' CD CD Contains No CBI ON NC TJ of t c i CD 4^ L4A^ ON Page 60 of 227 Contains No CBI -270- Project Number 454-116 Report E03-0583 J 3O ^ o S o OO .2 r a mo oo =iJl T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues ) Nu>O) i } CM CM CM ' C ->o pfl- *' o'o o in **- >- t- Tt cotm--rt- i ppp c o' o o' d o 'c r3 (o0D o0 t ( 1 i d o'o' < 2a - in w < - r-> r-~ r O3 5<1O o'd o i o oco O5lZ00o ' < I >I B^ OotNn <o ID >do'd o o o o o o o o o i o Oo ( Mo- o->r oM; tt 0 0. \hl :g1ja | a s o I- Ct--Mcr--m fc- i oaa a oa OOO1 tM (N CM CM CM r - r-- qqq qqq rc~m rc-m rrQOO l Qa ) O OO inin OOO cI> C<-3MD3r o> 0CtIDMt m<0CcMDm ioID CoD (oD CoM CoM oCM 5 CD CO |l tD CD CD I CM CM CM I 555i a a a 555 i CO 03 ro- ro) Q Q Q |Q Q 3M Environmental Laboratory Page 61 of 227 Report E03-0583 3M Environmental Laboratory Q uail S eru m . C ontrol G roun S a m p le File Name D1027031 .D 73104-2 D1027032.D 73104-3 D1027034.D 73105-2 D1027037.D 73105-3 D1027039.D 73106-2 D1027045.D 73107-2 D1027046.D 73107-3 D1027048.D 73108-2 D1027049.0 73108-3 D1027053.D 73109-2 D1027054.D 73109-3 D1027056.D 73110-2 D1027060.D 73110-3 D1027064.D 73111-2 D1027067.D 73111-3 D1027069.D 73112-2 01027070.0 73112-3 D1027075.D 73113-2 D1027076.D 73113-3 D1027078.0 73114-2 D1027079.D 73114-3 D1027083.D 73115-2 D1027084.D 73115-3 D1027085.D 73116-1 D1027086.D 73116-2 D1027090.0 73116-3 RT 3.489 3.489 Area C oncentration of PFBS in Extract, On-Column O n -co lu m n , D ilution ng/m L F a c to r 3477 3865 < LLOQ < LLOQ < LLOQ 0.1 0.1 0.1 0 0 < LLOQ 0 0 < LLOQ 0.1 0.1 3.489 7898 < LLOQ 0.1 Not enouqh samle fo r triplicate analysis Concentration of PFBS in Extract, W eight of serum (in n g /m L homogenate), g < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.2033 0.2033 0.2033 0.3147 0.3147 0.3147 0.0714 0.0714 W eight of ACN (in hom ogenate) W eight of , g Aliquot, g 4.180 4.180 4.180 6.344 6.344 6.344 1.483 1.483 0.7311 0.7611 0.7655 0.7715 0.7877 0.7320 0.7378 0.7420 Concentration o f PFBS in O riginal Serum Sample, Average, ug/g < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 3.489 0 2018 0 < LLOQ < LLOQ 0.1 0.1 0 0 < LLOQ 0 0 < LLOQ 0.1 0.1 0 3.489 0 8149 < LLOQ < LLOQ 0.1 0.1 3.489 3.489 197925 34771 60.18 9.327 0.1 0.1 0 0 < LLOQ 0 0 < LLOQ 0.1 0.1 3.409 3.496 2813 20016 < LLOQ < LLOQ 0.1 0.1 3.489 0 3445 0 < LLOQ < LLOQ 0.1 0.1 0 0 < LLOQ 0 0 < LLOQ 0.1 0.1 0 03.496 2144 3.489 3.489 108432 42757 < LLOQ < LLOQ 32.14 11.79 0.1 0,1 0.1 0.1 0.1 Not enough sample fo r triplicate analysis _Notenough sample fo r triplicate analysis < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.2558 6.0181 0.9327 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 3.214 1.179 < LLOQ ooo ODO 0.1846 0.1846 0.1846 0.3132 0.3132 0.3132 0.1903 0.1903 0.1903 0.2736 0.2736 0.2736 0.3435 0.3435 0.3435 0.2799 0.2799 0.2799 0.2189 0.2189 0.2189 0.1338 0.1338 0.1338 0.3293 0.3293 0.3293 0.2725 0.2725 0.2725 0.0342 3.741 3.741 3.741 6.286 6.286 6.286 3.904 3.904 3.904 5.551 5.551 5.551 6.952 6.952 6.952 5.668 5.668 5.668 4.558 4.558 4.558 2.552 2.552 2.552 6.553 6.553 6.553 5.462 5.462 5.462 0.660 0.7607 0.7645 0.7643 0.7136 0.7048 0.6962 0.6972 0.6688 0.7100 0.7310 0.6941 0.7640 0.7669 0.7549 0.7606 0.7184 0.7020 0.7091 0.7655 0.7631 0.7551 0.7757 0.7497 0.7467 0.7392 0.7488 0.7763 0.6652 0.7090 0.7668 0.6700 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0149 0.369 0.0520 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0162 0.191 0.0647 < LLOQ < LLOQ < LLOQ < LLOQ 0.1453 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0906 < LLOQ OJseid 1/2 LLOQ fo r one of the replicate calculations ~ U sed 1/2 LLOQ for two of the replicate calculations ***Used ULOQ for one of the replicate calculations LLOQ On- C olum n, LLOQ on Dilution Corrected fo r C o lu m n , F actor On D ilu tio n LLOQ in M atrix, Average, 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0,1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.5115 0.0302 0.0290 0.0281 0.0275 0.0302 0.0300 0.0286 0.0285 0.0302 0.0306 0.0316 0.0329 0.0298 0.0314 0.0283 0.0288 0 .0 3 0 3 0 .0 3 1 0 U.U3U/ 0.0292 0.0293 0.0265 0 .0 2 7 4 0.0289 0.0286 0.0324 0.0304 0.0310 0.0293 0 .0 2 8 4 0.0301 0.0285 0.0306 0.0318 0.0299 0 .0 2 8 6 0.0306 0.0293 0.0271 0 .0 2 8 3 0.0303 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS In Northern Bobwhite Tissues Page 62 of 227 Page 3 of 3 Contains No CBI L/i i On "o1 On Contains No CBI Report E03-0583 T-/485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page of 2 Q u ail S e ru m . 100 p p m D o se G ro u p 3M Environmental Laboratory Page 63 of 227 3M Environmental Laboratory Quail Serum. 100 ppm Dose Groun NJ U) > 73 T3 ~cun Sco CO Q . f CD i o0 cn' 2c! 1 C"-Dt 4L4^^/i Contains No CBI Page 64 of 227 Page 65 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Q u a il S erum . 300 ppm D ose G rou p File D0922030.D D0922031.D D0922033.D D0922035.D D0922037.D D0922038.D D0922043.D D0922044.D D0922046.D D0922047.D D0922050.D D0922051.D D0922053.D D0922057.D D0922059.D D0922060.D D0922063.D D0922064.D D0922066.D D0922067.D D0922072.D D0922073.D D0922075.D D0922077.D D0922079.D D0922080.D D0922085.D D0922086.D D0923030.D D0923031.D Sample Name 72912-2 72912-3 72913-2 72913-3 72914-2 72914-3 72915-2 72915-3 72916-2 72916-3 72917-2 72917-3 72918-2 72918-3 72919-2 72919-3 72920-2 72920-3 72921-2 72921-3 72922-2 72922-3 72923-2 72923-3 72924-2 72924-3 72926-2 72926-3 72927-2 72927-3 Concentra tion of PFBS In Extract, On- column, RT Area ng/mL 1656511 3.258 1453367 3.259 1631613 3.258 1811217 3.258 1881530 3.258 1955651 3.258 1205864 3.258 1176837 3.259 1199138 3.258 1355183 3.265 1452583 3.265 1415026 3.266 4067076 3.265 4156847 3.266 4041199 3.266 2641417 3.265 2548305 3.266 2585007 3.265 1847234 3.265 1746677 3.266 1843061 3.265 2699960 3.266 2734453 3.265 2725370 3.265 1505033 3.265 1491272 3.265 1443872 3.265 1609508 3.265 1701631 3.265 1704806 3.265 1066947 3.265 1056274 3.266 1019646 217.8 187.7 214.1 241.3 252.1 263.5 151.9 147.7 150.9 173.4 187.6 182.1 651.3 671.7 645.5 375.1 359.3 365.5 246.8 231.4 246.1 385.1 391.1 389.5 195.3 193.3 186.3 210.8 224.6 225.1 132.2 130.7 125.5 3.265 3.265 3.265 3.265 3.265 3.265 3.266 3.265 3.265 3.259 3.258 3.258 3421955 3281011 3413938 2780526 2654689 2777323 2456058 2388451 2343218 5131081 5014034 4859066 516.9 489.9 515.4 399.1 377.3 398.5 343.9 332.7 325.3 985.6 946.1 897.2 OnColumn Dilution Factor 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 Concentrai ion of Concentrati Weight of Weight of on of PFBS serum (in ACN (in in Extract, homogena hom ogenat Weight of ng/mL te), 9 e). 9 Aliquot, a 435.7 375.4 428.2 0.3886 0.3886 0.3886 8.114 8.114 8.114 0.7781 0.7039 0.7780 482,5 504.1 527.1 0.1403 0.1403 0.1403 2.841 2.841 2.841 0.7582 0.7679 PFBS in Original Serum Sample, 24.5 23.3 27.0 27.9 Average, 24.0 27.8 303.8 295.5 301.8 0.2200 0.2200 0.2200 4.511 4.511 4.511 0.7792 0.7714 16.8 16.5 16.7 346.8 375.2 364.2 0.1837 0.1837 0.1837 3.891 3.891 3.891 0.7577 0.7798 20.3 21.3 20.7 1303 1343 1291 0.4000 0.4000 0.4000 4.078 4.078 4.078 0.7903 0.8162 36.9 36.8 36.9 750.2 718.6 731.0 ^^956TM 0.3956 0.3956 7.903 7.903 7.903 0.7563 0.7527 41.6 40.1 40.5 493.6 462.9 492.3 770.2 782.1 779.0 390.6 386.6 372.6 421.6 449.3 450.2 0.3404 0.3404 0.3404 0.3971 0.3971 0.3971 0.2458 0.2458 0.2458 0.3831 0.3831 0.3831 6.804 6.804 6.804 8.143 8.143 8.143 4.915 4.915 4.915 7.751 7.751 7.751 0.7937 0.7858 0 7762 0.7642 0.7908 0.7733 0.7855 0.7469 0.7793 0.7893 26.1 24.7 43.4 42.5 21.2 20.7 23.0 24.2 25.8 42.9 20.9 23.8 264.3 261.3 251.0 1034 979.7 1031 798.1 754.7 797.0 0.2536 0.2536 0.2536 0.3995 0.3995 0.3995 0.3941 0.3941 0.3941 4.973 4.973 4.973 8.418 8.418 8.418 7.919 7.919 7.919 0.7739 0.7729 0 7111 0.7722 0.7724 0.7847 0.7877 0.7536 14.1 13.9 59.1 56.0 42.7 42.2 14.2 57.7 42.9 687.8 665.4 650.6 1971 1892 1794 0.3777 0.3777 0.3777 0.2529 0.2529 0.2529 7.719 7.719 7.719 5.065 5.065 5.065 0.7834 0.7766 0.7704 0.7788 0.7825 0.7694 37.6 36.7 106 102 98.1 36.9 102 LLOQ On- LLOQ on Column, ng/mL Column, Corrected Dilution for Factor On Diiution LLOQ in Matrix, 25.58 2 51.16 2.88 25.58 2 51.16 3.18 25.58 2 51.16 2.88 25.58 2 51.16 2.87 25.58 2 51,16 2.83 25.58 2 51.16 2.75 25.58 2 51.16 2.82 25.58 2 51.16 2.85 25.58 2 51.16 2.86 25.58 2 51.16 3.00 25.58 2 51.16 2.91 25.58 2 51.16 2.86 25.58 2 51.16 1.45 25.58 2 51.16 1.40 25.58 2 51.16 1.47 25.58 2 51.16 2.84 25.58 2 51.16 2.85 25.58 2 51.16 2.78 25.58 2 51.16 2.71 25.58 2 51.16 2.73 25.58 2 51.16 2.77 25.58 2 51.16 2.88 25.58 2 51.16 2.78 25.58 2 51.16 2.81 25.58 2 51.16 2.78 25.58 2 51.16 2.73 25.58 2 51.16 2.88 25.58 2 51.16 2.79 25.58 2 51.16 2.75 25.58 2 51.16 2.76 25.58 2 51.16 2.72 25.58 2 51.16 2.73 25.58 2 51.16 2.97 25.58 2 51.16 2.92 25.58 2 51.16 2.92 25.58 2 51.16 2.88 25.58 2 51.16 2.74 25.58 2 51.16 2.86 25.58 2 51.16 2.80 25.58 2 51.16 2.80 25.58 2 51.16 2.82 25.58 2 51.16 2.85 25.58 2 51.16 2.76 25.58 2 51.16 2.75 25.58 2 51.16 2.80 2.74 2 91 2 82 2 77 Page 1 of 3 Contains No CBI O-i L/i Os Page 66 of 227 3M Environmental Laboratory Quail Serum, 300 ppm Dose Group Fite D0923033.D D0923035.D D0923037.D D0923038.D D0923043.D D0923044.D D0923046.D D0923047.D D0923050.D D0923051.D D0923053.D D0923057.D D0923059.D D0923060.D D0923063.D D0923064.D D0923066.D D0923067.D D0924098.D D0924099.D D0917033.D D0917036.D D0917038.D D0917039.D D0917044.D D0917045.D D0917047.D D0917048.D D0917052.D D0917053.D D0917055.D D0917059.D Sample Name 72928-1 72928-2 72928-3 72929-2 72929-3 72930-2 72930-3 72931-2 72931-3 72932-2 72932-3 72933-2 72933-3 72934-2 72934-3 72935-2 72935-3 72936-2 72936-3 72937-2 72937-3 72938-2 72938-3 72939-2 72939-3 72940-2 72940-3 72941-2 72941-3 72942-2 72942-3 72943-2 72943-3 RT 3.258 3.259 3.258 3.258 3.259 Concentra tion of PFBS in Extract, On- Area c o lu m n , n g/m L 2073734 2003215 2002428 297.7 286.0 285.9 2383659 2366620 355.0 350.3 347.3 On- C olum n Dilution Factor 2.0 2.0 2.0 2.0 2.0 2.0 3.252 2678027 3.252 2485912 3.259 2842951 3.259 2759885 3.258 1261710 3.258 1254555 3.259 1618975 3.258 1618851 3.258 1758609 3.259 1763346 402.4 368.1 432.5 417.3 168.8 167.8 221.8 224.1 224.0 227.8 246.2 247,0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 3.258 1215428 3.258 1257222 3.258 3.258 3.272 3.273 3.258 3.258 1018120 1015994 4505185 4442993 4325491 5334315 7657976 5094578 3.258 4074881 3.258 3907006 3.259 3.259 Spilled 3.265 3.265 4774496 4805480 4530691 4338195 3.265 5407729 3.265 5295335 3.265 2474355 3.265 2406857 161.8 168.2 132.4 132.1 614.5 592.4 561.5 936.5 529.5 401.9 382.1 465.8 488.0 492.0 457.3 433.6 571.5 556.2 221.2 223.4 216.3 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 Concentrai ion of Concentrati Weight of W eight of on o f PFBS serum (in ACN (in in Extract, homogena hom ogenat W eight of ng/mL te). a e), g Aliquot, q 595.3 572.0 571.7 710.1 700.6 694.7 781.4 804.7 736.3 843.7 865.1 834.5 324.2 335.5 443.7 448.1 448.1 455.7 492.5 494.0 315.7 336,3 240,9 264 7 264.1 1253 1229 1185 561.5 936.5 529.5 383.8 401.9 382.1 465.8 488.0 492.0 0.3124 0.3124 0.3124 0.3914 0.3914 0.3914 0.3976 0.3976 0.3976 0.3867 0.3867 0.3867 0.2241 0.2241 0.3708 0.3708 0.3708 0.2009 0.2009 0.2009 0.3886 0.3886 0.3981 0.3981 0.3799 0.3799 0.3799 0.3956 0.3956 0.3956 0.3958 0.3958 0.3958 0.4513 0.4513 0.4513 6.243 6.243 6.243 7.905 7.905 7.905 7.931 7.931 7.931 7.732 7.732 7.732 4.826 4.826 4.826 7.479 7.479 7.479 4.086 4.086 4.086 7.817 7.817 7.817 7.904 7.904 7.904 7.512 7.512 7.512 8.029 8.029 8.029 8.028 8.028 8.028 8.963 8.963 8.963 0.7811 0.7366 0.7828 0.7816 0.7694 0.7681 0.7832 0.7856 0.7109 0.7593 0.7876 0.7880 0.7801 0.7639 0.7495 0.7848 0.7645 0.7653 0.7713 0.7792 0.7798 0.7652 0.7789 0.6895 0.7687 0.7789 0.7845 0.7493 0.7315 0.7815 0.7797 0.7635 0.7478 0.7758 0.7579 0.7724 0.7709 0.7821 PFBS in Original Serum Sample, ug/g 32.0 32.6 30.7 38.5 38.6 38.3 41.8 42.9 43.4 46.7 46.1 44.5 18.7 19.8 25.1 24.2 24.8 25.4 27.2 27.1 17.1 18.2 14.6 14.1 66.3 68.1 67.3 30.6 51.2 29.5 21.8 22.1 21.5 25.2 26.4 26.2 Average, ug/g 31.7 38.5 42.7 45.7 19.4 24.7 26.6 17.7 14.4 67.3 37.1 21.8 25.9 457.3 433.6 570.2 571.5 556.2 221.2 223 4 216.3 0.4008 0.4008 0.1351 0.1351 0.1351 0.3852 0.3852 8.048 8.048 2.706 2.706 2.706 7.678 7.678 7.678 0.7875 0.7898 0.7816 0.7744 0.7441 0.7651 0.7853 0.7266 24.5 23.1 30.7 31,0 31.4 12.1 12.5 23.8 31.1 12.2 Page 2 of 3 Report E03-0583 T-/485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues LLO Q on C o lu m n , LLOQ On- Column, Corrected Dilution for Factor On Dilution LLO Q in Matrix, 25.58 2 51.16 2.75 25.58 2 51.16 2.91 25.58 2 51.16 25.58 2 51.16 2.77 25.58 2 51.16 2.82 25.58 2 51.16 25.58 2 51.16 2.74 25.58 2 51.16 2.73 25.58 2 25.58 2 51.16 2.83 25.58 2 51.16 2.73 25.58 2 25.58 2 51.16 2.96 25.58 2 51.16 2.96 25.58 2 25.58 2 51.16 2.89 25.58 2 51.16 2.76 25.58 2 25.58 2 51.16 2.85 25.58 2 51.16 2.83 25.58 2 51.16 25.58 2 51.16 2.77 25.58 2 51.16 2.82 25.58 2 25.58 2 51.16 3.09 25.58 2 51.16 2.78 25.58 2 51.15 2 102.3 5.42 51.15 2 102.3 5.67 51.15 2 51.15 51.15 51.15 51.15 51.15 51.15 2.79 2.79 51.15 51.15 51.15 51.15 51.15 51.15 2.91 2.81 51.15 51.15 51.15 51.15 51.15 51.15 2.76 2.77 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51 15 51.15 51.15 61 16 2.74 2.75 2.78 2.80 2.73 2.80 2.81 2.83 2.76 2.98 2.83 2.83 2.79 2.87 5.63 2.81 2.86 2.75 2.73 2.81 2.82 Contains No CBI CD 4^ L/i i Q\ Page 67 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Quail Serum. 300 ppm Dose Groun File D0928029.D D0928030.D D0928031 .D D0928032D D0928033.D D0928036.D D0928037.D D0928038.D D0928039.D D0928040.D D0928044.D D0928045.D D0928046.D D0928047.D D0928048.D D0928051.D D0928052.D D0928053.D D0928054.D D0928055.D D0928059.D D0928060.D D0928061.D D0928062.D DO928063.D D0928066.D D0928067 D D0928068.D D0928069.D D0928070.D D0928074.D D0928075.D D0928076.D D0928077.D D0928078.D D0928081.D D0928082.D D0928083.D D0928084.D D0928085.D D0928089 D D0928090.D D0928091.D Sample Name Concentra tion of PFBS in Extract, On- column, RT Area ng/ml_ 73133-2 73133-3 73134-2 73134-3 3.258 3.258 3.258 3.258 37666 56369 r 1.J 83943 48280 12.47 19.54 29.97 16.48 73135-2 73135- 7313673136-2 73136-3 7313773137-2 73137- 7313873138-2 73138- 7313973139-2 73139- 7314073140-2 73140-3 3.258 3.3258 1 3.258 3.259 1 3.265 3.3266 1 3.265 3.3266 1 3.265 3.3273 1 3.272 3.272 43171 48428 30200 25750 18187 23421 25713 20796 18927 14579 24645 18283 14.55 16.54 9.647 7.965 5.107 7.085 7.951 6.093 5.387 2.558 7.547 5.143 73141-2 3.272 55241 19.11 Not enough sample for triplicates 73142-2 73142- 73143- 3.273 3.3273 1 17788 9422 6.417 2.558 2.558 73143-2 7314373144- 3.272 3.3279 1 26062 28843 8.083 9.134 73144- 2 3.279 6300 2.558 Not enough sample for triplicates 73145- 1 73145-2 73145- 73146- 3.279 3.3279 1 21410 18716 6.325 5.307 73146-2 73146- 73147- 3.279 30 1 19087 0 5.447 2.558 73147-2 3.28 20299 73147-3 3.286 20623 5.905 6.028 OnColumn Dilution Factor 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Concentrai ion of Concentrati Weight of Weight of on of PFBS serum (In ACN (in in Extract, homogena hom ogenat Weight of ng/mL te ). 9 e), g Aliquot, g 1.878 1.247 1.954 1.929 2.997 1.648 2.365 1.455 1.654 0.2151 0.2151 0.2151 0.3241 0.3241 0.3241 0.3158 0.3158 0.3158 4.503 4.503 4.503 6.509 6.509 6.509 6.318 6.318 6.318 0.7767 0.7791 0.7749 0.7739 0.7800 0.7635 0.7815 0.7603 0.7605 1.687 0.965 0.7965 0.0938 0.0938 0.0938 1.914 1.914 1.914 0.7183 0.7214 0'6916 0.4642 0.5107 0.7085 0.2937 0.2937 0.2937 5.933 5.933 5.933 0.7521 0.7802 0.7388 0.9294 0.7951 0.6093 0.2409 0.2409 0.2409 4.838 4.838 4.838 0.7796 0.7829 0.7765 0.5387 0.2558 1.268 0.7547 0.5143 1.586 1.911 0.3390 0.3390 0.3390 0.2797 0.2797 0.2797 0.0680 0.0680 6.813 6.813 6.813 5.949 5.949 5.949 1.465 1.465 0.7744 0.7738 0.7668 0.7764 0.7768 0.7435 0.7330 0.7614 PFBS in Original Serum Sample, ug/g 0.106 0.0702 0.111 0.105 0.162 0.0910 0.127 0.0804 0.0914 0.101 0.0572 0.0493 0.0262 0.0278 0.0407 0.0503 0.0428 0.0331 0.0322 0.0294 0.0141 0.0727 0.0433 0.0308 0.0975 0.113 Average, ug/g 0.0956 0.119 0.0996 0.0690 0.0315 0.0421 0.0252 0.0489 0.105 0.6417 0.2558 0.2558 0.7077 0.8083 0.9134 0.6072 0.2558 0.0962 0.0962 0.0962 0.1626 0.1626 0.1626 0.0466 0.0466 1.918 1.918 1.918 3.216 3.216 3.216 1.033 1.033 0.7817 0.7701 0.4225 0.7791 0.7794 0.7715 0.7605 0.2827 0.0344 0.0139 0.0253 0.0377 0.0431 0.0492 0.0370 0.0419 0.0245 0.0433 0.0395 0.6863 0.6325 0.5307 0.5427 0.5447 0.2558 0.2558 0.5905 0.6028 0.1742 0.1742 0.1742 0.0732 0.0732 0.0732 0.3121 0.3121 0.3121 3.436 3.436 3.436 1.651 1.651 1.651 6.361 6.361 6.361 0.7770 0.7786 0.7742 0.7700 0.7534 0.1621 0.7731 0.7574 0.7755 0.0366 0.0337 0.0284 0.0332 0.0341 0.0743 0.0141 0.0333 0.0332 0.0329 0.0472 0.0269 LLOQ On- Column, Corrected LLOQ on Dilution for LLOQ in Column, Factor On Dilution Matrix, 5.115 5.115 ug/g 0.1 0.5115 0.0289 0.1 0.5115 0.0288 5.115 5.115 0.0290 0.1 0.5115 0.0279 0.1 0.5115 0.0277 0.0282 5.115 0.1 0.5115 0.0275 5.115 5.115 0.1 0.5115 0.0283 0.0283 5.115 0.1 0.5115 0.0305 5.115 0.1 0.5115 0.0304 5.115 0.0317 57TT5 0.1 0.5115 0.0288 5.115 0.1 0.5115 0.0278 5.115 5.115 5.115 0.0294 0.1 0.5115 0.0277 0.1 0.5115 0.0275 5.115 5.115 5.115 5.115 5.115 5.115 0.1 0.5115 0.0279 0.1 0.5115 0.0279 0.0281 0.1 0.5115 0.0293 0.1 0.5115 0.0293 0.0306 0.1 0.5115 0.0315 0.1 0.5115 0.0303 5.115 5.115 5.115 5.115 5.115 5.115 5.115 0.1 0.5115 0.0274 0.1 0.5115 0.0278 U.U5/ 0.1 0.5115 0.0273 0.1 0.5115 0.0273 0.0276 0.1 0.5115 0.0312 0.1 0.5115 0.0838 5.115 5.115 5.115 5.115 5.115 5.115 0.1 0.5115 0.0273 0.1 0.5115 0.0272 0.0274 0.1 0.5115 0.0313 0.1 0.5115 0.0320 0.149 0.1 0.5115 0.0283 0.1 0.5115 0.0289 _ _ _____________________ Used 1/2 LLOQ for one of the replicate c a l c u l a t i o n s ------------------------------ u udaz " ---------------------------------------- ----------------------------------------------- " Used 1/2 LLOQ for two of the replicate calculations__________ ________________________ __ A verage, ug/g 0.0289 0.0279 0.0280 0.0308 0.0287 0.0277 0.0280 0.0298 0.0309 0.0353 0.0274 0.0575 0.0273 0.0707 0.0285 Page 3 of 3 Contains No CBI Os P a g e 68 of 227 3M Environmental Laboratory Quail Serum, 900 ppm Dose Groun File S am ple Name RT D0924030.D 72944-2 D0924031.D 72944-3 D0924033.D 72945-2 D0924035.D 72945-3 D0924037.D 72946-2 D0924038.D 72946-3 D0924043.D 72947-2 D0924044.D 72947-3 D0924046.D 72948-2 D0924047.D 72948-3 D0924050.D 72949-2 D0924051.D 72949-3 D0924053.D 72950-2 D0924057.D 72950-3 D0924059.D 72951-2 D0924060.D 72951-3 D0924063.D 72952-2 D0924064.D 72952-3 D1223022.D 72953-2 D1222031.D 72953-3 D0924072.D 72954-2 D0924073.D 72954-3 D0924075.D 72955-2 D0924077.D 72955-3 3.258 3.258 3.259 3.259 3.259 3.258 3.258 3.258 3.258 3.265 3.265 3.266 3.265 3.266 3.266 3.265 3.259 3.265 3.265 3.265 3.265 3.266 3.46 3.468 3.265 3.266 3.265 3.265 D0924079.D 72956-2 D0924080.D 72956-3 D0924085.D 72957-2 DQ9240B6.D 72957-3 D0924088.D 72958-2 D0924089.D 72958-3 3.265 3.266 3.265 3.265 3.265 3.265 3.266 D0924092.D 72959-2 D0924093.D 72959-3 3.266 3.266 D0925030.D 72960-2 D0925031.D 72960-3 3.258 3.258 C o n c e n tra ta n of PFBS in Area Extract, Onco lu m n , ng /m L O n -C o lu m n D ilution F a c to r 1419324 1438635 175.2 151.5 154.0 4 4 4 2923649 357.71 4 2928154 2788662 2782108 360.5 339.6 339.7 338.7 4 4 4 4 5408646 5330822 3440470 3191726 719.7 819.0 800.6 440.1 4 4 4 4 3513823 451.9 4 3513346 3523565 469.6 451.85 453.51 4 4 4 1775165 1877647 194.7 198.0 21 1.6 4 4 4 5437716 5546292 826.0 852.8 4 4 4 3503380 3433740 446.37 450.23 439.0 4 4 4 19113316 20726022 2330898 885.2 636.4 892.0 284.1 8 10 8 4 2269594 3231635 3375064 3376561 265.2 407.0 429.6 429.8 4 4 4 4 2554598 2492977 296.8 305.6 296.7 4 4 4 3416650 423.3 4 3430552 2595185 2575420 2577442 438.5 311.4 308.6 308.9 4 4 4 4 2241302 2239848 261.3 261.1 4 4 4 1775659 1782262 218.2 213.3 214.3 4 4 4 C o n ce n tra tio n o f PFBS in Extract, ng/mL 700.8 606.0 615.9 1431 1439 1442 1358 1359 1355 2879 3275 3202 1760 1603 1808 1878 1807 1814 778.7 791.8 846.5 3323 3304 3411 1785 1801 1756 7082 6364 7136 1136 1095 1061 1628 1718 1719 1187 1222 1187 1693 1745 1754 1246 1234 1235 1070 1045 1044 872.7 853.4 857.2 W eight of serum (in hom ogenate), 9 0.3903 0.3903 0.3903 0.3982 0.3982 0.4005 0.4005 0.4005 0.4420 0.4420 0.4420 0.3999 0.3999 0.3996 0.3996 0.3996 0.3599 0.3599 0.3599 0.3885 0.3885 0.3885 0.3953 0.3953 0.3953 0.3999 0.3999 0.3999 0.3090 0.3090 0.3966 0.3966 0.3966 0.3927 0.3927 0.3927 0.4010 0.4010 0.2078 0.2078 0.2078 0.2620 0.2620 0.2620 0.3976 0.3976 0.3976 W eight o f ACN (in hom ogenate), W eight of C o n ce n tra tio n o f PFBS in O riginal Serum 7.747 7.747 7 .7 4 7 7.901 7.901 0.7809 0.7838 0.7864 0.7642 37.4 32.2 75.8 78.5 8 .2 6 6 8 .2 6 6 8.488 8.488 8.009 8.009 0.7889 0.7827 0 .7 6 3 9 0.7794 0.7788 0 .7 0 6 7 74.5 75.1 152 170 95.1 95.4 3.002 8.00 2 0.7953 0.7747 99.3 98.1 7.190 7.190 7.190 7.767 7.767 0.7856 0.7833 0.7799 0 .7 9 0 5 41.6 42.4 179 175 8.096 8.096 8.096 8.142 8.142 8 .1 4 2 6.321 6.321 6.321 8.00 0 8.00 0 7.961 7.961 7.976 7.976 0.7756 0.7674 0.7919 0.7959 0.7863 0.7708 0.7949 0.7944 0.7646 0.7860 0.7750 0.7804 98.9 101 382 342 62.0 61.0 86.7 91.6 6 6 .1 66 .2 91.3 93.4 4 .1 4 2 4 .1 4 2 4.142 5.264 5.264 5.264 8.00 0 8.00 0 8.00 0 0.7709 0.7749 0 .7 7 5 5 0 .7 8 9 8 0.7580 0.7768 0.7871 0.7515 67.6 66.7 57.2 58.2 47.5 45.8 48.2 Average, 34.3 77.4 74.9 162 95.7 97.7 43.1 173 98.8 370 61.3 90.4 65.6 93.1 67.0 57.3 47.1 Page 1 of 3 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues L LO Q On- Colum n, LLOQ on C olum n, ng /m L 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 C orrected D ilution for Factor On D ilution Column Factor 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 204.6 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 8 409.2 10 511.5 8 409.2 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204,6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 4 204.6 LLO Q in M atrix, ug/g 10.9 10.5 11 .0 11 .2 11 .0 11 .2 11.3 10 .8 10.4 1 2 .2 11 .0 10 .8 10.9 11 .0 10.9 10.9 10 .8 11.3 11.3 22 .1 27.5 22 .1 11.4 11.7 10.9 11 .0 11.4 1 1 .1 1 1 .1 11 .0 11 .0 11 .0 1 1 .1 11 .0 10.9 1 1 .1 1 1 .1 11 .0 11.5 Average, ug/g 10.9 11 .0 Contains No CBI O U\ i On Page 69 of 227 3M Environmental Laboratory Q uail Serum. 900 ppm Dose Group File Sam ple Name D0925033.D 72961-2 D0925035.D 72961-3 RT 3.258 3.259 3.259 Area C o n ce n tra tio n o f PFBS in Extract, On- On-Column colum n, D ilu tio n ng /m L Factor 4763101 5005943 4781735 749.7 810.7 754.2 4 4 C o n ce n tra tio n o f PFBS in Extract, ng/mL 2999 3243 3017 D0925037.D 72962-2 D0925038.D 72962-3 D0925043.D 72963-2 D0925044.D 72963-3 D0925046.D 72964-2 D0925047.D 72964-3 D0925050.D 72965-2 D0925051.D 72965-3 D0925053.D 72966-2 D0925057.D 72966-3 D0925059.D 72967-2 D0925060.D 72967-3 D0925063.D 72968-2 D0925064.D 72968-3 D0925066.D 72969-2 D0925067.D 72969-3 D092SO72.D 72970-2 D0925073.D 72970-3 3.258 3.258 3.259 3.259 3.258 3.258 3.258 3.259 3.258 3.258 3.259 3.266 3.258 3.259 3.265 3.259 3.259 3.265 2443057 2493757 1598741 1696181 2154764 2202225 2189563 3016763 3099473 2975379 2139563 2110616 1848636 1857202 3224631 3315190 1910380 1860653 3164546 3086252 312.4 320.3 188.3 2 0 2 .0 275.8 273.9 404.6 418.5 397.7 266.4 262.1 223.8 225.0 439.9 455.7 232.7 225.5 429.6 416.3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 1250 1281 731.2 753.2 808.2 1075 1103 1096 1618 1674 1591 1077 1066 1048 874,4 895.3 900.2 1878 1760 1823 890.8 930.9 902.2 1741 1718 1665 D0925075.D 72971-2 D0925077.D 72971-3 D0925079.D 72972-2 D0925080.D 72972-3 D0925085.D 72973-2 D0925Q86.D 72973-3 D0925088.D 72974-2 D0925089.D 72974-3 D0925092.D 72975-2 D0925093.D 72975-3 D0930028.D 73148-2 D0930029.D 73148-3 73149-2 73149-3 3.266 3.265 3.265 3.265 3.265 3.265 3.265 3.265 3.265 3.265 3.279 3.28 2518719 2437423 1680946 1711185 1286592 1135970 2203172 2201375 1546939 1525737 93522 103382 85970 324.2 311.6 199.9 204.2 145.2 124.9 275.9 275.6 181.1 178.1 36.00 29.70 Not enough sample for triplicates Not enough sample for triplicates 4 4 4 4 4 4 4 4 4 4 0.1 0.1 0.1 0.1 1297 1246 792.4 799.5 816.7 573.0 580.9 499.7 1008 1104 1103 712.3 724.3 712.4 3.243 3.600 2.970 2.285 W eight of serum {in hom ogenate), g 0.4020 0.4020 0.4020 0.3742 0.3742 0.3742 0.3421 0.3421 0.3421 0.5518 0.5518 0.5518 0.3995 0 .3 9 9 5 0 .3 9 9 5 0 .2 3 3 8 0 .2 3 3 8 0 .2 3 3 8 0 .3 7 6 2 0 .3 7 6 2 0 .3 7 6 2 0 .3 7 7 6 0 .3 7 7 6 0 .3 7 7 6 0.22 12 0.22 12 0.22 12 0 .3 6 8 4 0 .3 8 8 4 0 .3 8 8 4 0.2629 0.2629 0.2629 0.3896 0 .3 8 9 6 0.3896 0.3116 0.3116 0.3116 0 .3 9 6 8 0 .3 9 6 8 0 .3 9 6 8 0 .4 7 7 5 0 .4 7 7 5 0 .4 7 7 5 0 .3 2 6 5 0 .3 2 6 5 0 .3 2 6 5 0 .0 2 8 6 W eight o f ACN (in hom ogenate), g 8.073 8.073 8.073 7.434 7.434 7.434 6.870 6.870 6.870 11.029 11.029 11.029 7.987 7.987 7.987 4.703 4.703 4.703 7.669 7.669 7.669 7.517 7.517 7.517 4.455 4.455 4.455 7.877 7.877 7.877 5.434 5.434 5.434 7.900 7.900 7.900 6.205 6.205 6.205 7.909 7.909 7.909 9.553 9.553 9.553 6.585 6.585 6.585 0.660 W eight of A liquot, g 0.7555 0.7979 0.7868 0.7666 0.7189 0.7460 0.6710 0.7535 0.7855 0.7581 0.7638 0.7584 0.7903 0.7758 0.7731 0.7774 0.7432 0.7709 0.7895 0.7890 0.7817 0.7538 0.7739 0.7467 0.7829 0.7892 0.7806 0.7791 0.7381 0.7761 0.7815 0.7806 0.7558 0.7810 0.7795 0.7849 0.7797 0.7013 0.7169 0.7747 0.7770 0.7818 0.7720 0.7541 0.7691 0.7484 0.7013 0.6530 C oncentration o f PFBS in O riginal Serum Sample, ug/q 167 171 162 115 72.5 71.7 45.9 42.1 59.5 60.6 60.6 8 6 .0 86 .1 58.8 57.9 59.6 48.5 48.5 48.8 100 97.6 98.5 50.4 50.3 48.3 94.9 93.9 96.0 71.3 71.9 69.2 44.6 43.6 44.6 30.5 31.2 29.8 58.9 59.6 59.4 38.3 39.4 39.7 0 .1 7 9 0 .2 0 4 0 .1 7 9 0.168 Average, 167 86.5 43.8 60.3 87.0 58.8 48.6 98.8 49.7 94.9 70.8 44.3 30.5 59.3 39.1 0.187 0.158 Page 2 of 3 LLOQ on C o lu m n , LLOQ On- Colum n, C orrected D ilution fo r Factor On D ilution LLO Q in M a trix, 51.15 4 204.6 11.4 51.15 204.6 10 .8 51.15 5.1S 8 409.2 22.3 51.15 4 204.6 11.9 Average, ug/g 11.1 15.2 51.15 4 204.6 12.9 51.15 4 204.6 11.4 11 .8 51.15 4 204.6 11.3 51.15 4 204.6 11 .2 11.3 51.15 4 204.6 51.15 4 204.6 10.9 10.9 51.15 4 204.6 11 .2 51.15 4 204.6 1 1 .1 11.3 51.15 4 204.6 11.4 51.15 4 204.6 1 1 .1 1 1 .2 51.15 4 204.6 10.9 51.15 4 204.6 11.3 11.1 51.15 4 204.6 11 .6 51.15 4 204.6 11 .0 1 1 .2 51.15 4 204.6 11 .2 51.15 4 204.6 11 .2 11.4 51.15 4 204.6 11.4 51.15 4 204.6 11.3 11.4 51.15 51.15 4 204.6 11.5 51.15 4 204.6 1 1 .1 11.3 51.15 51.15 4 204.6 10.9 51.15 4 204.6 11 .0 11.4 51.15 4 204.6 11.9 51.15 4 204.6 1 1 .1 11.3 51.15 4 204.6 11 .0 51.15 4 204.6 1 1 .1 1 1 .2 5.115 0.1 0.5115 0.0282 5.115 0.1 0.5115 0.0289 0.0293 5.115 0.1 0.5115 0.0377 "7CODDJ Contains No CBI NJ 00 a ft 2c Ia 4^ ON Report E03-0583 3M Environmental Laboratory Quail Serum. 900 ppm Dose Gronn File S am ple Name RT D0930036.D 73150-2 D0930037.D 73150-3 D0930038.D 73151-1 D0930042.D 73151-2 D0930043.D 73151-3 3.279 3.279 3.286 3.286 Area C o n c e n tra tio n o f PFBS in Extract, Oncolum n, ng /m L O n-Colum n D ilution F a c to r 149152 143607 67.94 52.56 50.55 0.1 0.1 0.1 46899 49668 15.57 16.57 0.1 0.1 D0930045.D 73152-2 D0930046.D 73152-3 3.28 3.279 23418 52954 7.077 17.76 0.1 0.1 D0930050.D 73153-2 DO930051 .D 73153-3 00930052.D 73154-1 D0930053.D 73154-2 D0930057.D 73154-3 D0930058.D 73155-1 D0930059.D 73155-2 D0930060.D 73155-3 3.286 3.287 3.286 3.286 3.286 3.286 106527 98675 102515 69858 45587 39806 37.13 34.29 35.68 23.87 15.09 13.00 0.1 0.1 0.1 0.1 0.1 0.1 DO930064.D 73156-2 D0930065.D 73156-3 30930066.D 73157-1 D0930067.D 73157-2 73157-3 3.286 3.287 46614 39707 15.46 12.97 3.293 8981 2.558 Not enouqh samle fo r triolicates 0.1 0.1 0.1 C o n ce n tra tio n o f PFBS in Extract, ng/mL 6.794 5.256 5.055 3.900 1.557 1.657 1.158 0.7077 1.776 5.583 3.713 3.429 3.568 2.387 1.509 1.300 0.5115 1.546 1.297 0.2558 D0930073.D 73158-2 D0930074.D 73158-3 30930075.D 73159-1 D0930076.D 73159-2 73159-3 D0930080.D 73160-1 D0930081.D 73160-2 73160-3 D0930083.D 73161-1 73161-2 73161-3 30930089.D 73162-1 D0930090.D 73162-2 D0930091.D 73162-3 3.286 3.287 40659 43071 13.31 14.18 3.286 26640 8.242 Not enouah sample for triplicates 3.286 3.286 24167 7.347 Not enouqh sample for triplicates Not enough sample for triplicates Not enouah sample fo r triplicates 3.286 3.286 32824 39321 10.48 12.83 0.1 0.1 0.1 0.1 0.1 0.1 0.1 1.743 1.331 1.418 0.8242 0.7347 0.9092 1.048 1.283 W eight of serum (in hom ogenate), g 0.1181 0.1181 0.1181 0.2370 0.2370 0.2370 0.2249 0.2249 0.2249 0.3524 0.3524 0.3524 0.2821 0.2821 0.2821 0.2769 0.2769 0.2769 0.2280 0.2280 0.2280 0.0533 0.0533 W eight o f ACN (in hom ogenate), g 2.464 2.464 2.464 5.101 5.101 5.101 4.650 4.650 4.650 7.085 7.085 7.085 5.659 5.659 5.659 5.620 5.620 5 .6 2 5.732 5.732 5.732 1 .1 1 S 1.119 W eight of Aliquot, g 0.7775 0.7771 0.7844 0 .7 8 4 2 0 .7 5 1 0 0.7807 0.7811 0.7547 0.7631 0 .7 9 2 4 0.7831 0.7527 0.7768 0.7796 0.7618 0.7568 0.7659 0.7456 0.7776 0.7783 0.7702 0.7804 0.3564 C o n ce n tra tio n o f PFBS in O rig in a l Serum Sample, uq/q 0.382 0.296 0.282 0.224 0.0934 0.0643 0.0407 0 .1 0 1 0.297 0 .2 0 0 0.192 0.184 0.193 0.132 0.0917 0.0839 0.0344 0.104 0.0880 0.0511 0.0316 0.2664 0.2664 0.2664 0.0583 0.0583 5.447 5.447 5.447 1.125 1.125 0.7535 0.7773 0.7784 0.7533 0.3949 0.0992 0.0735 0.0782 0.125 0.0847 0.0492 0.0492 1.084 1.084 0.7708 0.3305 0.132 0 .1 0 2 0.857 0.8669 0.147 0.1385 0.1385 0.1385 2.759 2.759 2.759 0.7779 0.7741 0.7099 0.0489 0.0566 0.0756 Average, 0.320 0.138 0.0686 0.230 0.170 0.0833 0.0754 0.0413 0.0836 0.105 0.117 0.147 0.0604 'U sed 112 LLOQ fo r one of the replicate calculations LL O Q On- C o lu m n , C o rre c te d LLOQ on D ilution fo r Column, Factor On D ilution LLO Q in M atrix, ug/g 5.115 0.1 0 .5 1 1 5 0.0288 5.115 0.1 0.5115 0.0288 U.6 I ID 5.115 0.1 0.5115 0.0294 5.115 0.1 0.5115 0.0307 0.0295 5.115 0.1 0.5115 0.0284 5.115 0.1 0.5115 0.0294 0.0291 5.115 0.1 0.5115 0.0272 5.115 0.1 0 .5 1 1 5 0.0276 Average, ug/g 0.0287 0.0299 0.0289 0.0278 5.115 0.1 0.5115 0.0277 5.115 0.1 0 .5 1 1 5 0.0276 0.0279 0.0283 5.115 0.1 0.5115 0.0288 5.115 0.1 0 .5 1 1 5 0 .0 2 8 4 0.0288 0.0292 5.115 0.1 0,5115 0.0344 5.115 0.1 0 .5 1 1 5 0 .0 3 4 4 0.0345 5.115 0.1 0.5115 0.0288 5.115 0.1 0,5115 0.0631 0.0460 5.115 0.1 0 .5 1 1 5 0.0291 5.115 0.1 0 .5 1 1 5 0.0282 0.0285 5.115 0.1 0.5115 0.0276 5.115 0.1 0 .5 1 1 5 0.0526 0.0401 5.115 0.1 0.5115 0.0306 5.115 0.1 0 .5 1 1 5 0.0713 0.0509 5.115 0.1 0.5115 0.0271 5.115 0.1 0.5115 0.0275 5.115 0.1 0.5115 0.0276 0.0284 5.115 0.1 0.5115 0.0301 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 70 of 227 Page 3 of 3 Contains No CBI O-* L/i i ON Report E03-0583 3M Environmental Laboratory Quail Liver Matrix Spikes File Sample Name D1018073.D D1018074.D D1018078.D D1022099.D D1022100.D D1022101.D LS-041005-1 LS-041005-2 LS-041005-3 LS-041019-1 LS-041019-2 LS-041019-3 D1018079.D D1018080.D D1018081.D D1021099.D D1021100.D D1021101.D MS-041005-1 MS-041005-2 MS-041005-3 MS-041019-1 MS-041019-2 MS-041019-3 D1103026.D D1103027.D D1103028.D D1021102.D D1021103.D D1021107.D HS-041005-1 HS-041005-2 HS-041005-3 HS-041019-1 HS-041019-2 HS-041019-3 D1108032.D D1108033.D D1108034.D D1108035.D D1108038.D D1108040.D D1108041.D D1108042.D D1108043.D D1108036.D D1108047.D D1108048.D EC-040927-10 EC-040927-25 EC-040927-50 EC-040927-75 EC-040927-250 EC-040930-10 EC-040930-25 EC-040930-50 EC-040930-75 EC-040927-100 EC-040930-100 EC-040930-250 300 ppm Dose Group; Low Spike; Liver; 05 Oct 04; MLACMC/MEE* 300 ppm Dose Group; Low Spike; Liver; 05 Oct 04; MLACMC/MEE+ 300 ppm Dose Group; Low Spike; Liver; 05 Oct 04; MLACMC/MEE, Low Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLACMC/MEEG Low Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLACMC/MEEH Low Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLACMC/MEEl 300 ppm Dose Group; Med Spike Liver; 05 Oct 04; MLACMC/MEE300 ppm Dose Group; Med Spike, Liver, 05 Oct 04; MLACMC/MEE. 300 ppm Dose Group; Med Spike; Liver; 05 Oct 04; MLACMC/MEE/ Med Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLACMC/MEEJ Med Spike; 900 ppm Dose Group; Liver, 19 Oct 04; MLACMC/MEEK Med Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLACMC/MEEL High Spike; 300ppm DG Liver; MLA/CMC/MEE; 05 Oct 04 High Spike; 300ppm DG Liver, MLA/CMC/MEE; 05 Oct 04 High Spike; 300ppm DG Liver MLA/CMC/MEE; 05 Oct 04 High Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE High Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE High Spike; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Extracted Curve; 10ppb; Liver; 27 Sep 04 Extracted Curve; 25ppb; Liver; 27 Sep 04 Extracted Curve; 50ppb; Liver; 27 Sep 04 Extracted Curve; 75ppb; Liver; 27 Sep 04 Extracted Curve; 250ppb; Liver; 27 Sep 04 Extracted Curve; 10ppb; Liver; 30 Sep 04 Extracted Curve; 25ppb; Liver; 30 Sep 04 Extracted Curve; 50ppb; Liver; 30 Sep 04 Extracted Curve; 75ppb; Liver; 30 Sep 04 Extracted Curve; 100ppb; Liver; 27 Sep 04 Extracted Curve; 100ppb; Liver; 30 Sep 04 Extracted Curve; 250ppb; Liver; 30 Sep 04 3.454 3.454 3.453 3.46 3.461 3.461 3.454 3.454 3.454 3.468 3.461 3.461 3.573 3.58 3.58 3.461 3.461 3.461 3.44 3.441 3.44 3.44 3.44 3.44 3.44 3.44 3.44 3.44 3.441 3.44 Area Concentration of PFBS in Extract, Oncolumn, ng/mL Recovery, % Average Recovery, % RSD, % 247534 232831 232994 69996 67506 69231 1819530 1827919 1805066 1378946 1402599 1290097 24.5 22.5 22.5 23.3 22.4 23.1 262 263 259 232 237 215 9886%% 88% 9881%% 90% 102% 110013%% 91% 93% 84% 90.2% 3615535 3577228 3683641 3038661 3265979 3089768 185755 431713 1701180421292 3206639 175251 442373 822165 967836 1028782 1093647 2571915 720 709 738 612 675 626 11.4 28.2 48.3 69.6 268 10.7 28.9 55.8 66.4 70.9 7250.18 94% 92% 96% 8880%% 82% 111110%% 94% 91% 105% 110143%% 109% 87% 69% 74% 79% Entire sample not extracted Failed Failed T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 71 of 227 Page 1 of 2 Contains No CBI C-Dt I On Report E03-0583 _________________________________________ Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues 3M Environmental Laboratory Quail Liver Matrix Spikes File Sam ple Name D1004089. D MB-041004-1 D1004090. D MB-041004-2 D1010096.D MB-040930-1 D101Q097.D MB-040930-2 D1010098.D MB-040930-3 D1010099.D MB-040930-4 D1014096.D MB-040930-1 D1014097.D MB-040930-2 D1014098.D MB-040930-3 D1014099.D MB-040930-4 D1018087.D MB-041005-1 D1018088.D MB-041005-2 D1018089.D MB-041005-3 D1021108.D MB-041019-1 D1021109.D MB-041019-2 D1021110.D MB-041019-3 11022028.D MB-041012-2 D1022102.D MB-041019-1 D1022103.D MB-041019-2 D1022107.D MB-041019-3 11023028.D MB-041012-2 D1024098.D MB-040930-1 D1024099.D MB-040930-2 D1024100.D MB-040930-3 D1024101.D MB-040930-4 11024095. D MB-040927-1 11024096. D MB-040927-2 11024097.D MB-040927-3 11024100.D MB-040927-4 11024028.D EC-041012b-750 D1101094.D MB-040927-1 D1101095.D MB-040927-2 D1101096.D MB-040927-3 D1101099.D MB-040927-4 S am ple Info Method Blank; 4 Oct 04; MLA/CMC/MEE Method Blank; 4 Oct 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE 100 ppm Dose Group; Liver, 30 Sept 04; MLA/CMC/MEE 300 ppm Dose Group; Method Blank; Liver; 05 Oct 04; MLA/CMC/MEE 300 ppm Dose Group; Method Blank; Liver; 05 Oct 04; MLA/CMC/MEE 300 ppm Dose Group; Method Blank; Liver; 05 Oct 04; MLA/CMC/MEE Method Blank; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Method Blank; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Method Blank; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Extracted Curve; Method Blank; Liver; 12 Oct 04 Method Blank; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Method Blank; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Method Blank; 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Method Blank; Liver; 12 Oct 04 100 ppm Dose Group; Method Blank; Liver; 30 sep 04; MLA/CMC/MEE 100 ppm Dose Group; Method Blank; Liver; 30 sep 04; MLA/CMC/MEE 100 ppm Dose Group; Method Blank; Liver; 30 sep 04; MLA/CMC/MEE 100 ppm Dose Group; Method Blank; Liver; 30 sep 04; MLA/CMC/MEE Method Blank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Method Blank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Method Blank; 0 ppm Dose Group; Liven 27 Sep 04; MLA/CMC/MEE Method Blank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Method Blank; Liver; 12 Oct 04 Method Blank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Method Blank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Method Blank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Method Biank; 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE RT 0 0 0 0 0 0 0 3.468 3.467 0 0 0 0 0 0 0 3.469 0 0 0 3.478 0 0 0 0 3.468 3.469 3.468 3.468 3.468 0 0 0 0 Concentration of PFBS in Extract, On- Area column, ng/mL 0 0 0 0 0 0 0 2092 4128 0 0 0 0 0 0 0 15465 0 0 0 8860 0 0 0 0 8256 6528 4795 11240 6081 0 0 0 0 0 0 0 0 0 0 0 < LLOQ < LLOQ 0 0 0 0 0 0 0 < LLOQ 0 0 0 < LLOQ 0 0 0 0 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0 0 0 0 Recovery, % Average Recovery, % RSD, T-7485: A Reproduction Study with the Northern Bobwhite Page 72 of 227 Page 2 of 2 Contains No CBI O-t L4^/l On 3M Environmental Laboratory Quail Liver. Control Group File 11023034. 