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i /[R 2 1 6 -0 4 5 H 3M Environmental Laboratory Final Report- Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6067, T-6068 and T-6069 in Rabbits In-Vivo Study Reference Number: HWI#6329-152 Study Number: A M D T-011095.1 Test Substance: L -13492 (T-6067, T-6068 and T-6069) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M Environmental Technology & Services 935 Bush Avenue St. Paul, MN 55106 Method Numbers and Revisions: A M D T-M -1-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver AM DT-M --2- 0, Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer AM DT-M ->4-0, Extraction o f Fluorochemicals from Rabbit Liver AM DT-M -5-0, Analysis o f Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry AM DT-M - 8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode AM D T-M -14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum Initiation Date: See attached protocol Author: James D. Johnson CQ2Q*^ 1.0 SUMMARY A t-butyl ammonium salt o f perfluorooctanoate in the form o f treated fabric and as a liquid formulation was applied dermally to rabbits. Liver samples were analyzed at 28 days post dose by combustion for total organic fluorine. The results from treated animals were the same as control values. All total organic values were below the practical quantitation limit. Serum levels were also below the practical quantitation limits o f the analysis for samples collected at day 1 and 2 after administration o f the mixture or the treated fabric. From the pharmacokinetic study (HW I#6329-151), it would be unlikely that any extent o f absorption could have been detected in this study. 2.0 INTRODUCTION This study was designed to provide information as to assess the systemic absorption and toxicity and relative skin irritancy o f L -13492 (T-6067, T-6068 and T-6069) when applied to the skin o f rabbits. In a pharmacokinetic study (HW I#6329-151), this compound showed little accumulation at 48 hours in liver. The biological half-life determined from serum levels in female rabbits was about 4 hours after an intravenous dose. A rabbit injected with 40 mg/kg died 5 minutes after injection. With this rapid half-life o f disappearance, there is almost no chance o f being able to see the material after a dermal dose when the first sampling interval is 24 hours for serum and 28 days for liver. However, if there is a sex difference and absorption it is possible that there will be some appearance o f the material in male rabbits. 3.0 TEST M ATERIALS 3.1 Test, Control, and Reference Substances and M atrices 3.1.1 A nalytical R eference Substance: FC-95, lot 161 or 171. They are equivalent. 3.1.2 A nalytical Reference M atrix: Bovine liver, bovine serum and rabbit serum 3.1.3 Analytical Control Substance: None 3.1.4 A nalytical Control M atrix: Bovine liver, bovine serum and rabbit serum 3.2 Source o f M aterials: 3M ICP/PCP Division for FC-95, bovine liver from grocery store, bovine serum from Sigma Chemical Company, rabbit serum from AM DT-110394.1 control animals wwrrnwW 2 3.3. Purity and Strength of Reference Substance: Responsibility o f Sponsor. 3.4 Stability o f Reference Substance: To be determined by Sponsor. 3.5 Storage Conditions for Test M aterials: Room temperature for FC-95. For biological samples the storage is -20+10 C. 3.6 D isposition o f Specimens: Biological tissues and fluids will be retained per GLP Regulation for the time period required for studies longer than 28 days. 4.0 EX PER IM EN TA L - Overview_________________________________________ The tissues from animals dosed as described (HW I#6329-152), were available for analysis for fluorine compounds. At the discretion o f the Study Director, a series o f analytical tests could be performed. The screening for fluoride in liver via combustion was the most likely analysis to present definitive data for absorption if the pharmacokinetic test (intravenous administration) was positive for fluorine in the liver. Other available tests were electrospray mass spectroscopy and gas chromatography/mass spectrometry. However, if the definitive results could be obtained with combustion analysis alone, only the liver samples would be analyzed and any other tests would be for confirmation. And, if no organic fluorine increase was detectable even just using meter readings below the practical quantitation limit o f the method, then further analysis o f liver for specific fluorinated compounds would not be performed. 