11023035. M 023036,D 11023037.D 11023040.D 11023042. 11023043. 11023048. 11023049. 11023051. 11023052. 11023055. 11023056. 11023057. 11023059.D I1023063.D 11023064. 11023065. 11023066. I1023067.D 11023070. 11023071. 11023072. 11023073. 11023074. 11023079. 11023080. 11023081. 11023082. I1 0 2 3 0 8 5 .D 11023086. 11023087. 11023088. 11023089.D 11023093. D 11023094. 11023096.D M 023097.D 11023101. D 11023102. S am ple Name 72976-1 7D2 9 7 6 - 2 7D2 9 7 6 - 3 72977-1 72977-2 72977-3 72978-1 7D2 9 7 8 - 2 7D2 9 7 8 - 3 72979-1 7D2 9 7 9 - 2 7D2 9 7 9 - 3 72980-1 7D2 9 8 0 - 2 7D2 9 8 0 - 3 7D2981-1 7D2 9 8 1 - 2 7D2 9 8 1 - 3 72982-1 72982-2 72982-3 7D2983-1 7D2 9 8 3 - 2 7D2 9 8 3 - 3 72984-1 7D2 9 8 4 - 2 7D2 9 8 4 - 3 7D2 9 8 5 - 1 7D2 9 8 5 - 2 7D2 9 8 5 - 3 72986-1 7D2 9 8 6 - 2 7D2 9 8 6 - 3 7D2987-1 7D2 9 8 7 - 2 72987-3 7D2988-1 7D2 9 8 8 - 2 7D2 9 8 8 - 3 72989-1 72989-2 7D2 9 8 9 - 3 72990-1 72990-2 72990-3 72991-1 72991-2 7D2 9 9 1 - 3 C oncentration of On- C o n c e n tra ti PFBS in Extract, C olum n on o f PFBS O n-colum n, D ilution in E xtract, RT Area ng/m L F a c to r ng/m L 3.489 3.509 3.498 23914 91983 40237 1.279 7.643 2.596 0.1 0.1279 0.1 0.7643 0 .1 0.2596 3.499 3.499 3.499 54121 44472 160075 3.948 3.008 14.31 0 .1 0.3948 0.1 0.3008 0 .1 1.431 3.498 3.499 0 59815 36722 0 4.503 1.279 1.279 0.1 0.4503 0.1 0.1279 0.1 0.1279 3.498 3.499 3.489 27965 18207 43623 1.279 1 .2 7 9 2.925 0 .1 0.1279 0.1 0.1279 0.1 0.2925 3.498 3.498 3.498 26206 33385 63566 1 .2 7 9 1.279 4.869 0.1 0.1279 0.1 0.1279 0.1 0.4869 3.498 3.498 3.498 57381 29163 24211 4.266 1 .2 7 9 1 .2 7 9 0.1 0.4266 0.1 0.1279 0.1 0.1279 S am ple not com pletely aspirated by th e R apid T race N o Data 3.498 23910 < LLOQ 0.1 < LLO Q 3.488 32588 < LLOQ 0.1 < LLO Q 3.489 58967 4.421 0.1 0.4421 3.489 20899 1 .2 7 9 0.1 0.1279 3.488 23277 1 .2 7 9 0.1 0.1279 3.488 3.488 3.489 47709 47396 30156 3.323 3.293 1.279 0 .1 0.3323 0.1 0.3293 0.1 0.1279 3.488 18453 < LLOQ 0.1 < LLO Q Sam ple not com pletely aspirated by the Rapid Trace, No Data 3.489 15975 < LLOQ 0.1 < LLO Q 3.488 3.489 3.488 23492 56274 13731 1.279 4.158 1.279 0.1 0.1279 0.1 0.4158 0.1 0.1279 3.488 3.488 3.489 37113 22931 41409 1.279 1.279 2.710 0.1 0.1279 0.1 0.1279 0.1 0.2710 3.488 3.489 3.489 3.489 3.488 3.489 3.488 3.488 3.488 21152 20907 18168 12338 13716 26259 19391 28107 18855 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0 3.489 3.488 0 29192 26216 < LLOQ < LLOQ < LLOQ 0.1 < LLOQ 0.1 < LLOQ 0.1 < LLOQ W eight of L ive r (in homogen ate), g 1.0629 1.0629 1.0629 1.1613 1.1613 1.1613 0.9087 0.9087 0.9087 0.9838 0.9838 0.9838 1.0032 1 .0 0 3 2 1.0032 1 .0 8 2 6 1 .0 8 2 6 1.0826 1 .0 4 4 8 1 .0 4 4 8 1 .0 1 8 8 1.0188 1.0188 0.9686 0.9686 0.9686 1.0108 1.0108 1.0228 1.0228 1.0228 1 .0 0 0 1 1 .0 0 0 1 1 .0 0 0 1 0.9930 0.9930 0.9930 0.9920 0.9920 0.9920 1.0194 1.0194 1.0194 1.0208 1 .0 2 0 8 1 .0 2 0 8 W eight of ACN (in hom ogen ate), g 21.1653 21.1653 21.1653 23.0000 23.0000 23.0000 17.9305 17.9305 17.9305 19.2484 19.2484 19.2484 20.0704 20.0704 20.0704 21.4246 21.4246 21.4246 19.7518 19.7518 20.0127 20.0127 20.0127 19.4546 19.4546 19.4546 19.7606 19.7606 19.8665 19.8665 19.8665 20.1062 20.1062 20.1062 19.4059 19.4059 19.4059 19.7244 19.7244 19.7244 20.3374 20.3374 20.3374 20.4674 20.4674 20.4674 W eight of A liq u o t, g 0.7678 0.7742 0.7811 0.7837 0.7869 0.7657 0.7705 0.7808 0 .7 1 3 1 0.7814 0.7923 0.7834 0.7630 0.7642 0.7629 0.7599 0.7766 0.7870 0.7222 0 .7 9 0 1 0 .7 8 3 1 0.7843 0.7790 0.7505 0,7764 0.7799 0.7709 0.7744 0.7756 0.7780 0.7494 0.7792 0.7901 0.7840 0.7855 0.7793 0.7834 0.7765 0.7939 0.7886 0.7798 0.7999 0.8022 0.7818 0.7779 0.7897 C oncentration o f PFBS in O rig in a l Liver Sam ple, uq/q 0.00697 0.0413 0.0139 0 .0 2 1 0 0.0159 0.0778 0.0242 0.00679 0.00744 0.00673 0.00664 0.0154 0.00704 0.00703 0.0268 0.0233 0.00685 < LLOQ 0.0233 0.00673 0.00678 0.0187 0.0179 < LLOQ < LLOQ 0.00674 0.0218 0.00693 0.00683 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ Average, 0.0207 0.0382 0.0128 0.00958 0.0136 0.0123 < LLOQ 0.0123 0.0145 < LLOQ 0.0118 0.00945 < LLOQ < LLOQ < LLOQ < LLOQ LLOQ on C olum n, ng/m L D ilution Factor On C olum n 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 LLOQ OnC olum n, C orrected fo r D ilu tio n F actor 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 L L O Q in M atrix, u g /g 0.0139 0.0138 0.0137 0.0136 0.0135 0.0139 0.0138 0.0136 0.0149 0.0135 0.0133 0.0134 0.0141 0.0141 0.0141 0.0140 0.0137 0.0135 A v e ra g e , ug/g 0.0138 0.0137 0.0141 0.0134 0.0141 0.0137 2.558 2.558 2.558 2.558 2.558 2.558 2.556 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.0141 0.0129 0.0135 0.0135 0.0136 0.0144 0.0139 0.0138 0.0136 0.0136 0.0135 0.0134 0.0139 0.0139 0.0137 0.0138 0.0134 0.0135 0,0134 0.0138 0.0135 0.0135 0.0137 0.0134 0.0134 0.0138 0.0138 0.0136 0.0135 0.0135 0.0140 0.0136 0.0136 0.0138 0.0134 0.0136 0.0138 70 CD O m o o en co co "O rr CD CD > CD o > 3 70 l- s cu o Se - n => n co CD c CO CL II: O 3* 3- 3" C . ZT S^ <0 d %o in ^ ws- NJ 0N0J Page 73 of 227 Project Number 454-116 Page 1 of 3 Contains No CBI Page 74 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Quail Liver. Control G ro u p Sample File Name I1023104.D 72992-2 11023108.D 72992-3 11023110.D 72993-2 11023111.D 72993-3 11022034. 7D2994-2 11022035. 7D2994-3 11022037.D 72995-2 1022040.D 72995-3 11022042. 11022043. 11022044.D 1022048.D 1022049. 1022050. 1022051, 1Q22052.D 1022055. 1022056. 1022057-D 1022058, 1022059. 1022063.D 7D2996-2 7D299672997- 72997-2 7D2 9 9 7 - 3 7D2998-1 7D2 9 9 8 - 2 72996-3 7D2999-1 7D2 9 9 9 - 2 72999- 3 T 3D0 0 0 -1 7D3 0 0 0 73000-3 11022065. 11022066. 7D3 0 0 1 - 2 7D3 0 0 1 - 3 11022070.D 11022071.D 73002-2 73002-3 11022073. 11022074. 7D3 0 0 3 - 2 7D3 0 0 3 - 3 11022079 D 11022080.D 73004-2 73004-3 H022082.D 11022085.D 73005-2 73005-3 11022087. 11022088. 7D3 0 0 6 - 2 7D3 0 0 6 - 3 11022093. 11022094. D 7D3 0 0 7 - 2 73007-3 RT 3.488 3.488 3.489 3.488 3.488 0 0 0 0 0 30 10 2 0 0 0 0 0 0 0 0 0 0 0 0 0 Concentrationof On- PFBS inExtract, Column On-column, Dilution Area ng/mL Factor 21749 21161 38480 < LLOQ < LLOQ <LLOQ 0.1 0.1 0.1 37987 28577 25349 <LLOQ <LLOQ <LLOQ 0.1 0.1 0.1 <LLOQ 0 <LLOQ 0 <LLOQ 0.1 0.1 0.1 < LLOQ 0 <LLOQ 0 <LLOQ 0.1 0.1 0.1 0 <LLOQ 0 <LLOQ 0 <LLOQ 0 LLOQ 0.1 0.1 0.1 < LLOQ LLOQ s LLOQ g LLOQ 0.1 0.1 ? LLOQ LLOQ g LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0 < LLOQ 0 < LLOQ 0.1 0.1 0.1 0 < LLOQ < LLOQ 0 < LLOQ 0.1 0.1 0.1 0 < LLOQ 0 < LLOQ 0 < LLOQ 0.1 0.1 0.1 0 < LLOQ < LLOQ 0 < LLOQ 0.1 0.1 0.1 < LLOQ 0 < LLOQ 0 < LLOQ 0.1 0.1 0.1 < LLOQ 0 < LLOQ 0 < LLOQ 0.1 0.1 0.1 0 < LLOQ 0 < LLOQ 0 < LLOQ 0.1 0.1 0.1 Concentrati on ofPFBS inExtract, Weightof Weightof Liver(in ACN (in homogen homogen Weightof Concentration ofPFBS in OriginalLiver ng/mL ate),g ate),g <LLOQ <LLOQ <LLOO 1.0502 21.4038 0.7769 1.0502 21.4038 0.7548 1.0502 21.4038 <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ <LLOQ 1.0863 1.0863 1.0863 1.0847 1.0847 1.0847 1.0193 1.0193 1.0193 21.6671 21.6671 21.6671 21.3220 21.3220 21.3220 20.2168 20.2168 0.7829 0.7430 0.7748 0.7864 0.7903 0.7871 0.7951 < LLOQ <LLOQ <LLOQ <LLOQ < LLOQ <LLOQ <LLOQ <LLOQ <LLOQ 0.9916 19.5261 0.7917 0.9916 19.5261 0.7982 0.9916 19.5261 0.7935 <LLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.9981 0.9981 1.0311 1.0311 1.0311 0.9794 0.9794 0.9794 1.0014 1.0014 1.0014 20.0549 20.0549 20.9015 20.9015 20.9015 19.8365 19.8365 19.8365 20.0331 20.0331 20.0331 0.7886 0.7797 0.7673 0.7920 0.7855 0.7735 0.7878 0.7648 0.7805 0.7805 0.7676 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ c LLOQ s LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 1.0046 1.0046 1.0046 1.0007 1.0007 1.0007 20.3044 20.3044 20.3044 20.0842 20.0842 20.0842 0.7700 0.7579 0 .7 8 5 1 0.7672 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.9778 0.9778 0.9778 19.7911 1 9 .7 9 1 1 1 9 .7 9 1 1 0.7860 0.7859 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 1.0095 1.0095 1.0095 1.0917 1.0917 1.0917 19.4813 19.4813 19.4813 21.0007 21.0007 21.0007 0.7213 0.7826 0.7774 0.7889 0.7715 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 1.0028 1.0028 1.0028 21.2599 21.2599 21.2599 0.7856 0.7849 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 1.0058 1.0058 1.0058 20.0358 20.0358 20.0358 0.7837 0 .7 2 9 1 0.7884 < LLOQ < LLOQ < LLOQ Average, <LLOQ <LLOQ <LLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ LLOQ on D ilution C olum n, F a c to r On ng/m L C olum n 2.558 2.558 2.558 0 .1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 0.1 2.558 0.1 0 .1 2.558 2.558 2.558 0 .1 0 .1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0 .1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0 .1 2.558 2.558 0 .1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 0.1 0.1 0 .1 2.558 2.558 2 .8 0.1 0.1 0.1 2.558 2.558 2.558 0 .1 0.1 0 .1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 2.558 2.558 0.1 0.1 0.1 2.558 2.558 2.558 0.1 0.1 0.1 LLOQ OnC olum n, C orrected fo r D ilu tio n Factor 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0,2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 LLO Q in M atrix, ug/g 0 .0 1 4 1 0.0145 0.0140 0.0137 0.0144 0.0138 0.0134 0.0134 0.0134 0.0135 0.0134 0.0136 0.0134 0.0133 0.0133 0.0137 0.0138 0.0141 0.0137 0.0139 0.0141 0.0138 0.0142 0.0139 0.0138 0.0140 0.0137 0.0141 0.0143 0.0140 0.0137 0 .0 1 4 1 0.0138 0.0138 0.0138 0.0140 0.0144 0.0133 0.0134 0 .0 1 3 1 0.0134 0.0131 0.0145 0.0145 0.0146 0.0137 0.0147 0.0136 0.0137 Page 2 of 3 Contains No CBI c0-*r Ui 1 Os Page 75 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Quail Liver. Control Group Sam ple File Name D 1101033.D 73163-2 D1101034.D 73163-3 I1024037.D 11024040,D 73164-2 73164-3 D 1101036.D 73164-2 D 1101039.D 73164-3 M 024042.D I1 0 2 4 Q 4 3 .D 73165-2 73165-3 D1101041.D D 1101042.D D1101043.D D1101047.D D1101048.D 73165-2 73165- 7316673166-2 73166-3 D1101049.D D1101050.D D 1 1 01051.D 73167-1 73167-2 73167-3 D 1 1 01054.D D 1 1 01055.D D 1 1 0 1 0 5 6 .D 73168-1 73168-2 73168-3 D1101057.D D1101058.D P 1 1 Q 1 0 6 2 .D 73169-1 73169-2 73169-3 D1101063.D D1101064.D D i 101065.D 73170-1 73170-2 7317Q-3 D1101066.D D1101069.D D 1 1 0 1 0 7 0 .D 73171-1 73171-2 73171-3 D 1 1 01071.D D 1 1 01072.D D 1 1 01073.D 73172-1 73172-2 73172-3 D 1 1 01077.D D 1 1 01078.D D 1 1 01079.D 73173-1 73173-2 73173-3 D11Q1080.D D 1101081.D D 1 1 01084.D 73174-1 73174-2 73174-3 D 1 1 01085.D D 1 1 01086.D D 1 1 0 1 0 8 7 .D 73175-1 73175-2 73175-3 D 1101088.D D1101092.D D1101093.D 73176-1 73176-2 73176-3 RT 3.503 3.503 3.479 3.478 3.478 3.503 3.503 3.503 3.478 3.478 3.503 3.503 33 . 5 0 3 1 3.503 3.496 0 3.503 3.503 3.503 3.503 3.503 3.51 3.503 3.503 3.51 3.503 3.51 3.51 3.51 3.51 3.51 3.51 3.51 003.51 3.517 03 .5 1 7 A re a C oncentration of PFBS in Extract, O n-colum n, ng/m L 0 3394 6909 282264 25703 59930 < LLOQ < LLOQ < LLOQ 27.09 2.558 2.558 8352 28925 45.22 1.279 7.588 18170 11236 219934 5893 1925 40.59 2.558 2.558 67.48 1 .2 7 9 1 .2 7 9 OnC olum n D ilution F a c to r 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 4470 2867 3544 2804 00_ 4017 3796 19987 3861 5051 10653 2090 03231 3820 3987 1742 1932 4645 4564 3308 10712 2432 7735 2941 02738 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 1 .2 7 9 4.823 1.279 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0,1 0.1 0.1 0,1 0.1 0.1 0.1 0.1 0.1 0.1 C oncentrati on of PFBS in E xtract, ng/m L < LLOQ < LLOQ < LLOQ 2.709 0.2558 0.2558 4.522 0.1279 0.7588 4.059 0.2558 0.2558 6.748 0.1279 0.1279 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.1279 0.4823 0.1279 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ W eight o f W eight of L iv e r (in AC N (in hom ogen hom ogen W eight of ate), g ate), a A liq u o t, q 0.9854 0.9854 0.9854 19.6739 19.6739 19.6739 0.7541 0.7912 0.6963 0.6963 0.6963 14.3016 14.3016 14.3016 0.7275 0.7693 0.6963 0.6963 0.6963 14.3016 14.3016 14.3016 0.7275 0.7693 0.7594 0.7594 0.7594 15.3754 15.3754 15.3754 0.7660 0.7910 0.7594 0.7594 0.7594 15.3754 15.3754 15.3754 0.7660 0.7910 0.7789 0.5995 0.5995 0.7069 0.7069 0.7069 0.9230 0.9230 0.9230 0.8248 0.8248 0.8248 0.9366 0.9366 0.9366 0.7040 0.7040 0.7040 0.6488 0.6488 0.6488 0.5965 0.5965 0.5965 0.5431 0.5431 0.5431 0.9380 0.9380 0.9380 0.7304 0.7304 0.7304 11.9290 11.9290 11.9290 _ 14.0154 14.0154 14.0154 18.4346 18.4346 18.4346 16.4307 16.4307 16.4307 _ 18.6926 18.6926 18.6926 14.1404 14.1404 14.1404 14.1818 14.1818 14.1818 11.8149 11.8149 11.8149 10.8355 10.8355 10.8355 17.8374 17.8374 17.8374 14.6274 14.6274 14.6274 0.7426 0.7763 0,7897 ~ 07846 0.7850 0.7912 0.7524 0.7858 0.7850 0.7837 0 .7 8 5 1 0.7849 0.7649 0.7799 0,7933 0.7842 0.7836 0.7697 0.7869 0.7856 0.7767 0.7839 0.7895 0,7862 0 .7 8 0 1 0.7626 0.7352 0.7896 0.7907 0 .7 9 3 1 0.7893 0.7837 0.7746 C oncentration o f PFBS in O riginal Liver Average, LLOQ on C olum n, < LLOQ < LLOQ < LLOQ ng/m L 2.558 2.558 0 .1 6 0 0.0143 0.0629 2.558 5.115 5.115 0.268 0.00716 0.0843 5.115 2.558 2.558 0.225 0.0137 0.0843 2.558 5.115 5.115 0.374 0.00687 0.00698 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.00683 0.0257 0.00682 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.129 < LLOQ < LLOQ < LLOQ 5.115 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 < LLOQ 2-558 2.558 __________________________ 2 .5 5 8 D ilution Factor On C olum n 0.1 0.1 LLOQ OnC olum n, C orrected fo r D ilu tio n F actor 0.2558 0.2558 0.2558 0.1 0.5115 0.1 0.5115 0.1 0.5115 0.1 0.2558 0.1 0.2558 0 .1 0.2558 0.1 0.5115 0.1 0.5 1 1 5 0 .1 0.5115 0.2558 0.2558 0.2558 0.1 0.2 5 5 8 0 .1 0.2558 0.1 0.2 5 5 8 0.1 0.2558 0.1 0.2558 0.1 0.2558 0.1 0 .2558 0.1 0.2558 0.1 0.2558 0.2558 0.1 0.2558 0.1 0.2558 0.1 0 .2558 0.1 0.2558 0.1 0.2558 0.1 0.2558 0.1 0.2 5 5 8 0.1 0.2 5 5 8 0.1 0.2558 0.1 0.2 5 5 8 0.1 0.2558 0.2558 0.1 0.2558 0.1 0.2558 0.2558 0.2558 0.2558 0.2558 00..11 0.2558 0.2558 0.1 0.2558 0.1 0 .2558 0.1__________ Q.2558 ^Used 1/2 LLO Q for one o f the replicate calculations Used 1 /2 LLO Q fo r tw o o f th e replicate calculations LLO Q in M atrix, Average, u g / g ______u g /g 0.0142 0.0136 0.0135 0.0138 0.0303 0.0286 0.0282 0.0151 0.0143 0.0141 0.0214 0.0218 0.0284 0.0275 0.0279 0.0279 0.0142 0.0137 0.0140 0.0140 0.0144 0.0138 0.0135 0.0139 0.0136 0.0136 0.0135 0.0135 0.0143 0.0137 0.0137 0.0137 0.0136 0.0136 0.0139 0.0136 0.0140 0.0137 0.0135 0.0138 0.0138 0.0138 0.0140 0.0149 0.0149 0.0151 0.0136 0.0135 0.0135 0.0135 0.0137 0.0141 0.0146 0.0130 0.0130 0.0129 0.0141 0.0129 0.0136 0.0137 0.0139 0.0137 Page 3 of 3 Contains No CBI aa L4*>.i ON Page 76 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Q uail Liver, 100 ppm D ose G ronn File D 1 0 0 9 Q 3 0 .D D1009031.D D1009033.D D1009036.D D1009038.D D1009039.D D1222088.D D1009045.D D1009047.D D1009048.D D1009052.D D1009053.D D1009055.D D1009059.D D1009061.D D1009062.D D1009066.D D1009067.D D1009069.D D1009070.D D1009075.D D10Q9076.D D1009078.D D1009081.D D1009083.D D1009084.D D1009089.D D1009090.D D1009092.D D1009093.D D1009097.D D1009098.D D1008030. D D1008031.D Sam ple Name 73008-2 73008-3 73009-2 73009-3 73010-2 73010-3 73011-2 73011-3 73012-2 73012-3 73013-2 73013-3 73014-2 73014-3 73015-2 73015-3 73016-2 73016-3 73017-2 73017-3 73018-2 73018-3 73019-2 73019-3 73020-2 73020-3 73021-2 73021-3 73022-2 73022-3 73023-2 73023-3 73024-2 73024-3 RT 3.439 3.439 3.439 3 .4 4 3.439 3.439 3.474 3 .4 4 3.439 3.44 3.439 3.439 3.446 3.446 3.446 3.439 3.439 3.446 3.446 3.447 3.446 3.446 3.446 3.446 3.446 3.446 3.447 3.446 3.447 3.447 3.446 3.446 3.447 3.447 3.447 3.446 Area C oncentration o f PFBS in Extract, On- colum n, ng/m L 666726 648877 669689 49.57 48.02 51.60 638884 604629 920409 591132 786685 44.05 47.17 44.29 74.24 43.17 50.51 2238418 837635 551350 514785 516539 625768 804963 507315 269222 250907 539758 55.68 98.66 65.52 39.93 37.03 37.17 46.06 62.28 36.44 19.01 17.76 39.01 1015412 664733 64.36 85.40 49.39 321120 715894 37.07 22.63 53.92 91778 75912 973108 1024135 594553 339699 7.260 6.252 73.13 80.25 86.50 42.05 43.45 23.95 159079 144213 11.61 10.64 190380 166462 465369 435890 471721 581257 559034 360952 259098 366929 13.68 1 2 .1 0 33.21 30.98 33.69 41.68 42.36 40.55 24.48 17.65 24.90 OnC olum n D ilution Factor 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 1 1 1 2 2 2 1 1 1 1 1 1 2 2 2 1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 1 1 1 C o n ce n tr ation of W eight of PFBS in Liver (in W eight of ACN (in Extract, hom ogen hom oge ng/m L ate), g nate), g 99.13 96.05 103.2 8 8 .1 0 94.34 88.57 148.5 86.34 1 2 1 .0 111.4 197.33 131.0 1.0762 1.0762 1.0762 1.0159 1.0159 1.0159 1.0227 1.0227 1.0227 1.0017 1.0017 1.0017 21.0955 21.0955 21.0955 20.3928 20.3928 20.3928 20.6157 20.6157 20.6157 20.2763 20.2763 20.2763 79.87 74.06 74.34 1.0054 1.0054 1.0054 20.0941 20.0941 20.0941 92.12 124.6 72.89 1.0205 1.0205 1.0205 20.5018 20.5018 20.5018 19.01 17.76 39.01 128.7 170.8 98.79 37.07 22.63 53.92 0 .9 4 4 2 0 .9 4 4 2 0 .9 4 4 2 0 .9 5 4 2 0 .9 5 4 2 0 .9 5 4 2 1.0398 1.0398 1.0398 1 9 .0 0 5 5 1 9 .0 0 5 5 1 9 .0 0 5 5 1 9 .3 9 4 2 1 9 .3 9 4 2 19.3942 20.9973 20.9973 20.9973 8 .0 1 1 7.260 6 .2 5 2 1.0549 1.0549 1.0549 21.5598 21.5598 21.5598 146.3 160.5 173.0 42.05 43.45 23.95 1.0208 1.0208 1.0208 1.0335 1.0335 1.0335 20.7196 20.7196 20.7196 20.6414 20.6414 20.6414 25.87 11.61 10.64 0 .9 9 6 2 0 .9 9 6 2 0.9962 20.3115 20.3115 20.3115 8.982 13.68 1 2 .1 0 1.0291 1.0291 1.0291 20.5171 20.5171 20.5171 33.21 30.98 33.69 83.36 84.72 81.10 0 .9 8 8 9 0.9889 0 .9 8 8 9 1.0050 1.0050 1.0050 1 9 .4 5 1 6 19.4516 1 9 .4 5 1 6 20.2467 20.2467 20.2467 24.48 17.65 24.90 0 .9 9 0 6 0 .9 9 0 6 0 .9 9 0 6 20.3074 20.3074 20.3074 W eight of A liq u o t, 9 0.7787 0.7908 0.7844 0.7383 0.7445 0.7777 0.7658 0.7734 0.7692 0.7019 0.7801 0.7853 0.7805 0.7794 0.7827 0.7610 0.7697 0.7760 0.7033 0.7759 0.7754 0.7618 0.7815 0.7537 0.7820 0.7267 0.7796 0.7730 0.7692 0.7879 0.7333 0.7719 0.7911 0.7814 0.7829 0.7854 0.7820 0.7807 0.7819 0.7725 0.7840 0.7809 0.7462 0.7681 0,7879 0.7804 0.7849 0.7701 0.7705 0.7893 0.7714 C oncentration o f PFBS in O riginal Liver S am ple, ug/g 5 .2 5 5.00 5.42 5.03 5 .3 4 4.80 8.2 0 4.72 6 .6 6 6.74 10.75 7.09 4.29 3.99 3.99 5.11 6 .8 3 3 .9 6 1.14 0.967 2 .1 3 7.21 9.32 5 .5 9 2 .0 1 1 .3 2 2.93 0.444 0.405 0.340 8 .5 0 8.8 6 9.31 2 .2 6 2 .3 3 1.28 1 .4 2 0.636 0.582 0.487 0.731 0.649 1.84 1.67 1.77 4.52 4.56 4.45 1.37 0.962 1.39 Average, ug/g 5 .2 2 5 .0 6 6.53 8.19 4.09 5.30 1.41 7.37 2.09 0.396 8.89 1.95 0.878 0.622 1.76 4.51 1.24 Page 1 of 3 LLOQ on C olum n, ng/m L D ilution F actor On C olum n LLOQ O n-Colum n, Corrected fo r 5.115 5 .1 1 5 5.115 2 2 2 10.23 10.23 5.115 5 .1 1 5 5 .1 1 5 2 2 10.23 10.23 5 .1 1 5 5 .1 1 5 5.115 2 2 2 10.23 10.23 5.115 5.115 5 .1 1 5 2 2 10.23 10.23 5.115 5.115 5.115 2 2 10.23 10.23 5.115 5.115 5 .1 1 5 2 2 10.23 10.23 5 .1 1 5 5.115 5.115 5.115 5.115 5 .1 1 5 5 .1 1 5 5.115 2 2 10.23 10.23 5.115 5 .1 1 5 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 2 2 2 10.23 10.23 5.115 5.115 5.115 1 1 1 5.115 5.115 5.115 5.115 5.115 5.115 1 1 1 5.115 5.115 5.115 5.115 5.115 1 1 1 5.115 5.115 5.115 5.115 5.115 1 1 5.115 5.115 5.115 5.115 5.115 2 2 2 10.23 10.23 5.115 5.115 5.115 1 1 1 5.115 5.115 5.115 LLO Q in M atrix, 0.541 0 .5 3 3 0.584 0.579 0 .5 6 5 0 .5 6 0 0.619 0.557 0.550 0.551 0 .5 6 7 0.561 0 .3 0 7 0 .2 7 9 0 .5 7 3 0 .5 5 8 0 .2 7 7 0.298 0.284 0,285 0.594 0.565 0.275 0.274 0.280 0.280 0.277 0.273 0.283 0.275 0.554 0.551 0.285 0.279 0.285 Average, 0 .5 3 7 0.573 0 .5 6 3 0 .5 7 7 0 .5 5 0 0.561 0 .2 8 8 0 .5 7 0 0 .2 8 5 0.282 0 .5 7 0 0 .2 7 4 0 .2 8 0 0 .2 7 5 0.276 0.556 0.283 Contains No CBI O1 L/i t ON Page 77 of 227 3M Environmental Laboratory Q uail Liver, 100 ppm Dose Group File D1008033.D D1008036.D D1008038.D D1008039.D D1008044.D D1008045.D D1008047.D D1008048.D D1008052.D D1008053.D D1008055.D D 1 0 Q 8 0 5 9 .D D1008061.D D1008062.D D1008066.D D1008067.D D1008069.D D1008070.D D1008075.D D1008076.D D1008078.D D1008081.D D1008083.D D1Q08084.D D1008089.D D1008090.D D1008092.D D1008093.D D1008097.D D1008098.D D1008100.D D1008104.D D1024032.D D1024033.D Sam ple Name 73025-2 73025-3 73026-2 73026-3 73027-2 73027-3 73028-2 73028-3 73029-2 73029-3 73030-1 73030-2 73030-3 73031-2 73031-3 73032-1 73032-2 73032-3 73033-1 73033-2 73033-3 73034-2 73034-3 73035-1 73035-2 73035-3 73036-1 73036-2 73036-3 73037-2 73037-3 73038-1 73038-2 73038-3 73039-2 73039-3 73040-2 73040-3 73177-2 73177-3 RT 3.44 3.44 3.44 3.439 3.439 3.44 3 .4 4 6 3 .4 4 6 3.446 3.44 3.439 3.439 3.446 3.446 3.446 3.446 3.446 3.446 3 .4 4 6 3 .4 4 6 3 .4 4 7 3.446 3.447 3.447 3.446 3.446 3.446 3.439 3.446 3.446 3.447 3.447 3.446 3.447 3.446 3.447 3.446 3.447 3.446 3 .4 4 6 3.447 3.517 3.482 3 .4 7 5 Area C oncentration o f PFBS in Extract, On- colum n, ng/m L 557683 568328 515009 38.73 39.55 35.51 759566 556678 842882 618719 595363 587983 663628 661454 823646 596029 515499 390664 55.28 38.66 62.89 43.49 41.65 41.07 47.12 46.94 61.08 41.70 35.54 26.54 199582 198292 208993 263937 269955 979836 801411 818649 403156 332248 543445 543007 977498 531877 598369 567198 522343 378966 526389 16.77 13.80 13.72 14.41 17.97 18.37 76.84 59.03 60.62 20.09 27.42 22.53 37.65 37.61 76.59 36.77 41.89 39.46 36.06 25.73 36.36 593633 589140 433814 474315 498093 41.90 41.51 41.16 29.59 32.51 34.25 1052120 772489 1037784 896580 876582 5767 8308 10781 83.22 85.20 56.42 83.47 68.32 66.14 < LLOQ < LLOQ < LLOQ OnC olum n D ilution F a c to r 2 2 2 2 2 2 1 1 1 1 1 1 2 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0.1 0.1 0.1 C o n ce n tr ation of PFBS in Extract, ng/m L 77.46 79.10 71.02 1 1 0 .6 77.31 125.8 43.49 41.65 41.07 47.12 46.94 61.08 83.41 71.09 53.08 16.77 13.80 13.72 14.41 17.97 18.37 76.84 59.03 60.62 20.09 27.42 22.53 37.65 37.61 76.59 73.54 83.77 78.93 36.06 25.73 36.36 41.90 41.51 41.16 29.59 32.51 34.25 83.22 85.20 56.42 83.47 68.32 66.14 OOOi QO O W eight o f Live r (in homogen ate), g 1 .0 1 7 2 1.0172 1 .0 1 7 2 1 .0 5 1 9 1.0519 1.0519 1.0279 1.0279 1.0279 1.0280 1.0280 1.0280 1.0628 1.0628 1.0628 1.0822 1.0822 1.0822 1.0512 1.0512 1.0512 1.0468 1.0468 1.0468 1.0186 1.0186 1 .0 1 8 6 0.9914 0.9914 0.9914 1.0651 1.0651 1.0651 1 .0 0 2 8 1 .0 0 2 8 1.0028 1.0478 1.0478 1 .0 4 7 8 1 .0 1 8 2 1.0182 1.0182 0.9838 0.9838 0.9838 0.9973 0.9973 0.9973 0.9321 0.9321 0.9321 W eight of ACN (in bomoge nate), g 20.4815 20.4815 20.4815 21.6740 21.6740 21.6740 20.4170 20.4170 20.4170 20.2589 20.2589 20.2589 21.3860 21.3860 21.3860 21.8000 21.8000 21.8000 2 1 .1 2 2 1 2 1 .1 2 2 1 2 1 .1 22 1 2 1 .0 2 0 2 2 1 .0 2 0 2 2 1 .0 2 0 2 20.4425 20.4425 20.4425 19.9288 19.9288 19.9288 21.6735 21.6735 21.6735 2 0 .0 3 4 1 20.0341 20.0341 2 0 .7 4 9 1 20.7491 2 0 .7 4 9 1 20.4022 20.4022 20.4022 19.7285 19.7285 1 9 .7 2 8 5 20.0126 20.0126 20.0126 1 8 .6 6 1 3 18.6613 18.6613 W eight of A liquot, 9 0 .7 8 7 6 0 .7 8 5 9 0.7761 0.7848 0.7851 0 .7 9 0 2 0 .7 6 9 0 0 .7 7 7 5 0 .7 5 2 5 0.7556 0 .7 8 7 2 0 .7 8 5 2 0 .7 7 8 5 0 .7 7 3 0 0 .7 2 6 0 0.7891 0.7410 0.7861 0.7645 0.7792 0.7816 0.7502 0.7732 0.7103 0.7386 0.7839 0.7799 0.7901 0 .7 9 2 8 0.7880 0 .7 8 6 5 0 .7 8 1 9 0 .7 8 4 3 0.7671 0 .8 0 0 2 0.7711 0 .7 8 7 7 0.7902 0 .7 9 1 7 0.7622 0.7839 0.7907 0 .7 9 5 5 0 .7 8 1 9 0.7981 0 .7 5 5 0 0 .7 8 6 5 0.7874 0.7712 0.7883 0.7859 C oncentration o f PFBS in O riginal Liver Sam ple, ug/g 4 .1 6 4 .2 5 3.87 6.09 4.25 6.8 8 2.36 2.23 2.28 2.58 2.47 3 .2 2 4 .5 3 3.88 3.09 0.899 0.788 0.738 0.795 0.973 0.991 4 .3 2 3.22 3.60 1.15 1.47 1 .2 2 2.01 2 .0 0 4 .1 0 3.99 4.57 4.30 1.97 1.35 1.98 2 .2 1 2.19 2 .1 6 1.63 1.74 1.82 4 .4 0 4 .5 9 2 .9 8 4 .6 6 3.66 3.54 < LLOQ < LLOQ < LLOQ Average, ug/g 4.09 5.74 2.29 2.76 3.83 0.808 0.920 3.71 1.28 2.71 4.29 1.77 2.19 1.73 3.99 3.95 < LLOQ Page 2 of 3 LLOQ on Colum n, ng/m L D ilution Factor On Colum n LLOQ O n-C olum n, C orrected fo r 5.115 5.115 5.115 2 2 2 10.23 10.23 10.23 5.115 5.115 5.115 2 2 2 10.23 10.23 5.115 5.115 5.115 5 .1 1 5 5 .1 1 5 5.115 5.115 5.115 5 .1 1 5 5 .1 1 5 5.115 5.115 5 .1 1 5 2 2 2 10.23 10.23 10.23 5.115 5.115 5.115 5.115 5.115 5.115 1 1 1 1 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5 .1 1 5 5.115 5.115 5.115 1 1 1 1 1 1 5.115 5.115 5.115 5.115 5.115 5.115 5.115 5.115 1 1 5.115 5.115 5.115 5.115 5.115 2 2 2 10.23 10.23 5.115 5.115 5.115 5.115 5.115 5.115 1 1 1 1 1 5.115 5 .1 1 5 5 .1 1 5 5 .1 1 5 5 .1 1 5 5.115 5.115 5.115 1 1 1 5 .1 1 5 5 .1 1 5 5.115 5.115 5.115 5.115 1 1 1 5.115 5 .1 1 5 5.115 5.115 5.115 1 1 1 5 .1 1 5 5 .1 1 5 5.115 2.558 2.588 2.588 0.1 0.1 0.1 0.2558 0.2588 0.2588 LLO Q in M atrix, Average, 0.549 0.550 0.563 0.563 0.278 0.274 0.552 0.562 0.279 0.280 0.269 0.555 0.559 0.274 0.292 0.282 0.277 0.273 0.570 0.280 0.278 0.287 0.279 0.292 0.275 0.290 0.281 0.273 0.272 0.273 0.555 0.559 0.557 0.280 0.268 . 0.275 0.270 0.269 0.269 0.282 0.275 0.276 0 .2 7 1 0.275 0.285 0.274 0.0139 0.0138 0.0138 0.272 0.278 0.0139 y<oO0 o m o CO Ul CD CO Contains No CBI boOo ON *0 ,o oo' 2 Io 4^ i ON Page 78 of 227 3M Environmental Laboratory Q uail Liver, 100 ppm Dose G roup File D1024035.D D1024038.D D1024040.D D1024041.D D1024046.D D1024047.D D1014040.D D1014044.D D1014045.D D1024049.D D1024050.D D1024054.D D1024055.D D1024057.D D1024061.D D1024063.D D1024064.D D1024068.D D1024069.D D1024071.D D1Q24072.D D1024077.D D1024078.D D1024080.D D1024083.D D1024085.D D1024086.D D1024091.D D1024092.D D1014092.D D1014093.D D1024093.D D1024094.D D1024095.D Sam ple Name 73178-2 73178-3 73179-2 73179-3 RT 3.475 3.475 3.475 3.475 3.475 3.475 73180-2 73180-3 73180-1 73180-2 73180-3 3.482 3.475 3.454 3.453 3.461 73181-2 73181-3 3.482 3 .4 8 2 73182-2 73182-3 3.475 3.475 73183-2 73183-3 3.474 3 .4 8 2 73184-2 73184-3 3 .4 8 2 3 .4 8 2 73185-2 73185-3 3 .4 8 2 3 .4 8 2 73186-2 73186-3 73187-2 73187-3 3.482 3 .4 8 2 3 .4 8 2 3 .4 8 2 3.482 73188-2 73188-3 3.482 3 .4 8 2 73189-2 73189-3 3.482 3.482 73190-2 73190-3 3.482 3.482 73191-2 73191-3 73191-1 73191-2 73191-3 3.468 3.468 3.482 3.482 3 .4 8 2 ' Area C oncentration o f PFBS in Extract, On- colum n, ng/m L 8342 6807 9938 < LLOQ < LLOQ < LLOQ 10282 9339 6580 4703 34438 8349 6347 43286 8576 7146 5317 6271 6541 6540 7886 6967 < LLOQ < LLOQ < LLOQ 1.279 10.81 1.279 1.279 10.47 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 10479 10766 6443 4966 5117 4727 5588 4318 2208 3072 4563 3188 7690 7538 7249 8947 21206 24835 20460 16894 19570 1.279 1.279 < LLOQ < LLOQ <LLOQ < LLOQ <LLOQ < LLOQ < LLOQ < LLOQ <LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 6.473 5.890 6.636 5.742 4 .4 5 0 5.420 OnC olum n D ilution F a c to r 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 C oncentr ation of PFBS in Extract, ng/m L W eight of Live r (in homogen ate), g W eight o f ACN (in homoge nate), g < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0 .1 2 7 9 0 .1 2 7 9 1.081 0 .1 2 7 9 0 .1 2 7 9 1.047 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0 .2 9 6 7 0 .1 2 7 9 0 .1 2 7 9 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0 .6 4 7 3 0 .5 8 9 0 0 .6 6 3 6 0 .5 7 4 2 0 .4 4 5 0 0 .5 4 2 0 0 .8 1 0 7 0 .8 1 0 7 0 .8 1 0 7 0 .5 8 0 4 0.5804 0.5804 0 .8 9 1 8 0.8918 0.8918 0 .8 9 1 8 0.8918 0 .8 9 1 8 0 .6 5 5 6 0 .6 5 5 6 0 .6 5 5 6 0.7327 0 .7 3 2 7 0.7327 0 .6 2 9 2 0 .6 2 9 2 0.6292 0.6671 0.6671 0.6671 0 .9 4 3 7 0 .9 4 3 7 0 .9 4 3 7 0 .8 1 7 8 0.8178 0 .8 1 7 8 0 .6 8 2 6 0 .6 8 2 6 0 .6 8 2 6 0 .8 1 0 6 0.8106 0.8106 0.7743 0 .7 7 4 3 0 .7 7 4 3 0.8306 0.8306 0.8306 1.1828 1.1828 1.1828 1.1828 1.1828 1.1828 1 6 .2 8 3 7 16.2837 1 6 .2 8 3 7 1 1 .6 5 8 7 1 1 .6 5 8 7 11.6587 17.7711 17.7711 17.7711 17.7711 17.7711 17.7711 14.4844 14.4844 1 4 .4 8 4 4 14.5823 1 4 .5 8 2 3 1 4 .5 8 2 3 12.5301 12.5301 12.5301 1 3 .3 4 9 5 13.3495 1 3 .3 4 9 5 18.8321 18.8321 18.8321 16.3612 1 6 .3 6 1 2 1 6 .3 6 1 2 1 3 .7 1 2 8 1 3 .7 1 2 8 1 3 .7 1 2 8 16.3642 16.3642 16.3642 1 5 .4 9 1 4 1 5 .4 9 1 4 1 5 .4 9 1 4 1 6 .6 5 2 6 1 6 .6 5 2 6 16.6526 22.9655 22.9655 22.9655 22.9655 22.9655 22.9655 W eight of A liquot, g 0.7852 0.791 0.7757 0.7713 0.7831 0.7947 0.7938 0.7845 0.7905 0.7938 0.7845 0.7905 0.7831 0.7921 0.7900 0.7817 0.7906 0.7506 0.7904 0.7804 0.7935 0.7841 0.7818 0.7942 0.7823 0.7881 0.7507 0.793 0.7807 0.7822 0.7409 0.7724 0.7753 0.7889 0.7917 0.7468 0.7193 0.7152 0.7749 0.7786 0.7913 0.7776 0.7785 0 .7 8 3 3 0 .7 8 5 5 0.7785 0 .7 8 3 3 0.7855 ' C oncentration o f PFBS in O riginal Liver S am ple, ug/g < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.00674 0.00682 0.0572 0.00674 0.00682 0.0555 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0159 0.00687 0.00677 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.0340 0.0307 0.0345 0.0301 0.0232 0.0282 Average, ug/g < LLOQ < LLOQ 0 .0 2 3 3 < LLOQ < LLOQ < LLOQ 0.00985 < LLOQ < LLOQ LLOQ on D ilu tio n LLO Q O n-C olum n, LLO Q in C olum n, F a cto r On Corrected for M atrix, 2.588 2.588 2.588 2.588 2.588 2.588 2.588 2 .5 8 8 2 .5 8 8 2 .5 8 8 2.588 2.588 2.588 2 .5 8 8 2 .5 8 8 2.588 2 .5 8 8 2.588 2.588 2 .5 8 8 2.588 2.588 2.588 2.588 2 .5 8 8 2.588 2.588 2.588 2 .5 8 8 0.1 0.1 0.1 0.1 0.1 0-1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0-1 0-1 0.1 0.1 0.1 0-1 0.1 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0.2588 0 .2 5 8 8 0.2588 0 .2 5 8 8 0.2588 0 .2 5 8 8 0.2588 0.2588 0 .2 5 8 8 0.0139 0.0138 0.0142 0.0139 0.0136 0.0138 0.0137 0.0136 0.0138 0.0153 0.0151 0.0138 0.0137 0.0137 0.0139 0.0139 0.0139 0.0139 0.0138 0.0137 0.0139 < LLOQ 2.588 2.588 2 .5 8 8 0.1 0.1 0.2588 0 .2 5 8 8 < LLOQ 2.588 2.588 2.588 0.1 0.1 0 .2 5 8 8 0.2588 < LLOQ 2.588 2.588 2 .5 8 8 0.1 0.1 0 .2 5 8 8 0 .2 5 8 8 < LLOQ 0.0301 2.588 2 .5 8 8 2.588 2.588 2 .5 8 8 2.588 2.588 2.588 2.588 0-1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.2588 0 .2 5 8 8 0.2588 0 .2 5 8 8 0 .2 5 8 8 0.2588 0.2588 0.2588 Used 1/2 L L O Q fo r one o f the re p lica te calculations Used 1/2 L L O Q fo r tw o o f th e re p lica te calculations 0.0147 0.0141 0.0139 0.0139 0.0151 0.0152 0.0140 0.0138 0.0136 0.0135 0.0135 0.0136 0.0135 A verage, 0.0139 0.0139 0.0137 0.0137 0.0152 0.0140 0.0137 0.0138 0.0140 0.0138 0.0143 0.0141 0.0148 0.0139 0,0135 0.0135 73 CD -o O m o CO o cn CO CO "O zr 03 <0 > 03 o' ^ |3 CD 3^ 03 O =r. q . Oc 3q 2,o "0 3 " T1 C73 CD c __ < II if o5 & CD - 3 8 - '? 1 CD CD V> Page 3 of 3 Contains No CBI Koo> 02 . ft 2c 1 O L/i Os Q uail Liver, 300 ppm Dose G roiin 3M Environmental Laboratory File D1015033.D D1015034.D D1015036.D D 1 0 1 5 0 3 9 .D D 1 0 1 5 0 4 1 .D D 1 0 1 5 0 4 2 .D D1015047.D D1015048.D D1015050.D D1015051.D D1015055.D D1015056.D D1015058.D D1015062.D D1015064.D D1015065.D D1015069.D D1015070.D D1018069.D D1018070.D D1101101.D D1101102.D D1101103.D D1015077.D D 1 0 1 5 0 7 8 .D D 1 0 1 5 0 7 9 .D D 1 0 1 5 0 8 0 .D D 1 0 1 5 0 8 1 .D D1015084.D D1015085.D D1015086.D D1015087.D D1015088.D D1015092.D D1015093.D Sample Name 73041-2 73041-3 73042-2 73042-3 73043-2 73043-3 73044-2 73044-3 73045-2 73045-3 73046-2 73046-3 73047-2 73047-3 73048-2 73048-3 73049-2 73049-3 73050-2 73050-3 73050-1 73050-2 730507305173051-2 730517305273052-2 73052-3 73053-1 73053-2 73053-3 73054-1 73054-2 73054-3 3 1 3 1 RT 3.446 3.446 3.447 3.446 3.446 3.446 3.446 3.447 3.446 3.446 3 .4 4 6 3 .4 4 6 3.446 3 .4 5 3 3 .4 5 4 3 .4 5 3 3.454 3.453 3 .4 5 4 3 .4 5 4 3.461 3.461 3.503 3.503 3.503 3.454 3.453 3.454 3.454 3.454 3.454 3.453 3.453 3 .4 5 3 3 .4 5 3 3.454 Area 1353885 1305893 1332931 811247 799364 1014721 856895 820375 1495479 1643668 1452869 1940492 1948718 1973144 1226618 1178086 1117033 1566478 1587265 1758639 1002214 1040659 1199895 1128369 1132582 481632 424099 198713 171951 166073 2695473 2628884 1848947 1929357 1796924 563559 564224 504801 1159675 1517681 1325612 C o n ce n tra tio n of PFBS in Extract, On- column, ng/m L 183.3 176.1 180.1 100.3 102.7 1 0 1 .0 132.3 109.3 104.1 205.3 228.8 198.6 277.5 278.9 283.1 163.9 156.6 147.4 216.5 219.8 247.4 125.3 130.4 136.1 159.8 149.1 149.7 56.83 48.81 60.75 52.29 50.44 413.7 400.9 262.3 275.7 253.7 67.75 67.84 59.60 153.8 208.8 178.9 OnC o lu m n D ilu tio n Factor C oncentratio W eight o f W eight of n o f PFBS in L ive r (in ACN (in Extract, ng/m L homogen homogen ate), g W eight of C o n c e n tra tio n o f PFBS in O riginai Liver 183.3 176.1 180.1 100.3 102.7 1 0 1 .0 1.0668 1.0668 1.0668 0.9979 0.9979 0.9979 21.4399 21.4399 21.4399 19.9432 19.9432 0.7873 0.7882 0.7448 0.7876 9.82 9.43 5.65 5.48 132.3 109.3 104.1 1.0213 1.0213 1.0213 20.4160 20.4160 0.7862 0.7962 7.06 5.76 205.3 228.8 198.6 277.5 278.9 283.1 1 .0 0 2 2 1 .0 0 2 2 1 .0 0 2 2 1.0013 1.0013 1.0013 20.0134 20.0134 20.1890 20.1890 0.7923 0.7881 0.7568 0.7820 10.9 12 .2 15.5 15.1 163.9 156.6 147.4 ' 0.9966" 0.9966 0.9966 20.0956 20.0956 0.7788 0.7880 8.91 8.41 216.5 219.8 247.4 1.0562 1.0562 1.0562 21.3913 21.3913 0.7730 0.7857 11.9 11.9 1 125.3 0.9745 19.5102 0.7928 6.64 1 130.4 0.9745 19.5102 0.7814 7.02 1 136.1 0 .9 7 4 5 1 159.8 1.0247 20.5287 0.7679 8.76 1 149.1 1.0247 20.5287 0.7737 8.11 1 149.7 1.0247 0.5 28.42 1.0097 20.0977 0.7721 0.5 24.41 1.0097 20.0977 0.7730 0.5 20.0977 0.7873 0.5 30.38 1.0097 20.0977 0.7721 0.5 26.15 1.0097 20.0977 0.7730 0.5 25.22 1.0097 20.0977 0.7873 423.5 413.7 400.9 0.9845 0.9845 0.9845 19.8891 19.8891 19,6891 0.7858 0.7955 0.7744 262.3 275.7 253.7 0.9980 0.9980 0.9980 19.9726 19.9726 19.9726 0.7866 0.7769 0.7871 67.75 67.84 59.60 1.0696 1.0696 1.0696 21.5577 21.5577 21.5577 0.7724 0.7793 0.7593 153.8 208.8 178.9 1.0792 1.0792 1.0792 21.5161 21.5161 21.5161 0.7791 0.7778 0.7890 1.54 1.32 1.24 1.64 1.41 1.34 22.9 22.1 22.0 14.0 14.9 13.5 3.71 3.68 3.32 8.27 11.2 9.50 Average, 9.63 5.51 6.18 11.3 15.4 8.42 12.4 6.99 8.47 1.42 LLOQ on C olum n, ng /m L 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 2.558 2.558 2.558 2.558 2.558 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 D ilution Factor On C o lu m n 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0.5 0.5 0.5 0.5 0.5 1 1 1 1 1 1 1 1 1 1 1 LLOQ O n-Colum n, Corrected fo r D ilution Factor 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 1.279 1.279 1.279 1.279 1.279 1.279 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 M a trix, ug/g 1.35 1.34 1.34 1.41 1.34 1.34 1.34 1.32 1.38 1.34 1 .3 6 1.36 1.37 1.36 1 .3 5 1 .3 5 1.36 1.36 1.35 1.35 1.37 1.43 0.0693 0.0692 0.0679 0.0692 0.0679 1.34 1.37 1.36 1 .3 4 1.38 1.36 1.40 1.35 1 .3 5 1.33 1.34 1.36 1.35 1.34 1.38 1.36 1.37 1.34 1.39 0.0688 0 .0 6 8 8 1.36 1.35 1.38 1.34 >0 K1 153 '4^^5 CD O 3 o!o= ac. CD <T> 3? wd <f CO 3^ Page 1 of 3 Contains No CBI NooJ 00 n> a c cot* L/i Os Page 79 of 227 Quail Liver, 300 ppm Dose Group 3M Environmental Laboratory File D1015095.D 01015096.0 D1015100.D D1015101.D D1015102.D D1015103.0 D1015107.D D1015108.D D1015109.D D1015110.D D1015111.D D1015114.D D1015115.D D1015116.D D1015117.D D1015118.D D1015122.D D1015123.D D1015124.D D1015125.D D1015126.D D1015129.D D1015130.D D1015131.D D1015132.D D1018032.D D1018O33.D D1018034.D D1018036.D D1018039.D D1018041.D D1018042.D D1018043.D D1018047.D D1018048.D D1018049.D D1018050.D D1018051.D D1018054.D D1018055.D D1018056.D D1018057.D D1018058.D D1018062.D D1018063.D D1018064.D D1018065.D Sam ple Name 73055-2 73055-3 73056-2 73056-3 73057-2 73057-3 73058-2 73058-3 73059-2 730597306073060-2 730607306173061-2 73061-3 73062-2 730627306373063-2 73063-3 73064-2 73064-3 73065-2 73065-3 3 1 3 1 3 1 73066-2 73066- 7306773067-2 73067-3 3 1 73068-2 73068-3 73069-2 73069-3 73070-2 73070-3 73071-2 73071-3 C o n ce n tra tio n On- Concentratio W eight of W eight o f o f PFBS in Colum n n o f PFBS in Liver (in ACN (in Extract, On- D ilution Extract, homogen homogen RT Area colum n, ng/m L Factor ng/m L ate), g ate), q 3.454 3.454 2624231 2873243 3.453 3.453 1042448 1031898 3.46 3.453 1638790 1709593 3.454 3.454 1565730 1565862 3.453 3.454 1615650 1627958 3.454 3.46 624954 606389 438.5 400.0 449.0 136.4 134.8 228.0 239.4 216.4 216.4 224.3 226.3 76.33 73.73 438.5 400.0 449.0 134.7 136.4 134.8 233.0 228.0 239.4 225.6 216.4 216.4 235.3 224.3 226.3 59.50 76.33 73.73 1.0659 1.0659 1.0659 1.0620 1.0620 1.0620 1.0783 1.0783 1.0783 1.0033 1.0033 1.0033 1.0959 1.0959 1.0959 0.9996 0.9996 0.9996 21.3918 21.3918 21.3918 21.2292 21.2292 21.2292 21.8042 21.8042 21.8042 20.1370 20.1370 20.1370 21.8765 21.8765 21.8765 20.6673 20.6673 20.6673 3.454 3.453 1164835 1205992 3.454 3.453 1180597 1185068 154.6 160.8 156.9 157.6 154.6 160.8 146.8 156.9 157.6 0.9628 0.9628 0.9628 1.0214 1.0214 1.0214 19.3156 19.3156 19.3156 20.4711 20.4711 20.4711 3.454 3.454 902039 872126 3.447 3.454 593808 570710 3.447 2180537 3.454 2190632 3.454 3.454 1033257 1005938 115.6 111.5 72.63 69.36 334.1 325.0 326.9 137.3 136.7 132.6 115.8 111.5 0.9248 0.9248 0.9248 18.4382 18.4382 18.4382 65.12 1.0551 21.1857 1 72.63 1.0551 21.1857 1 69.36 1.0551 21.1857 1 334.1 0.9927 19.9459 1 325.0 0.9927 19.9459 1 326.9 0.9927 19.9459 1 137.3 0.9507 19.5267 1 136.7 0.9507 19.5267 1 132.6 0.9507 19.5267 3.454 3.454 935078 966050 3.454 3.454 1031002 1012157 1 2 2 .1 126.7 136.4 133.6 1.0807 21.6405 1 1 2 2 .1 1.0807 21.6405 1 126.7 1.0807 21.6405 133.3 0.9895 19.8414 1 136.4 0.9895 19.8414 1 133.6 0.9895 19.8414 3.454 1071140 142.4 1 142'4 Sample not completely aspirated bv the Rapid Trace No nata 1 .0 1 1 2 1 .0 1 1 2 20.3534 20.3534 W eight of Aliquot, q 0.7853 0.7635 0.7558 0.7763 0.7558 0.7475 0.7591 0.7944 0.7951 0.7971 0.7720 0.7949 0.7459 0.7560 0.7566 0.8017 0.7892 0.7993 0.8029 0.7868 0 .7 8 6 9 0 .7 7 9 2 0 .7 8 8 9 0 .8 0 5 3 0 .7 7 9 7 0 .7 9 5 2 0 .7 8 7 0 0 .7 4 7 6 0 .7 8 8 4 0 .7 7 7 7 0 .7 7 7 0 0 .7 8 7 9 0 .7 8 8 8 0 .7 5 3 8 0.7888 0 .7 8 7 7 0 .7 5 2 8 0 .7 6 8 2 0 .7 6 1 5 0 .7 4 4 2 0 .7 8 5 6 0.7952 0.7819 0.7694 C o n c e n tra tio n o f PFBS in O riginal Liver Sam ple, uq/q 23.5 22 .1 25.0 7 .2 9 7.57 7.57 13.0 12 .2 12 .8 11.9 11 .8 11.5 13.2 12.4 12.5 3.22 4 .1 9 4 .0 0 8.74 8.28 8.61 7.93 8.37 8.24 5.79 6 .1 0 5 .9 3 3 .6 7 3 .8 8 3 .7 6 18.1 17.4 7 .8 5 7 .4 7 7 .2 5 7 .0 8 6.6 8 6 .9 9 7,54 7.31 7 .0 7 7.87 7 .8 2 Average, 23.5 7 48 12.7 11.7 12.7 3.80 8.54 8.18 5.94 3.77 17.7 7.52 6.92 7.31 7.85 3.454 3.454 1321833 1285166 3.454 3.454 668110 637801 180.9 175.2 83.22 78.89 1 166.0 1 .0 1 0 0 20.2035 0.7904 8 .8 2 1 180.9 1 .0 1 0 0 20.2035 0.7928 9.59 1 175.2 1 .0 1 0 0 20.2035 0.7442 9.89 1 78.23 1.0482 20.8266 0.7916 4 .1 2 1 83.22 1.0482 20.8266 0.7878 4.41 1 78.89 1.0482 20.8266 0.7443 4 .4 2 9.43 4.32 LLOQ on D ilution LLOQ On-Coiumn, C olum n, Factor On Corrected fo r 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 LLOQ in M a trix , ug/g 1.35 1.39 1.40 1.36 1.39 1.41 1.40 1.34 1.34 1.33 1.37 1.33 1.41 1.39 Average, ug/g 1.38 1.39 1.36 1.34 1.40 25.11 25.11 25.11 25.11 25.11 25.11 25.11 1 1 1 1 25.11 25.11 25.11 25.11 25.11 25.11 1.36 1.38 1.36 1.32 1.34 1.34 1.36 1.34 1.37 1.34 1.34 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 1 1 1 1 1 1 1 1 1 1 1 1 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 25.11 1.35 1.32 1.34 1.42 1.34 1.36 1.36 1.34 1.34 1.43 1.37 1.40 1.37 1.39 1.42 1.35 1.34 1.37 1.35 1 .3 9 1.39 1.37 25.11 25.11 1 1 25.11 25.11 1.36 1.38 1.37 25.11 25.11 25.11 25.11 25.11 1 1 1 1 1 25.11 25.11 25.11 25.11 25.11 1.33 1.33 1.42 1.32 1.33 1.41 1 .3 6 1.35 Page 2 of 3 Contains No CBI 00 VO o a ? cor Ui i Os Page 80 of 227 Page 81 of 227 3M Environmental Laboratory Report E03-0583 Q uail L iver, 300 ppm Dose Group File D1018131.D D1018132.D D1018133.D D1018134.D D1018135.D D1018138.D D1018139.D D1018140.D D1018141.D D1018142.D D1018146.D D1018147.D D1018148.D D1018149.D D1018150.D D 0 1 8 1 5 3 .D D1018154.D D1Q181S5.D D1018156.D D1018157.D D1018161.D D1018162.D D1018163.D D1018164.D D1018165.D D1018168.D D1018169.D D1018170.D D1018171.D D1018172.D D1018176.D D1018177.D D1018178.D D1018179.D D1018180.D D1018183.D 018184.D D1018185.D D1018186.D D1018187.D D1018191.D D1018192-D D1018193.D D1018194.D D1018195,D 73192-1 73192-2 73192-3 73193-1 73193-2 731937319473194-2 73194-3 73195-1 73195-2 73195-3 73196-1 73196-2 73196-3 73197-1 73197-2 73197-3 73198-1 73198-2 73198-3 73199-1 73199-2 73199-3 73200-1 73200-2 73200- 3 ~ 73201-1 7320173201-3 73202-1 73202-2 73202-3 73203-1 73203-2 73203- 3 ~ 73204-1 73204- 73205-1 73205-2 73205-3 73206-2 73206-3 3 1 2 2 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 _ 3~461 3.461 3.461 3.461 3.461 3.461 3.461 3.468 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 C o n ce n tra tio n of PFBSin Extract, On- column, ng/m L 34195 24105 24763 18754 19967 27176 37038 20627 23012 29916 20645 15161 8.594 5.627 5.820 4.055 4.411 6.529 9.431 4.605 5.305 7.335 4.610 7206 6679 15863 13684 13841 7601 8561 6698 14443 __ 7250 11948 11672 14950 9173 9166 11076 11683 8169 9548 " ~99593 106181 106471 22235 23309 < LLOQ < LLOQ < LLOQ 3.206 2.566 2.613 < LLOQ < LLOQ < LLOQ 1.279 2.789 1.279 1.279 1.279 2.938 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 27.94 29.90 29.99 5.077 5.393 OnC o lu m n D ilu tio n Factor 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 C oncentratio W eight of W eight of n o f PFBS in L iv e r (in ACN (in Extract, hom ogen homogen ng/m L ate), g ate), g 0.8594 0.5627 0.5820 0.4055 0.4411 0.6529 0 .7 6 6 3 0 .7 6 6 3 0 .7 6 6 3 0 .7 5 6 0 0 .7 5 6 0 0.7560 15.7395 15.7395 15.7395 15.1584 15.1584 15.1584 0.9431 0 ,4 6 0 5 0 .5 3 0 5 0 .7 3 3 5 0 .4 6 1 0 0 .3 0 0 0 < LLOQ < LLOQ < LLOQ 0.3206 0.2566 0 .2 6 1 3 < LLOQ 0 .1 2 7 9 0 .2 7 8 9 0.1279 0 .1 2 7 9 0 .1 2 7 9 0 .2 9 3 8 < LLOQ LLOQ g LLOQ < LLOQ < LLOQ < LLOQ 2.794 0.5077 0.5393 OO DO 1 .1 8 8 3 1 .1 8 8 3 1.1883 0 .6 1 9 8 0 .6 1 9 8 0 .6 1 9 8 1.1793 1.1793 1 .1 7 9 3 0.9090 0.9090 0.9090 1.0027 1.0027 1.0027 0.8798 0.8798 0,8798 0.9467 0.9467 0.9467 1.1064 1.1064 1.1064 1.1679 1.1679 1.1679 0.7751 0.7751 0.7751 0 .9 5 3 3 0 .9 5 3 3 23.7018 23.7018 23.7018 13.34 13.34 2 3 .5 9 1 9 2 3 .5 9 1 9 2 3 .5 9 1 9 19.4172 19.4172 19.4172 20.0548 20.0548 2 0 .0 5 4 6 17.5238 17.5238 17.5236 22.398 22.398 22.398 22.4925 22.4925 2 2 .4 9 2 5 23.2014 23.2014 23.2014 15.6588 15.6588 15.6588 19.0808 19.0808 W eight of A liquot, g 0.7845 0.7604 0.7916 0.7531 0.7709 0.7909 0.7798 0.7829 0.7822 0.7795 0.7808 0.7748 0.7114 0.7801 0.7948 0.7582 0.7841 0.7879 0.7324 0.7809 0.728 0.7707 0.7866 0.7473 0.825 0.7811 0,7686 0.7782 0.7624 0.7869 0.7707 0.7556 0.7875 0.859 0.8547 0.8305 0.7474 0.774 C o n c e n tra tio n o f PFBS in Original Liver Sample, ug/g 0.0472 0.0319 0.0317 0.0227 0.0241 0.0348 0.0507 0.0246 0.0284 0.0424 0.0266 0.0174 < LLOQ < LLOQ < LLOQ 0.0189 0.0146 0.0148 < LLOQ c LLOQ < LLOQ 0.00694 0.0148 0 ,0 0 7 1 6 0 .0 0 7 6 5 0 .0 0 8 0 8 0.0189 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ c LLOQ 0.138 0.148 0.153 Average, ug/g 3.461 3.461 3.468 21593 20814 17726 4.889 4.660 3.753 0.9533 19.0808 0.7683 0.0335 0.1 0.4889 0.9813 19.6369 0.7759 0.0265 0.1 0.1 0.4660 0.3753 0.9813 0.9813 19.6369 0.7805 0.0251 0.0240 3.461 3.461 7067 6979 368157 1.279 1.279 109.5 0.1 0.1279 1.1576 23.1342 0.7736 0.00694 0.1 0.1 0.1279 10.95 1 .1 5 7 6 1 .1 5 7 6 23.1342 23.1342 0.7505 0.7833 0.00715 0.587 0.20 0 LLOQ on C olum n, ng/m L 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 D ilution Factor On C o lu m n 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 LLOQ On-C olum n, C orrected fo r D ilution Factor 0.2558 0,2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0 .2 5 5 8 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 LLOQ in M atrix, ug/g 0.0140 0.0145 0.0139 0.0143 0 .0 1 4 0 0.0136 0.0137 0.0137 0 ,0 1 3 7 0.0148 0.0148 0.0149 0.0151 0.0138 0.0135 0.0151 0 .0 1 4 6 0,0145 0.0147 0.0138 0.0148 0.0139 0.0136 0 .0 1 4 3 0 .0 1 5 3 0 .0 1 6 2 0.0164 0.0140 0.0143 0.0139 0.0139 0.0141 0.0136 0.0126 0.0127 0.0131 0.0144 0.0139 0.0140 0.0139 0.0138 0.0140 0.0139 0.0143 0.0137 Average, ug/g JJsed 1/2 LLOQ for one of the replicate calculations - ~ Used 1/2 LLOQ for two of the replicate calculations T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 3 of 3 Contains No CBI O U4i^t Os Page 82 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Quail Liver. 900 ppm Dose Groun D1023032.D D1023033.D D1023034.D D1023035.D D1023036.0 D1023039.D D1023040.D D1023041.D D 1 0 2 3 Q 4 2 .D D1023043.D D1023047.D D1023048.D D1023049.D D1023050.D D1023051.D D1023054.D D1023055.D D1023056.D D1023058.D D1023062.D D1023064.D D1023065.D D1023069.D D1023070.D D 1 102030.D D1102031.D D1023078.D D1023079.D D1023081.D D1023084.D D1023086.D D1023087.D D1023092.D D1023093.D D1023095.D D1023096.D D1023100.D D1023101.D D1023103.D D1023107.D Sam ple Name 73072-1 73072-2 73072-3 73073-1 73073-2 73073-3 73074-1 73074-2 730747307573075-2 73075-3 73076-1 73076-2 73076-3 73077-1 73077-2 73077-3 73078-2 73078-3 73079-2 73079-3 73080-2 73080-3 73081-2 73081-3 73082-2 73082-3 73083-2 73083-3 73084-2 73084-3 73085-2 73085-3 73086-2 73086-3 73087-2 73087-3 73088-2 73088-3 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.461 3.4361 3.4161 3.461 3.461 3.461 3.461 3.461 Area C oncentration o f PFBS in Extract, On- colum n, ng/mL 492162 440276 481240 46.98 41.65 45,86 1120075 1120641 1088897 116.8 116.9 113.1 493596 47.13 434812 41.09 547404______ 52.73 2224594 281.1 2289851 294.1 1877559 220.4 705200 966286 69.52 98.69 O n-C olum n D ilution Factor C oncentrati on ofPFBS in Extract, ng/m L 187.9 166.6 183.4 467.2 467.5 452.3 188.5 164.4 210.9 1124 1176 881,5 278.1 394.8 3.461 3.461 3.461 1325101 1203305 1220545 142.2 126.9 129.1 4 568.8 4 507.7 4 516.2 3.461 3.468 495304 571533 659573 47.31 55.26 64.61 4 189.2 4 22 1.0 4 258.4 3.468 3.468 3.468 3.461 3 .4 6 8 1680803 1751223 1961725 712252 700481 190.6 200.9 234.0 97.84 70.29 69.01 4 762.2 4 803.7 4 936.1 4 391.4 4 281.1 4 276.0 .o I 3U2/U82 3.51 2978244 476.7 467.0 3 1430 3 1401 Sam ple not com pletely aspirated bv the R aoiri T ra n * No Data 3 .4 6 8 3.467 1025739 862298 729362 105.6 86.85 72.14 4 422.4 4 347.4 4 288.6 3.468 3.468 3.468 1134866 1076289 939403 118.6 1 1 1 .6 95.60 4 474.3 4 446.3 4 382.4 3 .4 6 7 3 .4 6 8 3.468 3 .4 6 8 3 .4 6 8 3 .4 6 7 3.468 3.468 3.468 921074 946085 929333 1497741 1590473 1321973 705690 708740 93.50 96.36 94.44 164.9 177.7 141.8 63.50 69.57 69.90 4 374.0 4 385.5 4 377.8 4 659.6 4 710.7 4 567.2 4 254.0 4 278.3 4 279.6 3.468 3.468 721859 758871 771907 71.33 75.37 76.80 4 285.3 4 301.5 4 307.2 S am ple no t completely aspirated by the R apid Trace, No Data 3.468 577659 55.90 4 ' 223.6 3.468 614949 59 85 4 239.4 W eight of W eight o f L iv e r (in ACN (in hom ogenat hom ogenate), e). g 0 .9 7 3 9 0 .9 7 3 9 0 .9 7 3 9 20.3185 20.3185 20.3185 1.0826 1.0826 1.0826 21.5632 21.5632 21.5632 0.9945 0.9945 0 .9 9 4 5 20.3631 20.3631 20.3631 0 .9 6 8 5 0 .9 6 8 5 0 .9 6 8 5 19.7817 19.7817 19.7817 1.0054 1.0054 20.1221 20.1221 1.0341 1.0341 1.0341 2 0 .1 22 1 20.4995 20.4995 1.0186 1.0186 1.0186 2 0 .1 6 9 9 20.1699 1.0873 1.0873 1.0873 21.7807 21.7807 1.0057 1.0057 1.0057 20.1040 20.1040 1.0057 1.0057 20.1040 20.1040 0.9947 0.9947 0.9947 0.9981 0.9981 0.9981 1.0305 1.0305 1.0305 1.0834 1.0834 1.0834 0 .9 8 1 6 0 .9 8 1 6 0 .9 8 1 6 1.008 1.008 1.008 1 9 .6 5 4 2 19.6542 1 9 .9 3 9 3 1 9 .9 3 9 3 20.796 20.796 20.796 21.7103 21.7103 21.7103 19.6313 1 9 .6 3 1 3 1 9 .6 3 1 3 20.1478 20.1478 1.0335 1.0336 20.6683 20.6683 W eight of A liquot, 9 0.7452 0 .7 5 3 1 0.7540 0.7633 0.7668 0.7590 0.7524 0.7580 0.7214 0.7635 0.7642 0.7592 0.7695 0.7731 0.7790 0.7665 0.7653 0.7627 0.7747 0.7635 0.7580 0.7782 0.7567 0.7782 0.7567 0.7856 0 .7 8 6 1 0.7477 0.7543 0.7649 0.7587 0.7701 0.7686 0.7282 0.7342 0.7306 0.7320 0.7664 0.7693 C oncentration o f PFBS in O riginal Liver Sam ple, ug/g 11.0 9.67 10.6 25.6 25.5 10.8 9.31 63.1 66 .0 49.8 15.2 21.5 2 0 .8 30.9 27.6 10.3 11.9 42.0 44.6 2 1 .1 15.6 77.1 77.7 22.3 18.3 26.6 24.8 20.7 21.5 36.0 38.9 14.7 15.9 16.4 17.3 12.3 13.1 Average, ug/g 10.4 28.9 12 .0 46.1 17.3 77.4 18.7 24.3 2 1 .1 35.2 15.5 17.2 12.7 LLOQ on C olum n, ng/m L 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 51.15 51.15 D ilution LLOQ On-C olum n, F actor On Corrected fo r C olum n D ilution Factor 4 40.92 40,92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 40.92 4 40.92 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 4 40.92 3 153.45 3 153.45 LLO Q in M atrix, ug/g 2.40 2.38 2 .3 7 2 .2 4 2.23 2.26 2.32 2.44 2.30 2.29 2.31 2.23 2 .2 1 2 .2 2 2.23 2.23 2.23 2.2 0 2 .2 0 2.25 2.27 2.26 2 .2 1 2 .2 7 2.25 8.51 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 10.23 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 40.92 40.92 40.92 40.92 40.92 40.92 40.92 40.92 40.92 40.92 40.92 40,92 40.92 40.92 40.92 40.92 40.92 40.92 2.16 2.17 2.30 2 .2 8 2.29 2.27 2.28 2 .2 8 2 .2 4 2.24 2 .2 2 2.36 2.33 2 .3 5 2.35 2 .3 7 10.23 4 4 40.92 40.92 2 .2 4 2.23 ug/g 2.24 8.40 Page 1 of 3 Contains No CBI O -t 4^ ON Page 83 of 227 3M Environmental Laboratory Quail Liver, 900 ppm Dose Groun File D1023109.D D1023110.D D1021033.D D1021034.D D1021036.D D1021039.D D1021041.D D1021042.D D1021047.D D1021048.D D1021050.D D1021051.D D1021055.D D1021056.D D1021058.D D1021062.D D1021064.D D1021065.D D1021069.D D1021070.D D1021072.D D1021073.D D1021078.D D1021079.D D1021081.D D1021084.D D1021086.D D1021087.D D1021092.D D1021093.D D1022033.D D1022034.D Sam ple Name 73089-2 73089-3 73090-2 73090-3 73091-2 73091-3 73092-2 73092-3 73093-2 73093-3 73094-2 73094-3 C oncentration C oncentrati o f PFBS in O n-Colum n on o f PFBS Extract, On- D ilution in E xtract, RT Area colum n, ng/m L F a c to r ng/m L 3 .4 6 8 3 .4 6 8 3 .4 6 8 3.461 3.461 3.461 3.4b1 2016250 1982025 2150156 1521210 1388149 1439516 14/8214 243.2 237.4 266.9 261.0 234.2 244.5 252.3 4 972.7 4 949.6 4 1068 1 261.0 1 234.2 1 244.5 1 252.3 Sam ple not com pletely aspirated by the Rapid Trace. No Data 3.461 1501557 257.0 1 7^7 n 3.461 3.461 3.461 3.461 3.4b I 1740153 1701971 1662021 2297525 2397823 2126545 16494/5 306.1 298.2 289.9 428.3 451.6 389.6 266./ 1 306.1 1 298.2 1 289.9 1 428.3 1 451,6 1 389.6 1 2 66.7 3.461 1615122 280.2 1 280^2 Sam ple not com pletely aspirated bv the Raoid Trace No Data W eight o f W eight o f Live r (in ACN (in hom ogenat hom ogenate), e ),g g 1.0929 1.0929 1.0929 0.9748 0.9748 0.9748 0.9969 2 1 .4 5 2 8 2 1 .4 5 2 8 21.4528 19.9677 19.9677 19.9677 2 0 .8 4 4 0 0.9969 0.9853 0.9853 0.9853 0.9759 0.9759 0.9759 0.9863 0.9863 20.8440 19.7607 19.7607 19.7607 19.4285 19.4285 19.4285 19.6777 19.6777 W eight of A liq u o t, 0 .7 6 7 6 0 .7 7 4 0 0 .7 6 0 7 0.7671 0 .7 6 1 0 0 .7 5 8 5 0,7441 0 .7 4 8 4 0 .7 7 1 0 0 .7 7 4 4 0.7718 0.7851 C o n c e n tra tio n o f PFBS in O riginal Liver 52.3 50.6 14.7 13.1 14.5 17.3 16.8 23.2 24.4 14.5 15.0 73095-2 73095-3 73096-2 73096-3 73097-2 73097-3 73098-1 73098-2 73098-3 73099-2 73099-3 73100-2 73100-3 73101-2 73101-3 73102-2 73102-3 73103-2 73103-3 73207-2 73207-3 Sam ple not com pletely aspirated b y the Rapid Trace No Data 3.461 1547102 266.2 1 266 2 3.461 1427555 242.1 1 3.461 3.461 2149084 2331622 2307313 394.6 436.2 430.6 1 394.6 1 436.2 1 430.6 3.461 3.461 1513189 1476891 1512863 259.4 252.0 259.3 1 259.4 1 252.0 1 259.3 oam pie not com pletely aspirated by the Rapid Trace, No Data Sam ple no t com pletely aspirated by the Rapid Trace, No Data 3.461 2320063 433.5 1 4^ s 3.461 3.461 3.461 3.468 3.4b8 1026917 1064164 1081142 805294 727806 670934 1806219 164.0 171.1 174.3 122.4 108.1 97.73 320.0 1 164.0 1 171.1 1 174.3 1 122.4 1 108.1 1 97 73 1 320 Q Sam ple no t com pletely aspirated by the Rapid Trace, No Data Sam ple no t com pletely aspirated by the Rapid Trace. No Data 3.461 3.461 3.461 3.461 5.4b8 1457719 1350945 1330409 1245980 1285062 1228795 9 6 2 /3 248.2 226.8 22 2.8 206.2 213.8 2 0 2 .8 32.65 1 248.2 1 226.8 1 2 2 2 .8 1 206.2 1 213.8 1 2 0 2 .8 0.1 3.265 3.468 115065 39.32 0.1 3.932 -S a m p le no t com pletely aspirated bv the Raoid Trace. No Data 1.0959 1.0959 0.9885 0.9885 0.9885 0.9963 0.9963 0.9963 0.9945 0.9912 0.9912 0.9912 1.0388 1.0388 1 .0 3 8 8 1 .0 7 1 3 1 .0 1 8 9 1 .0 1 8 9 1 .0 1 8 9 1.0050 1.0050 1.0050 1.0556 1.0556 2 1 .7 9 1 7 2 1 .7 9 1 7 19.8832 19.8832 1 9 .8 8 3 2 2 0 .0 8 3 8 2 0 .0 8 3 8 2 0 .0 8 3 8 0 .7 4 9 4 0.7473 0.7644 0.7388 0.7356 19.8518 19.9755 19.9755 19.9755 23.2071 23.2071 23.2071 2 1 .6 7 8 7 0 .7 2 5 0 0 .7 4 1 7 0.7506 0 .7 6 3 8 0 .7 7 8 5 21.3543 21.3543 21.3543 2 1 .5 4 8 9 2 1 .5 4 8 9 2 1 .5 4 8 9 2 1 .1 2 8 6 2 1 .1 2 8 6 0 .7 5 8 5 0.7571 0.7674 0.7710 0.7781 0.7476 14.8 22.3 24.1 14.9 14.5 9.57 9.76 7.61 6.61 17.5 14.4 13.2 1 2 .1 12.4 0.176 0 .2 2 1 73208-2 73208-3 Sam ple no t com pletely aspirated by the Rapid Trace, No Data 3.468 3.468 64785 81013 21.48 27.23 0.1 2.148 0.1 2.723 0.679 0.679 13.5593 13.5593 0.7651 0.7648 0.118 0.149 Average, 53.4 13.9 14.7 16.8 22.8 14.7 14.1 23.5 14.7 23.7 9 .7 3 6 .7 3 17.46 13.5 1 2 .1 0.199 LLOQ on C olum n, ng/m L 10.23 10.23 51.15 51.15 51.15 51.15 D ilution Factor On C olum n 4 4 4 LLOQ On-C olum n, C orrected for D ilution Factor 40.92 40.92 40.92 51.15 51.15 51.15 51.15 LLO Q in M atrix, ug/g 2.2 0 2 .1 8 2 .1 9 2 .8 7 2 .8 7 2.95 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 2.95 2 .8 9 2.8 8 2 .8 9 2 .7 7 2 .7 6 2.73 2.78 2 .7 3 2 .1 9 2 .8 8 2 .9 5 2.89 2 .7 6 2.75 70 CD O O CO cn CD 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 2.558 2.558 0.1 0.1 2.558 0.1 0.1 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 0.2558 0.2558 0.2558 0.2558 2 .8 5 2.83 2 .8 9 2 .8 3 2 .8 7 2 .9 3 2 .9 4 2.91 2 .8 0 2 .9 8 2 .9 2 2 .9 0 3 .1 8 3.12 2 .7 9 2.96 2 .9 7 2.96 2 .9 9 2.98 2 .9 9 0.0138 0.0144 0.0140 0.0140 2 .8 4 2 .8 6 2.93 2.93 3.14 2.79 2 .9 6 2.99 0.0141 0.0140 X CD CD > < T* 0 S2. CO |> 5 '^ 0c ?3 X C/D CO Cl _ . >< O =T 3= CD 3z 03 . 3- 3 s-' " H S' CO CO 3T CD <3 Page 2 of 3 Contains No CBI NJ NO NJ 0 a 2: c 1 o 4^ U 1 ON Report E03-0583 3M Environmental Laboratory Quail Liver. 900 ppm Dose Group File D1022041.D D1022042.D D1022047.D D1022048.D D1022050.D D 1 0220 5 1 .D D1022055.D D1022056.D D1022058.D D1022062.D D1022064.D D1022065.D D1022069.D D1022070.D D1022072.D D1022073.D D1022078.D D1022079.D D1022081.D D1022084.D D1022086.D D1022087.D D1022092.D D1022093.D D1022095.D D1022096.D Sam ple Name RT 73209-2 73209-3 3.468 3 .4 6 8 73210-2 73210-3 3.468 3.468 73211-2 73211-3 73212-2 73212-3 73213-2 73213-3 73214-2 73214-3 73215-2 73215-3 3.468 3.468 3.468 3.468 3.468 3.468 3.468 3.468 3.468 3.468 3.468 73216-2 73216-3 73217-2 73217-3 73218-2 73218-3 73219-2 73219-3 3.468 3.468 3.468 3.468 3.468 3.468 3.468 3.468 3.468 73220-2 73220-3 73221-2 73221-3 3.475 3.475 3.468 3.475 Area C o n c e n tra tio n o f PFBS in Extract, On- colum n, ng/m L 68738 62300 11870 19928 35498 30217 174380 165763 33621 31491 33213 35030 92890 88902 25641 30048 30097 32198 53003 55574 105015 118444 32889 32060 53627 57707 2 2 .8 8 20.60 7.869 2.732 5.584 9.866 1 1 .1 0 9.227 60.42 57.35 10.43 9.679 10.29 10.93 30.46 31.45 30.03 7.607 9.168 8.371 9.185 9.929 17.30 18.21 36.16 35.75 40.52 10.17 9.680 17.52 18.97 O n-C olum n D ilution F a c to r 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 C o n ce n tra ti on o f PFBS in Extract, ng/m L 1.593 2.288 2.060 0.7869 0.2732 0.5584 0.9866 1 .1 1 0 0.9227 6.042 5.735 0.6933 1.043 0.9679 1.029 1.093 3.046 3 .1 4 5 3 .0 0 3 0.9163 0.7607 0.9168 0.8371 0.9185 0.9929 2 .0 4 7 1.730 1.821 3 .6 1 6 3 .5 7 5 4 .0 5 2 0.8390 1.017 0.9880 1.501 1.752 1.897 W eight o f W eight o f Liver (in ACN (in hom ogenat homogenate), e) > 9 g 0.6675 0.6675 0.6675 1 3 .3 0 1 4 1 3 .3 0 1 4 1 3 .3 0 1 4 0.6669 0.6669 1 4 .6 8 4 8 1 4 .6 8 4 8 1 4 .6 8 4 8 0.7753 0.7753 0.7753 15.5411 15.5411 15.5411 0.7186 0.7186 0.7186 1 4 .2 5 8 2 14.2582 14.2582 1.1755 1.1755 1.1755 23.5564 23.5564 23.5564 0.8396 0.8396 0.8396 0.9961 0.9961 0.9961 16.7557 16.7557 1 6 .7 5 5 7 19.1102 1 9 .1 1 0 2 1 9 .1 1 0 2 0.8449 0.8449 0.8449 1 7 .0 1 3 4 17.0134 1 7 .0 1 3 4 0.8130 0.8130 0.8130 1 6 .2 9 0 5 1 6 .2 9 0 5 1 6 .2 9 0 5 0.8599 0.8599 0.8599 0.9651 0.9651 0.9651 0.7836 0.7836 0.7836 17.2397 17.2397 1 7 .2 3 9 7 1 9 .4 2 7 8 1 9 .4 2 7 8 1 9 .4 2 7 8 1 5 .7 5 0 7 15.7507 1 5 .7 5 0 7 0.7175 0.7175 0.7175 14.4181 14.4181 14.4181 W eight of A liquot, g 0.7262 0.7725 0.7697 0.7637 0.5967 0.7662 0.7709 0 .7 7 4 0.7753 0.7759 0 .7 8 0.7905 0.7828 0.7827 0.7855 0.7661 0.7579 0.6398 0.7639 0.7711 0.7585 0 .7 6 5 0.6963 0.7632 0.7747 0.7734 0.7700 0.7126 0.7523 0.7516 0.7627 0.7651 0.7722 0.7854 0.7875 0.7831 0.7728 0 .7 7 6 0.7741 C oncentration o f PFBS in O riginal Liver Sam ple, ug/g 0 .0 9 1 8 0.124 0.11 2 0 .0 4 7 4 0 .0 2 1 1 0.0336 0 .0 5 3 9 0 .0 6 0 4 0.05Q1 0.313 0.323 0.302 0.0373 0.0561 0.0518 0 .0 5 6 5 0 .0 5 6 9 0.161 0.165 0.160 0.0506 0.0462 0 ,0 4 5 5 0 .0 5 0 0 0 .0 5 4 3 0 .1 2 1 0 .0 9 6 8 0.10 2 0.20 0 0.197 0.22 2 0.0451 0 .0 5 4 5 0.0532 0 .0 8 2 0 0 .0 9 5 3 0.103 Average, ug/g 0.109 0.0340 0.0548 0.313 0.0484 0 .0 6 1 7 0.162 0.0492 0.0499 0.107 0.207 0.0509 0.0935 L L O Q o n D ilu tio n LLOQ O n-C olum n, LLO Q in C olum n, Factor On C orrected fo r M atrix, A verage, 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2 .5 5 8 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2.558 2 .5 5 8 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.2558 0.2558 0.2558 0.2550 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.2558 0.0147 0.0139 0.0154 0.0197 0.0140 0.0139 0.0137 0.0137 0.0138 0.0138 0.0140 0 .0 1 4 1 0.0135 0.0134 0 .0 1 4 1 0.0155 0.0142 0.0139 0.0139 0.0151 0.0143 0.0142 0 .0 1 4 1 0.0137 0.0137 0.0140 0.0139 0.0139 0.0142 0.0168 0.0139 0.0136 0.0137 0.0150 0.0135 0.0146 0.0139 0.0146 0.0141 0.0137 0.0139 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 84 of 227 Page 3 of 3 Contains No CBI Cl? 4^ L fx 4^ ON Report E03-0583 3M Environmental Laboratory Egg Matrix Spikes and Blank* Sample Name M B -041202-1 M B-041202-2 M B-041202-3 M B-041202-4 L S -0 4 1 202-1 L S -0 4 1202-2 L S -0 4 1 2 0 2 -3 M S-041202-1 M S-041202-2 M S-041202-3 H S -041202-1 H S -041202-2 H S-041202-3 RT 0 0 0 0 3.467 3.468 3.467 3.439 3.439 3.44 3.439 3.439 3.439 Area 0 0 0 0 403810 401475 395378 2209971 2380481 2207419 8680863 8541773 7794102 Concentration of Injection PFBS in Extract, On- Volume. column, ng/mL uL < LLOQ < LLOQ < LLOQ < LLOQ 84.39 83.88 82.56 102.7 1 1 1 .6 1 0 2 .6 513.1 501.9 4 4 4 .2 20 20 20 20 6 6 6 4 4 4 4 4 4 Recovery% 110% 109% 108% 100% 109% 100% 100% 98% 87% Average Recovery, % 109% 103% 95% %RSD 1.13% 4.89% 7.60% LLOQ on Dilution Column, Factor On ng/mL Column 25.58 2 5 .5 8 25.58 25.58 51.15 51.15 51.15 25.58 25.58 25.58 25.58 25.58 25.58 0.2 0.2 0.2 0.2 0.33 0.33 0 .3 3 1 1 1 1 1 1 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 85 of 227 Page 1 of 1 Contains No CBI L/1 ON , rjBTTcyntr 3M Environmental Laboratory a tio n of PFBS in E xtra ct, Oncolum n, _____ F ile______ S a m p le Name_______________________ S a m p le Info_________________________ n g /m L D1207077.D 01207078.0 D1207079.D 72577-1 72577-2 72577-3 0 ppm Dose Group: Egg; 02 D ec 04; C M C /M L/V M E eT 0 ppm Dose Group; Egg; 02 D ec 04; CMC/M LA/M EE 0 ppm Dose Group; Egg; 02 D ec 04; C M C /M LA/M EE _ <LL0Q <LLOQ < LLOQ D1207082.D D1207083.D D1207084-D 72578-1 72578-2 72578-3 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 0 ppm Dose Group; Egg; 02 D ec 04; CMC/M LA/M EE 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE < LLOQ <LLOQ <LLOQ D12070S5.D D 1 2 07 08 6.D D 1 2 07 08 8.D 72579-1 72579-2 72579-3 0 ppm Dose Group; Egg; 02 D ec 04; CMC/MLA/MEE 0 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 0 ppm Dose Group; Egg; 02 D ec 04; C M C /M LA/M EE <LLOQ <LLOQ <LLOQ D1207089.D D1207090.D D1207091.D 72580-1 72580-2 72580-3 Q ppm Dose Group; Egg; 02 D ec 04; CMC/M LA/M EE 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 0 pptn Dose Group; Egg; 02 D ec 04; CMC/MLA/MEE <LLOQ <LLOQ <LLOQ D1207092.D D 12 07 09 5.D D1207096.D 72581-1 72581-2 72581-3 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 0 ppm Dose Group; Egg; 02 Dec 04; CM C/MLA/M EE <LLOQ <LLOQ < LLOQ D1207097.D D1207098.D D1207099.D D 1 2 08 03 0.D D1208031.D D1208032-D 72582-1 72582-2 72582-3 72583-1 72583-2 72583-3 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 100 ppm Dose Group; Egg; 02 Pec 04; CMC/MLA/M EE <LLOQ <LLOQ <LLOQ 105.6 99.99 101.8 D1208033,D D 12 08 03 4.D D 1 2 0 8 0 3 6 .D 72584-1 72584-2 72584-3 100 ppm Dose G roup; Egg; 0 2 D ec 04; CMC/M LA/M EE 100 ppm Dose G roup; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 D ec 04; CMC/MLA/MEE 115.1 105.3 97 .2 2 D1208037.D D 12 08 03 8.D D 12 08 03 9.D 72585-1 72585-2 72585-3 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 0 2 D ec 04; CMC/M LA/M EE 174.7 173.9 189.3 D 12 08 04 0.D 0 1 2 0 8 0 4 3 .0 D1208044. D 72586-1 72586-2 72586-3 100 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 99.41 95 .7 2 101.8 D 1 2 0 80 45 .D D 1 2 08 04 6.D D 1 2 08 04 7.D D 1 2 08 04 9.D D1208050.D D1208051 ,D D1208052.D D 12 08 05 3.D D 12 08 05 6.D 72587-1 72587-2 72587-3 72588-1 72588-2 72588-3 725B9-1 72589-2 72589-3 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 0 2 D ec Q4; C M C /M LA/M EE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 100 ppm Dose G roup; Egg; 02 D e c 04; CMC/MLA/MEE 80 .9 4 BC.53 76 .6 2 90 .7 2 93 .0 4 99 .0 6 "TeaT 156.1 162.0 D1208057.D D 12 06 05 8.D D1208059.D 72590-1 72590-2 72590-3 300 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 300 ppm Dose Group; Egg; 02 Pec 04; CMC/MLA/MEE 292.6 299.7 278.6 D1208060.D 72591-1 300 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 640.8 " 0 D1208062. D 72591 -2 CD CQ D1208063.D 72591-3 <D D 1208064.D 72592-1 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 300 ppm Dose Group; Egg; 02 Pec 04; CMC/M LA/M EE 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 596.9 630.6 249.0 CO D1208065.D 72 592-2 CT1 D 1208066.D 72 592-3 o D1208069.D 72593-1 300 ppm Dose Group; Egg; 02 Dec 04; C M C /M LA/M EE 300 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 300 ppm Dose G roup; Egg; 02 Dec 04; CMC/M LA/M EE 232.1 249.5 469.0 D1208070.D 72593-2 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 493.7 D12Q807^Q___72593^3____a__300_ggmDose!G ro u g ^^ |^=0 2 D e c ^4 ^ C M C /M ^/M E E _ _ s_ ^ = _474;7= D1208072.D D 12 08 07 3.D D1208075.D 72594-1 72594-2 72594-3 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 300 ppm Dose Group; Egg; 02 Dec 04; CMC/M LA/M EE 300 ppm Dose Group; Egg; 02 Pec 04; CMC/MLA/MEE 239.8 229.8 253.9 D 12 08 07 6.0 0 1 2 0 8 0 7 7 .0 D1208078.D 72595-1 72595-2 72595-3 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 485.3 300 ppm Dose Group; Egg; 02 D ec 04; CMC/MLA/MEE 483.5 300 ppm Dose Group; Egg; 02 Pec 04; C M C /M LA/M EE_________ 481.8 Quail Eggs. Lots B anrl R OnC o lu m n D ilution Factor 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 weign r C o n c e n tr ation of PFBS in E xtra ct, ng/m L W eight of Egg (in homoge nate), g o f E gg W eight W eight Hom oge o f ACN of W ater nate(in (in W eight (in so lve n t solve nt of hom oge p re cip ita precipita Aliquot, nateb g te), g te), g q Concentration of PFBS in O riginal Egg Sam ple, ug/g iOOC lOOC 8 o o ooo o o o ODD c LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 100.0 101.8 105.3 97.22 174.7 173.9 189.3 95 .7 2 101.8 80 .5 3 76 .6 2 93 .0 4 99 .0 6 156.1 162.0 299.7 278.6 596.9 630.8 6.0012 6.0012 6.5650 6.5650 6 .5 6 5 0 6.5424 6.5424 6.5424 5.6951 5.6951 6.7825 6.7825 6 .5 1 6 7 6 .5 1 6 7 5.9543 5.9543 6.2879 6.2879 7.0978 7.0978 5 .8 4 1 1 5 .8 4 1 1 5.8411 6.6303 6.6303 6.6303 6.4766 6.4766 6.4766 5.8621 6.0183 5.8621 6.0183 5.9660 6.1055 5.9660 6.1055 6.7819 _ 7.0869 6.7819 7.0869 6.7819 7.0869 5.5413 5.5413 6.1868 6.1868 6.1868 6.0890 6.0890 6.4026 6.4026 5.3528 5.3528 5.3528 5 .4 9 2 1 5.4921 6.0769 6.0769 6.0769 5.9833 5.9833 5.9833 6.2746 6.2746 6.2746 5.2959 5.2959 6.0191 6.0191 5.9767 5.9767 09980 0.9980 0.8758 0.8758 0.8758 0.9079 0.9079 0.9079 0.9713 0.9713 0.9713 0.9615 0.9615 0.9615 0.8704 0.8704 0.7157 0.7157 0.8508 0.8508 0.8508 0 .9 5 1 1 0 .9 5 1 1 0.8773 0.8773 0.9380 0.9380 0.9380 0.9069 0.9069 0.9768 0.9768 0.9476 0.9476 0.9476 1 .0 0 0 8 1 .0 0 0 8 9.8838 9.8838 9.8838 9.8846 9.8846 9.8846 9.8874 9.8874 9.8874 1 0 .7 9 7 7 1 0 .7 9 7 7 1 0 .7 9 7 ? 