5.0 EXPERIM ENTAL - METHODS 5.1 A M DT-M -1-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver 5.2 A M D T -M -2-0, Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer 5.3 AM DT-M -4-0, Extraction o f Fluorochemicals from Rabbit Liver 5.4 A M DT-M -5-0, Analysis o f Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry O O O tK J 3 G 0265# 5.5 AM DT-M -8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 5.6 AM DT-M -14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum 6.0 DA TA ANALYSIS______________________________________________________ The data are attached. There appears to no difference in total organic fluorine in whole liver between the controls and the 4, 16, 40 mg/kg groups. The dose was with respect to the T-numbered compound. In addition, there appears to be no difference between either fabric treatment and the controls. The values are all below the practical limit o f quantitation and the comparisons are relative and based on meter reading. Very little total organic fluorine was observed in serum at 1 and 2 days post dose in either sex. The method did not work because the time interval o f sampling was too late. The material if present would have been cleared before the samples were collected. Other data was collected using electrospray mass spectrometry (see appendices). This data, although supportive, in the opinion o f the Study Director is not required to reach the conclusion stated here and therefore is not discussed in detail. 6.1 Circumstances that M ay Affect the Quality o f the Data: The possible problem with this analysis is that there is not a good marker at the times samples were collected to measure dermal absorption if it did occur. No conclusion can be drawn from this data as to the extent o f dermal absorption. 7.0 C O N C LU SIO N ________________________________________________________ In this study, no conclusion can be drawn from this data as to the extent o f dermal absorption. 8.0 M AIN TEN A N C E OF RA W DA TA AND RECORDS__________________ 8.1 Raw Data and Data: Raw data, approved protocol, approved final report, appropriate specimens, and electronic data will be maintained in the AMDT archives. WTtTTT/TFT h CQ265JL* 9.0 APPENDICES 9.1 Protocol and Am endm ents 9.1.1 Protocol and Final Report HW I#6329-152 "Single-Dose Dermal Absorption/Toxicity Study o f T-6067, T-6068 and T-6069 in Rabbits" (Protocol type TP3016.AB for dosing o f animals, tissue collection, etc.) 9.1.2 Analytical protocol A M D T-011095.1 9.1.3 Amendment to analytical protocol AM DT-011095.1 9.2 Signed Reports from Individual Scientists: None 9.3 Q uality Assurance Unit Statement: See attached 9.4 K ey Personnel Involved in the Study: See attached 9.5 M aterials and Equipment: See methods 9.6 Solutions, Reagents, and Standards: See methods 9.7 Sample Preparation: See methods 9.8 Q uality Control Practices: See methods 9.9 Test M ethods: See Protocol AM DT-011095.1 9.10 Instrum ent Settings: See methods 9.11 Data: See attached. 9.11.1 Summary and raw data; ug F' in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. 9.11.2 Summary and raw data; analysis o f liver extracts using electrospray mass spectrometry. 5 C 02S5J 9.11.3 Summary and raw data; ppm F` in serum as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. 9.11.4 Summary and raw data; ppm F' in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. 6 CQZGSf 9.1.1 Protocol and Final Report HWI#6329-152 "SingleDose Dermal Absorption/Toxicity Study of T-6067, T-6068 and T-6069 in Rabbits" (Protocol type TP-3016.AB for dosing o f animals, tissue collection, etc.) HW IASZCLOENTSOI NN ('[>, : i I i c i h ' ' -I M A D I ,i J rI >,\ 1 / ') ' !>!>, Sponsor: 3M St. Paul, Minnesota CORNING 'li. i ij; ' c.\ \ !.. : t FINAL REPORT Study Title: Single-Dose Dermal Absorption/Toxicity Study of T-6067, T-6068, and T-6069 in Rabbits Author: Steven M. Glaza Study Completion Date: May 26, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: HWI 6329-152 Pnone 60d 24l-44 7l E X P R E S S -MAIL DE LIV ER Y Page 1 of 39 3 3 0 1 KI NS M AN lil.7 0 Fa/ o 0 o L 1 /3 MADISON. VY I 3 3 ' Q-I AX I O M S Page 2 of 39 HWI 6329-152 QUALITY ASSURANCE STATEMENT This report has been reviewed by the Quality Assurance Unit of Hazleton Wisconsin, Inc., in accordance with the Food and Drug Administration (FDA) Good Laboratory Practice Regulations, 21 CFR 58.35 (b) (6) (7). The following inspections were conducted and findings reported to the Study Director and management. Written status reports of inspections and findings are issued to Hazleton management monthly according to standard operating procedures. Inspection Dates From To Date Reported to Date to Phase_________ Study Director Management 11/28/94 12/08/94 02/20/95 05/24/95 05/25/95 11/28/94 12/08/94 02/22/95 05/24/95 05/25/95 Protocol Review Dose Preparation Data/Report Review Report Rereview Report Rereview 11/28/94 12/08/94 02/22/95 05/24/95 05/25/95 12/10/94 01/10/95 03/10/95 06/10/95 06/10/95 Cecilia M. Danner (J Representaittiivvee, Quality Assurance Unit sii, Date Page 3 of 39 STUDY IDENTIFICATION Single-Dose Dermal Absorption/Toxicity Study of Study of T-6067, T-6068, and T-6069 in Rabbits HWI 6329-152 Test Materials 1. T-6067 2. T-6068 3. T-6069 Sponsor 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.0. Box 33220 St. Paul, MN 55133-3220 Sponsor's Representative John L. Butenhoff, PhD 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.0. Box 33220 St. Paul, MN 55133-3220 (612) 733-1962 Study Director Steven M. Glaza Hazleton Wisconsin, Inc. P.0. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Study Location Hazleton Wisconsin, Inc. Building No. 3 3802 Packers Avenue Madison, WI 53704 Study Timetable Study Initiation Date Experimental (In-life) Start Date In-1ife End Date Experimental Termination Date Study Completion Date December 6, 1994 December 8, 1994 January 5, 1995 May 26, 1995 May 26, 1995 C0265g Page 4 of 39 HWI 6329-152 KEY PERSONNEL Acute Toxicology Laboratory Animal Medicine Steven M. Glaza Study Director Manager Cindy J. Cary, DVM Dipl ornate, ACLAM Supervisor Francis (Bud) W. McDonald Study Coordinator Anatomical Pathology Patricia Padgham In-life Supervisor Thomas E. Palmer, PhD Anatomical Pathologist Rose M. Bridge Report Supervisor Quality Assurance Sherry R. W. Petsel Manager Jack Serfort/ Deborah L. Pirkel Supervisors Necropsy Anne Mosher Supervi sor Pathology Data C026O Page 5 of 39 CONTENTS Quality Assurance Statement Study Identification Key Personnel Summary Objective Regulatory Compliance Test and Control Materials Test System Procedures Results Discussion Signature Reference Pathology Report Table 1 Individual and Mean Body Weights (g) 2 Individual Clinical Signs 3 Individual Dermal Irritation Scores 4 Individual Pathology Comments Appendix A Protocol TP3016.AB HWI 6329-152 Page 2 3 4 6 8 8 8 g 10 13 13 13 14 15 16 18 20 26 28 29 r\t \r\L-2Z 00266# Page 6 of 39 SUMMARY HWI 6329-152 This study was done to assess the systemic absorption/toxicity and relative skin irritancy of T-6067, T-6068, and T-6069 when applied to the skin of rabbits. The study was conducted using three male and three female acclimated rabbits of the Hra:(NZW)SPF strain for each treatment group. Group Dose Level Number of Animals Test Material (mq/kq) Males Females 1 (Control) 2 3 4 5 5 Distilled water T-6067 T-6067 T-6067 T-6068 T-6069 0a 4 16 40 b b 3 3 3 3 3 3 3 3 3 3 3 3 a Administered at a dose volume of 2.0 mL/kg. b Administered as a 10-cm'x 10-cm piece of test material (fabric). The back of each rabbit was clipped free of hair and a single dose of the respective material at the indicated dose level was administered to the skin of the rabbits. The treatment sites remained intact. The area of application was covered with a gauze bandage secured with paper tape around all edges and overwrapped with Saran Wrap and Elastoplast tape to provide an occlusive dressing for a 24-hour exposure period. Clinical observations were conducted predose and at approximately 1, 2.5, and 4 hours after test or control material administration. Additional clinical observations and twice a day mortality checks were conducted daily thereafter for 28 days. Body weights were determined on Day -8 or Day -7 for randomization purposes, before test or control material administration (Day 1), and at in-life termination (Day 29). The initial dermal irritation reading was made before test or control material administration (recorded as the Day 1 reading). Subsequent readings of dermal irritation were made approximately 30 minutes after bandage removal (Day 2) and on Days 4 and 8. Blood samples were collected from a marginal ear vein of the animals before in-life initiation (Day 1), approximately 24-hours postdose (Day 2), on Days 4, 8, 15, and 22. In addition, at the time of necropsy on Day 29, approximately 20 mL of blood was obtained from each animal. All samples were centrifuged and separated into serum and cellular fractions. All animals were euthanized at termination of the in-life phase and necropsied. The whole liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from one male and one female in each group were collected at necropsy. The blood samples (serum and cellular fractions), livers, bile, dermal application sites, and kidneys were sent frozen to the Sponsor after termination of the in-life phase. 0026SJ Application of T-6067, T-6068, and T-6031 did not result in any test material-related changes in body weight gain or macroscopic findings at necropsy. All animals appeared clinically normal throughout the study. The control material did not produce any dermal irritation. Test material T-6067 produced very slight irritation in a few of the animals at all three dose levels (4, 16, and 40 mg/kg). Both fabric test materials, T-6068 and T-6069, produced slight to moderate erythema and slight edema and/or desquamation reactions. 