9.8743 9.8743 9.3743 9.9441 9.9441 9.9441 9.9449 9.9449 9.9449 9 .9 7 8 9 9 .9 7 8 9 9.9789 9.9929 9 .9 9 2 9 9 .7 9 4 8 9.7948 9.7948 9 .5 0 9 3 9 .5 0 9 8 9 .5 0 9 8 9 .6 0 8 7 9.6087 9 .6 0 8 7 9 .7 8 3 2 9 .7 8 3 2 9 .7 8 3 2 9.5770 9.5770 9 .5 7 7 0 9 .9 0 5 9 9 .9 0 5 9 9.9059 0.6924 0.7300 0.6480 0.6780 0.7114 0.5110 0.7505 0.7425 0.8127 0.7086 0.7268 0.7929 0.7468 0.7507 0.7051 0.7811 0.7860 0.6184 0.7303 0.7405 0.8072 0.7355 0.7714 0.7911 0.7871 0.7593 0.7991 0 ,6 9 1 7 0.6563 0.6964 0.7746 0.6862 0 .6 7 9 6 0.6956 0 .7 9 8 5 0.7215 0.7738 0.6868 0 .7 3 4 7 0.8085 0.7132 0.7586 0.7533 0 .7 1 9 2 0.7516 < LLOQ < LLOQ < LLOO < LLOQ < LLOQ < LLOO 8.73 8.15 7.61 8.06 7.03 6.33 10.4 10.8 11.2 6.96 7.07 7.08 4.61 5.18 4.98 6.00 5.36 6.31 9.23 9.91 9.62 16.0 18.6 16.2 36.9 36.0 36.5 OOO OOO ooo ooO| 232.1 249.5 493.7 474.7 229.8 253.9 485.3 483.5 481.8 6.6361 6.6361 6.1262 6.1262 6.9918 6.9918 6.1803 6.1803 6.2336 6.2336 6.0186 6.0186 6.9682 6.9682 6 .1 5 5 1 6.1551 6 .1 5 5 1 0.8661 0.8661 0.8771 0.8771 0.8771 0.9742 0.9742 0.9742 1 .0 2 3 3 1.0233 1.0233 9 .6 4 9 2 9 .6 4 9 2 9.6492 9.7310 9 .7 3 1 0 9 .7 3 1 0 9 .8 4 5 6 9 .8 4 5 6 9 .8 4 5 6 9 .7 0 1 4 9 .7 0 1 4 9 .7 0 1 4 0 .7 2 8 0 0.7216 0 .7 4 8 9 0.8044 0.8055 0 .7 9 9 2 0 .8 0 1 7 Q.S01S 0.8040 0 .8 0 0 0 0.8034 0.8038 16.1 15.1 15.7 28.0 29.4 28.5 13.3 12.7 14 0 25.4 25.2 25.1 A v e ra g e , u g /g < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 0.17 7.14 10,8 7.04 4.92 5.89 9.59 16.9 36.5 15.6 28.6 13.3 25.2 LLOQ On LLOQ on Colum n, ng/m L D ilution F a c to r On- C olum n, C orrete d for D ilu tio n LLOQ in M atrix, A v e ra g e , 25.58 0.2 5.116 1.60 25.58 0.2 5.116 1.52 1.61 25.58 0.2 5.116 1.82 25.58 0.2 5.116 1.73 1.98 25.58 0.2 5.116 1.69 25.58 0.2 5.116 1.71 1.65 25.58 0.2 5.116 1.77 25.58 0.2 5.116 1.73 1.69 25.58 0.2 5.116 1.53 25.58 0.2 5.116 1.52 0.2 25.58 0.2 5.116 1.62 25.58 0.2 5.116 1.61 0.2 51.15 51.15 51 .1 5 51.15 51.15 4.23 4.17 51 .1 5 51 .1 5 51.15 51.15 51.15 51.15 3.58 3.42 51 .1 5 51.15 3.06 51.15 1 51.15 3.17 1 51.15 1 51.15 3.58 51 .1 5 1 51.15 3.78 1.55 1.76 4.07 3.44 3.08 3.64 51 .1 5 51.15 2.92 51.15 51.15 3.29 3.18 1 51 .1 5 1 51 .1 5 3.38 51.15 1 51.15 2.95 3.20 51.15 1 51.15 2.88 51.15 1 51.15 3.25 3.06 51 .1 5 1 51 .1 5 2.80 51.15 1 51 .1 5 3.17 2.98 1 51 .1 5 51 .1 5 1 51.15 2.95 1 51.15 3.09 3.00 1 51.15 51 .1 5 51.15 3.31 1 51.15 3.34 3.29 51 .1 5 1 51 .1 5 3.05 51.15 1 51.15 3.05 3.05 51 15 1 51 .1 5 2.83 51.15 1 51.15 2.83 2.83 51 .1 5 51 .1 5 1 51.15 2.67 1 51 .1 5 2.66 2.67 1 73 CD -o o m o co Ol co co "O 37 ai CD > aO > | 73 M =r. Q . OC CJ O -o -T1 3 03 Cl __ > < It- o =5>r 31 I o tS- %3 <z> <_ en CD <*> ir> Page 1 of 3 Contains No CBI 295 l 1 H 3. a & z | cr a - 4^ I >-- * 0\ Page 87 of 227 3M Environmental Laboratory Quail Eggs. Lots B and K F ile D 12 08 07 9.D D 12 08 08 2.D D1208083.D D1215028.D D 1 2 15 02 9.D D121S030.D D 1 2 15 03 1.D D 1 2 15 03 2.D D1215035.D D1215036.D D 1 2 15 03 7.D D 1 2 15 Q 38 .D D 1 2 15 03 9.D D1215043.D D 1 2 15 04 4.D 01215045.D D1215046.D D121S047.D D 12 15 05 0.D D 1 2 1 5 0 5 1 .D D 1 2 1 5 0 5 2 .D D 1 2 1 5 0 5 3 .D D 1 2 1 5 0 5 4 .D D 1 2 1 5 0 5 8 .D D 1 2 1 5 0 5 9 .D D 1 2 1 5 0 6 0 .D D 1 2 1 5 0 6 1 .D D1207046.D D 1 2 0 7 0 4 7 .D D 12 07 04 9.D D1207050.D D1207051.D D1207052.D D1207053.D D1207056.D D1207057.D D1207058.D D1207059.D D1207060D D1207062.D D1207063.D D12P7064-D D 1 2 07 06 5.D D1207066.D D 12 07 06 9.D D 12 07 07 0.D D 12 07 07 1.D D 12 07 07 2.D D 1 2 0 7 0 7 3 .D D 12 07 07 5.D D1207076-D D 12 09 02 8.D D 12 09 02 9.D D 1 2 0 9 0 3 0 .D D 1 2 Q 9 0 3 1 .D D 1 2 0 9 0 3 2 .D D1209034-D D 12 09 03 5.D D1209036.D D1209037.D Sam ple Name Sam ple Info 72596-1 72596-2 72596-3 72597-1 72597-2 72597-3 72598-1 72598-2 72598-3 72599-1 72599-2 72599-3 72600-1 ' 72600-2 72600-3 72601-1 ' ^ 72601-2 72601-3 72602-1 72602-2 72602-3 72603-1 ' 72603-2 72603-3 300 ppm D ose Group; Egg; 02 Dec 04; CMC/M LA/M EE 300 ppm D ose Group; Egg; 02 Dec 04; CMC/M LA/M EE -----2-=-gg21--::O|e_G--r-a-u-';=-E-3-g;=-02--_MD-ewco_w0-4r,t WC/VMI^/CIV^IU^/^VIEVIECC 900 PPm D ose Group; Egg; 02 D ec 04; CMC/M LA/M EE 900 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE ---.-9-0--0--pp m Do--s--e--G--r-o--u-vp-;r,Eogivgi;w0iv2tLD^eVcry0tc4r; C M C /M IA /M EF 990000 podpimti DO nocsee G(Orrnoiuipn-; ECngig>; 'm02 rD^e7c7^0^4=; SCfM7^C7/fMrrL=A?:/TM=SESE== 900 ppm D ose Group; Egg; 02 Dec 04; CMC/M LA/M EE _ 9 0 0 ppm D ose G r o u D ^ o n M l T ^ n ov-r-j.T^Avi^wrrui.irut M/'v i^v^icic^ 900 ppm D ose G roup; Egg; 02 D ec 04; CMC/M LA/M EE 900 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE --__ 9-0-^-p-p--m--D-o--se G--r-o-utp-';tr.Ei.ov-oT,; 0WI2VIDWeIVcIL0/4V;IVCIMEEC/M LA/M FF 990000 nPopmm fuTonseee G roup; Egg; 02 Dec 0 4 ; CMC/M LA/M EE 900 ppm D ose Group; Egg; 02 D ec 04; CMC/M LA/M EE _=9=_00 ip! p=ms ^D^ ^o-s-5e^G^cro^u_p^;a BEgi_g^;-0y 2rLDg yerc_0y4^;' wCMiviCw/iMvnL^Av/iMyiEeEe Q9n0n0 Pn Pnmm Dfl--ons.e,, G_ro_u_p; Eg g; 02 D ec 04; CMC/M LA/M EE ' 900 ppm D ose G roup; Eg g; 02 D ec 04; CMC/M LA/M EE _--.9o-0-n-0e-pr--,np|\--rmvn rDv-o-.-s^e- G-- rou_p_;r~E_uq_n_ai; 02 D ec v0-4v,- OclmVIcur/imViILa/V/ImVIpCce 900 ppm D ose G roup; Egg; 02 D ec 04; CMC/M LA/M EE 900 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE ==*Q9'n0--n0 Pr-.Pn-rm-vi--FD-ino---?,.e-aGr~n_r'o_u**p.-;1Eaa: 02 Dec X0T4.: ^CiMwoC<i/vMiiI->am/vmirp-rp900 ppm D ose Group; Eg g; 02 D ec 04; CMC/M LA/M EE 900 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE _ 900 ppm D ose G roup; Egg; 02 Dec 04:J2MC/M LA/MEE 72604-1 72604-2 72604-3 72727-1 72727-2 72727-3 72728-1 72728-2 72728-3 900 ppm D ose Group; Egg; 02 D ec 04; CMC/M LA/M EE 9 0 ppm D ose Group; Eg g; 02 Dec 04; CMC/M LA/M EE J22=`EmD--ose_--G:TOUj_|gg;~_--02--_wDw|cw0-4r.^sSC^IVMIwr^IVMU_n^UIMVIC| ^ _ 0 Ppm u o s e G roup; Egg; 0 2 Dec 0 4 ; CMC/M LA/M EE 0 ppm D ose G roup; Egg; 0 2 Dec 04; CMC/M LA/M EE =J =gm=Dose_Groui Jgg^2_D ec04j_C M ^M L^M EE 0 ppm D ose G roup; Egg; 02 D ec 04; CMC/M LA/M EE 0 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE -- P-PPm D ose G roup; Egg; 02 D ec 04: CMC/M LA/M EE 72729-1 72729-2 72729-3 0 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE 0 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE 72730-1 72730-2 72730-3 72731-1 72731-2 72731-3 0 ppm D ose G roup; Egg; 02 D ec 04; CMC/M LA/M EE 0 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE ==^ =2= ! e =Grou4=Egg;J)2_DecJ>4jaC M O M ^M E E 0 ppm Dose G roup; Egg; 02 Dec 0 4; CMC/M LA/M EE 0 ppm D ose Group; Egg; 02 Dec 04; CMC/M LA/M EE n '""^ ^ g S ^ U j_ ifla ij3 2 _ D e M 3 4 ; CMC/M LA/M EE 72732-2 72732-3 72733-2 7273372734- 72735-1 72735-2 72735-3 72736-1 72736-2 72736-3 72737-1 72737-2 72737-3 0 ppm Dose G roup; Egg; 0 2 D ec 04; CMC/M LA/M EE 0 ppm Dose G roup; Egg; 0 2 D ec 04: CMC/M I.A/M FF 0 ppm Uose Group; Egg; 0 2 Dec 04; CMC/M LA/M EE 0 ppm Dose G roup; Egg; 02 Dec 04; CMC/M LA/M EE .... 0 P3Pm D ose G roup; Egg; 02 Dec 04: CMC/MI A/MPP 0 p1pm D ose G roup: Egg; 0 2 Dec 04; CMC/M LA/M EE . i_yy, u i. uet, UH, GMG/M LA/M EE ^O j^jm -D o s e-G ^up ; --Egaga;i 0 2 D^ec0v-4,;wCivMnC-rt/iMvnL-jA-m/MvitEriE- 100 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE- 100 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE ^ 100 ppm i1n0n0 pmpmm Dose Dose Group; Egg; rG-_ro_u_pn; ~Egg; Q2 02 Dec Dec 0v-4r;, 04; C^vMiwCfi/vMiuL^Ay/iMviEc cF CMC/M LA/M EE 100 ppm D ose G roup; Egg; 02 D ec 04; CMC/M LA/M EE ^_ 1 0 0 ppm Dos e G roup; Eg g; 02 D ec 0w4-r;, C^mM\Cw/iMviiL,^Av/iMviEc Fc -inn r.rNm /--___ : r-__ 100 PPm D ose G roup; Eg g; 02 D ec 04; CMC/M LA/M EE 100 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE 100 ppm D ose G roup; Egg; 02 Dec 04; CMC/M LA/M EE ' Lunesntr alien o f PFBS in E x tra c t, C o lu m n C oncentr PFBSm --------- --- W eight W eight '| EB9 wwm u------------------------------------------------- ----- ----------------- o f Egg W eight Homoge o f ACN ' ' a te " ' ' " " W `>'9h ' C n o . n . m t i,, ,, o f 462.0 511.5 467.7 406.1 388.0 410.4 1 462.0 511.5 467,7 812.2 776.1 820.6 6 .9 6 1 4 6.9614 6.9614 6.4044 6.4044 6.4044 6.8616 6.8616 6.8616 6 .4 8 7 0 6.4870 6 .4 8 7 0 0.8775 0,8775 0,8775 0.9059 0.9059 0,9059 9.8844 9.8844 9.8844 9.7535 9.7535 9.7535 0.8012 0.0005 0.7973 0,8006 0.8084 0.8100 359.5 360.3 353.5 393.6 416.2 397.1 325.6 323.0 335.2 320.0 321.4 316.3 493.3 523.3 4H9 7 320 3~ 308 1 -- ' 718.9 720.6 707,0 787.3 832.4 794,2 651.1 645.9 670,5 640.0 642.8 632.5 1973 2093 1959 640.7 616.3 6 .5 1 6 0 6 .5 1 6 0 6 .5 1 6 0 5 .8 7 6 2 5 .8 7 6 2 5 .8 7 6 2 5 .9 8 5 0 5.9850 5 .9 8 5 0 6 .9 9 6 0 6 .9 9 6 0 7.0453 7.0453 7 .0 4 5 3 6 .2 0 9 4 6 .2 0 9 4 6.5751 6.5751 6.5751 6 .0 8 4 4 6 .0 8 4 4 6 .0 8 4 4 6 .1 9 4 2 6 .1 9 4 2 6 .1 9 4 2 7 .1 5 0 9 7 .1 5 0 9 7 .1 5 0 9 7.1335 7.1335 7 .1 3 3 5 6 .5 8 6 4 6 .5 8 6 4 0.9503 0.9503 0.9503 _0.8634 0.8634 0.8634 0.9589 0.9589 0.9589 0.9637 0.9637 0.9637 0.9192 0.9192 0.9192 9.8197 9.8197 9.8197 10.0666 10.0666 100666 9.5706 9.5706 9.5706 10.2208 10.2208 10.2208 9.8949 9.8949 9.8949 9.7780 9.7780 0 .8 0 3 1 0.8027 0.8000 0.8006 0.8028 0.8098 0.8038 0.8066 0.8000 0.8051 0.8091 0 .8 0 2 1 0.8012 0.8027 0.8057 0.7980 0.8060 298.8 293.5 278.4 < LLOQ 2 2 2 0.2 597.6 587.0 556.0 < LLOQ 6.4870 6 .4 8 7 0 6 .4 8 7 0 6 .5 8 6 4 6.5151 6.5151 6.5151 0.8938 0.9313 0.9313 0.9313 9.7780 9.8760 9.8760 9.8760 0.8039 0.8002 0.7964 0.8019 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ <LLOQ < LLOQ < LLOQ 0.2 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ 5.3847 5 .3 8 4 7 5 .3 9 3 7 5.3937 5.3937 4.8310 4.8310 4.8310 7.1333 7.1333 7 .1 3 3 3 5.4336 5 .4 3 3 6 5 .4 3 3 6 6 .0 6 3 7 6.0637 6.1000 6.1000 6,1000 5.7115 5.7115 5.7115 6.1762 6.1762 6 .1 7 6 2 5 .8 0 7 7 5 .8 0 7 7 5 .8 0 7 7 1.2811 1.2811 0.9772 0.9772 0.9772 1.1197 1.1197 1.1197 1.0097 1.0097 1.0097 1.0236 1.0236 1.0236 9.6918 9.6918 9.5684 9.5684 9.5684 9.6822 9.6822 9.6822 10.0317 10.0317 10.0317 9.3654 9.3654 9.3654 0 .7 7 9 1 0.7989 0.8017 0.7918 0.8084 0.8010 0.8030 0 .7 9 6 1 0.8005 0.7964 0.7912 0.8000 0.7178 OOC DOO OOO O O D; 0.2 0.2 0.2 5 .1 3 4 0 5.1340 5 .8 3 6 0 5.8360 1 .3 0 9 3 1 .3 0 9 3 9.3674 0.7771 9.3674 0.7818 < LLOQ < LLOQ < LLOO < LLOQ 0.2 0.2 0.2 0.2 < LLOQ < LLOQ < LLOO < LLOQ 6 .6 3 4 5 5 .6 3 4 5 5 .6 3 4 5 5 .0 9 4 6 5 .0 9 4 6 5 .0 9 4 6 1.1424 1.1424 1.1424 9.2504 9.2504 9.2504 0.7981 0.8030 0.7989 < LLOQ 247.8 < LLOQ < LLOQ 247.8 6 .2 5 5 6 6 .2 5 5 6 5 .7 0 8 6 5 .5 6 2 8 5.5628 5 .8 3 8 5 1.3371 1.3371 1.0697 9.3602 9.3602 9.6358 0 .7 9 7 1 0.7993 233.3 5.7086 5.8385 1.0697 9.6358 247.8 248.5 270.1 I l I 247.8 248.5 220 1 5.7086 5.2914 5.2914 5.2914 5.8385 5 .6 7 2 0 5 .6 7 2 0 5 .6 7 2 0 1 .0 6 9 7 1.0491 1.0491 1.0491 9.6358 9.4826 9.4826 9.4826 0.8125 0.7997 0.8060 266.0 254.0 l 1! 1I 251.4 266.0 254.0 5 .0 2 9 5 5 .0 2 9 5 5.0295 5 .5 2 3 2 5 .5 2 3 2 5 .5 2 3 2 1.0820 1.0820 1.0820 9.5708 9.5708 9.5708 0.8007 0.8082 0.8146 28.1 31.1 28.6 48.1 45.5 48.0 40.8 40.9 40.2 50.7 53.4 50.5 36.2 35.8 37.5 37.3 37.3 37.0 117 123 115 39.5 34.7 34.3 32.3 < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOQ < LLOO < LLOQ < LLOQ < LLOO < LLOQ < LLOQ 12.5 12.9 12.8 13.0 13.6 12.9 29.3 33.8 12.5 13.1 LLOQ on C olum n, ng/m L 51 .1 5 51 .1 5 51.15 51.15 51.15 51 .1 5 51.15 51 .1 5 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51 .1 5 51 .1 5 51 15 51.15 25.58 25.58 25.58 25 .5 B 25 .5 8 25.58 25 .5 8 25 .5 8 25.58 25 .5 8 25.58 25.58 25.58 25 .5 8 25.58 25.58 25.58 25.58 25.58 25.58 25.58 25.58 25.58 25.58 51.15 51.15 51.15 51 .1 5 51.15 51.15 51.15 51.15 D ilu tio n Factor OnC olum n 1 1 1 2 2 2 2 2 2 2 2 2 2 2 2 2 4 44 2 2 2 2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 1 1 1 1 1 1 1 1 1 LLOQ On C olum n, C orrecte d for D ilu tion Factor 51.15 51.15 51.15 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 102.3 204.6 204.6 204.6 102.3 102.3 102.3 102.3 102.3 102.3 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 5.116 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 L L O Q in M atrix, u g /g 3.11 3.11 3.12 3.00 2.99 2.90 2.90 2.91 3.29 3.28 3.26 2.84 2.83 2.86 2.98 2.97 2.99 3.02 3.02 3.01 3.12 3.13 2.97 2.99 2.97 1.20 1.17 1.49 1.46 1.34 1.35 1.32 1.34 1.35 1.15 1.20 1.1 1 1.11 0.976 0.970 0.967 2.60 2.60 2.55 2.64 2.76 2.64 2.61 2.59 ug/g 3.12 3.01 2.90 3.28 2.98 3.02 1.22 1.35 1.31 1.40 0.971 2.69 2.62 X O m 0 W 0 m go or CD CP D QJ_ 1 O t*. -- CD O > X CL 0 Z3 CD wC zs 7 3 CD !3 cr ~ CD 0 H T c CD V) Contains No CBI K> Os CD a c 3 ct* CD L /l 4^ Os Page 88 of 227 3M Environmental Laboratory Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Quail Eggs, Lots B and fi Sam ple Name ; CMC/M LA/M EE ; CMC/M LA/M EE f; CMC/M LA/M EE I; CMC/M LA/M EE D1209047.D D1209048.D D1209049. D D1209058.D D1209060.D 0 1 2 0 9 0 6 1 .D D1209070.D D1209071.D D1209073-D D1209074.D D1209075.D D1209076.D 72741-1 72741-i 727417274272742-2 72742-3 72744-1 72744-2 72744-3 72745-1 72745-2 72745-3 72746-1 72746-2 7274672747-1 72747-2 72747-3 k CMC/MLA/MEE k CMC/MLA/MEE uu pum u ose uroup; I k CMC/MLA/MEE 100 ppm Dose Group; ! k CMC/MLA/MEE 100 ppm Dose Group; i __10:0 ppm Dose Group: I k CMC/MLA/MEE k CMC/MLA/MEE 3010 ppm Dose Group; 'Egg; 0 2 Dec 04~; C M C /M LA /M E E 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE _300griyDoseG roug^Egg; 02 Dec 04: CMC/MLA/MEE 300 ppm Dose Group; l 300 ppm Dose Group; E _300_gm_Dojy|:G ro u g y i; CMC/MLA/MEE k CMC/MLA/MEE >; CMC/M LA/M EE 300 ppm Dose Group; E 300 ppm Dose Group; E 300 ppm Dose Group: ( k CMC/MLA/MEE ; CMC/M LA/M EE CMC/MLA/MEE 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE 300 D om P o s e G ro u ^ Egg; 02 Dec 04: CMC/MLA/MEE 300 ppm C 300 ppm C _3003_ggm__C 300 ppm C 300 ppm C _300_m m C k CMC/MLA/MEE k CMC/MLA/MEE n Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE n Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE n Dose Group: Egg; 02 Dec 04: CMC/MLA/MEE D1215098.D D1215099.D D1215100.D 72751-2 72751-3 D1215106.D D1215107.D D1215108.D D1215109.D D1215113.D D1215114.D I; CMC /M LA/M EE k CMC/MLA/MEE 900 ppm i; CMC/MLA/MEE 900 ppm Dose Group; [ 900 Dose Group; i 900 ppm C 900 ppm C _ ^ 0 0 _ggrnC i; CMC/M LA/M EE k CMC/MLA/MEE i; CM C /M LA/M EE 900 ppm C 900 ppm C __900j)gm C >; Egg; 02 Dec 04; CM C /M LA/M EE j; Egg; 02 Dec 04; CMC/M LA/M EE >; Egg; 02 Dec 04; CMC/M LA/M EE n Dose Group; E n Dose Group; E n Dose Group; I k CMC/MLA/MEE k CMC/MLA/MEE -; CMC/M LA/M EE n Dose Group; n Dose Group; I n Dose Group; I k CMC/MLA/MEE i; CMC/M LA/M EE k CMC/MLA/MEE >; Egg; 02 Dec 04; CMC/M LA/M EE ; Egg; 02 Dec 04; CMC/M LA/M EE i; 02 Dec 04; CMC/M LA/M EE 900 ppm Dose Group; [ 900 ppm Dose Group; [ 900 ppm D ose Group; k CMC/MLA/MEE k CMC/MLA/MEE k CMC/MLA/MEE On- C o lu m n D ilu tio n Factor "i Concentr W eight aton of o f Egg PFBS in (in Extract, hom oge ng/m L nate), 'vveignr o f Egg W eight W eight H om oge o f ACN o f W ater nate(in (in (in s o lv e n t so lv e n t hom oge p re cip ita p re cip ita t nate). te), te), g Concentration of PFBS in Original Egg Sample, ug/g Average, u g /g 9.9761 9.9761 9.9761 487.6 430.1 436.0 350.1 337.7 350.0 612.2 665.3 596.0 ~ 586.3 588.8 564.1 6.3395 6.3395 6.3395 5.3769 5.3789 5.3789 1.1076 1.1076 1.1076 0.7970 0 .7 9 1 6 0.8038 0 .8 0 4 9 0 .7 9 9 0 0 .7 9 4 6 5.9287 5.9287 5.9267 0.9626 0.9626 0,9626 9.4078 9.4078 9.4078 627.3 642.3 619.4 643.6 589.1 576.9 533.9 553.6 532.2 2509 2569 2478 1068 1107 1064 1287 1255 1256 5.0125 5.0125 50125 1.1070 1 .1 0 7 0 1.1070 0 ,7 9 7 9 0 .7 8 8 3 0,7637 LLOQ on Colum n, ng/m L 51 .1 5 51.15 51.15 51.15 51.15 51 .1 5 51 .1 5 51 .1 5 51 .1 5 51 .1 5 51.15 51.15 51 .1 5 51.15 51.15 51.15 51.15 51.15 51 .1 5 51 .1 5 51.15 51.15 51.15 51.15 51.15 51.15 51 .1 5 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51.15 51 .1 5 51.15 51.15 51 .1 5 51.15 51.15 51 .1 5 51.15 51 .1 5 51 .1 5 51 .1 5 51 .1 5 51 .1 5 51.15 51.15 51.15 51.15 LLOQ On D ilu tio n Factor OnC olum n C olum n, C o rrecte d for D ilution Factor LLO Q in M atrix, ug/g 51 .1 5 51 .1 5 51 .1 5 51 15 51 .1 5 51 .1 5 51 .1 5 51 .1 5 51 .1 5 2.07 2.08 2.05 2.20 2.19 2.20 2.06 2.06 2.08 51.15 2.85 51 .1 5 51.15 2.86 2.85 51.15 2.21 51.15 51.15 2.27 2.23 51 .1 5 2.48 51 .1 5 51 .1 5 2.50 2.46 102.3 2.09 102.3 2.10 2 102.3 2.21 1 51.15 2.36 1 51.15 2.38 1 51.15 2.39 1 51 .1 5 2.27 1 51 .1 5 2.26 1 51.15 2.27 1 51 .1 5 2.56 1 51 .1 5 2.56 1 51.15 2.55 1 51.15 3.01 1 51.15 3.02 1 51.15 3.00 1 51.15 o no 1 51 .1 5 2.86 1 51.15 2.86 2 102.3 2.25 2 102.3 2.27 2 102.3 2.27 4 204.6 2.88 4 204.6 2.89 4 204.6 2.87 4 204.6 2.51 4 204.6 2.48 4 204.6 2.51 4 204.6 2.54 4 204.6 2.53 4 204.6 2.51 2 102.3 2.46 2 102.3 2.46 2 102.3 2.55 2 102.3 2.45 2 102.3 2.48 2 102.3 2.56 2 102.3 2.46 2 102.3 2.43 2 102.3 2.56 4 204.6 2.19 4 204.6 2.18 4 204.6 2.20 A v e ra g e , u g /g 2.07 2.49 Contains No CBI CD Ut 4^ Os Extracted Serum C alibration File_____________ S am ple Nam e______________ Sam ple Info D0902016.D D0902017.D D0902018.D D0902019.D D0902020.D D0902021.D D0902022.D D0902023.D D0902024.D D0902025.D D0907016.D D0907017.D D0907018.D D0907019.D D0907020.D D0907021.D D0907022.D D0907023.D D0907024.D D0907025.D D0908016.D D0908017.D D0908018.D D0908019.D D0908020.D D0908021 .D D0908022.D D0908023.D D0908024.D D0908025.D CD CD O 040901-5ppb 040901-10ppb 040901-25ppb 040901-50ppb 040901-75ppb 040901-100ppb 040901-250ppb 040901-500ppb 040901-750ppb 040901-1000ppb 040901-5ppb 040901-10ppb 040901-25ppb 040901-50ppb 040901-75ppb 040901-100ppb 040901-250ppb 040901-500ppb 040901-750ppb 040901-1000ppb 040901-5ppb 040901-10ppb 040901-25ppb 040901-50ppb 040901-75ppb 040901-100ppb 040901-250ppb 040901-500ppb 040901-750ppb 040901-1000ppb EC-040901; 5 ppb extracted curve EC-040901; 10 ppb extracted curve EC-040901; 25 ppb extracted curve EC-040901; 50 ppb extracted curve EC-040901; 75 ppb extracted curve EC-040901; 100 ppb extracted curve EC-040901; 250 ppb extracted cuive EC-040901; 500 ppb extracted cu ve EC-040901; 750 ppb extracted cuve EC-040901; 1000 ppb extracted cu v e EC-040901; 5 ppb extracted cu ve EC-040901; 10 ppb extracted c u v e EC-040901; 25 ppb extracted cu ve EC-040901; 50 ppb extracted cu ve EC-040901; 75 ppb extracted cu ve EC-040901; 100 ppb extracted cu ve EC-040901; 250 ppb extracted cu ve EC-040901; 500 ppb extracted cu ve EC-040901; 750 ppb extracted c u v e EC-040901; 1000 ppb extracted c u v e EC-040901; 5 ppb extracted curve EC-040901; 10 ppb extracted curve EC-040901; 25 ppb extracted curve EC-040901; 50 ppb extracted curve EC-040901; 75 ppb extracted curve EC-040901; 100 ppb extracted curve EC-040901; 250 ppb extracted curve EC-040901; 500 ppb extracted curve EC-040901; 750 ppb extracted cuive EC-040901; 1000 ppb extracted curve Page 1 of 3 Compound Name RT Area Amount, ng/mi. Recovery, % PFB S (299) PFB S (299) PFB S (299) PFBS (299) P FB S (299) PFB S (299) PFB S (299) PFBS (299) PFB S (299) PFB S (299) 1.915 1.922 1.922 1.915 1.929 1.929 1.922 1.929 1 .9 2 2 1.901 164156 316861 741202 1384835 1912327 2703477 6042810 10004778 13129533 15905851 5 .2 5 8 1 0 .8 9 2 6 .7 3 5 1 .3 2 7 2 .0 2 1 0 4 .1 257 .0 5 1 1 .1 >ULOQ >ULOQ 103% 106% 104% 100% 94% 102% 100% 100% "T c TC rc c c c c P F B S (299) P F B S (299) P FB S (299) P FB S (299) P FB S (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFB S (299) PFBS (299) PFB S (299) PFB S (299) PFBS (299) P F B S (299) P F B S (299) P F B S (299) P FB S (299) PFB S (299) PFBS (299) 3 .2 7 9 3 .2 7 9 3 .2 7 9 3.279 3.28 3.279 3 .2 7 9 3 .2 7 9 3 .2 7 9 3 .2 7 9 244112 465475 1093039 1983810 2706774 3840497 8365781 13726732 18090283 22046841 3.28 3.279 3 .2 7 9 3 .2 7 9 3 .2 7 9 3 .2 7 9 3.28 3.279 3 .2 7 9 3 .2 7 9 202099 382592 905873 1652395 2291043 3206354 6997586 11563346 15318841 18769611 4 .5 1 7 10.37 2 7 .1 9 5 1 .6 8 72.11 1 0 5 .3 255 .8 5 1 1 .4 > ULOQ > ULOQ 4 .4 5 8 1 0 .1 8 2 6 .9 7 5 1 .5 3 7 3 .1 4 1 0 5 .2 255 .4 5 1 1 .6 > ULOQ > ULOQ 88% 101% 106% 101% 94% 103% 100% 3" Q) 100% CD > 3 CD 1 o2 87% 99% 105% o it 5T : a 101% ) c 95% oc 3C 103% OT100% 3 > 100% CO L 3' 2! O r 5 =r 3? CD J3 ss =! S' W <_ w 2! CD S ' to voc> Project Number 454-116 Contains No CBI F ile D0910016.D D0910017.D D0910018.D D0910019.D D0910020.D D0910021.D D0910022.D D0910023.D D0910024.D D0910025.D D0914016.D D0914017.D D0914018.D D0914019.D D0914020.D D0914021.D D0914022.D D0914023.D D0914024.D D0914025.D D0917016.D D0917017.D D0917018.D D0917019.D D0917020.D D0917021.D D0917022.D D0917023.D D0917024.D D0917025.D D0917105.D D0917106.D D0917107.D D0917108.D D0917109.D D0917110.D D0917111.D D0917112.D D0917113.D D0917114.D S am p le N a m e 040901 -5ppb 040901 -10ppb 040901 25ppb 040901 50ppb 040901 75ppb 040901 100ppb 040901. 250ppb 040901. 500ppb 040901750ppb 040901 IQOOppb 040901 5ppb 040901 -10ppb 040901 25ppb 040901 50ppb 040901 75ppb 040901 100ppb 040901250ppb 040901500ppb 040901750ppb 040901- IQOOppb EC-040901 5ppb EC-040901 10ppb EC-040901 -25ppb EC-040901 -50ppb EC-040901 75ppb EC-040901 100ppb EC-040901 250ppb EC-040901 -500ppb EC-040901 -750ppb EC040901- IQOOppb EC-040901 -5ppb EC-040901 10ppb EC-040901 25ppb EC-040901 50ppb EC-040901 -75ppb EC-040901 - 100ppb EC-040901 250ppb EC-040901 -500ppb EC-040901 -750ppb EC040901 IQOOppb E xtracted S e ru m C alihratinn S a m p le Info EC-040901 5 ppb extracted curve EC-040901; 10 ppb extracted curve EC-040901; 25 ppb extracted curve EC-040901; 50 ppb extracted cuive EC-040901; 75 ppb extracted curve EC-040901; 100 ppb extracted curve EC-040901; 250 ppb extracted curve EC-040901; 500 ppb extracted curve EC-040901; 750 ppb extracted curve EC-040901 1000 ppb extracted curve EC-040901 5 ppb extracted curve EC-040901; 10 ppb extracted curve EC-040901; 25 ppb extracted curve EC-040901; 50 ppb extracted curve EC-040901; 75 ppb extracted curve EC-040901 ; 100 ppb extracted curve EC-040901; 250 ppb extracted curve EC-040901; 500 ppb extracted curve EC-040901; 750 ppb extracted curve EC-040901; 1000 ppb extracted curve Extracted Curve; 5ppb; Serum; 01 Sept 04 Extracted Curve; 10ppb; Serum; 01 Sept 04 Extracted Curve; 25ppb; Serum; 01 Sept 04 Extracted Curve; 50ppb; Serum; 01 Sept 04 Extracted Curve; 75ppb; Serum; 01 Sept 04 Extracted Curve; 100ppb; Serum; 01 Sept 04 Extracted Curve; 250ppb; Serum; 01 Sept 04 Extracted Curve; 500ppb; Serum; 01 Sept 04 Extracted Curve; 750ppb; Serum; 01 Sept 04 Extracted Curve IQOOppb; Serum; 01 Sept 04 Extracted Cuive; 5ppb; Serum; 01 Sept 04 Extracted Curve; 10ppb; Serum; 01 Sept 04 Extracted Curve; 25ppb; Serum; 01 Sept 04 Extracted Curve; SOppb; Serum; 01 Sept 04 Extracted Curve; 75ppb; Serum; 01 Sept 04 Extracted Curve; 10Oppb; Serum; 01 Sept 04 Extracted Curve; 250ppb; Serum; 01 Sept 04 Extracted Curve; 500ppb; Serum; 01 Sept 04 Extracted Curve; 750ppb; Serum; 01 Sept 04 Extracted Curve; IQOOppb; Serum; 01 Sept 04 Page 2 of 3 Compound Name RT PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) 130592 244823 584310 1074139 1501286 2111367 4652274 7826002 10298002 12673814 3.258 3.258 3.258 3.258 3.258 3.259 3.258 3.258 3.259 3.258 108064 203150 484794 886733 1246916 1755168 3860219 6411109 8514373 10501469 3.259 3.258 3.258 3.258 3.259 3.259 3.259 3.258 3.258 3.258 74738 143950 351259 665138 936286 1319943 2955185 4955921 6568909 8141449 3.272 3.273 3.273 3.272 3.272 3.273 3.272 3.273 3.272 3.272 68746 135430 330925 610635 854927 1214751 2714174 4533966 5992791 7353470 < LLOQ < LLOQ 23.95 50.04 73.27 107.3 262.2 501.9 772.5 > ULOQ < LLOQ < LLOQ 23 23 49.39 73.34 108.0 264 7 498.5 774 2 > ULOQ < LLOQ < LLOQ < LLOQ 44.32 69 92 106 8 274.7 511 3 742.7 1040 5.238 10.72 27.00 50.81 72.15 104.6 256.7 511.2 ULOQ > ULOQ 91% 97% 96% 106% 103% 97% 101% 87% 91% 104% 107% 100% 97% 102% 3< T| <I 102% 105% 106% 99% 94% 102% 100% 100% S $ Contains No CBI NJ \D \o <3. 2 c <y 4^ L /i i Os 3M Environmental Laboratory E xtracted Seru m C alibration F ile S am p le N am e S am p le Info Compound Name RT A rea A m ou nt, n g /m L R ecovery, % D0921016.D D0921017.D D0921018.D D0921019.D D0921020.D D0921021.D D0921022.D D0921023.D D0921024.D D0921025.D EC-040901 -5ppb Extracted Curve; 5ppb; Serum; 01 Sept 04 EC-040901 -10ppb Extracted Curve; 10ppb; Serum; 01 Sept 04 EC-040901 -25ppb Extracted Curve; 25ppb; Serum; 01 Sept 04 EC-040901 -50ppb Extracted Curve; 50ppb; Serum; 01 Sept 04 EC-040901 -75ppb Extracted Curve; 75ppb; Serum; 01 Sept 04 EC-040901 -100ppb Extracted Curve; 100ppb; Serum; 01 Sept 04 EC-040901 -250ppb Extracted Curve; 250ppb; Serum; 01 Sept 04 EC-040901 -500ppb Extracted Curve; 500ppb; Serum; 01 Sept 04 EC-040901 -750ppb Extracted Curve; 750ppb; Serum; 01 Sept 04 EC040901- IQOOppb_________ Extracted Curve; IQOOppb; Serum; 01 Sept 04 D0922016.D D0922017.D D0922018.D D0922019.D D0922020.D D0922021.D D0922022.D D0922023.D D0922024.D D0922025.D EC-040901-5ppb Extracted Curve; 5ppb; Serum; 01 Sept 04 EC-040901-10ppb Extracted Curve; 10ppb; Serum; 01 Sept 04 EC-040901-25ppb Extracted Curve; 25ppb; Serum; 01 Sept 04 EC-040901-50ppb Extracted Curve; 50ppb; Serum; 01 Sept 04 EC-040901-75ppb Extracted Curve; 75ppb; Serum; 01 Sept 04 EC-040901-100ppb Extracted Cuive; 100ppb; Serum; 01 Sept 04 EC-040901-250ppb Extracted Curve; 250ppb; Serum; 01 Sept 04 EC-040901-500ppb Extracted Curve; 500ppb; Serum; 01 Sept 04 EC-040901-750ppb Extracted Curve; 750ppb; Serum; 01 Sept 04 EC040901-IQOOppb_________ Extracted Curve; IQOOppb; Serum; 01 Sept 04 Results Summary PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299). 3.258 3.258 3.259 3.258 3.258 3.258 3.258 3.258 3.258 3.252 3.259 3.258 3.258 3.258 3.258 3.258 3.258 3.258 3.258 3.258 47780 91783 220434 407707 570808 802705 1827741 3082655 4119449 5103755 54465 103369 247708 451778 633838 892808 2003093 3386705 4465383 5416611 EC-040901-5ppb EC-040901-10ppb EC-040901-25ppb EC-040901-50ppb EC-040901-75ppb EC-040901-100ppb EC-040901-250ppb EC-040901-500ppb E C -0 4 0 9 0 1-7 5 0 p p b EC040901-1000ppb %RSD Over 9 Calibrations (Listed Above) Extracted Curve; 5ppb; Serum; 01 Sept 04 q n% Extracted Curve; 10ppb; Serum; 01 Sept 04 Extracted Curve; 25ppb; Serum; 01 Sept 04 Extracted Curve; 50ppb; Serum; 01 Sept 04 Extracted Curve; 75ppb; Serum; 01 Sept 04 Extracted Curve; 100ppb; Serum; 01 Sept 04 Extracted Curve; 250ppb; Serum; 01 Sept 04 Extracted Curve; 500ppb; Serum; 01 Sept 04 Extracted Curve; 750ppb; Serum; 01 Sept 04 Extracted Cuive; 1000ppb; Serum; 01 Sect 04 3 1% 8 .8 % 4 7% 1 4% 1 4% 2.7% 1 1% 2 1% 0.015% < LLOQ < LLOQ 24.62 50.15 72.82 105.8 264.3 500.7 772.6 > ULOQ < LLOQ < LLOQ 21.18 48.05 72.44 107.9 270.9 510.1 745.7 1040 96% 98% 95% 103% 103% 98% 101% Tc C c cc 83% 94% 94% 105% 106% 100% 97% 102% ? - 1 2 8> & g9 D ' 11 =s: =5 C o ; Tl C CD c 3Z ;: 3- u> o o Page 91 of 227 Project Number 454-116 Page 3 of 3 Contains No CBI Extracted Liver C alibration F ile ____________S a m p le N a m e S a m p le Info Compound Name CO 2 D1004014.D EC-041004-5ppb Extracted Curve; 5ppb; Liver; 4 Oct 04 PFBS (299) 3 D1004015.D EC-041004-1 Oppb Extracted Curve; 10ppb; Liver; 4 Oct 04 PFBS (299) D1004016.D EC-041004-25ppb Extracted Curve; 25ppb; Liver; 4 Oct 04 PFBS (299) 3 D1004017.D EC-041004-50ppb Extracted Curve; 50ppb; Liver; 4 Oct 04 PFBS (299) O D1004018.D EC-041004-75ppb Extracted Curve; 75ppb; Liver; 4 Oct 04 PFBS (299) Si D1004019.D EC-041004-1 OOppb Extracted Curve; 100ppb; Liver; 4 Oct 04 PFBS (299) i-- 05 D1004020.D EC-041004-250ppb Extracted Curve; 250ppb; Liver; 4 Oct 04 PFBS (299) CT O D1004021.D EC-041004-500ppb Extracted Curve; 500ppb; Liver; 4 Oct 04 PFBS (299) J D1004022.D EC-041004-750ppb Extracted Curve; 750ppb; Liver; 4 Oct 04 PFBS (299) < D1004023.D EC-041004-1 ppm Extracted Curve; 1ppm; Liver; 4 Oct 04 PFBS (299) D1008016.D D 1 008017.D D1008018.D D1008019.D D1008020.D D1008021.D D1008022.D D1008023.D D 1 008024.D D1008025.D EC-041004-5ppb EC-041004-1 Oppb EC-041004-25ppb EC-041004-50ppb EC-041004-75ppb EC-041004-1 OOppb EC-041004-250ppb EC-041004-500ppb EC-041004-750ppb EC041004-1 OOOppb Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 10Oppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 750ppb; Liver; 04 Oct 04 Extracted Curve; 10OOppb; Liver; 04 Oct 04 PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) D1009016.D D1009017.D D1009018.D D1009019.D D1009020.D D1009021 ,D D1009022.D D1009023.D D1009024.D D1009025.D EC-041004-5ppb EC-041004-1 Oppb EC-041004-25ppb EC-041004-50ppb EC-041004-75ppb EC-041004-1 OOppb EC-041004-250ppb EC-041004-500ppb EC-041004-750ppb EC041004-1 OOOppb Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 10Oppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 750ppb; Liver; 04 Oct 04 Extracted Curve; 10OOppb; Liver; 04 Oct 04 PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) D1014016.D D1014017.D D1014018.D D1014019.D D1014020.D D1014021.D D1014022.D D1014023.D D1014024.D D1014025.D EC-041004-5ppb EC-041004-1 Oppb EC-041004-25ppb EC-041004-50ppb EC-041004-75ppb EC-041004-1 OOppb EC-041004-250ppb EC-041004-500ppb EC-041004-750ppb EC041004-1 OOOppb Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 10Oppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 75Qppb; Liver; 04 Oct 04 Extracted Curve; 10OOppb; Liver; 04 Oct 04 PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) Page 1 of 3 Amount PFBS, RT Area ng/mL Recovery, % 3.265 3.335 3.265 3.265 3.265 3.272 3.265 3.265 3.265 3.273 43498 86532 237796 478465 708799 806519 1924274 3170525 4757997 6070909 < LLOQ < LLOQ 23.90 53.27 82.37 95.03 257.3 511.2 > ULOQ >ULOQ 3.439 3.446 3.439 3.439 3.439 3.447 3.439 3.447 3.446 3.439 63549 140546 372023 716092 978098 1177030 2602783 3985908 5681577 6937696 5.358 10.08 25.25 51.51 76.65 102.2 > ULOQ > ULOQ > ULOQ > ULOQ 3.447 3.447 3.446 3.44 3.446 3.446 3.446 3.44 3.439 3.439 61758 134849 357898 692330 937737 1137124 2518554 3911666 5506036 6783621 5.357 10.03 25.25 51.81 76.17 102.5 > ULOQ > ULOQ > ULOQ > ULOQ 3.446 3.447 3.447 3.447 3.446 3.446 3.446 3.446 3.446 3.439 17953 41902 110637 220423 302345 378902 898613 1470759 2136736 2698945 5.223 10.18 25.17 52.11 75.77 102.7 > ULOQ > ULOQ > ULOQ > ULOQ 93% 104% 107% 93% 101% 100% 105% 99% 99% 101% 100% 100% 105% 98% 99% 101% 99% 100% 102% 100% 98% 102% 99% 100% 73 3. m -> ; 73 U) O Project Number 454-116 Contains No CBI 3M Environmental Laboratory Extracted Liver C alibration File S am ple Nam e Sam ple Info Compound Name D1015016.D D1015017.D D1015018.D D1015019.D D1015020.D D1015021.D D1015022.D D1015023.D D1015024.D D1015025.D D1015026.D D1015027.D D1015028.D EC-041012d-2.5 EC-041012d-5 EC-041012d-10 EC-041012d-25 EC-041012d-50 EC-041012d-75 EC-041012d-100 EC-041012d-175 EC-041012d-250 EC-041012d-375 EC-041012d-500 EC-041012d-750 EC041012d-1000 Extracted Curve;2.5ppb; Liver; 12 Oct 04 Extracted Curve; 5ppb; Liver; 12 Oct 04 Extracted Curve; 10ppb; Liver; 12 Oct 04 Extracted Curve; 25ppb; Liver; 12 Oct 04 Extracted Curve; 50ppb; Liver; 12 Oct 04 Extracted Curve; 75ppb; Liver; 12 Oct 04 Extracted Curve; 10Oppb; Liver; 12 Oct 04 Extracted Curve; 175ppb; Liver; 12 Oct 04 Extracted Curve; 250ppb; Liver; 12 Oct 04 Extracted Curve; 375ppb; Liver; 12 Oct 04 Extracted Curve; 500ppb; Liver; 12 Oct 04 Extracted Curve; 750ppb; Liver; 12 Oct 04 Extracted Curve; 10OOppb; Liver; 12 Oct 04 PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) D1018016.D D1018017.D D1018018.D D1018019.D D1018020.D D1018021.D D1018022.D D1018023.D D1018024.D D1018025.D D1018026.D D1018027.D D1018028.D EC-041012d-2.5 EC-041012d-5 EC-041012d-10 EC-041012d-25 EC-041012d-50 EC-041012d-75 EC-041012d-100 EC-041012d-175 EC-041012d-250 EC-041012d-375 EC-041012d-500 EC-041012d-750 ECQ41012d-1000 Extracted Curve;2.5ppb; Liver; 04 Oct 04 Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 10Oppb; Liver; 04 Oct 04 Extracted Curve; 175ppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 375ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 750ppb; Liver; 04 Oct 04 Extracted Curve; 10OOppb; Liver; 04 Oct 04 PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) D1018115.D EC-041012d-2.5 Extracted Curve;2.5ppb; Liver; 04 Oct 04 PFBS (299) D1018116.D EC-041012d-5 Extracted Curve; 5ppb; Liver; 04 Oct 04 PFBS (299) D1018117.D EC-041012d-10 Extracted Curve; 10ppb; Liver; 04 Oct 04 PFBS (299) D1018118.D EC-041012d-25 Extracted Curve; 25ppb; Liver; 04 Oct 04 PFBS (299) CD D1018119.D EC-041012d-50 Extracted Curve; 50ppb; Liver; 04 Oct 04 PFBS (299) CD D1018120.D EC-041012d-75 Extracted Curve; 75ppb; Liver; 04 Oct 04 PFBS (299) D1018121.D EC-041012d-100 Extracted Curve; 100ppb; Liver; 04 Oct 04 PFBS (299) D1018122.D EC-041012d-175 Extracted Curve; 175ppb; Liver; 04 Oct 04 PFBS (299) ro -> i D1018123.D EC-041012d-250 Extracted Curve; 250ppb; Liver; 04 Oct 04 PFBS (299) D1018124-D EC-041012d-375 Extracted Curve; 375ppb; Liver; 04 Oct 04 PFBS (299) D1018125.D EC-041012d-500 Extracted Curve; 500ppb; Liver; 04 Oct 04 PFBS (299) D1018126.D EC-041012d-750 Extracted Curve; 750ppb; Liver; 04 Oct 04 PFBS (299) D1018127.D EC041012d-1000 Extracted Curve; 10OOppb; Liver; 04 Oct 04 PFBS (299) |D1021016.D EC-041012d-2.5 Extracted Curve; 2.5ppb; Liver; 04 Oct 04 PFBS (299) Page 2 of 3 Amount PFBS, RT_____ Area_____ ng/mL Recovery, % 3.454 3.453 3.454 3.454 3.453 3.454 3.453 3.454 3.453 3.447 3.447 3.447 3.446 33986 66347 101188 219291 451082 606824 810741 1347993 1852981 2348046 3216160 4141918 5145502 < LLOQ < LLOQ < LLOQ 20.72 52.19 73.79 1 0 2 .7 182.4 2 6 2 .9 3 4 8 .6 5 2 1 .6 >ULOQ > ULOQ 83% 102% 99% 100% 103% 105% 94% 102% 3.454 3.454 3.454 3.454 3.454 3.454 3.454 3.454 3.454 3.454 3.453 3.454 3.454 32118 65488 100601 217705 449938 602456 799451 1334009 1833669 2297312 3152930 4051681 5033558 < LLOQ < LLOQ < LLOQ 20.47 52.41 73.86 102.2 1 8 2 .8 2 6 4 .2 3 4 6 .4 5 2 2 .6 > ULOQ > ULOQ 82% 102% 99% 100% 103% 105% 93% 102% 3.454 3.454 3.447 3.454 3.454 3.454 3.447 3.454 3.447 3.447 3.447 3.447 3.447 13361 25921 39174 86164 181055 250217 342790 588622 838048 1078273 1579022 2041229 2583329 2.472 10.06 23.95 52.31 73.25 101.7 179.3 2 6 1 .7 345.1 5 3 4 .5 7 3 5 .4 1025 97% Point excluded 98% 95% 102% 98% 99% 101% 104% 93% 105% 99% 100% 3.46 29280 < LLOQ 7J CD T3 O 8. O > q* U) o Project Number 454-116 Contains No CBI Page 94 of 22 / 3M Environmental Laboratory Report E03-0583 File D1021017.D D1021018.D D1021019.D D1021020.D D1021021.D D1021022.D D1021023.D D1021024.D D1021025.D D1021026.D D1021027.D D1021028.D Sam ple Name EC-041012d-5 EC-041012d-10 EC-041012d-25 EC-041012d-50 EC-041012d-75 EC-041012d-100 EC-041012d-175 EC-041012d-250 EC-041012d-375 EC-041012d-500 EC-041012d-750 EC041012d-1000 Results Summary E xtracted Liver C alibration S am ple Info Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 10Oppb; Liver; 04 Oct 04 Extracted Curve; 175ppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 375ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 750ppb; Liver; 04 Oct 04 Extracted Curve; IQOOppb; Liver; 04 Oct 04 Compound Name PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) PFBS (299) RT 3.461 3.461 3.461 3.461 3.461 3.46 3.461 3.461 3.461 3.461 3.461 3.461 Area 57454 86951 187133 382908 520081 691379 1142466 1550926 1954983 2688300 3464107 4292500 Amount PFBS, ng/mL < LLOQ < LLOQ < LLOQ 46.09 70.48 101.5 186.1 267.0 351.9 521.5 733.1 1030 Recovery, % 90% 95% 99% 105% 106% 95% 102% 99% 101% EC-041004-5ppb EC-041004-10ppb EC-041004-25ppb EC-041004-50ppb EC-041004-75ppb EC-041004-100ppb EC-041004- 250ppb EC-041004- 500ppb EC-041004--750ppb EC-041004- 1ppm EC-041012d-2.5 EC-041012d-5 EC-041012d-10 EC-041012d-25 EC-041012d-50 EC-041012d-75 EC-041012d-100 EC-041012d-175 EC-041012d-250 EC-041012d-375 EC-041012d-500 EC-041012d-750 EC041012d-1000 Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 100ppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 750ppb; Liver; 04 Oct 04 Extracted Curve; 1000ppb; Liver; 04 Oct 04 Extracted Curve; 2.5ppb; Liver; 04 Oct 04 Extracted Curve; 5ppb; Liver; 04 Oct 04 Extracted Curve; 10ppb; Liver; 04 Oct 04 Extracted Curve; 25ppb; Liver; 04 Oct 04 Extracted Curve; 50ppb; Liver; 04 Oct 04 Extracted Curve; 75ppb; Liver; 04 Oct 04 Extracted Curve; 100ppb; Liver; 04 Oct 04 Extracted Curve; 175ppb; Liver; 04 Oct 04 Extracted Curve; 250ppb; Liver; 04 Oct 04 Extracted Curve; 375ppb; Liver; 04 Oct 04 Extracted Curve; 500ppb; Liver; 04 Oct 04 Extracted Curve; 750ppb; Liver; 04 Oct 04 Extracted Curve; IQOOppb; Liver; 04 Oct 04 < LLOQ < LLOQ 23.90 53^7 82.37 95 03 257 3 511.2 > ULOQ > ULOQ < LLOQ < |_LOQ < LLOQ 20 72 52.19 73 79 102.7 182.4 262 9 348.6 521 6 > ULOQ > ULOQ 5.358 10.08 25.25 51.51 76.65 102.2 > ULOQ > ULOQ > ULOQ > ULOQ 5.357 10.03 25.25 51.81 76.17 102.5 > ULOQ > ULOQ > ULOQ > ULOQ < LLOQ < LLOQ < LLOQ 20.47 52.41 73.86 102.2 182.8 264.2 346.4 522.6 > ULOQ > ULOQ 2.472 10.06 23.95 52.31 73.25 101.7 179.3 261.7 345.1 534.5 735.4 1025 5.223 10.18 25.17 52.11 75.77 102.7 > ULOQ > ULOQ > ULOQ > ULOQ < LLOQ < LLOQ < LLOQ < LLOQ 46.09 70.48 101.5 186.1 267.0 351.9 521.5 733.1 1030 %RSD 1.5% 0.8% 2.7% 1.5% 4.0% 3.7% 8.9% 6.1% 2 .2 % 0 .5 % 1 .5 % 0 .9 % 0 .9 % 1 .2 % 0 .2 % 0 .3 % T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 3 of 3 Contains No CBI c*1t Lb 0\ E x tra c te d E gg C alib ratio n F ile ________ S a m p le N a m e S a m p le In fo CO m D1207016.D EC-041202-10 Extracted Curve; 10ppb; Egg; 02 Dec 04 a C3 D 1207017.D EC-041202-50 Extracted Curve; 50ppb; Egg; 02 Dec 04 3 D12Q7018.D EC-041202-50 Extracted Curve; 50ppb; Egg; 02 Dec 04 03 03 S D 1207019.D EC-041202-75 Extracted Curve; 75ppb; Egg; 02 Dec 04 D 1207020.D EC -041202-100 Extracted Curve; 100ppb; Egg; 02 Dec 04 O03' O D1207021.D EC-041202-175 Extracted Curve; 175ppb; Egg; 02 Dec 04 3 D1207022.D EC-041202-250 Extracted Curve; 250ppb; Egg; 02 Dec 04 5T 3 D 1207023.D EC -041202-375 Extracted Curve; 375ppb; Egg; 02 Dec 04 D 1207024.D EC-041202-500 Extracted Curve; 500ppb; Egg; 02 Dec 04 D 1207025.D EC-041202-750 Extracted Curve; 750ppb; Egg; 02 Dec 04 D 1207026.D E C 041202-1000 Extracted Curve; IQOOppb; Egg; 02 Dec 04 D1208016.D EC-041202-10 Extracted Curve; 10ppb; Egg; 02 Dec 04 D1208017.D EC-041202-50 Extracted Curve; 50ppb; Egg; 02 Dec 04 D12Q8Q18.D EC -041202-50 Extracted Curve; 50ppb; Egg; 02 Dec 04 D1208019.D EC-041202-75 Extracted Curve; 75ppb; Egg; 02 Dec 04 D1208020.D EC -041202-100 Extracted Curve; 100ppb; Egg; 02 Dec 04 D 1208021.D EC-041202-175 Extracted Curve; 175ppb; Egg; 02 Dec 04 D1208022.D EC-041202-250 Extracted Curve; 250ppb; Egg; 02 Dec 04 D1208023.D EC-041202-375 Extracted Curve; 375ppb; Egg; 02 Dec 04 D1208024.D EC -041202-500 Extracted Curve; 500ppb; Egg; 02 Dec 04 D 1208025.D EC -041202-750 Extracted Curve; 750ppb; Egg; 02 Dec 04 D 1208026.D EC 041202-1000 Extracted Curve; IQOOppb; Egg; 02 Dec 04 D1209016.D EC-041202-50 Extracted Curve; 50ppb; Egg; 02 Dec 04 D 1209017.D EC-041202-75 Extracted Curve; 75ppb; Egg; 02 Dec 04 D 1209018.D EC -041202-100 Extracted Curve; 100ppb; Egg; 02 Dec 04 D1209019.D EC-041202-175 Extracted Curve; 175ppb; Egg; 02 Dec 04 D 1209020 .D EC-041202-250 Extracted Curve; 250ppb; Egg; 02 Dec 04 D1209021.D EC-041202-375 Extracted Curve; 375ppb; Egg; 02 Dec 04 D 1209022.D EC-041202-500 Extracted Curve; 500ppb; Egg; 02 Dec 04 D1209023.D EC-041202-750 Extracted Curve; 750ppb; Egg; 02 Dec 04 D1209024.D EC04122-1000 Extracted Curve; 1000ppb; Egg; 02 Dec 04 D1215016.D EC-041202-50 D1215017.D EC-041202-75 D1215018.D EC-041202-100 D1215019.D EC-041202-175 Extracted Curve; 50ppb; Egg; 02 Dec 04 Extracted Curve; 75ppb; Egg; 02 Dec 04 Extracted Cun/e; 100ppb; Egg; 02 Dec 04 Extracted Curve; 175ppb; Egg; 02 Dec 04 Page 1 of 2 Amount, Recovery, RT Area ng/mL % 3.44 3.439 3.439 3.439 3.44 3.44 3.439 3.44 3.439 3.439 3.439 364846 595848 1174019 1648807 2264284 3559585 4918180 7159838 8187975 11187829 13631574 < LLOQ 21.81 50.17 73.97 105.6 175.1 253.4 398.4 474.0 759.1 > ULOQ 3.44 3.433 3.439 3.439 3.439 3.432 3.439 3.439 3.433 3.432 3.432 311298 515324 1002521 1413158 1930812 3065203 4223798 6230573 7172449 9883555 12149887 4.515 16.63 45.94 71.08 103.4 176.6 255.5 404.3 480.8 737.5 1033 3.44 3.44 3.439 3.439 3.44 3.44 3.439 3.439 3.44 845627 1196684 1636757 2623719 3564783 5236990 6043432 8295804 10118338 45.52 70.90 103.3 178.9 255.0 402.5 480.9 738.6 1033 85% 98% 99% 103% 99% 101% 107% 93% 102% 90% 96% 101% 100% 102% 109% 94% 99% 101% 89% 95% 101% 101% 102% 108% 94% 99% 101% 3.481 3.481 3.481 3.482 1002161 1401761 1865902 2837849 41.94 70.76 105.1 180.1 82% 95% 103% 102% O Project Number 454-116 Contains No CBI E x tracted Egg C a lib ra tio n w m F ile S a m p le N a m e S a m p le Info " f 2 0 '0 EC-041202-250 E x tra c te d C u rve ; 2 5 0 p p b ; E gg; 02 D e c 04 Amount, R e c o v e ry , RT Area ng/mL % o 3 3 o D D12152 1.D EC-041202-375 E x tra c te d C urve ; 3 7 5 p p b ; E gg; 0 2 D e c 04 D1215022.D EC-041202-500 Extracted C urve; 500ppb; E g g ' 02 D e c 04 D 215023.D EC-041202-750 E xtra cte d C urve ; 7 5 0 ^ E gg; 02 D e c 04 3 .4 8 2 3.481 3 .4 7 4 3795760 5385045 6106476 2 5 8 .9 4 0 3 .6 4 7 6 .9 103% 108% 93% S' D1215024.D EC041202-1000 E x tra c te d C urve; IQOOppb; E gg; 02 D e c 04 3.481 8193832 7 3 4 .5 99% su 3.474 9769277 1041 102% cr o 5 D1215086.D EC-041202-50 Extracted Curve; 50ppb; Egg; 02 D ec 04 3 .4 0 7 O 3 D1215087.D EC-041202-75 D1215088.D EC-041202-100 D1215089.D EC -041202-175 D1215090.D EC-041202-250 D1215091.D EC-041202-375 D1215092.D EC -041202-500 D1215093.D EC-041202-750 Extracted Curve; 75ppb; Egg; 02 Dec 04 Extracted Curve; 100ppb; Egg; 02Dec 04 Extracted Curve; 175ppb; Egg; 02Dec 04 Extracted Curve; 250ppb; Egg; 02D ec 04 Extracted Curve; 375ppb; Egg; 02Dec 04 Extracted Curve; 500ppb; Egg; 02Dec 04 Extracted Curve; 750ppb; Egg; 02Dec 04 3.467 3.468 3.468 3 467 3 .4 6 7 3 467 3 .4 6 7 943077 1311200 1755813 2678405 3558425 5085658 5796261 4 2 .8 3 7 0 .8 4 105.5 180.4 2 5 6 .4 4 0 1 .9 4 7 7 .2 84% 95% 103% 102% 102% 108% 93% D 1215094.D E C 041202-1000 E xtracted C urve; IQOOppb; E gg; 02 D ec 0 4 3 .4 6 7 7842100 9342717 7 3 9 .9 1035 99% 101% D 1 2 1 5 1 4 5 .D D 1 2 1 5 1 4 6 .D D 1 2 1 5 1 4 7 .D D 121 514 8.D D 1 2 1 5 1 4 9 .D D 1 2 1 5 1 5 0 .D D 1 2 1 5 1 5 1 .D D 1 2 1 5 1 5 2 .D D 1 2 1 5 1 5 3 .D E C -0 4 1 2 0 2 -5 0 EC-041202-75 EC -041202-100 E C -0 4 1 2 0 2 -1 7 5 E C -0 4 1 2 0 2 -2 5 0 E C -041 202 -37 5 E C -0 4 1 2 0 2 -5 0 0 E C -0 4 1 2 0 2 -7 5 0 E C 0 4 1 2 0 2 -1 0 0 0 Extracted Curve; 50ppb; Egg; 02 D ec 04 Extracted Curve; 75ppb; Egg; 02 Dec 04 Extracted Curve; 100ppb; Egg; 02 Dec 04 Extracted Curve; 175ppb; Egg; 02 D ec 04 Extracted Curve; 250ppb; Egg; 02 Dec 04 Extracted Curve; 375ppb; Egg; 02 Dec 04 Extracted Curve; 500ppb; Egg; 02 D ec 04 Extracted Curve; 750ppb; Egg; 02 D ec 04 E xtracted C urve; IQOOppb; E gg; 02 D e c 04 3 .461 3.461 3.461 3.461 3.46 3 46 3.4 6 3 461 3.461 248214 348880 492082 794277 1129776 1727260 2000362 2878879 3527453 5 0 .8 0 7 2 .4 6 103.7 171.8 2 5 0 .7 4 0 2 .5 4 7 7 .6 7 5 5 .5 1022 99% 97% 101% 97% 100% 108% 93% 102% 100% Results Summary "D QJ (Q %RSD Over 6 Calibrations (Listed Above) EC -041202-50 Extracted Curve; 50ppb; Egg; 02 D ec 04 7.9% CD CO EC-041202-75 E xtracted Curve; 75ppb; Egg; 02 D ec 04 1.8% O) O EC -041202-100 Extracted Curve; 100ppb; Egg; 02 D ec 04 1.0% EC -041202-175 E xtracted Curve; 175ppb; Egg; 02 D ec 04 1.9% EC-041202-250 E xtracted Curve; 250ppb; Egg; 02 D ec 04 1.1% EC-041202-375 Extracted Curve; 375ppb; Egg; 02 Dec 04 0.5% EC -041202-500 E xtracted Curve; 500ppb; Egg; 02 D ec 04 0.5% EC-041202-750 E xtracted Curve; 750ppb; Egg; 02 D ec 04 1.4% E C041202-1000 E xtracted Curve; IQOOppb; Egg; 02 D ec 04 0.7% T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Report E03-0583 1 Page 2 of 2 Os Contains No CBI i neoreticai concentration, ng/mL 0.2558 0.5115 0.9974 1.535 2.302 5.115 Area Counts 68175 82118 121253 188329 205039 406370 SUMMARY OUTPUT R e g r e s s io n S ta tis tic s Multiple R 0.9940 R Square 0.9881 Adjusted R Square 0.9851 Standard Error 15197 Observations 6 AN OVA D 0 5 0 1 19_Serum .xls LOD 1 LOQ 0.732 - | 2.20 Page 1 of 1 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Report E03-0583 Contains No CBI CD >1 U/< Os D050119Jiver.xls 1h e o re tic a l c o n c e n tra tio n , n g /m L 0.2558 0.5115 0.9974 1.535 2.302 5.115 A re a C o u n ts 27240 33869 49507 73037 100344 246855 SUMMARY OUTPUT R e g r e s s io n Multiple R R Square Adjusted R Square Standard Error Observations S ta tis tic s 0.9957 0.9914 0.9893 8505 6 AN OVA Regression Residual Total LO D 0.620 LO Q 1.86 SS 3.342E+10 2.893E+08 1 ^71 Cxin MS 3.342E+10 7.233E+07 4.621 E+02 S ig n ific a n c e F 2.770E-05 Page 1 of 1 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS In Northern Bobwhite Tissues Report E03-0583 Contains No CBI a> L /i 4^ Page 99 of 227 3M Environmental Laboratory D050119_Egg.xls 1heoretical concentration, ng/mL 0 .2 5 5 8 0 .9 9 7 4 1 .5 3 5 2 .3 0 2 5 .1 1 5 Area Counts 21169 60742 91911 123258 181415 SUMMARY OUTPUT Regression Statistics M ultiple R 0 .9 6 6 4 R Square 0 .9 3 3 9 A djusted R S quare 0 .9 1 1 9 Standard E rror O bservations 18113 5 ANOVA R egression R esidual Total df LOD 1 .9 1 7 SS 1.391E +10 9.842E +08 1 .4 9 0 F + m MS 1.391E +10 3.281 E +08 LOQ 5 .7 6 42.41 Significance F 7.355E -03 Page 1 of 1 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Report E03-0583 Contains No CBI n i -F I ON -309- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Attachment C: Sample Chromatograms 3M Environmental Laboratory Page 100 of 227 Contains No CBI -310- Project Number 454-116 D a ta F & e b t:E o :V ^ ^ s ta rg e t\T a rg e t\c h e m \d u T c J d f^ .4 e ty I9 # 4 iM )$ $ y W ^ M tW B T O ^ ? W ti )a g e 1 R e p o r t D a t e - 0 4 - J a n - 2 0 0 5 0 6 - 4 5 Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues 3M E n v ir o n m e n t a l L a b o r a t o r y E 0 3 - 0 5 8 3 , Q u a n ta tio n o f PFBS in Q u a il D a ta f i l e : \ \ E t s t a r g e t \ T a r g e t \ c h e m \ d u d e j r . i\ D 0 4 1 0 2 5 . b \ D 1 0 2 5 0 3 2 . D L a b S nip I d : 7 2 8 4 8 - 1 I n j D a te : 2 5 -O C T - 2 0 04 1 8 :2 8 O p e ra to r : m la In s t ID : d u d e jr . i Smp I n f o : 0 p p m D o s e G r o u p ; S e ru m ; 3 0 A u g 0 4 ; M L A /C M C /M E E M is e I n f o : Com m ent : S e ru m , 0 pp m D o s e G ro u p , C o n t r o l S a m p le s M e th o d : \ \ E t s t a r g e t \ t a r g e t \che m \du dej r .i\D 0 4 1 0 2 5 .b \D 0 4 1 0 2 5 .m M e th D a te : 0 4 - J a n - 2 0 0 5 0 6 :4 2 c a r l s o n Q u a n t T y p e : ESTD C a l D a te : 2 5 - O C T - 2 0 04 1 7 :1 5 C al F i l e : D 1025025.D .A l s b o t t l e : 1 4 D i l F a c t o r . 1 00000 In te g ra to r : F a lc o n Com pound S u b l i s t : a l l . s u b T a rg e t V e rs i on: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) Compound Not Detected. 3M Environmental Laboratory Page 101 of 227 Contains No CBI -311 - Project Number 454-116 3M Environmental Laboratory Page 102 of 227 Contains No CBI -312- Project Number 454-116 3M Environmental Laboratory E03-0583, Quantation of PFBS in Quail Data file : \\Etstarget\Target\chem\dudej r.i\D041025.b\Dl025039.D Lab Smp Id: 72849-1 Inj Date : 25-OCT-2004 19:41 Operator : ml a Inst ID: dudejr.i Smp Info : 0 ppm Dose Group; Serum; 30 Aug 04; MLA/CMC/MEE Misc Info : Comment : Serum, 0 ppm Dose Group, Control Samples Method : \\Etstarget\target\chem\dudejr.i\D041025.b\D041025.m Meth Date : 04-Jan-2005 06:42 carlson Quant Type: ESTD Cal Date : 25-OCT-2004 17:15 Cal File: D1025025.D Als bottle: 19 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.475 3.468 0.007 6607 2785.82 27 9 0 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 103 of 227 Contains No CBI -313 - Project Number 454-116 Report EQ3-0583 . . T-7485: A Reproduction Study with the Northern Bobwhite D-ata F ile : N ^ E ts ta rg e tS T a rg e t\c h e m S d u d e jr*iS D 0 4 ^ f^ ^ ^ |^ ^ g ^ ^ g termjna{jon 0f PFBS in Northern Bobwhite Tissues Pag 2 3M Environmental Laboratory Page 104 of 227 Contains No CBI -314- Project Number 454-116 Data FHefc<to^gtst arge t \Targe t\chem\diM4fiA.fie^rISffe1^iirt^l5y, W i <2tiP?^ltv itePage 1 E g p g * 0 4 - Jan- 20 0 5 0 6 ' 4 5 Phsse: AnalyticaiDetermination of PFBS inNorthern Bobwhite Tissues 3M Environmental Laboratory E03-0583, Quantation of PFBS in Quail Data file : \\Etstarget\Target\chem\dudej r .i\D041025.b\D1025052.D Lab Smp Id: 72851-3 Inj Date : 25-OCT-2004 21:58 Operator : mla Inst ID: dudejr.i Smp Info : 0 ppm Dose Group; Serum; 30 Aug 04; MLA/CMC/MEE Misc Info : Comment : Serum, 0 ppm Dose Group, Control Samples Method : \\Etstarget\target\chem\dudejr .i\D041025.b\D041025.m Meth Date : 04-Jan-2005 06:42 carlson Quant Type: ESTD Cal Date : 25-OCT-2004 17:15 Cal File: D1025025.D Als bottle: 27 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.401 3.468 0.013 126S2 1.83292 1 . 8 3 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 105 of 227 Contains No CBI -315 - Project Number 454-116 3M Environmental Laboratory Page 106 of 227 Contains No CBI -316- Project Number 454-116 Data FfteletrEO^-O^ia;starget\Target \chem\jgttjjtjjigg<ji|)q!)r^bv$itepaae n Report Date: 04-Jan-2005 06:58 phase:Analytical Determination of PFBS In Northern Bobwhite tissues a 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr .i\D040910.b\D0910029 D Lab Smp Id: 72882-1 ' Inj Date : 10 -SEP-2004 16:01 Operator : m^a Inst ID: dudejr.i Smp Info : 100 ppm group, Serum, 9/1/2004 MLA/CMC/MEE Misc Info : Comment : Serum, 100 ppm Dose Group Method : \\Etstarget/Target\chem\dudejr.i\D040910.b\D040910.m Meth Date : 04-Jan-20 05 06:55 carlson Quant Type: ESTD Cal Date : 10 -SEP-2004 15:20 Cal File: D0910025.D Als bottle: Dil Factor: 1 .00000 Integrator: Falcon Target Versi on: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN RT EXP RT DLT RT RESPONSE (ng/mL) FINAL (ng/mL) 3-272 3.272 0.000 9525142 672.921 673 3M Environmental Laboratory Page 107 of 227 Contains No CBI -317 - Project Number 454-116 Data t o T K ^ M ^ e t X T a r g e t S o h e ^ d u d e jr . iS D O io s io . t o S D o ii^ if Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Page 2 1 PFBS 3M Environmental Laboratory Page 108 of 227 Contains No CBI -318- Project Number 454-116 Data Flebetfo^Ststarget\ T a r g e t \ c h e m \ d u T d M 5 r A jgtt|B0StittW3)b\itepaqe 1 Report Date: 04-Jan-2005 06:58 Phase: Analytical Determination of PFBS in Norlhern Bobwhite tissues a 3M Environmental Laboratory , E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D040910,b\D0910032 D Lab Smp Id: 72883-1 ' Inj Date : 10-SEP-2004 16:32 Operator : mla Inst ID: dudejr.i Smp Info : 100 ppm group, Serum, 9/1/2004 MLA/CMC/MEE Misc Info : Comment : Serum, 100 ppm Dose Group Method : \\Etstarget\Target\chem\dudejr.i\D040910.b\D040910.m Meth Date : 04-Jan-2005 06:55 carlson Quant Type: ESTD Cal Date : 10-SEP-2004 15:20 Als bottle: 14 Cal File: D0910025 D Dil Factor: 1.00000 Integrator: Falcon Target Version: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS RT EXP RT DLT RT RESPONSE ON-COLUMN (ng/mL) FINAL (ng/mL) 3.265 3.272 -0.007 8782325 592.285 592 3M Environmental Laboratory Page 109 of 227 Contains No CBI -319- Project Number 454-116 Report E03-0583 . T-7485: A Reproduction Study with the Northern Bobwhite Data Filet S\Etstar'get\Target\chem\dudejr', i\D 04fll$eb W f^ j)< g ^ g term|na|ion of PFBS in Northern Bobwhite Tissues Page 2 1 PFBS <29SO 3M Environmental Laboratory Page 110 of 227 Contains No CBI -320- Project Number 454-116 Data M e j t e t f Report Date: s target\Target\chem\d u rd ' 4 * p r A. q t i Q r g ^ u 04-Jan-2005 07:01 Phase: AnalyticarDetermination of jiy$ j PFBS tf in ogg Northern Bobwmte 'age tissues 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D040927,b\D0927029 D Lab Smp Id : 73118-1 Inj Date 27 -SEP-2004 15:58 Operator ; cmc Inst ID: dudejr.i Smp Info : 100 ppm Dose Group, Serum, 9/1/2004 MLA/CMC/MEE Mise Info : Comment : Serum, 100 ppm Dose Group Offspring Method : \\Etstarget\Target\chem\dudejr.i\D040927,b\D040927.m Meth Date : 04-Jan-2005 06:59 carlson Quant Type: ESTD Cal Date : 27 -SEP-2 004 15:16 Cal File: D0927025.D Als bottle: 11 Dii Factor: 1 . 00000 Integrator : Falcon Target Vers ion: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.265 3.258 0.007 17958 5.22483 S.22(M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 111 of 227 Contains No CBI -321 - Project Number 454-116 r\ejjuu cuo-uuoo i - / h o u . n r \ c ( J i uuuuli um o i u u y w i n i m e inui ui ci n d u u w i m i c Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\EtstargetMarget\chemSdudejr*iSD040927*b\D0927029*D Page 2 3M Environmental Laboratory Page 112 of 227 Contains No CBI -322 - Project Number 454-116 r\epuf l cuo-uuoo i -i h o u . m r \ e p i u u u u . i u n o i u u y n u n m e inui m e m d u u w i i h c Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D040927.b\D0927032 D Paqe 1 Report Date: 04-Jan-2005 07:01 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D040927.b\D0927032 D Lab Smp Id : 73119-1 Inj Date : 27-SEP-2004 16:29 Operator :: cmc Inst ID: dudej r .i Smp Info : 100 ppm Dose Group, Serum, 9/1/2004 MLA/CMC/MEE Misc Info : Comment : Serum, 100 ppm Dose Group Offspring Method : \\Etstarget/Target\chem\dudej r .i\D040927.b\D040927.m Meth Date : 04-Jan-2005 06:59 carlson Quant Type: ESTD Cal Date : 27-SEP-2004 15:16 Cal File: D0927025.D Als b o t t l e : 14 Dil Factor: 1 . 0 0 0 0 0 Integrator: Falcon Target Vers ion: 4.12 Compound Sublist: all.sub Compounds 1 PFBS 1299) QUANT SIG MASS 299 CONCENTRATIONS RT EXP RT DLT RT RESPONSE ON-COLUMN (ng/mL) FINAL (ng/mL) 3.265 3.258 0.007 17804 5 .16393 5 . 1 6 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 113 of 227 Contains No CBI - 323 - Project Number 454-116 r\epuu cuo-uuoo I-/HOJ. r\ep iu u u o u u n oiuuy wmi m e hui m ei n o u u w im e Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: N\Etstarget\Target\cherfi\dudejr*i\D040927+b\D0927032*D Pag 2 3M Environmental Laboratory Page 114 of 227 Contains No CBI -324 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File; \\Etstarget\Target\chem\dudej r.i\D040922.b\D0922029.D Page Report Date: 04-Jan-2005 07:04 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudejr.i\D040922.b\D0 922029.D Lab Smp Id 72 912 -1 Inj Date 22 -SEP-2004 18:52 Operator : Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Serum; 07 Sept 04; MLA/CMC/MEE Misc Info : Comment : Serum, 300 ppm Dose Group Method : \\Etstarget\target\chem\dudej r.i\D040922,b\D040922.m Meth Date : 04-Jan-2005 07:03 carlson Quant Type: ESTD Cal Date : 22 -SEP-2004 18:11 Cal File: D0922025.D Als b o t t l e : 11 Dil Factor: 1 . 0 0 0 0 0 Integrator: Falcon Target Vers i o n : 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 218 3M Environmental Laboratory Page 115 of 227 Contains No CBI -325 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File; WEtstargetSTargetSchemSdudejr, i\D040922,LAD0922029.D Page 2 2*42+3-i 2+22.1: YsEtstarget\TargetSchem\dude,jr+i\D040922+b\D0922029+D ' 1+91+81+7-' 1+6: 1+5- 1.4- i?' H 1.2- i*i-: > 1+0: 0+8-; A r^j : 0+4-; 0 3- 11 0+2 o*i-: ; --- l--L \ _iL A t 1_ 1 U U .1--11 1--------- ------------- -- - !1 0,5 1,0 1.5 2,0 2,5 3,0 3.5 4.0 4,5 5,0 5.5 6.0 6^5 ?|o 7^5 8lo 8ls Min 3M Environmental Laboratory Page 116 of 227 Contains No CBI -326 - Project Number 454-116 Report E03-0583 . T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r.i\D040922,b\D0922032.D Page Report Date: 04-Jan-2005 07:04 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D040922,b\D0922032.D Lab Smp Id: 72913-1 Inj Date : 22 -SEP -2004 19:24 Operator : mla Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Serum; 07 Sept 04; MLA/CMC/MEE Mise Info : Comment : Serum, 300 ppm Dose Group Method : \\Etstarget\target\chem\dudej r .i\D040922.b\D040922. m Meth Date : 04 -Jan-20 05 07:03 carlson Quant Type: ESTD Cal Date : 22 -SEP -2004 18:11 Cal File: D0922025.D Als bottle: 14 Dii Factor: 1.00000 Integrator : Falcon Compound Sublist: all.sub Target Versi on: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.250 3.253 0.000 1811217 241.268 241 3M Environmental Laboratory Page 117 of 227 Contains No CBI -327 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File? \\Etstarget\Target.\ohero\dudejr,i\D040922+b\D0922032,D Pag 2 \\Etstarget\TargetSchem\dudejr* i\D040922,b\D0922032,D 2*4-; 2,3 2 ?.' 2 *1 -; 2 , 0 -: 1,9 1,81*7-; 1,6-; i*5-i 1*4-1 1,3-; 1,2-i i* i- j l* 0 -: 0,9-; 0 , 8 -; 0,7 -; 0 ,6 -; 0 , 5 -; 0,4-; 0*3- 0 +2 -: 0*1-; 1 PFBS <2950 3M Environmental Laboratory Page 118 of 227 Contains No CBI - 328 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D040928,b\D0928029.D Page Report Date: 04-Jan-2005 07:07 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r.i\D040928,b\D0928029.D Lab Smp Id: 73133-1 Inj Date : 28-SEP-2004 11:47 Operator : cmc Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Serum; 07 Sept 04; MLA/CMC/MEE Misc Info : Comment : Serum, 300 ppm Dose Group Offspring Method : \\Ets target \Target Vchem'v dude jr.i\D040928.b\D040928.m Meth Date : 04-Jan-2005 07:06 carlson Quant Type: ESTD Cal Date : 28-SEP-2004 11:06 Als bottle: 11 Cal File: D0928025.D Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Processing Host: W50352 Compounds 1 PFBS (299) QUANT SIG MASS CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.258 .3.252 0.006 54344 QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 119 of 227 Contains No CBI -329 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr*i\D040928*bSD0928029*D Page 2 3M Environmental Laboratory Page 120 of 227 Contains No CBI - 330 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D040928.b\D0928032.D Page Report Date: 04-Jan-2005 07:07 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D040928,b\D0928032.D Lab Smp Id: 73134-1 Inj Date : 28-SEP-2004 12:19 Operator : emc Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Serum; 07 Sept 04; MLA/CMC/MEE Mise Info : Comment : Serum, 300 ppm Dose Group Offspring Method : \\Etstarget\Target\chem\dudej r.i\D040 92 8 .b\D040928.m Meth Date : 04-Jan-2005 07:06 carlson Quant Type: ESTD Cal Date : 28 -SEP-2004 11:06 Als bottle: 14 Cal File: D0928025.D Dii Factor: 1.00000 Integrator: Falcon Target Version: 4.12 Compound Sublist: all.sub Processing Host: W50352 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.258 3.252 0.006 55715 19.2936 1 9 . 3 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 121 of 227 Contains No CBI -331 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudevir.iSD040928+b\D0928032.B Page 2 3M Environmental Laboratory Page 122 of 227 Contains No CBI - 332 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D040924.b\D0924029.D Page Report Date: 04-Jan-2005 06:37 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D040924.b\D0924029.D Lab Smp Id: 72944-1 Inj Date : 24-SEP-2004 18:13 Operator : mla Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Serum; 13 Sept 04; MLA/CMC/MEE Misc Info : Comment : Serum, 900 ppm Dose Group Method : \\Etstarget\Target\chem\dudejr.i\D040924,b\D040924.m Meth Date : 04-Jan-2005 06:36 carlson Quant Type: ESTD Cal Date : 24-SEP-2004 17:31 Cal File: D0924025.D Als b o t t l e : 11 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT S IG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/rnL) (ng/mL) 3.2S8 3.258 0.000 1602357 175.206 175 3M Environmental Laboratory Page 123 of 227 Contains No CBI - 333 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data F i l e j S S E ts ta rg e t\T arg et\ch em \d u d ej r * i\D040924+b\D0924029*D Page 2 \\E ts ta rg e tM a rg e tS c h e m \d u d e jr+i\D 040924,b\D0924029*D 1,9-j 1.51 , 7 :: 1.61*5-1 1,4- l*3-j ^ 1* 2is> o 1*1- H 1,0- 0,5 0,4 0,3- 0* 1- : . LtO ^ , y . , 0,5 1,0 1,5 2,0 1 PFBS <2950 ^ l . - - . r L ^ L - U t : I I \r l- - - !5 4 * 0 415 5 . 0 Hin 5^5 -- 6*0 3M Environmental Laboratory Page 124 of 227 Contains No CBI -334 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase; Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr .i\D040924.b\D0924032.D Page Report Date: 04-Jan-2005 06:37 3M Environmental Laboratory , E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D040924.b\D0924032 D Lab Smp Id: 72945-1 Inj Date : 24-SEP-2004 18:44 Operator : mla Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Serum; 13 Sept 04; MLA/CMC/MEE Misc Info : Comment : Serum, 900 ppm Dose Group Method : \\Etstarget/Target\chem\dudej r.i\D040924.b\D040924.m Meth Date : 04-Jan-2005 06:36 carlson Quant Type: ESTD Cal Date : 24-SEP-2004 17:31 Als bottle: 14 Cal File: D0924025.D Dil Factor: 1.00000 Integrator: Falcon Target Version: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.258 3.258 0.000 2909643 357.710 358 3M Environmental Laboratory Page 125 of 227 Contains No CBI -335 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File* SSEtstarget\Target\chem\dudejr*iSD040924*b\D0924032*D Page 2 3*8- WEtstarget\TargetScheroSdudeJri\D040924*b\D0924032D 3*43*2- 2*8- 2*6: 2*4- 2*2- 6<6 2*0-- V< 1.8- > 1*6- 1*4- 1*2- i *0-j 0,8- .- 0*4- \ V. i i 111 -1 M i . 0*S 1*0 1*5 2*0 2*5 3*0 3*5 4*0 4*5 5*0 5*5 6*0 6*5 7*0 7*5 3*0 S*5 Hin 1 PFBS 3M Environmental Laboratory Page 126 of 227 Contains No CBI -336 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D040930.b\D0930027.D Page Report Date: 04-Jan-2005 06:41 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D040930.b\D0930027.D Lab Smp Id: 73148-1 Inj Date :30-SEP-2004 18:44 Operator : cmc Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Serum; 13 Sept 04; MLA/CMC/MEE Mise Info : Comment Method : Serum, 900 ppm Dose Group Offspring : \\Etstarget\Target\chem\dudej r .i\D040930.b\D040930.m Meth Date :04-Jan-200506:40 carlson Quant Type: ESTD Cal Date :30-SEP-2004 18:02 CalFile:D0930023.D Als bottle : 11 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: a l l .sub Target Version: 4.12 Compounds 1 PFBS (2 99) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.272 3.265 0.007 93522 32.4297 32.4 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 127 of 227 Contains No CBI - 337 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filet \\Etstarget\Target\chero\dudejr,i\D040930.bSD0930027.D Page 2 3M Environmental Laboratory Page 128 of 227 Contains No CBI - 338 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr .i\D040930.b\D0930030 D Paqe Report Date: 04-Jan-2005 06:41 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D040930,b\D0930030 D Lab Smp Id: 73149-1 Inj Date : 3 0-SEP-2 004 19:15 Operator : cmc Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Serum; 13 Sept 04; MLA/CMC/MEE Misc Info : Comment : Serum, 900 ppm Dose Group Offspring Method : \\Etstarget\Target\chem\dudej r .i\D040930.b\D040930.m Meth Date : 04-Jan-2005 06:40 carlson Quant Type: ESTD Cal Date : 30-SEP-2004 18:02 Cal File: D0930023.D Als b o t t l e : 14 Dil Factor: 1 . 0 0 0 0 0 Integrator: Falcon Target Vers: on: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS RT EXP RT DLT RT RESPONSE ON-COLUMN (ng/mL) FINAL (ng/mL) 3.279 3.265 0.014 67034 22.8488 2 2 . 8 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 129 of 227 Contains No CBI - 339 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filer '\SEtstarget\Target\chem\ductejr* 1SIl040930.b\B0930030.D Page 2 1 PFBS i2SS> 3M Environmental Laboratory Page 130 of 227 Contains No CBI -340 - Project Number 454-116 ncoi Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite phase: Ana|yticg| Deterrnination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\itchy.i\I041023.b\I1023033 .D Page 1 Report Date: 04-Jan-2005 06:51 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\itchy.i\I041023,b\I1023033.D Lab Smp Id: 72976-1 Inj Date : 23-OCT-2004 19:04 Operator : mla Inst ID: itchy.i Smp Info : 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Mise Info : Comment : Liver, Control Group Method : \\Etstarget\Target\chem\itchy.i\I041023.b\I041023.m Meth Date : 04-Jan-2005 06:49 carlson Quant Type: ESTD Cal Date : 23-OCT-2004 18:23 Cal File: 11023029.D Als bottle: 21 Dil Factor: 1 . 0 0 0 0 0 Integrator: Falcon Compound Sublist: all.sub Target Vers ion : 4.12 Compounds 1 PFBS (299) QUANT 5IG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 488 3.478 0.010 23914 1.00677 1 . 0 1 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 131 of 227 Contains No CBI -341 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination ofPFBS in Northern Bobwhite Tissues Data File: S\Etstarget\TargetSchemSitchy,i\I041023.b\I1023033*D Page 2 1 PFES (299) 3M Environm ental Laboratory Page 132 of 227 Contains No CBI -342 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\itchy.i\104102 3.b\1102 3036.D Page Report Date: 04-Jan-2005 06:51 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\itchy.i\I041023.b\I1023036.D Lab Smp Id 72977-1 Inj Date 23 -OCT-2004 19:35 Operator mla Inst ID: itchy.i Smp Info 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Mise Info Comment Liver, Control Group Method \\Etstarget/Target\chem\itchy.i\l041023.b\l041023.m Meth Date 04-Jan-2005 06:49 carlson Quant Type: ESTD Cal Date 2 3 -OCT-20 04 18:23 Cal File: 11023029.D Als bottle 24 Dii Factor 1 . 0 0 0 0 0 Integrator Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS . 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.498 3.478 0.020 54121 3.94820 3 . 9 5 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 133 of 227 Contains No CBI -343 - Project Number 454-116 Report E03-0583 T-/485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chen\itchy*iM 041023.b\11023036*D Page 2 3M Environmental Laboratory Page 134 of 227 , Contains No CBI -344- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\itchy.i\I041023.b\I1023041.D Page Report Date: 04-Jan-2005 06:51 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\itchy.i\I041023.b\11023041.D Lab Smp Id: 72978-1 Inj Date : 23-OCT-2004 20:27 Operator : nnla Inst ID: itchy, i Smp Info : 0 ppm Dose Group; Liver; 27 Sep 04; MLA/CMC/MEE Mise Info : Comment : Liver, Control Group Method : \\Etstarget\Target\chem\itchy.i\l041023,b\I041023.m Meth Date : 04-Jan-2005 06:49 carlson Quant Type: ESTD Cal Date : 23-OCT-2004 18:23 Cal File: 11023029.D Als b o t t l e : 27 Dil Factor: 1.00000 Integrator: Falcon . Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.490 3.478 .020 59815 4.50326 4 . 