0Q26& Page 8 of 39 OBJECTIVE HWI 6329-152 The objective of this study was to assess the systemic toxicity/absorption and relative skin irritancy of test materials when applied to the skin of rabbits. REGULATORY COMPLIANCE This study was conducted in accordance with the U.S. Food and Drug Administration's Good Laboratory Practice Regulations for Nonclinical Laboratory Studies, 21 CFR 58. All procedures used in this study are in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study did not unnecessarily duplicate any previous work. TEST AND CONTROL MATERIALS Identification The test materials were identified and described as follows: Identification Physical Description T-6067 T-6068 T-6069 Clear, colorless liquid White web cloth White web cloth The control material was distilled water and was described as a clear, colorless liquid. Purity and Stability The Sponsor assumes responsibility for test material purity and stability determinations (including under test conditions). The purity and stability of the control material were considered to be adequate for the purposes of this study. Storage and Retention The test and control materials were stored at room temperature. A reserve sample of each test and control material was taken and will be retained in a freezer set to maintain a temperature of -20C 10C for 10 years in accordance with Flazleton Wisconsin (HWI) Standard Operating Procedure (SOP). Any unused test material was returned to the Sponsor after completion of all in-life testing according to HWI SOP. Any remaining control material is retained for other testing and will not be discarded after issuance of the final report. Qozsqg Page 9 of 39 Safety Precautions HWI 6329-152 The test and control material handling procedures were according to HWI SOPs and policies. TEST SYSTEM Test Animal Adult albino rabbits of the Hra:(NZW)SPF strain were procured from HRP, Inc., Kalamazoo, MI, on November 16, 1994 and maintained at the Hazleton Wisconsin facility at 3802 Packers Avenue, Madison, Wisconsin. Housing After receipt, the animals were acclimated for a period of at least 7 days. During acclimation and throughout the study, the animals were individually housed in screen-bottom stainless steel cages in temperature- and humiditycontrolled quarters. Environmental 'controls for the animal room were set to maintain a temperature of 19 to 23C, a relative humidity of 50% 20%, and a 12-hour 1ight/12-hour dark lighting cycle. In cases where variations from these conditions existed, they were documented and considered to have had no adverse effect on the study outcome. Animal Diet The animals were provided access to water ad libitum and a measured amount of Laboratory Rabbit Diet HF #5326, PMI Feeds, Inc. The feed is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. Samples of the water are periodically analyzed by HWI. There were no known contaminants in the feed or water at levels that would have interfered with or affected the results of the study. Selection of Test Animals The animals were identified by animal number and corresponding ear tag and were placed into study groups using a stratified body weight randomization program. The randomization body weights were determined on Day -8. The weight variation of the animals for each group of each sex selected for the study did not exceed 2 standard deviations of the mean weight, and the mean body weights for each group of each sex were not statistically different at the 5% probability level. 002664^ Page 10 of 39 Study Design HWI 6329-152 Animals weighing from 2,377 to 2,796 g at initiation of treatment were placed into the following study groups: GrouD Dose Level Number of Animals Test Material (mq/kq) Males Females 1 (Control) 2 3 4 5 6 Distilled water T-6067 T-6067 T-6067 T-6068 T-6069 0a 4 16 40 b b 3 3 3 3 3 3 3 3 3 3 3 3 a Administered at a dose volume of 2.0 mL/kg. b Administered as a 10-cm x 10-cm section of test material (fabric). Justification for Species Selection Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. PROCEDURES Preparation of Exposure Area On the day before test material application, the back and, if necessary (to obtain unblemished skin), the flanks of each rabbit was clipped free of hair. The clipped area made up approximately 20% of the total body surface area. The test sites (intact skin) were inspected for interfering lesions, irritation, or defects that would preclude the use of any of the animals. The animals were clipped on Days 8 and 29 to aid in visualizing the application sites. Dose Administration All animals received a single administration of the respective test or control material. The day of treatment was designated