5 0 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 135 of 227 Contains No CBI - 345 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File? SSEtstargetSTargetScheroSitchy*i\I041023*bM1023041*D Page 2 3M Environmental Laboratory Page 136 of 227 Contains No CBI -346- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r.i\D041009,b\D1009029.D Page Report Date: 04-Jan-2005 07:13 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D041009.b\Dl009029.D Lab Smp Id: 73008-1 Inj Date : 09 -OCT-2004 17:58 Operator : mla Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE Misc Info : Comment : Liver, 100 ppm Dose Group Method : \\Etstarget\target\chem\dudejr.i\D041009,b\D041009.m Meth Date : 04-Jan-2005 07:13 carlson Quant Type: ESTD Cal Date : 09-OCT-20 04 17:16 Cal File: D1009025.D Als b o t t l e : 11 Dil Factor: 1.00000 Integrator: Falcon Target Versi on: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.439 3.439 0.000 666726 49.5659 49.6 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 137 of 227 Contains No CBI -347- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filej SSEtstargetSTargetNchemSdude.jr,i\D041009.b\D1009029.D Page 2 1,4- S\Etst.arget\Target\chemSdudejr* i\D041009.b\D1009029,D 1* 2- 1,1- 1* 00,90.8 0.70*6- '050,4- ' 1,0 1.5 2.0 1 PFBS <299) , l-i^WYv-i 4.0 4,5 5.0 Min 6.0 6.5 7.0 8.5 3M Environmental Laboratory Page 138 of 227 Contains No CBI - 348 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D041009.b\D1009032.D Page Report Date: 04-Jan-2005 07:13 1 3M Environmental Laboratory E03-0583 Phase.- Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D041009.b\Dl009032.D Lab Smp Id: 73009-1 Inj Date : 09-OCT-2004 18:29 Operator : mla Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Liver; 30 Sept 04; MLA/CMC/MEE Mise Info : Comment : Liver, 100 ppm Dose Group Method : \\Etstarget\target\chem\dudejr.i\D041009,b\D041009 .m Meth Date : 04-Jan-2005 07:13 carlson Quant Type: ESTD Cal Date : 09-OCT-20 04 17:16 Cal File: D1009025.D Als bottle: 14 Dil Factor: 1.00000 Integrator : Falcon Target Versi o n : 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) .1.439 3.439 0.000 601802 44.0515 44.1 3M Environmental Laboratory Page 139 of 227 Contains No CBI -349 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D ata F ile ; S \E tstarget\T arget\chem S dude,jr.i\D 041009.b\D 1009032.D Page 2 1*3- 1+2 1.1 1,0- \\E ts ta r g e t\ T a r g e t S c h e r n \d u d e .i r i SD041009 . b\D 1009032 *D " 0 9 - 0.3 % 0.7 - 0.1ols ilo i l s 2I0 1 PFBS (2 9 9 ) i A u A A - \ i i J 1~J------ .5 3 .0 3 .5 4*0 4I5 5I0 s is &lo I_I si0 sis 3M Environmental Laboratory Page 140 of 227 Contains No CBI - 350 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D041024,b\D1024087.D Page Report Date: 04-Jan-2005 07:17 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041024,b\D1024087.D Lab Smp Id: 73190-1 Inj Date : 2 5 -OCT-2004 07:09 Operator : ml a Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Offspring; Liver; 30 sep 04; MLA/CMC/MEE< Mise Info : Comment : Liver, 100 ppm Dose Group, Offspring Method : \\Etstarget\target\chem\dudejr.i\D041024,b\D041024.m Meth Date : 04-Jan-2005 07:16 carlson Quant Type: ESTD Cal Date : 2 4 -OCT-20 04 20:49 Cal File: D1024028.D Als bottle: 60 Dil Factor: 1.00000 Integrator : Falcon Compound Sublist: a l l .sub Target Versi on: 4.12 . Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS O N -COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.482 3.460 0.022 9834 1.89137 1 . 8 9 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 141 of 227 Contains No CBI -351 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Fil e i \\Etstarget\Target\chem\dudejr.iSB041024.b\D1024087,D 3M Environmental Laboratory Page 142 of 227 Contains No CBI -352 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041024.b\D1024093.D Page Report Date: 04-Jan-2005 07:17 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D041024.b\D1024093.D Lab Smp Id: 73191-1 Inj Date : 25-OCT-2004 08:12 Operator : mla Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Offspring; Liver; 30 sep 04; MLA/CMC/MEE? Misc Info : Comment : Liver, 100 ppm Dose Group, Offspring Method : \\Etstarget\target\chem\dudejr.i\D041024.b\D041024 .m Meth Date : 04-Jan-2005 07:16 carlson Quant Type: ESTD Cal Date : 24-OCT-2004 20:49 Als b o t t l e : 63 Cal File: D1024028.D Dil Factor: 1.00000 Integrator: Falcon Target Version: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.481 3.460 0.021 20460 5.74229 5. 7 4 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 143 of 227 Contains No CBI - 353 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: SSEtstargetSTargetSchemSdudejr,i\D041024+b\D1024093*D 3M Environmental Laboratory Page 144 of 227 Contains No CBI -354- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041015.b\D1015032 D Page Report Date: 04-Jan-2005 07:21 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041015.b\D1015032.D Lab Smp Id : 73041-1 Inj Date : 15-OCT-2004 15:04 Operator : mla Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Liver; 05 Oct 04; MLA/CMC/MEE Mise Info : Comment : Liver, 300 ppm Dose Group Method : \\Etstarget\target\chem\dudejr.i\D041015,b\D0410 1 5 .m Meth Date : 04-Jan-20 05 07:19 carlson Quant Type: ESTD Cal Date : 15-OCT-2004 14:23 Cal File: D1015028.D A l s bottle: 21 Dii Factor: 1.00000 Integrator : Falcon Target Vers ion: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3 .446 3.446 0.000 1353865 183 .282 1 8 3 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 145 of 227 Contains No CBI - 355 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D-ata F i l e : W E ts ta rg e tS T a rg e t\c h e m \d u d e jri SD041015*bSD10150324D Page 2 3M Environmental Laboratory Page 146 of 227 Contains No CBI -356- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041015.b\D1015035.D Page Report Date: 04-Jan-2005 07:21 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej .i\DO41015.b\D1015035.D Lab Smp I d : 73042-1 Inj Date : 15-OCT-2004 15:36 Operator : ml a Smp Info : 300 ppm Dose Group; Liver; Mise Info : nst ID: dudejr.i Oct 04; MLA/CMC/MEE Comment : Liver, 300 ppm Dose Group Method : \\Etstarget\target\chem\dud Meth Date : 04-Jan-2005 07:19 carlson Cal Date : 15-OCT-2004 14:23 Als bottle: 24 Dii Factor: 1.00000 r.i\D041015.b\D041015 .m Quant Type : ESTD Cal File: D1015028.D Integrator : Falcon Target Versi on: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/inL) (ng/mL) 3.446 3.446 0.000 794000 100.276 1 0 0 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 147 of 227 Contains No CBI -357- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D ata F i le } \\E ts ta r g e t\T a r g e t\c h e m \d u d e j r * iSD041015*b\D1015035*D Page 2 1 +7- 1*6-;' 1 +5 1 +4 1 +3^ 1+21* 1-' 1*0-; S \E ts ta r g e t\T a r g e t\c h e n i's d u d e jr + i\D 0 4 1 0 1 5 +b \M 0 1 5 0 3 5 +D t! 0+8- 0+7-; 0+60*5-' 0.4 0 +3 0+2 0+1- 1+0 1+5 1 PFBS ( 29 9 ) ij ._L-Jv_ 4+0 4+5 5+0 Min 8+0 3M Environmental Laboratory Page 148 of 227 Contains No CBI - 358 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041018b .b\D1018131. D Page Report Date: 04-Jan-2005 07:24 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041018b.b\D1018131.D Lab Smp Id: 73192-1 Inj Date : 19-OCT-2004 11:37 Operator : mla Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Liver; 05 Oct 04; MLA/CMC/MEE Misc Info : Comment : Liver, 300 ppm Dose Group Offspring Method : \\Etstarget\target\chem\dudej r .i\D041018b.b\D041018b.m Meth Date : 04-Jan-2005 07:23 carlson Quant Type: ESTD Cal Date : 19-OCT-2004 10:55 Cal File: D1018127.D Als bottle: 54 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE ing/mL) ing/mL) 3.461 3.447 0.014 34195 8.59423 8. 5 9 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 149 of 227 Contains No CBI - 359 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filer \SEtstargetNTargetSchenSdudejr.i'\D04101Sb,b\D1018131*D Page 2 3M Environmental Laboratory Page 150 of 227 Contains No CBI -360 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041018b.b\D1018134.D Page Report Date: 04-Jan-2005 07:24 1 3M Environmental Laboratory E03-05 83 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041018b.b\D1018134.D Lab Smp Id : 73193-1 Inj Date : 19-OCT-2004 12:09 Operator : mla Inst I D : dudej r .i Smp Info : 300 ppm Dose Group; Liver; 05 Oct 04; MLA/CMC/MEE Mise Info : Comment : Liver, 300 ppm Dose Group Offspring Method : \\Etstarget\target\chem\dudej r .i\D041018b.b\D041018b.m Meth Date : 0 4 -Jan-2005 07:23 carlson Quant Type: ESTD Cal Date : 19-OCT-2004 10:55 Cal File: D1018127.D Als bottle: 57 Dii Factor: 1.00000 Integrator : Falcon Target Versi on: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299} QUANT SIG MASS CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.460 3 .447 0.013 1A7S4 4 0546 4 05 fMl QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 151 of 227 Contains No CBI -361 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \SEtstarget\Target\chero\dudejr+i\D041018b*b\D1018134*D Page 2 3M Environmental Laboratory Page 152 of 227 Contains No CBI -362 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS In Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041023.b\D1023032.D Page Report Date: 04-Jan-2005 07:27 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041023 ,b\D1023032.D Lab Smp Id: 73072-1 Inj Date : 23-OCT-2004 18:46 Operator : mla Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Misc Info : Comment : Liver, 900 ppm Dose Group Method : \\Etstarget\target\chem\dudejr.i\D041023 .b\D041023 .m Meth Date : 04-Jan-2005 07:27 carlson Quant Type: ESTD Cal Date : 23-OCT-2004 18:04 Cal File: D1023028.D Als bottle: 21 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.461 3.468 -0.007 492162 46.9848 4 7 . 0 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 153 of 227 Contains No CBI - 363 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D ata F ile : S\Etstarget\TargetSchem \dudejr*iSD 041023*b\D 1023032*D Page 2 1*0- \\E ts ta r g e t\T a r g e t\c h e n \d u d e ,jr * i\D041023*b\D1023032*D 0*9- 0*7- 0*6- o< 0*4 0*3 0*2 0*1- u . i ( t ois l'o lis 1 PFBS <299) IV i h .J y 4 -.__ I*Q J J 3*0 4*0 4*5 7*0 8*0 8*5 3M Environmental Laboratory Page 154 of 227 Contains No CBI -364 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D041023.b\D1023035.D Page Report Date: 04-Jan-2005 07:27 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041023.b\Dl023035.D Lab Smp Id: 73073-1 Inj Date : 23 -OCT-2004 19:17 Operator : mla Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Liver; 19 Oct 04; MLA/CMC/MEE Mise Info : Comment : Liver, 900 ppm Dose Group Method : \\Etstarget\target\chem\dudejr.i\D041023.b\D041023.m Meth Date : 04-Jan-2005 07:27 carlson Quant Type: ESTD Cal Date : 23 -OCT-2004 18:04 Cal File: D1023028.D Als bottle : 24 Dil Factor: 1.00000 Integrator : Falcon Compound Sublist: all.sub Target Versi on: 4.12 Compounds 1 PFBS (2 99) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.460 3.468 -0.008 1120075 116.795 1 1 7 (M) QC Flag Legend M - Compound response manually integrated. 3M Environmental Laboratory Page 155 of 227 Contains No CBI -365 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D ata F i 1e W E t s t a r g e t\T a r g e tS c h e n \d u d e jr +i\D 0 4 1 0 2 3 b \D 1 0 2 3 0 3 5 D Page 2 SSEtstargetSTargetXchemSdudejr,i\D041023+b\B1023035+D 2*1 2*0 1+9 1.8 1*7 1*6 1*5 1+4 1+3 isO 1*2 1+1 1+0 0*9 0+8 0+7 0+6 0+5 0+4 0+3 0+2 0+1 1 PFBS <299> 3M Environmental Laboratory Page 156 of 227 Contains No CBI - 366 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr .i\D041022.b\D1022032.D Page Report Date: 04-Jan-2005 07:30 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D041022.b\D1022032.D Lab Smp Id: 73207-1 Inj Date : 22-OCT-2004 19:47 Operator : mla Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Offspring; Liver; 19 Oct 04; MLA/CMC/MEES Misc Info : Comment : Liver, 900 ppm Dose Group Offspring Method : \\Etstarget\target\chem\dudej r .i\D041022.b\D041022.m Meth Date : 04-Jan-2005 07:28 carlson Quant Type: ESTD Cal Date : 22-OCT-2004 19:05 Cal File: D1022028.D Als bottle : 21 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.468 3.454 0.014 96273 32.6494 32 .6 3M Environmental Laboratory Page 157 of 227 Contains No CBI -367 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filet \\Etstarget\Target\chem\duclejr*iSD041022*bSD1022032*D Page 2 1 PFBS <299) 3M Environmental Laboratory Page 158 of 227 Contains No CBI - 368 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041022 .b\D1022036 .D Page Report Date: 04-Jan-2005 07:30 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudejr.i\D041022.b\D1022036 .D Lab Smp Id 73208-2 Inj Date 22 -OCT-2004 20:29 Operator mla Inst ID: dudejr.i Smp Info 900 ppm Dose Group; Offspring; Liver; 19 Oct 04; MLA/CMC/MEET Mise Info Comment Liver, 900 ppm Dose Group Offspring Method \\Etstarget\target\chem\dudej r.i\D041022.b\D041022.m Meth Date 04-Jan-2 005 07:28 carlson Quant Type: ESTD Cal Date 2 2 -OCT-2004 19:05 Cal File: D1022028.D Als bottle 25 Dil Factor 1.00000 Integrator Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.467 .3.454 0.013 64785 21.4773 21.5 3M Environmental Laboratory Page 159 of 227 Contains No CBI -369- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Et.starget\Target.\cheMSdudejr*i\DO41O22.b\BlC'22036+D Page 2 1 PFBS <299> 3M Environmental Laboratory Page 160 of 227 Contains No CBI -370- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041207.b\D1207046.D Page Report Date: 04~Jan-2005 07:34 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041207.b\D1207046.D Lab Smp Id: 72727-1 Inj Date : 07-DEC-2004 18:42 Operator : rsk Inst ID: dudejr.i Smp Info : 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Misc Info : Comment : Egg Samples, Control Group Method : \\Etstarget\Target\chem\dudejr.i\D041207 ,b\D041207.m Meth Date : 04-Jan-2005 07:32 carlson Quant Type: ESTD Cal Date : 07-DEC-2004 15:15 Cal File: D1207026.D Als bottle : 41 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) Compound Not Detected. 3M Environmental Laboratory Page 161 of 227 Contains No CBI -371 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS In Northern Bobwhite Tissues Data Filei Y .Etstarget\Target\chem\dude,]r,i\D041207,lo\D1207046+D Page 2 1 PFBS <299) (Undetected) 3M Environmental Laboratory Page 162 of 227 Contains No CBI -372- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: ' Etstarqet\Tarqet\chem\dudejr.i\D041207.b\D1207050.D Page Report Date: 04-Jan-2005 07:34 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041207.b\D1207050.D Lab Smp Id: 72728-1 Inj Date : 07-DEC-2004 19:23 Operator : rsk Inst ID: dudejr.i Smp Info : 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE M i s c .Info : Comment : Egg Samples, Control Group Method : \\Etstarget\Target\chem\dudejr.i\D041207.b\D041207.m Meth Date : 04-Jan-2005 07:32 carlson Quant Type: ESTD Cal Date : 07-DEC-2004 15:15 Cal File: D1207026.D Als bottle: 44 Dil Factor: 1.00000 Integrator: Falcon Compound Sublist: all.sub Target Vers i o n : 4.12 Compounds 1 PFBS (299) QUANT SIC MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.453 3.439 0.014 167503 1.22748 1.23 3M Environmental Laboratory Page 163 of 227 Contains No CBI -373 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filet S\Etstarget\Target\chern\dudejr*i\D041207+b\D1207050+D Page 2 3M Environmental Laboratory Page 164 of 227 Contains No CBI -374- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D041207 ,b\D1207077.D Page Report Date: 04-Jan-2005 07:34 3M Environmental Laboratory E03 -0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041207.b\D1207077.D Lab Smp Id: 72577-1 Inj Date : 0 8 -DEC-2004 00:05 Operator : rsk Inst ID: dudejr.i Smp Info : 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples, Control Group Method : \\Etstarget\Target\chem\dudejr.i\D041207.b\D041207.m Meth Date : 04-Jan-20 05 07:32 carlson Quant Type: ESTD Cal Date : 07 -DEC-2004 15:15 Cal File: D1207026.D Als bottle: 71 Dil Factor: 1 . 0 0 0 0 0 Integrator : Falcon Compound Sublist: all.sub Target Vers ion : 4.12 Compounds 1 PFBS ( 2 9 3 ) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.454 3.439 0.015 144822 0 .14700 0.147 3M Environmental Laboratory Page 165 of 227 Contains No CBI -375 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filei \SEtstarget\Target\chero\dudejr+iXD041207*b\D1207077+D Page 2 1 F'FES <299; 3M Environmental Laboratory Page 166 of 227 Contains No CBI -376- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041207.b\D1207082.D Page Report Date: 04-Jan-2005 07:34 1 3M Environmental Laboratory E03 -0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudejr.i\D041207,b\D1207082.D Lab Smp Id 72578-1 Inj Date 08 -DEC-2004 00:57 Operator rsk Inst ID: dudejr.i Smp Info 0 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info Comment Egg Samples, Control Group Method Meth Date \\Etstarget\Target\chem\dudejr.i\D041207.b\D041207.m 04-Jan-2005 07:32 carlson Quant Type: ESTD Cal Date 07-DEC-20 04 15:15 Cal File: D1207026.D Als bottle 74 Dil Factor 1.00000 Integrator Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.460 3.439 0.021 204316 2.98319 2.98 3M Environmental Laboratory Page 167 of 227 Contains No CBI -377 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data F i l e j \\E ts ta rg e t\T a rg e t\c h e m \d u d e ji i\D 041207*bSE1207082.D Page 2 1 PFBS <299> 3M Environmental Laboratory Page 168 of 227 Contains No CBI - 378 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041208.b\D1208030.D Page Report Date: 04-Jan-2005 07:38 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041208.b\D1208030.D Lab Smp Id: 72583-1 Inj Date : 08-DEC-2004 17:19 Operator : vjt Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples, 100 and 300 ppm dose groups, lot B Method : \\Etstarget\Target\chem\dudejr.i\D041208.b\D041208.m Meth Date : 04-Jan-2005 07:37 carlson Quant Type: ESTD Cal Date : 08-DEC-2004 16:37 Cal File: D1208026.D Als bottle: 21 Dii Factor: 1.00000 Integrator : Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.432 3.432 0.000 1966789 105.636 106 3M Environmental Laboratory Page 169 of 227 Contains No CBI -379- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D a ta F i l e Y s E ts ta rg e tS T a rg e t\c h e m S d u d e jr * i SD041208 b\D 1208030 D Page 2 1 PFBS <299> 3M Environmental Laboratory Page 170 of 227 Contains No CBI - 380 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r.i\D041208.b\D1208033.D Page Report Date: 04-Jan-2005 07:38 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041208,b\D1208033.D Lab Smp Id: 72584-1 Inj Date : 08-DEC-2004 17:49 Operator : vj t Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples, 100 and 300 ppm dose groups, lot B Method : \\Etstarget\Target\chem\dudejr .i\D041208,b\D041208.m Meth Date : 04-Jan-2005 07:37 carlson Quant Type: ESTD Cal Date : 08-DEC-2004 16:37 Cal File: D1208026.D Als bottle : 24 Dil Factor: 1.00000 Integrator : Falcon Compound Sublist: all.sub Target Versi on: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.439 3.432 0.007 2115573 115.057 115 3M Environmental Laboratory Page 171 of 227 Contains No CBI -381 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chemlsdudejr*i\D041208*b\D12Q8033*D Page 2 4*6-; 4*4-; 4*2:' 4*0-; 3*8-; 3*63*4- SSEtstargetSTargetSchern'sdudejr*i\B041208*b\D1208033*D 3*0- 2*6-; 2*4x 2*2- 2*0^ 1 * 8-; 1+ 61*41*2 1 * 00*80*60*4- i PFBS <299 > .iJ. 'Nr--L- 4*0 4* Min I' ------- _ I I 7*5 8* 8^5 3M Environmental Laboratory Page 172 of 227 Contains No CBI -382 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr.i\D041209.b\D1209028.D Page Report Date: 04-Jan-2005 07:41 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041209.b\D1209028.D Lab Smp Id: 72735-1 Inj Date : 09-DEC-2004 18:25 Operator : rsk Inst ID: dudejr.i Smp Info : 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples, 100 and 300 ppm dose groups, lot G Method : \\Etstarget\Target\chem\dudej r .i\D0412 09.b\D0412 09.m Meth Date : 04-Jan-2005 07:39 carlson Quant Type: ESTD Cal Date : 09-DEC-20 04 17:44 Cal File: D1209024.D Als bottle: 21 Dil Factor: 1.00000 Integrator : Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.439 3.440 -0.001 3478355 247.787 248 3M Environmental Laboratory Page 173 of 227 Contains No CBI - 383 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filai WEtstargetSTarget'schemSdudejriSD041209.bSD1209028.D Page 2 7.2- \\Etstarget\Target\che-m\dudejr. i\D041209.b\D1209028.D 6.66.3- 5.75.45.14.84.54.2io 3.9< 1 3.6: 3.33.02.72.42.11.81.5-' 1.20.90.6- 11 O'15 1^0 l t5 1 PFBS <299) j V. .1 1 ~ 1 1_J- 1 1 1.......... . ..... 1. 1 2.0 25 3*0 3 5 4*0 4^5 5.0 55 6.0 6.5 7.0 7.5 8.0 8.5 Min 3M Environmental Laboratory Page 174 of 227 Contains No CBI -384- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Et starget\Target\chem\dudej r .i\D041209.b\D1209031.D Page Report Date: 04-Jan-2005 07:41 1 3M Environmental Laboratory EO 3 -058 3 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudejr.i\D041209.b\D1209031.D Lab Smp Id 72736-1 Inj Date 09-D E C -2004 18:56 Operator rsk Inst ID: dudejr.i Smp Info 100 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info Comment Egg Samples, 100 and 300 ppm dose groups, lot G Method \\Etstarget\Target\chem\dudejr.i\D041209.b\D041209.m Meth Date 04 -Jan-2005 07:39 carlson Quant Type: ESTD Cal Date 09-DEC-2004 17:44 Cal File: D1209024.D Als bottle 24 Dii Factor 1.00000 . Integrator Falcon Compound Sublist: all.sub Target Vers i o n : 4.12 Compounds 1 PFBS (2 99) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.439 3.440 -0.001 3478839 247.827 248 3M Environmental Laboratory Page 175 of 227 Contains No CBI - 385 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Fi le i \\E ts ta rg e t\T a rg e t\c h e n l\d u d e jr,i\D 041209t b\D1209031,D Page 2 ? 5- V \E ts ta rg e t\T a rg e t\c h e m \d u d e ,jr +iSD041209+b\B1209031+B 6 +6 6 3 - 6 *0 5 ,75*4- 5*1- 4*8- 4*5- 4*2to 3 * 9 - H 3 * 6 7 7_ > 3*0- 2*42 *1 1 ,81 ,5 1 *2 0 .9 0*8- u "V i a 1 11 1 1! 1 1 II 0 * 5 1 *0 1 * 5 2 +0 2 *5 3 *0 3 5 4 +0 4 +5 5 +0 5 +5 8 +0 8 +5 7 +0 7 +5 S +0 8 * 5 Min 1 PFBS <299> 3M Environmental Laboratory Page 176 of 227 Contains No CBI - 386 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Targe.t\chem\dude jr .i\D0412 08 .b\D12 08057 .D Page Report Date: 04-Jan-2005 07:38 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D0412 08 .b\D1208057.D Lab Smp Id: 72590-1 Inj Date : 08-DEC-2004 21:56 Operator : vjt Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples, 100 and 300 ppm dose groups, lot B Method : \\Etstarget\Target\chem\dudej r .i\D041208.b\D041208.m Meth Date : 04-Jan-2005 07:37carlson Quant Type: ESTD Cal Date : 08-DEC-20 04 16:37 Cal File: D1208026.D Als bottle: 42 Dil Factor: 1.00000 Integrator : Falcon Compound Sublist: all.sub Target Versi on: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.439 3.432 0.007 4745356 292.605 293 3M Environmental Laboratory Page 177 of 227 Contains No CBI -387 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chero\dudejr*i\D041208+bSD1208057+D Page 2 1 PFBS (299) 3M Environmental Laboratory Page 178 of 227 Contains No CBI - 388 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041208.b\D1208060.D Page Report Date: 04-Jan-2005 07:38 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudejr.i\D041208.b\Dl208060.D Lab Smp Id 72591-1 Inj Date 08-DEC-2004 22:27 Operator vjt Inst ID: dudejr.i Smp Info 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Misc Info Comment Egg Samples, 100 and 300 ppm dose groups, lot B Method \\Etstarget\Target\chem\dudejr.i\D041208,b\D041208.m Meth Date 04-Jan-2005 07:37 carlson Quant Type: ESTD Cal Date 08-DEC-2004 16:37 Cal File: D1208026.D Als bottle 45 Dil Factor 1.00000 Integrator Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.439 3.432 0.007 8942646 640.768 641 3M Environmental Laboratory Page 179 of 227 Contains No CBI -389 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues D ata F i l e } W E ts t arg e t\T a rg e t\c h e m S d u d e jr+i SD041208*b\D1208060*D Page 2 1 PFBS <299> 3M Environmental Laboratory Page 180 of 227 Contains No CBI -390- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041209.b\D1209055.D Page Report Date: 04-Jan-2005 07:41 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041209.b\D1209055 .D Lab Smp Id: 72742-1 Inj Date : 09-DEC-2004 23:03 Operator : rsk Inst ID: dudejr.i Smp Info : 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples, 100 and 300 ppm dose groups, lot G Method : \\Etstarget\Target\chem\dudejr.i\D041209.b\D041209.m Meth Date : 04-Jan-2005 07:39 carlson Quant Type: ESTD Cal Date : 09-DEC-20 04 17:44 Cal File: D1209024.D Als bottle: 42 Dii Factor: 1 . 0 0 0 0 0 Integrator : Falcon Compound Sublist: all.sub Target Versi on: 4.12 Compounds l PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.446 3.440 0.006 5018844 382.196 382 3M Environmental Laboratory Page 181 of 227 Contains No CBI -391 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Fi1e \\Etstarget\TargetSchemSdudejr*iSD041209*b\D1209055*D Page 2 \SEtstarget\Target\chem\dudejr* i\D041209*b\D1209055*D 1 0*9- 0*8- 0*6- i ^ . i a .1 -^i- ; __________ *5 1*0 1*5 2*0 3*0 4*0 45 5*0 55 lo ! 7*0 7*5 8*0 8*5 H in 1 PFBS (299) 3M Environmental Laboratory Page 182 of 227 Contains No CBI -392 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r .i\D041209.b\D1209058.D Page Report Date: 04-Jan-2005 07:41 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudej r .i\D041209.b\D1209058.D Lab Smp Id 72743-1 Inj Date 09-DEC -2004 23:34 Operator rsk Inst ID: dudejr.i Smp Info 300 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info Comment Egg Samples, 100 and 300 ppm dose groups, lot G Method \\Etstarget\Target\chem\dudej r.i\D0412 0 9 .b\D041209 .m Meth Date 04-Jan-2005 07:39 carlson Quant Type: ESTD Cal Date 09-DEC-20 04 17:44 Cal File: D1209024.D Als bottle 45 Dii Factor 1.00000 Integrator Falcon Compound Sublist: all.sub Target Vers ion : 4.12 Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN RT EXP RT DLT RT RESPONSE (ng/mL) FINAL (ng/mL) 3.446 3.440 0.006 6110226 487.607 488 3M Environmental Laboratory Page 183 of 227 Contains No CBI - 393 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filet \\Etstarget\Target\chem\dudejr* i\D041209*b\D1209058*D Page 2 3M Environmental Laboratory Page 184 of 227 Contains No CBI -394 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudej r.i\D041215a.b\D1215028 .D Page Report Date: 04-Jan-2005 07:43 1 3M Environmental Laboratory , E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudej r .i\D041215a.b\D1215028 D Lab Smp Id: 72597-1 Inj Date : 15-DEC-2004 12:33 Operator :cmc Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples 900 ppm Dose Group Method Meth Date Cal Date Als bottle: Dil Factor: :\\Etstarget\Target\chem\dudej r.i\D041215a.b\D041215a.m :04-Jan-2005 07:43 Carlson Quant Type: ESTD :15-DEC-2004 11:52 Cal File: D1215024.D 11 1.00000 Integrator: Falcon Target Version: 4.12 Compound Sublist: all.sub Compounds 1 PFBS (299) QUANT SIG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/mL) (ng/mL) 3.474 3.474 0.000 5410682 406.101 406 3M Environmental Laboratory Page 185 of 227 Contains No CBI - 395 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File* S\Etstarget\TargetSchem\dudejr* i\D041215a+b\D1215028*D Page 2 3M Environmental Laboratory Page 186 of 227 Contains No CBI -396- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \ \Etstarget\Target\chem\dudej r .i \ D 0 4 1 2 1 5 a .b \ D 1 215031.D Page Report Date: 04-Jan-2005 07:43 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file \\Etstarget\Target\chem\dudej r.i\D041215a.b\Dl215031. D Lab Smp Id 72598-1 Inj Date 15-DEC -2 004 13:04 Operator cmc Inst ID: dudejr.i Smp Info 900 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Misc Info Comment Egg Samples 9 0 0 ppm Dose Group Method \\Etstarget\Target\chem\dudejr.i\D041215a.b\D041215a.m Meth Date 0 4 -Jan-20 05 07:43 carlson Quant Type: ESTD Cal Date 1 5 -DEC-2 004 11:52 Cal File: D1215024.D Als bottle 14 Dil Factor 1.00000 Integrator Falcon Compound Sublist: all.sub Target Version: 4.12 Compounds 1 PFBS (299) QUANT S IG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/m L ) (ng/m L ) 3.474 3.474 0 .0 0 0 4923631 3 5 9 .4 5 4 359 3M Environmental Laboratory Page 187 of 227 Contains No CBI -397 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: S'sEtstargetSTargetSchemSdudejr* i\D041215a*h\Bi215031D Page 2 1 PFBS 3M Environmental Laboratory Page 188 of 227 Contains No CBI - 398 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \\Etstarget\Target\chem\dudejr .i\D041 2 1 5 a .b \ D 1 2 15062.D Page Report Date: 04-Jan-2005 07:43 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\d u d e j r .i\D041 2 1 5 a .b \ D 1 2 1 5 062 .D Lab Smp Id: 72750-1 Inj Date : 1 5 -DEC-2004 18:24 Operator : emc Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Egg; 02 Dec 04; C M C /M L A /MEE Mise Info : Comment : Egg Samples 900 ppm Dose Group Method : \\Etstarget\Target\chem\dudej r.i\D041215a.b\D041215a.m Meth Date : 04-Jan-2005 07:43 carlson Quant Type: ESTD Cal Date : 1 5 -DEC-20 04 11:52 Cal File: D1215024.D Als bottle : 41 Dii Factor: 1 . 0 0 0 0 0 Integrator : Falcon Compound Sublist: a l l .sub Target Versi on: 4.12 Compounds 1 PFBS (299) QUANT S I C MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/m L ) (ng/m L ) 3 .4 6 0 3.474 -0.014 8074161 7 1 7 .0 2 9 717 3M Environmental Laboratory Page 189 of 227 Contains No CBI -399- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filei SSEtstargetSTargetSchemSduclejr* i\D041215abSD1215062*D Page 2 1 PFBS 3M Environmental Laborator/ Page 190 of 227 Contains No CBI -400- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data File: \ \Etstarget\Target\chem\dudej r .i \ D 0 4 1 2 1 5 a .b \ D 1 2 15067.D Page Report Date: 04-Jan-2005 07:43 1 3M Environmental Laboratory E03-0583 Phase: Quantitation of PFBS in Quail Tissues Data file : \\Etstarget\Target\chem\dudejr.i\D041 2 1 5 a .b \ D 1 2 15 067 .D Lab Smp Id: 72751-1 Inj Date : 15-DEC -2004 19:15 Operator : emc Inst ID: dudejr.i Smp Info : 900 ppm Dose Group; Egg; 02 Dec 04; CMC/MLA/MEE Mise Info : Comment : Egg Samples 900 ppm Dose Group Method : \\Etstarget\Target\chem\dudejr.i\D041215a.b\D041215a.m Meth Date : 0 4 -Jan-2005 07:43 carlson Quant Type: ESTD Cal Date : 1 5 -DEC-2 0 04 11:52 Cal File: D1215024.D Als bottle : 44 Dil Factor: 1 . 0 0 0 0 0 Integrator : Falcon Compound Sublist: all.sub Target Versi on: 4.12 Compounds 1 PFBS QUANT S IG MASS 299 CONCENTRATIONS ON-COLUMN FINAL RT EXP RT DLT RT RESPONSE (ng/m L ) (ng/m L ) 3 .4 6 0 3.474 -0.014 7202747 6 0 1 .4 8 4 601 3M Environmental Laboratory Page 191 of 227 Contains No CBI -401 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Data Filej SSEtstargetSTargetNchemSdudejr i\D041215a*b\D1215067,D Page 2 1* 2- Y\EtstargetSTarget\chem\dudejr*i\D041215a*b\D1215067*D 1* 1- 1* 0- 0*9 0*8 0*7 so t 0*5 0*4- 0*3- 0*2- 0* 1- 0*5 1*0 1*5 1 PFBS <293> -I__ I__-I I-I. i 2*5 3*0 3*5 4*0 4*5 5*0 5*5 6*0 6*5 7*0 7*5 8*0 8^5 ________________________________ Mx n ixlO^) 3M Environmental Laboratory Page 192 of 227 Contains No CBI Report E03-0583 -402- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analyticai Determination of PFBS in Northern Bobwhite Tissues Attachment D: Study Protocol and Protocol Amendments 3M Environmental Laboratory Page 193 of 227 Contains No CBI Report E03-0583 -403 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues PROTOCOL T-7485: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE FIFRA Subdivision E, Section 71-4 ' OECD Guideline 206 3M Environmental Lab Project No. E03-0583 Submitted to 3M Corporation Environmental Laboratory P.O.Box 33331 St, Paul, Minnesota 55133 Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 2160] (410) 822-8600 July 14, 2003 3M Environmental Laboratory Page 194 of 227 Contains No CBI Report E03-0583 -404 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd. - 2- T-7485: A REPRODUCTION STUDY WITH THE NORTHERN BOBWHITE SPONSOR: 3M Corporation Environmental Laboratory 935 Bush Avenue St. Paul, Minnesota 55106 SPONSOR'S REPRESENTATIVE: Dr. John Newsted Entrix ' East Lansing Office 4295 Olcemos Road, Suite 101 Olcemos, MI 48864 TESTING FACILITY: Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 STUDY DIRECTOR: Sean P. Gallagher Senior Avian Biologist LABORATORY MANAOEMENT: Joann B. Beavers Director of Avian Toxicology Proposed Dales: Experimental Start Date: Project No.: Test Concentrations: Test Substance No.: FOR LABORATORY USE ONLY \\ ' I n d i t i T> Experimental Termination Date: Xi o | 16 1 )*4 '4 1-- 11 in Study Room: O -H Xe 1 \ o o \ o o f1oo /Xu 1 ____ ( n *-V0 I Reference Substance No. (inapplicable): *------ PROTOCOL APPROVAL STUDY DIRECTOR 4 '!> QZ DATE ____ . X LABORATORYMANAGEMENT ______________________ DATE iA SPONSOR'S REPRESENTATIVE _____ 9 / W a t DATE PROTOCOL NO.: 454/07140.VQR/SITI454 3M Environmental Laboratory 3M LAB PROJECT NO. 03-0583 Page 195 of 227 Contains No CBI Report E03-0583 -405 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd. -3 - OBJECTIVE The objective of this study is to evaluate the effects of dietary exposure to T-74S5 upon adult northern bobwhite ( Colinus virginianus) over a five-month period. Effects on adult health, weight gain and feed consumption will be evaluated. In addition, the effects of adult exposure to the test substance on the number of eggs laid, normal development of eggs, viability of the embryos, percent hatchability, offspring survival and egg shell thickness will be evaluated. SUMMARY Three treatment groups, each containing 16 pairs of Northern bobwhite, will be fed diets containing T-7485 at selected concentrations. A control group containing 16 pairs will be maintained concurrently. The bobwhite will be 16 to 36 weeks old and approaching their first breeding season. The birds will be fed the test diets for approximately ten weeks prior to the expected onset of egg laying, as well as during an egg laying period of at least 8 weeks. During the study, all birds will be monitored for toxicological responses. In addition, each of the treatment groups will be compared to the controls in order to detect statistically significant differences in body weight, feed consumption, egg production, fertility, hatching success, shell thickness and survivorship of offspring. Samples o f sera, liver, and egg contents will be collected for potential analysis. Additionally, samples of liver, kidney and gonad will be collected for histopatbological examination. The approximate duration of the study is six months. ' MATERIALS AND METHODS The methods, species used and route of administration described in this protocol are based upon procedures specified in Section 71-4 of the Environmental Protection Agency's Registration Guidelines, Pesticide Assessment Guidelines, FIFRA Subdivision E Hazard Evaluation: Wildlife and Aquatic Organisms (1); ASTM "Standard Practice for Conducting Reproductive Studies with Avian Species" (2); and upon OECD Guideline 206, Avian Reproduction Test (3), In order to control bias, adult birds will be randomized to pens. No other potential sources of bias are expected to affect the results of the study. PROTOCOL NO.: 454./071403/QR/$l:a i5 4 3M Environmental Laboratory 3M I..A.3 PROJECT NO. E03-0583 Page 196 of 227 Contains No CBI Report E03-0583 -406- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd. -4 - The test substance will be administered in the diet. This route of administration was selected because it represents the most likely route of exposure to avian species in the environment. Test Substance The test substance will be T-7485. Information on the characterization of test, control or reference substances is required by Good Laboratory Practice (GLP) Standards and Principles. The Sponsor is responsible for providing Wildlife International, Ltd. verification that the test substance has been characterized according to GLPs prior to its use in the study. If verification of GLP test substance characterization is not provided to Wildlife International, Ltd., it will be noted in the compliance statement of the final report The Sponsor is responsible for all information related to the test substance and agrees to accept any unused test substance and/or lest substance containers remaining at the end of the study. Treatment Groups There will bo at least three treatment groups and a control group. Each treatment and the control group will consist of 16 pairs of birds, housed with one male and one female per pen. Control birds will receive diet identical to the treated birds, but without the addition of the test substance. The test concentrations were established based upon results of a pilot reproduction study (Wildlife International, Ltd. Project No. 454-115). Wildlife International, Ltd. consulted with the Sponsor prior to establishing the test concentrations. Experimental Group (nmn a.i.) Control (0) 100 300 900 Number of Pens Per Concentration 16 16 16 16 BIRDS PER PEN MALES' FEMALES 11 11 11 11 Duration of Test The primary phases of the test and their approximate durations are: 1, Acclimation - At least 2 weeks. 2. Prc-photostimulation - 7 to 10 weeks. PROTOCOL NO.: 45-! V 71103;QP,'SUB454 3M Environmental Laboratory 5M LA 13 PROJECT NO. 1503-0585 Page 197 of 227 Contains No CBI Report E03-0583 -407 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd. -5- 3. Pre-egg laying (with photostirnulation) - Generally 1 to 4 weeks depending on response of birds, 4. Egg laying - At least 8 weeks. 5. Post-adult termination (final incubation, hatching, and 14-day offspring rearing peiiod) - Approximately 6 weeks. Test Species The Not them bobwhite represents an ecologically significant and widely distributed species in the United States. The bobwhite has demonstrated sensitivity to the effects produced by known toxic chemicals and has proven to be a good laboratory species from which a large amount of baseline data has been gathered. All bobwhite will be 16-36 weeks of age and apparently healthy at initiation of the test (first day of exposure to diets containing the test substance). All test birds will be from the same hatch. Selection of the appropriate weight range will be in accordance with Wildlife International, Ltd. Standaid Operating Procedures. Test birds will be approaching their first breeding season at the time of test initiation. Sex of the birds will be determined by a visual examination of tire plumage. Birds will be obtained fiomIC & L Quail Farm, Oroville, California or from another reputable supplier. All test biids will be acclimated to the caging and facilities for at least fourteen days prior' to initiation of the test. Extra birds from the same lot will be acclimated and may be used as replacements prior to the initiation of administration of test diets. Birds will be replaced if a pair appears to be incompatible or if a bird is deemed unsuitable due to injury, poor health or abnormal behavior. Identification , Adult biids will be identified by individual leg bands and will be randomly assigned to a pen and treatment group. Each pen will be identified by a unique number, and groups of pens will be identified by project number and concentration. All eggs laid during the study will be marked with a soft lead pencil or permanent ink for identification. Hatchlings will be identified by leg banding. Animal Diet All adult test birds will be fed a game bird ration formulated to Wildlife International, Ltd. speculations (Table ]). Five percent (w/w) limestone will be added lo the ration of adult birds to PROTOCOL NO.: 454/071403'QR,'SU.0454 3M Environmental Laboratory 3M LAB PROJECT NO, E0.5-05S3 Page 198 of 227 Contains No CBI Report E03-0583 -408 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues ife International, Ltd. - 6- provide a calcium source. Water will be provided by the town of Easton public water supply. Feed and water will be analyzed periodically according to Wildlife International, Ltd. Standard Operating Procedures. Neither the adults nor offspring will receive any form of antibiotic medication in the diet during the test without prior approval from the Sponsor. Specifications for acceptable levels of contaminants in game bird ration for avian species have not been established. However, there are no known levels of contaminants reasonably expected to be present in the diet that are considered to interfere with the purpose or conduct of the study. The adult birds will receive the appropriate treated or control diet for at least ten weeks prior to the anticipated onset of egg production. Exposure will continue until the adult termination. Appropriate feed and water will be provided ad libitum during acclimation and during the test. All offspring also will receive game bird ration (Table 1), but without the addition of limestone. The test substance will not be mixed into the diet of the offspring. Offspring will receive a watersoluble vitamin mix in their water. Feed and water will be provided to the offspring ad libitum during rearing. Diet Preparation Test diets will be prepared by mixing the test substance directly into the feed. Diet promtes will be prepared in a Hobart mixer (Model Number AS200T). If not used immediately, the premix will be frozen until used. Premixes will be prepared as frequently as necessary to assure stability of the test substance. Once each week, aliquots of the premix will be blended into bulle quantities of the ration to give the desired dietary concentrations of the test substance. Bulle diet mixing will be done in a Hobart mixer or a Patterson-Kelley twin shell dry blender. Excessive feed consumption may necessitate more frequent preparation of the diet. All test substance calculations will be based on the purity of the test substance as received or corrected to 100% active ingredient based on the information provided by the Sponsor. . f'T.O TO C O f NO.: MACO M O I/O R/SO IA;A 3M Environmental Laboratory AM LAB PROJECT NO. EOAOAA Pa9e 199 f 227 Contains No CBI Report E03-0583 -409- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd. -7 - Diet Sampling Samples of the experimental diets will be collected for chemical analysis to determine the homogeneity and stability of the test substance in the avian diet and to verify/ineasure test concentra tions. All samples will be placed in uniquely identified polypropylene jars. Samples will be collected on Day 0 of Week 1, at diet preparation, to determine homogeneity, measure test concentrations, and establish Day 0 values for evaluating stability. On Day .7 of Week 1, samples will be collected from the feeders containing the experimental diets to assess stability of the test substance under test conditions. Additionally, verification samples will be collected at specified intervals to measure/verify test concentrations. If a separate study is conducted with a different species (e.g., mallard), at the same test concentrations, diets that are prepared may be divided between the two studies. Therefore, samples collected for homogeneity and verification will be common to both studies. However, stability samples (e.g., Week 1 - Day 7) will be unique to that respective study. Stability samples will be collected by compositing diet from feeders presented to the birds, and taking samples from the composited feed. The diet sampling scheme is summarized below: ESTIMATED HUMBER OF SAMPLES . E xperim ental G roup C ontrol 100 ppui 300 ppm 900 ppm T o tals W eek 1 Day 0 Day 7 1 `1 62 2 62 2 62 2 19 7 W eek A I 2 2 2 7 W eek It W eek 12 I1 22 22 2 2 77 W eek 16 W eek 20 11 22 22 22 77 GRAND TOTAL =61 'S a m p le s c o llu d e d d u rin g W c cl:s 4 through 20 w ill be taken on D ay 0 o f th e c o rre s p o n d in g w eek. S a m p le s collected from the left and right sides o f the top, m iddle and bottom layers o f feed in the m ixing vessel to determ ine hom ogeneity. The above numbers of samples represent those collected from the test and do not include quality control (QC) samples such as matrix blades and fortifications prepared and analyzed during the analytical chcmistiy phase of the study. If the number of treatment groups is increased, the number of samples collected will increase proportionately. PROTOCOL NO.' 474/071407/QR/5 UB 3M Environmental Laboratory .1 HA PRO.IHt.TNO. FT0.TH Page 200 of 227 Contains No CBI Report E03-0583 -410- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues W ildlife international, Ltd. Diet Analyses Samples of the experimental diets will be stored in a freezer until prepared and/or extracted and analyzed. Chemical analyses of diets will be performed by Wildlife International, Ltd using liquid chromatography-mass spectrometry. The methodology used to analyze the test samples will be documented in the raw data and summarized in the final report. Housing and Environmental Conditions Tile adult birds will be housed in battery breeding pens manufactured by GQF Manufacturing Company (Model No. 206 or 0330) or equivalent. Each pen has floor space that measures approxi mately 25 X SI cm. Floors are sloping so that ceiling height will range from approximately 20 to 26 cm. External walls, ceilings and floors of each breeder pen are constructed of wire mesh while side walls are constructed of galvanized sheeting. Each pen will house one male and one female. During the test all birds will be maintained at an ambient temperature of approximately 15 to 30C. Ambient temperature and relative humidity will be recorded at least twice per tlay during the test. During the first seven weeks, lire photoperiod for the adults will be 8 hours of light per day. The photoperiod will then be increased to 17 hours of light per day to induce egg laying. The average light intensity for each test concentration will be at least sixty-five lux. Light intensity will be measured at least once during the test in accordance with Wildlife International, Ltd. Standard Operating Procedures. The average will be based upon light intensity measurements in each pen at bird height Lighting will be provided by fluorescent lights which closely approximate noon-day sunlight (noon-day sun - 4370, Chroma 50 or equivalent - 5000 Kelvin). Hatchlings will be placed in batteries of thermostatically, controlled brooding pens manufactured by the Beacon Steel Company (Model B735Q) or equivalent. Each pen measures approximately 72 X 90 X 23 cm high. The external walls and ceilings of each pen are constructed of wire mesh and galvanized sheeting. Floors are of wire mesh. The brooding compartment temperature will be maintained at approximately 38"C and will be recorded once per day. Acceptable ranges for brooding compartment temperatures will be specified in Wildlife International, Ltd. Standard Operating Procedures. Ambient room temperature will be maintained at approximately 16 to 32C. Ambient temperature and relative humidity will be recorded at least twice daily throughout the test. Photoperiod for the offspring will be )6 hours of light per day. PROTOCOL MO.: d5d/071d03/QR,'Sl,B/5d 3M Environmental Laboratory 3M LAB PROJECT NO. 03-0583 Page 201 of 227 Contains No CBI Report E03-0583 -411 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues W ildlife International, Ltd. -9- Housing and husbandry practices will be conducted so as to adhere to the guidelines established by the National Research Council (4). Incubation and Hatching Eggs will be collected daily, and marked with a soft lead pencil or permanent ink according to the pen from which they were collected. Eggs then will be stored at an average temperature of approximately 10 to 16C and an overall relative humidity of approximately 40 to 95%. Temperature and relative humidity in the cold room will be recorded twice per day during egg storage. All eggs collected will be placed in order by pen number and counted each week. Eggs selected for egg shell thickness measurements then will be removed. All remaining eggs will be candled to detect egg shell cracks or internal abnormalities. Cracked or abnormal eggs will be recorded and discarded. All eggs not selected for egg shell thickness measurement or discarded as cracked or abnormal will be fumigated with formaldehyde gas lo reduce the possibility of pathogen contamination. Following fumigation the eggs will be incubated. Temperature and relative humidity in the incubator will average approximately 37.2-37.8C anti 50-60%, respectively. The eggs will be candled between Days 10-12 for embryonation, and between Days 20-22 for embryo survival. Between Days 20-22 of incubation the eggs will bo removed from the incubator and placed in a hatcher. Temperature and humidity in the hatcher will average approximately 36.9-37.5"C and 60-80%, respectively. Dividers will be placed in the hatching trays to separate hatchlings by parental pen. Wet and dry bulb temperatures in the incubator and hatcher will be recorded at least twice daily during incubation and hatching. Hatchlings will be removed from the hatcher over an approximately 24 hour period beginning on approximately Day 25. Hatchlings will be leg banded so that they can be identified by parental pen. Offspring will be observed over a Id day period beginning when birds are first removed from the hatcher. Hie observation period will be extended if late mortality occurs which appears to be treatment related. PROTOCOL NO.: 454/071403/Q1RSUR45-I 3M Environmental Laboratory M LAB PROJECT NO. 1503-0583 Page 202 of 227 Contains No CBI Report E03-0583 -412- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues ' Wildlife International, Ltd. - 10- Observations All adults and offspring will be observed at least once daily for mortality, general condition, overt signs of toxicity and abnormal behavior. Animal Body Weigh ts/Fccd Consumption Individual body weights of the adults will be taken at the initiation of the test, at the end of test weeks 2, 4, 6, 8 and at adult termination. Group body weights of offspring by parental pen will be recorded at hatching and on Day 14 post-hatch. With the exception of tire last interval, feed consumption of the adults will be recorded by pen for a seven-day period each week throughout the test. The length of the last feed consumption interval will be determined by the scheduling of adult termination, Feed consumption will be measured by weighing the freshly Filled feeder on Day 0 of each week, recording any additional diet: added during the week, and weighing the feeder and remaining feed at the end of the seven day period. The accuracy of feed consumption values may be affected by unavoidable wastage of feed by birds. No attempt will be made to quantify the amount of wasted feed as it is normally scattered and mixed with water and Reproductive Parameters The following reproductive parameters will be measured and recorded: 1. eggs laid 2. eggs cracked 3. eggs incubated 4. infertile or dear eggs ' 5. dead embryos (to Day 10-12) 6. viable embiyos 7. dead embiyos (to Day 20-22) 8. live three week embiyos 9. unhatchcd eggs 10. hatchlings 1 1. 14-day old offspring survival PROTOCOL NO.: 454/07J4OT.'QR/SU345-1 3M Environmental Laboratory CM 1..AJ3 PROJECT NO, K0S 0 W Page 203 of 227 Contains No CBI Report E03-0583 -413 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues W ildlife international, Ltd. -n - 12. offspring body weights, day 0 and 14 13. egg shell thickness Egg Shell Thickness Measurements Each week eggs will be selected from those eggs laid during that week for egg shell thickness measurement. Normally, one egg will be collected from each of the odd numbered pens during odd numbered weeks (1,3,5, etc.), and onp egg will be collected from each of the even numbered pens during even numbered weeks (2,4,6, etc.). Prior to being opened, eggs will be rinsed with water to remove any excreta or feed adhering to the shell. Each egg will be cut open, the contents will be placed in an appropriate labeled container and stored frozen for potential analysis. The empty shell will be rinsed with tap water, and the shell air dried for at least one week. The thickness of the dried shell, including membranes, will be measured to the nearest 0.002 mm at five points around the equator of the egg with a micrometer. Egg contents that arc collected will be shipped to the Sponsor's designate and analyzed at the Sponsor's discretion. Necropsy and Tissue Collection All adult test birds that die during the course of the test and all adults remaining at the termination of the adult portion of the test will be subjected to a gross necropsy. Adult birds that arc not found dead during the course of the study will have blood drawn prior to euthanasia. The blood will be separated and the serum collected for potential analysis. Livers from each bird will be weighed and collected. A portion of the liver will be collected for histopathological examination, and the remaining liver will be stored frozen for potential analysis. Samples of kidney and gonad also will be collected for histopathological examination. All histopathological samples will be fixed in 10% buffered formalin and sent to EPL in Herndon, Virginia for histopathology. Additionally, similar samples will be collected from a single indiscriminately selected offspring from each pen (if available) at 14 days of age. Offspring will be sampled from the last lot of offspring produced. All samples collected for potential analysis will be placed in labeled polypropylene jars or other suitable containers and stored frozen until shipped to the Sponsor's designate. Blood and tissue samples will be analyzed at the Sponsor's discretion. PROTOCOL NO.: 4544014(1 TOILS L A ! V! 3M Environmental Laboratory 3 M LAB PRO.IECT NO. E()3-0Vo Page 204 of 227 Contains No CBI Report E03-0583 -414- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues 'Wildlife International, Ltd. - 12- Disposition of Test Birds At test termination, test birds will be euthanized by using carbon dioxide gas, cervical dislocation or other appropriate methods. The method used will be documented in the raw data. All birds wi ll be disposed of by incineration or other appropriate methods. Statistical Analyses Each of the treatment groups, will be compared to the control group using Dunnett's Multiple Comparison Procedure (5,6). The sample unit will be the individual pens within each experimental group, except for adult body weights where the sample unit will be the individual bird. Data expressed as a percentage will be arcsine transformed prior to using Dunnett's method. Nonpararnctric statistics may be used, if appropriate. Any pen in which au adult mortality occurs, including the on the day the study is terminated, will not be used in statistical comparisons of the reproductive data Analyses will be performed on each of the following parameters: 1. adult body weight 2. adult feed consumption 3. eggs laid per pen per day (eggs laid/maximum laid) 4. eggs cracked of eggs laid 5. viable embryos of eggs incubated 6. live 3-week embryos of viable embryos 7. hatchability of 3-weelt. embryos 8. fourteen-day old survivors of normal hatchlings 9. normal hatchlings of eggs incubated ' 10. nor mal hatchlings per pen per day (liatchlings/maximmn set) 11. fourteen-day old survivors of eggs incubated 12. fourteen-day old survivors per pen per day (14-day olds/maxinium set) 13. hatchling body weights 14. fourteen-day old survivor body weights 15. egg shell thickness A.OTOCOL, MO.: 454/07!403/Q1VSUB454 3M Environmental Laboratory I CAR i'CO.'CCT NO. 1704-05;:. Page 205 of 227 Contains No CBI Report E03-0583 -415 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS In Northern Bobwhite Tissues Wildlife International, Ltd.*1234567890 -13 - RECORDS TO BE MAINTAINED Records to be maintained for data generated by Wildlife International, Ltd. will include: 1. A copy of the signed protocol. 2. Identification and characterization of the test substance, if provided by the Sponsor. 3. Date of initiation, critical phases, and termination of the test, 4. Animal history. 5. Husbandly and environmental conditions. 6. Dietary concentration calculations and diet preparation. 7. Individual body weight measurements of adults and group body weight measurements of offspring. 8. Feed consumption measurements of adults. 9. Daily observations. 10. Necropsy findings. 11. Records of all reproductive parameters detailed in this protocol. 12. Analytical chemistry methods, results and chromatograms, if applicable. 13. Statistical calculations, if applicable. 14. A copy of the final report. FINAL REPORT A final report of the results of the study will be prepared by Wildlife International, Ltd. The report will include, but not be limited to, the following: 1. Name and address of the facility performing the study. 2. Experimental start and experimental termination dates and study completion date. It is the responsibility of the Sponsor to provide the final date that data are recorded for chemistry, pathology and/or supporting evaluations that may be generated at other laboratories. 3. A statement of compliance signed by the Study Director addressing any exceptions to Good Laboratory Practice Standards. 4. Objectives and procedures stated in the approved protocol, Including any changes in the original protocol. 5. Statistical methods employed for analyzing the data, when applicable. PROTOCOL NO.: -154,,071403/QR/SllB454 3M Environmental Laboratory 3M LAB PROJECT NO. LU3TLT3 Page 206 of 227 Contains No CBI Report E03-0583 -416- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd. - 14- 6. The test, control and reference substances identified by name, chemical abstracts number or code number, strength, purity, and composition or other appropriate characteristics, if provided by the Sponsor. 7. Stability and, when relevant to the conduct of the study, the solubility of the test, control and reference substances under the conditions of administration, if provided by the Sponsor. 8. A description of the methods used. 9. A description of the test system used. Where applicable, the final report shall include the number of animals used, sex, body weight range, source of supply, species, age, and procedure used for identification. 10. A description of the dosage, dosage regimen, route of administration, and duration. 11. A description of all circumstances that may have affected the quality or integrity of the data. 12. The name of the Study Director, the names of other scientists or professionals, and the names of all supervisory personnel, involved in the study. 13. A description of the transformations, calculations, or operations performed on the data, a summary and analysis of the data, and a statement of the conclusions drawn from the analysis. 14. The signed and dated reports of each of the individual scientists or other professionals involved in the study, if applicable. ' 15. The location where all specimens, raw data, and the final report are to be stored. 16. A statement prepared by the Quality Assurance Unit listing the dates that study inspections and audits were made and the dates of any findings reported to the Study Director and Management. 17. If it is necessary to make corrections or additions to a final report after it has been accepted, such changes shall be in the form of amendment by the Study Director. The amendment should clearly identify the part of the final report that is being added to or corrected and the reasons for the correction or addition. Amendments shall be signed and dated by the Study Director. CHANGING OF PROTOCOL Planned changes to the protocol will be in the form of written amendments signed by the Study Director and approved by the Sponsor's Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. Any other changes will be in the form of written deviations signed by the Study Director and filed with the raw data. All changes to the protocol will be indicated in the final report. PROTOCOL NO.: 45-1'071403'QR'Sl.:B-1.54 3M Environmental Laboratory Uvl LAB PBO J CT KO. E0.3-0583 Page 207 of 227 Contains No CBI Report E03-0583 -417 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues - 15 - GOOD LABORATORY PRACTICES This study will be conducted in accordance with Good Laboratory Practice Standards for EPA (40 CFR Part 160); OECD Principles of Good Laboratory Practices (ENV/MC/CPIEM (98) 17); and Japan MAFF (11 NohSan, Notification No. 6283, Agricultural Production Bureau, 1 October 1999). Each study conducted by Wildlife International, Ltd. is routinely examined by the Wildlife International, Ltd. Quality Assurance Unit for compliance with Good Laboratory Practices, Standard , , ildlife International, Ltd.Operating Procedures and the specified protocol. A statement of compliance with Good Laboratory Practices will be prepared for all portions of the study conducted by Wildlife International, Ltd. The Sponsor will be responsible for compliance with Good Laboratory Practices for procedures performed by other laboratories (e.g., residue analyses or pathology). Raw data for all work performed at Wildlife International, Ltd. and a copy of the final report will be filed by project number in archives located on the Wildlife International, Ltd. site, or at an alternative location to be specified in the final report. P14OT0 0 (U N O .: 454/071405 /Q R/Sl ' B4.44 3M Environmental Laboratory j M LAB PROJECT NO. E03 05:L Page 208 of 227 Contains No CBI Report E03-0583 -418 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Wildlife International, Ltd.*123456 -16- REFERENCES 1 U.S. Environmental Protection Agency. 1982. P esticide A ssessm ent Guidelines, F IF RA Subdivision F, H azard E valuation: Wildlife a n d A q u a tic O rganisms, subsection 714, Environmental Protection Agency, Office of Pesticide Programs. Washington, D.C. 2 American Society for Testing and Materials. 1986. Standard P ractice fo r C onducting R eproductive Studies with A vian Species, ASTM Standard El 062-86. Annual Book of ASTM Standards. Vol. 11.04. Philadelphia, PA. 15 pp. 3 Organization for Economic Cooperation and Development. 1984. A vian Reproduction le s t. OECD Guideline for Testing of Chemicals. Guideline 206, Paris. 4 National Research Council. 1996. Guide f o r the Care a n d Use o f I.aboraloiy Anim als. Washington, DC. National Academy Press. 125 pp. 5 Dumiett, C.W. 1955. A Multiple Comparison's Procedure for Comparing Several Treatments with a Control. Jour. Arner. Statis. Assoc. SO: 1096-1 121. 6 Dunnett, C.W. 1964. New Tables for Multiple Comparisons with a Control. Biom etrics 20: 482-491. PROTOCOL NO.: 454/07M03/QR/SUB754 3M Environmental Laboratory 3M LAB PROJECT NO. CO2-05S0 Pa9e 209 f 227 Contains No CBI Report E03-0583 -419- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues TABLE 1: DIET FORMULATION WILDLIFE INTERNATIONAL, LTD. GAMEBIRD RATION1 INGREDIENTS Pine Coro Meal Soy Bean Meal, 48% Protein Wheat Midds Protein Base Agway Special, 60% Protein Alfalfa Meal, 20% Protein _ Dried Whey Ground Limestone Eastman CalPhos Methionine Premix + Liquid Vitamin and Mineral Premix (see below) GL Perm (Fermalco)2 Salt Iodized Total PERCENT (%) 44.83 30,65 6.50 6.00 4.00 3.00 2.50 0.90 0,61 0.34 0.32 0,25 0.10 100.00 VITAMIN AND MINERAL PREMIA AMOUNT ADDED PER TON Vitamin D3 Vitamin A Riboflavin Niacin Pantothenic Acid Vitamin B |2 Folic Acid Biotin Pyridoxine Thiamine Vitamin E Vitamin K (Menadione Dimethylpyrimidiuol Bisulfite) Manganese Zinc Copper Iodine Eon Selenium 2.000. 0 001.C.U. 7.000. 0001.0, 6 grams 40 grains 10 grams 8 mgs 600 mgs 64 mgs 1.2 grams 1.2 grams 2 0 , 0 0 0 1.U. 5.8 grams 102 grams 47 grams 6.8 grams 1,5 grams 51 grams 182 mgs 1 The guaranteed analysis is a minimum o f 27% protein, a minimum of 2.5% crude fat and a maximum of 5% crude fiber. 2 Fermentation By-Products (Source ofUnidentified Growth Factors) PROTOCOL NO.: 454/071403/QR/SUB45-! 3M Environmental Laboratory 5\ i ! .AB PROJECT 5. EOT (UN 3 Page 210 of 227 Contains No CBI Report E03-0583 -420- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Lab Request Number E03-0583 PROTOCOL Amendment #1 S t u d y T itle T-7485: A reproduction Study w ith the N orthern Bobw hite D a ta R e q u i r e m e n t 40 CFR Part 160 A m e n d m e n t T itle P hase: A n aly tical D eterm in atio n of PFB S in N o rth e rn B obw hite T issues A m e n d m e n t D a te : Upon Signing P e r fo r m in g L a b o r a to r y 3M Environmental Health and Safety Operations 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 L a b o r a to r y P r o je c t Id e n tific a tio n 3M Environmental Lab Project No. E03-0583 W ildlife International, Ltd. Project No. 454-116 3M Environmental Laboratory 3M Environmental Laboratory Page 1 of 9 Page 211 of 227 Contains No CBI Report E03-0583 -421 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 Study Identification T-7485: A reproduction Study with the Northern Bobwhite Phase: Analytical Determination o f PFBS in Northern Bobwhite Tissues T e s t S u b s ta n c e Perfluorobutanesulfonate (PFBS) S p o n so r S p o n s o r R e p r e s e n ta tiv e S tu d y D ir e c to r P r in c ip a l A n a ly tic a l I n v e s tig a to r , A n a ly tic a l P h a s e T e s tin g F a c ility M a n a g e m e n t, A n a ly tic a l P h a se 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 Dr. John Newsted ENTRIX, Inc. East Lansing Office 4295 Okemos Road, Suite 101 Okemos, MI 48864 Sean P. Gallagher, Senior Avian Biologist Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 (410) 822-8600 Mark E. Ellefson 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 William K. Reagen 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 S tu d y L o c a tio n Testing Facility, In-Life Phase Testing Facility, Analytical Phase Wildlife International, Ltd. 8598 Commerce Drive Easton, Maryland 21601 3M Environmental Laboratory 935 Bush Avenue, Building 2-3E-09 St. Paul, MN 55106 P r o p o s e d S tu d y T im e ta b le Experimental Start Date Experimental Termination Date September 10, 2003 September 30, 2004 3M Environmental Laboratory 3M Environmental Laboratory Page 2 of 9 Page 212 of 227 Contains No CBI Report E03-0583 -422 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 1. S t u d y T-7485: A reproduction Study with the Northern Bobwhite; Phase: Analytical Determination of PFBS (T-7485) in Northern Bobwhite Tissues. 2. P u r p o se In this study, Northern Bobwhite quail were regularly exposed to nominal doses o f T-7485 (Perfluorobutanesulfonate, potassium salt or PFBS) in their feed in order to evaluate the effects o f dietary exposure upon adults over a five-month period. The purpose of this amendment is to address the analytical phase of the study in order to quantify PFBS in the liver, sera and eggs of both control and dosed Northern Bobwhite quail. 3. RegulatoryCompliance The analytical phase of this study will be conducted in accordance with Good Laboratory Practice Standards for EPA (40 CFR Part 160), OECD Principles of Good Laboratory Practices (ENV/MC/CHEM (98) 17); and Japan MAFF (11 NohSan, Notification No. 6283, Agricultural Production Bureau, 1 October 1999). The quality control samples prepared in this study will cumulatively constitute a cross validation of method ETS-8-231, "Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry." 4. QualityAssurance The 3M Environmental Laboratory Quality Assurance Unit will audit the study conduct, raw data, and final report to determine compliance with Good Laboratory Practice Regulations, this protocol, and 3M Environmental Laboratory Standard Operating Procedures for this phase o f the study. The Quality Assurance Unit will report all findings to the Study Director and the Principal Analytical Investigator. 3M Environmental Laboratory 3M Environmental Laboratory Page 3 of 9 Page 213 of 227 Contains No CBI Report E03-0583 -423 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 5. Test, Control, andReferenceSubstance Table 1: Test Substance Test Substance Perfluorobutane sulfonate** Acronym and Structure PFBS C4F9CO2 Identification Number TCR-282 Source 3M S ta b ility in Water y I____ i R9 R I pecial` y mg/mL 97'3% Chemicals | a ( Storage I Expiration Conditions I Date I Frozen 1/17/2007 * According to EOO-1877, "Determination of the Solubility o f Ammonium Perfluorobutanesulfonate (PFBS) in Water" **The vapor pressure and octanol/water partition coefficient are not available. 5 .1 S ta b ility The stability of PFBS has been investigated in the following studies: E00-2188, "Screening Studies on the Aqueous Photolytic Degradation of Perfluorobutanesulfonate (PFBS)" EOO-2170, "Hydrolysis Reactions of Perfluorobutanesulfonate (PFBS)" 5 .2 R e s e r v e S a m p le s A retention sample of the test substance has been taken and stored frozen at -20C 5C. The sample will be archived for at least the period required by the EPA GLPs, Section 792.195. 5 .3 D is p o s itio n The disposition of the test substance, if any, will be conducted and recorded according to ETS-3-8, "Disposition of Test, Control, and Reference Substances Received for GLP Studies." 5 .4 S a fe ty P r e c a u tio n s Wear appropriate personal protective equipment. Consult the MSDS located in the TCR database in Lotus Notes for more specific information. 5.5 L o c a t i o n o f M e t h o d D o c u m e n t a t i o n f o r S y n t h e s i s Contact 3M Specialty Materials for information regarding the synthesis/origins o f this lot of PFBS. 6 . TestSystem The test system is Northern Bobwhite quail. Northern Bobwhite serum, liver and egg samples were collected by Wildlife International, Ltd. 6 .1 J u s tific a tio n o f th e S e le c tio n o f th e T e s t S y s te m : The Northern Bobwhite quail represents wild bird populations and is an EPA recommended species. The use o f these matrices for this study is to determine the levels of test substance in the liver, serum and eggs of mating adults that were fed PFBS. Analysis of liver and serum from hatchlings will determine if PFBS was passed on to the offspring of these mating adults. 3M Environmental Laboratory Page 4 of 9 3M Environmental Laboratory Page 214 of 227 Contains No CBI Report E03-0583 -424- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 6 .2 D o s in g o f th e T e s t S y s te m The dosing o f the test system involved adding the test substance to the animal feed, and was described in the study protocol. This phase of the study involves only the analytical determination of PFBS in tissue samples. The sample identification scheme for this phase of the study will be as follows: Each study sample, each control sample, each control blank and all of their aliquots will have a unique identifier throughout the course o f the analytical phase of the study. (Sample numbers will be generated by the 3M Environmental lab LIMS system, and will reference the sample numbers generated at Wildlife International, Ltd.) When the study samples are received, they will be given a unique identification number in the LIMS system. This number will stay with the tissue sample throughout this phase o f the study. Homogenates, extracts, and diluted extracts will be designated as such on the label. 7. SpecimenReceipt Serum, liver and eggs were collected by Wildlife International, Ltd. and shipped on dry ice to the 3M Environmental Laboratory for analysis. Upon arrival in the lab, the samples will be maintained frozen except when removed for extraction and analysis as described in the method. The samples will be kept isolated from the test substance during storage. The following type and number of birds were euthanized at the 20-week endpoint in this study: 32 control adult birds (16 mating pairs) unexposed to PFBS 32 experimental adult birds (16 mating pairs) dosed at 100 ppm PFBS 32 experimental adult birds (16 mating pairs) dosed at 300 ppm PFBS 32 experimental adult birds (16 mating pairs) dosed at 900 ppm PFBS First generation offspring birds from each test group Sample receipt conditions were recorded on the external chain o f custody form that arrived with the samples. Sample movement within the 3M Environmental Lab will be tracked using an internal chain of custody. Both fonns will be archived with the study data at the end of the study. 8. Experimental Design 8 .1 P r e p a r a to r y M e th o d s Preparation and analysis will be conducted using ETS-8-231, "Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry" with the following modifications: 8.1.2 A different make/model of centrifuge, tissue grinder, vortex mixer, and analytical column may be used. 8.1.3 An Agilent MSD detector will be used in place of the Micromass instrument. The 3M Environmental Laboratory Page 5 of 9 3M Environmental Laboratory Page 215 ot 227 Contains No CBI Report E03-0583 -425 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 8. 1.4 8 .1 .5 8 .1 .6 8 .1 .7 8 .1 .8 conditions used for analysis will be recorded in the raw data. Modifications to the conditions in the method will be made as needed. All sample measurements and calculations will be based on weight, not volume (volumes need not be recorded). Sample weights other than 1 g + 0.100 g may be used. Dilutions other than 1/6 will be allowed. Samples will be homogenized in acetonitrile rather than water. Consequently, the protein precipitation step followed by shaking will be omitted. Acetonitrile will then also be used for the method blanks, etc. The homogenized samples will be extracted using a Zymark RapidTrace Automated SPE System, as described in ETS-9-29. Modifications to the conditions in the method will be made as needed. Extracted standards may be used for the system suitability standards in place of unextracted standards. 8 .3 M e th o d s fo r C o n tr o l o f B ia s Control o f bias is accomplished using analytical spikes (as indicators o f sample recovery and accuracy) and matrix blanks (for evaluation o f possible contamination of the matrix). Solvent blanks will also be analyzed (possible sample contamination during the dilution process, carryover between analyses, and instrument contamination). Samples will be prepared in triplicate and used as indicators of precision. 9. Da t a Q u a l ity O b je c t iv e s 9 .1 C a lib r a tio n S ta n d a r d s . Samples will be bracketed by a calibration curve and passing CCVs. Calibration standards used to generate a calibration curve should be prepared in the same manner as the study samples. The number of calibration standards and concentration levels should be sufficient to encompass the expected concentrations of the study samples or dilutions thereof. The coefficient of determination (r2) of the standard curve must be equal to or greater than 0.990. If the calibration curve residuals are greater than 15% deviation (LLOQ 80%-120%) from the theoretical value, quadratic curve fitting and/or dropping low/high curve points may be required if data review shows this to be a consistent and more accurate representation of the instniment response. 9 .2 C o n tin u in g C a lib r a tio n V e r ific a tio n (C C V ). Analyze a mid-range calibration standard after a maximum of every ten samples. Acceptable CCV values are 85-115% of true value. 9 .3 S o l v e n t b la n k . Solvent blanks are run at a minimum before and after every calibration curve, and after every CCV. Solvent blanks may be run before and after matrix and control blanks if contamination is noted. Acceptable values for the blanks are values below 50% o f the limit of quantitation (LLOQ). If analyte carryover is a problem, use back-to-back solvent blanks and use the last solvent blank to evaluate carryover. 3M Environmental Laboratory 3M Environmental Laboratory Page 6 of 9 Page 216 of 227 Contains No CBI Report E03-0583 -426- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 9 .4 Q u a lity C o n tr o l S a m p le s Quality control samples (QCS) will be prepared according to method ET8-8-231. The QCS will be spiked at three levels as described in the method. The analyst shall accept percent spike recoveries between 85% and 115% of theoretical value. A minimum o f 2/3 of all QCS in any one analytical run must pass; otherwise, the sample set must either be reanalyzed or re-extracted. 9 .5 L o w e r L im it o f Q u a n tita tio n (L L O Q ). The lowest concentration that can be reliably measured within specified limits of accuracy during routine laboratory operating conditions. The LLOQ is defined as the lowest non-zero standard (80%-120%) in the calibration curve that is greater than or equal to 2 times the level of the matrix blank. Sample LLOQ are highly matrix-dependent. 9 .6 D e m o n s tr a tio n o f S p e c ific ity . The analytical technique o f HPLC/MS provides for chromatographic separation of targeted materials and detection/quantitation of selected ions which are characteristic of the targeted compounds. 9 .7 S y s te m S u ita b ility Three injections of a mid-level standard will be run prior to the calibration curve and after the last sample. The area counts should have a relative standard deviation <5% and the retention times should be <2% when evaluated independently. 10. StatisticalAnalysis Statistical methods for the analytical results will be limited to the calculations of means, standard deviations, relative percent difference, and percent recoveries (as appropriate). Calculations will be performed as described in the applicable methods. 11 Report A report of the results of the analytical phase of this study will be prepared by 3M Enviromnental Laboratory. The report will include, but not be limited to, the following, when applicable: Name and address of the facility perfomiing the analysis, Dates upon which the analysis was initiated and completed. A statement o f compliance by the Study Director addressing any exceptions to Good Laboratory Practice Standards. Objectives and procedures as stated in this protocol amendment. The test substance identification by name, chemical abstracts number or code number, strength, purity, and composition or other appropriate characteristics, if provided by the Sponsor. Stability and the solubility o f the test substances under the conditions of administration, if provided by the Sponsor. A description o f the methods used to conduct the test(s). A description of the test system. A description of any circumstances that may have affected the quality or the integrity of the data. The name of the Study Director and the names of other scientists, professionals, and supervisory personnel involved in the study. 3M Environmental Laboratory Page 7 of 9 3M Environmental Laboratory Page 217 of 227 Contains No CBI Report E03-0583 -427- Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 A description of the transformations, calculations, or operations performed on the data, a summary and analysis of the analytical chemistry data, and a statement o f the conclusions drawn from the analyses. Statistical methods used to evaluate the data, if applicable. The signed and dated reports of each o f the individual scientists or other professionals involved in the study, if applicable. The location where raw data and the phase report are to be stored. A statement prepared by the Quality assurance unit listing the dates that study inspections and audits were made for this phase o f the study, and the dates o f any findings reported to the Study Director, the Principal Analytical Investigator and Management. If it is necessary to make corrections or additions to a phase report after it has been accepted, the changes will be made in the form of an amendment issued by the Study Director. The amendment will clearly identify the part of the phase report that is being amended, the reasons for the amendment, and will be signed by the Study Director. 12. L o c atio n o f R a w Da t a , R e c o r d s, a n d P h a s e R e p o r t Original data or copies thereof, will be available at 3M Environmental Laboratory to facilitate audits o f the study during its progress and before acceptance o f the final report. When the final report is completed, all original paper data, including those items listed below will be retained in the archives o f 3M Environmental Laboratory for a period of not less than 10 years following signing o f the final report. The following raw data and records will be retained in the study folder in the archives according to 3M Environmental Laboratory Standard Operating Procedures. 1 2 .1 .1 Approved protocol and amendments 1 2 .1 .2 Study correspondence 1 2 .1 .3 Shipping records 1 2 .1 .4 Raw data 1 2 .1 .5 Approved phase report (original signed copy) 1 2 .1 .6 Electronic copies o f data The following supporting records will be retained separately from the study folder in the archives according to 3M Environmental Laboratory Standard Operating Procedures: 1 2 .1 .7 Training records 1 2 .1 .8 Calibration records 1 2 .1 .9 Instrument maintenance logs 1 2 .1 .1 0 Standard Operating Procedures, Equipment Procedures, and Methods 1 2 .1 .1 1 Appropriate specimens 13. Data/ SampleRetention A portion o f the test substance used in this study will be retained for a period not less than 10 years following the application of the final test rule. The extracts will only be retained so long as their quality affords evaluation. 3M Environmental Laboratory 3M Environmental Laboratory Page 8 of 9 Page 218 of 227 Contains No CBI Report E03-0583 -428 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 14. Protocol Amendmentsand deviations Planned changes to the protocol will be in the form of written amendments signed by the Study Director and the Sponsor's Representative. Amendments will be considered as part o f the protocol and will be attached to the final protocol. Any other changes (unplanned) will be in the form of written deviations, signed by the Study Director and filed with the raw data. All changes to the protocol and the reason for the changes will be indicated in the final report. 15.Attachments N one Signatures S e a n P . G a lla g h e r , Study Director Date Date Date 3M Environmental Laboratory 3M Environmental Laboratory Page 9 of 9 Page 219 of 227 Contains No CBI -429- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues A u s 20 2 00 4 2 : 2 5PM ENTRIX EAST LANSING N o . 2720 P. 2 Laboratory Request Num ber E03-0583 14.ProtocolAmendmentsanddeviations Planned changes to the protocol will be in the form o f written amendments signed by the Study Director and the Sponsor's Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. Any other changes (unplanned) will be in the form of written deviations, signed by the Study Director and filed with the raw data. All changes to the protocol and the reason for the changes will be indicated in the final report. 15.Attachments None 1 I Signatures | S e a n P G a lla g h e r, Study Director Date ! J o ty p N e w s te d , Sponsor Representative . M a r k E Elle.faon, Principal Analytical Investigator Date I ............ | W illia m K . R ea g a n , 3M Environmental Laboratory Management Date 3M Environmental Laboratory 3M Environmental Laboratory Page a of 9 Page 220 of 227 Contains No CBI -430 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues 08/20/04 16:53 FAX 410 822 0632 WILDLIFE INTERNATIONAL 0)002 Laboratory Request Num ber E03-0583 14. ProtocolAmendments anddeviations Planned changes to the protocol will be in the form of written amendments signed by the Study Director and the Sponsor's Representative. Amendments will be considered as part of the protocol and will be attached to the final protocol. Any other changes (unplanned) will be m the form o f written deviations, signed by the Study Director and filed with the raw data. All changes to the protocol and the reason for the changes will be indicated in the final report. 15.Attachments None Signatures S e a n P. G allagher, Study Director 0^ Date J o h n N ew sted, Sponsor Representative Date M a r k E . E ilefson, Principal Analytical Investigator W illiam K. R eagen, 3M Environmental Laboratory Management Date Date f 3M Environmental Laboratory 3M Environmental Laboratory Page 9 o f 9 Page 221 of 227 Contains No CBI Report E03-0583 -431 - Project Number 454-116 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Lab Request Number E03-0583 PROTOCOL Amendment #2 Study Title T-7485: A Reproduction Study with the Northern Bobwhite Data Requirement 40 CFR Part 792 Amendment Title Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Amendment Date: Upon Signing Performing Laboratory 3M Environmental Health and Safety Operations 3M Environmental Laboratory 935 Bush Avenue St. Paul, MN 55106 Laboratory Project Identification 3M Environmental Lab Project No. E03-0583 Wildlife International, Ltd. Project No. 454-116 3M Environmental Laboratory 3M Environmental Laboratory Page 1 o f 6 Page 222 of 227 Contains No CBI -432- Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 Protocol Reads (from Protocol Amendment #1): 3. Regulatory Compliance The analytical phase o f this study will be conducted in accordance with Good Laboratory Practice Standards for EPA (40 CFR Part 160), OECD Principles of Good Laboratory Practices (ENV/M C/CHEM (98) 17); and Japan MAFF (11 NohSan, Notification No. 6283, Agricultural Production Bureau, 1 October 1999). The quality control samples prepared in this study will cumulatively constitute a cross validation of method ETS-8-231, "Solid Phase Extraction and Analysis o f Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry." Amend to Read: 3. Regulatory Compliance The analytical phase of this study will be conducted in accordance the United States Environmental Protection Agency Good Laboratory Practice Regulations, 40 CFR 792. The quality control samples prepared in this study will cumulatively constitute a cross validation o f method ETS-8-231, "Solid Phase Extraction and Analysis of Perfluorinated Acids, Alcohols, Amides, Sulfonates and other Fluorinated Compounds by High Performance Liquid Chromatography/Mass Spectrometry." Reason: The previous regulations were not appropriate for this type o f study. Protocol ReadsTPage 15): This study will be conducted in accordance with Good Laboratory Practice Standards for EPA (40 CFR Part 160); OECD Principles o f Good Laboratory Practices (ENV/MC/CHEM (98) 17); and Japan MAFF (11 NohSan, Notification No. 6283, Agricultural Production Bureau, 1 October 1999). Amend to Read: This study will be conducted in accordance with Good Laboratory Practice Standards for EPA (40 CFR Part 792). Reason: The previous regulations were not appropriate for this type o f study. Protocol Reads (from Protocol Amendment #11: 5. Test, Control, and Reference Substance Table 1: Test Substance Test Substance Acronym and Structure Identification Number ,, Source Solubility in Water' Purity Storage Conditions Expiration Date Perfluorobutane sulfonate** PFBS C 4F9C O 2 TCR-282 3M I 52 6 ChPeemdiacaltyls | mga/mL 97.3% Frozen 1/17/2007 3M Environmental Laboratory 3M Environmental Laboratory Page 2 o f 6 Page 223 of 227 Contains No CBI -433 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 Amend to Read: 5. Test and Reference Substance The test substance in this study is identified in the protocol as T-7485. This number is traceable to the substance PFBS, which was provided to the test facility by the sponsor. The lot number o f the test substance was not provided to the principal analytical investigator for the analytical phase o f the study. Table 1: Reference Substance R e fe re n c e S ubstance A c ro n y m an? S tru c tu re identification I Q N um ber Source S olubility I p . . 1 S torage 1 E xpiration in W ater* 1 uri ^ C o n d itio n s I D a te P erfluorobutane s u lfo n a te * * PFBS _ T C R -2 8 2 C4F9SO3 1 3M S pecialty 1 ch e m ica ls 52/ 6 . m g/m L 97.3% Frozen Reason: Incorrect molecular formula. Change "test" to "reference" substance. 1 /1 7 /2 0 0 7 Protocol Reads (from Protocol Amendment #1. Section 8.1.7): The homogenized samples will be extracted using a Zymark RapidTrace Automated SPE System, as described in ETS-9-29. Modifications to the conditions in the method will be made as needed. Amend to Read (Section 8.1.7): The homogenized samples will be extracted using a Zymark RapidTrace Automated SPE System, as described in ETS-9-29. Modifications to the conditions in the method will be made as needed. In the event o f incomplete aspiration of the sample by the RapidTrace, the amount left in the sample tube will be noted in the raw data. If more than 0.5 mL is left unextracted, that sample will be discounted. If less than or equal to 0.5 mL is left, the sample may be used without correction. Reason: Clarify sample usage in the event o f an incomplete extraction o f a sample. Protocol Reads (from Protocol Amendment #1, Section 10): Statistical methods for the analytical results will be limited to the calculations o f means, standard deviations, relative percent difference, and percent recoveries (as appropriate). Calculations will be performed as described in the applicable methods. Amend to Read (Section 10): Statistical methods for the analytical results will be limited to the calculations o f means, standard deviations, relative percent difference, and percent recoveries (as appropriate). Calculations will be performed as described in the applicable methods. In the event that one (or more) o f the replicate samples is below the LLOQ and one (or more) is above the LLOQ a result equal to the LLOQ will be used for the calculation o f an average value for that sample. Reason: Clarify calculations used in the analytical phase o f the study. Protocol Reads (from Protocol Amendment #1, Section 9.4): Quality control samples (QCS) will be prepared according to method ETS-8-231. The QCS will be spiked at three levels as described in the method. The analyst shall accept percent spike 3M Environmental Laboratory 3M Environmental Laboratory Page 3 o f 6 Page 224 of 227 a Contains No CBI -434 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 recoveries between 85% and 115% of theoretical value. A minimum of 2/3 o f all QCS in any one analytical run must pass; otherwise, the sample set must either be reanalyzed or re-extracted. Amend to Read (Section 9.4): Q uality control samples (QCS) will be prepared according to method ETS-8-231. The QCS will be spiked at three levels as described in the method. The analyst shall accept percent spike recoveries between 85% and 115% o f theoretical value. A minimum of 2/3 of all QCS from anyone extraction day must pass; otherwise, the sample set must either be reanalyzed or re extracted. Reason: Clarify data quality objectives actually used in the study. Protocol Reads: No previous section. Amend to Read (Section 9.8. Protocol Amendment #1): 9 .8 M ethod Validation The method will be validated as described below. 9.8.1 Specificity Specificity will be demonstrated by the acceptable recovery of quality control samples prepared during the course o f this study, and by the examination o f chromatographic resolution provided by the instrument. The acceptance limits will be the same as described in the protocol for the QCS samples (2/3 o f spike recoveries between 85% and 115% o f theoretical value and a precision o f 15%). 9.8.2 Selectivity Selectivity will be demonstrated by the evaluation o f the 0 ppm dose group samples. Specimens with endogenous levels of analyte greater than 50% o f the LLOQ will be noted in the phase report and will not be used as blank matrix in the remainder o f the study. 9.8.3 Calibration The calibration curves generated during the course o f the study will be evaluated according to the acceptance criteria described by the analytical method (Any level outside 85% - 115% of nominal must be deactivated, and regression re-calculated, except the LLOQ which must be w ithin 20% o f nominal. All levels must show a response greater than twice that o f the blank.). For information purposes only, relative standard deviation (RSD) will be calculated for each concentration over the several days' data, only using the values that were kept activated. 9.8.4 Limit of Detection (LOD) and Limit of Quantitation (LOQ) For each matrix, six matrix spikes will be prepared ranging in concentration from 0.25 ng/mL to 5.0 ng/mL. The instrument response will be evaluated against the theoretical concentration using a linear regression tool in Excel. The LOD and LOQ will be calculated using the resulting standard error (oo) and slope (called "X Variable 1" in the spreadsheet): L O D = 3.3 x <70/ S and L O Q ^ l O x a J S 9.8.5 Method Precision and Accuracy Quality control samples will be prepared according to the requirements of the method. These samples will be evaluated according to the acceptance limits described in the protocol to demonstrate precision and accuracy. 9.8.6 Absolute Recovery and Matrix Effect Blank matrix will be spiked at three levels and evaluated against a solvent curve. The results will be presented in the phase report. There is no acceptance limit for this evaluation. 3M Environmental Laboratory 3M Environmental Laboratory Page 4 o f 6 Page 225 of 227 Contains No CBI -435 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 9.8.7 Ruggedness Several analysts will participate in the preparation and analysis of the samples in this study. Since it is a long preparation and analysis procedure, it is not practical to have a totally separate group o f personnel perform the procedure from beginning to end. If the QC samples pass within the requirements o f the method (2/3 o f spike recoveries between 85% and 115% of theoretical value and a precision o f 15%) the ruggedness o f the method will be judged sufficient. 9.8.8 Robustness No experiments will performed to evaluate this parameter. As the sample calculations will be performed using weights rather than volumes individual technique is not expected to play a significant role in the generation of results. If the evaluation of the data quality objectives described in the protocol yields acceptable results, the method will be considered sufficiently robust. 9.8.9 C arryo ver During the course o f the study solvent blanks immediately following high level standards and samples will be evaluated for evidence of instrument carry over. Solvent blanks should show peak areas no more than 50% o f peak areas in the LLOQ standard. If carry-over is more than 50% o f LLOQ, additional steps must be taken to alleviate the carry-over (e.g., an additional blank analyzed after the high standard, re-injection of a LLOQ-level sample that follows a high sample, etc.). 9.8.10 Range o f Quan tita tion The range o f analyte concentrations for which all components o f the validation meet their respective acceptance criteria will be evaluated and discussed in the phase report. 9.8.11 System Suitability The test results for all the previous components will be evaluated. Parameters that may be used for documenting the proper operation o f the analytical equipment will be established and discussed in the phase report. 9.8.12 Dilution Scheme It is expected that samples may occasionally exceed the validated upper limit o f the range o f quantitation. The reference dilution scheme, which does not need to be validated, is to take a new aliquot o f the original matrix, dilute it with blank matrix, and extract the diluted sample. If instead the injection volume is varied to produce an on-column dilution (or concentration) then quality control samples will be evaluated using the same technique. They must pass within the same acceptance limits described in the protocol (2/3 o f spike recoveries between 85% and 115% o f theoretical value and a precision o f 15%). 9.8.13 Stability Testing The stability o f the test substance in the test system (tissues) and in the homogenate is not known and will not be investigated during the course o f this study. Stability o f the test substance in the final extract (stored in autovials) will be evaluated over some reasonable length of storage (approximately one month). Three levels of quality control samples will be run soon after extraction, and then again approximately one month later against a newly extracted curve. The acceptance criteria will be the same as discussed in the protocol for quality control samples (2/3 o f spike recoveries between 85% and 115% o f theoretical value and a precision of 15%). If the samples do not have acceptable recoveries an investigation of extract holding times will be made and described in the report. Also, any sample results obtained with extracts beyond this holding time will be evaluated. Either the samples will be re-extracted or the data flagged to indicate possible bias o f the data. Results will be presented in the phase report. 3M Environmental Laboratory 3M Environmental Laboratory Page 5 o f 6 Page 226 of 227 Contains No CBI -436 - Project Number 454-116 Report E03-0583 T-7485: A Reproduction Study with the Northern Bobwhite Phase: Analytical Determination of PFBS in Northern Bobwhite Tissues Laboratory Request Number E03-0583 9.8.14 Uncertainty After all other phases of the validation are complete the measurement uncertainty will be calculated using the model provided as Attachment A in ETS-8-231.2. The same factor will be used for estimating the uncertainty o f the liver subsample homogeneity as was used in that reference. The egg and serum samples do not require this additional factor, as the entire egg will be used to make the homogenate and the serum is expected to be homogeneous. Reason: Clarify the scheme used to validate the method for the current study. 3M Environmental Laboratory 3M Environmental Laboratory Page 6 of 6 Page 227 of 227 Contains No CBI ,W ildlife International, Ltd -437- Project Number 454-116 Appendix XVIII Diagram of Test Layout b - Chroma lights, four-foot Diagram not to scale. Room approximately 15 x 9 ft. (4.6 x 2.7 m) Contains No CBI Wildlife International, Ltd. -438 - Project Number 454-116 Appendix XIX Changes To Study Protocol This study was conducted in accordance with the study protocol signed on September 5, 2003 and the following amendments and deviations: 1. The protocol was amended to indicate sampling of offspring would be conducted when the offspring were 16 days of age. 2. The protocol was amended to change the description of the basal diet formulation for Wildlife International Ltd. Game Bird Ration in Table 1. Prior to test initiation the formulation for the game bird ration was modified to replace one protein source with another protein source. 3. The protocol was amended to add the description of the diet formulation for diet used for approximately the last quarter of the study. The source of Choline Chloride added to the diet was changed during the test. However, the overall nutritional balance of the diet did not change. 4. The protocol was amended to indicate the spleen from the adult female in Pen 1655 of the 900 ppm a.i. test concentration would be collected for histopathological examination. The spleen for this hen was noted to be slightly enlarged during gross necropsy. 5. The protocol was amended to indicate fresh diets would not be prepared and feed consumption would not be measured for the final day (Week 22, Day 1) of the adult in-life portion of the test. Instead, birds were fed diets remaining from Week 21 on the final day prior to adult termination. 6. A blood sample was inadvertently not collected from the female in Pen 1639 of the 300 ppm a.i. treatment group, prior to euthanasia. This female was euthanized in Week 14 of the test due to its moribund condition. 7. The liver for the adult male in Pen 1654 of the 900 ppm a.i. concentration was inadvertently not weighed prior to collection for histology and residue analysis. 8. Statistical comparison of liver weights were made by Dunnett's t-test, using TOXSTAT software. The protocol did not require statistical comparison of liver weights. 9. The protocol was amended to change the EPA regulations under which the study was conducted to Good Laboratory Practice Standards for EPA (40 CFR Part 792). Contains No CBI Wildlife International, Ltd. -439 - Project Number 454-116 Appendix XX Personnel Involved in Study The following key personnel were involved in the conduct or management of this study: Avian Toxicology (1) Mark Jaber, Wildlife Toxicologist (2) Joann B. Beavers, Director of Avian Toxicology (3) Linda R. Mitchell, Manager of EcoTox Operations (4) Diana Temple, Supervisor, Avian Toxicology (5) Sean P. Gallagher, Senior Biologist Chemistry (1) Willard B. Nixon, Ph D., Director of Chemistry (2) Raymond L. Van Hoven, Ph.D., Scientist Contains No CBI