as Day 1. Group 1 . An individual dose (2.0 mL/kg) was calculated and measured based on each animal's body weight on the day of treatment. The control material (distilled water) was applied evenly to the test site at a rate of approximately 0.05 mL/cm . m cf Page 11 of 39 HWI 6329-152 Groups 2, 3, and 4 . The test material (T-6067) was administered undiluted to the test sites of the Groups 2, 3, and 4 animals (4, 16, or 40 mg/kg, respectively) using the average bulk density of 1.04 g/mL to determine the dose volume for each dose level. An individual dose of the test material was calculated for each animal based on its body weight on the day of treatment. The area of exposure for the 4-, 16, and 40 mg/kg dose levels was 9, 9, and 16 cm2, respectively. The approximate rate of application ranged from 0.001 to 0.006 mL/cm2. Groups 5 and 6 . The test material (T-6068 or T-6069) was applied to the respective test site as a 10-cm x 10-cm section of material that was moistened with distilled water. Each area of application was covered with a 10-cm x 10-cm gauze bandage secured with paper tape around all edges and overwrapped with Saran Wrap and Elastoplast tape to provide an occlusive dressing. Collars were used to restrain the animals during the 24-hour exposure period. Approximately 24 hours after test or control material application, the restraining collars and bandages were removed and the residual test material was removed with tap water and disposable paper towels. Reason for Route of Administration The dermal route is a potential route of exposure in humans. Observations of Animals Clinical observations were conducted predose and at approximately 1, 2.5, and 4 hours after test or control material administration. Additional clinical observations and twice a day mortality checks (morning and afternoon) were conducted daily thereafter for 28 days. Body weights were determined for randomization purposes on Day -8 or Day -7, before test material administration (Day 1), and at in-life termination (Day 29). The initial dermal irritation reading was made before test or control material administration according to the Draize1 technique (recorded as the Day 1 reading). Subsequent readings of dermal irritation were made approximately 30 minutes after bandage removal (Day 2) and on Days 4 and 8. 00266 Page 12 of 39 HWI 6329-152 Sample Collections Blood samples (approximately 4 mL) were collected from a marginal ear vein of all animals before experimental initiation (Day 1). Subsequent collection of blood was conducted 24-hours postdose (Day 2), and on Days 4, 8, 15, and 22. In addition, at the time of necropsy on Day 29, approximately 20 mL of blood was obtained from the posterior vena cava of each animal. All samples were centrifuged and separated into serum and cellular fractions. These samples were then stored in a freezer set to maintain a temperature of -20C 10C until shipped to the Sponsor. Pathology At termination of the experimental phase (Day 29), animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, exsanguinated, and necropsied in random order. The sites of test and control material application were washed with lukewarm tap water before the necropsy procedure. All animals were subjected to an abbreviated gross necropsy examination and any abnormalities were recorded. The whole liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from the first male and female in each group were collected and immediately placed on dry ice, then placed in a freezer set to maintain a temperature of -20C 10C. After necropsy, the animals were discarded. Shipment of Blood. Bile, and Tissues After experimental termination, the blood samples (serum and cellular fractions), livers, bile, dermal application sites, and kidneys were sent frozen (on dry ice) to the Sponsor (James D. Johnson, 3M E.E. & P.C., Bldg. 2-3E-09, 935 Bush Avenue, St. Paul, MN, 55106). The Sponsor is responsible for the retention and disposition of the samples. HWI does not accept any responsibility for the analysis of the tissue samples collected in this study nor are these results presented in this report. Statistical Analyses No statistical analyses were required by the protocol. Location of Raw Data. Records, and Final Report The raw data, records, and an original signed copy of the final report will be retained in the archives of HWI in accordance with HWI SOP. C02SS$r Page 13 of 39 RESULTS HWI 6329-152 Body Weights Individual and mean body weights are in Table 1. All animals exhibited body weight gains from Day 1 to Day 29. Clinical Observations Individual clinical signs are in Table 2. All animals appeared normal throughout the study. Dermal Irritation Individual dermal irritation scores are in Table 3. The control material produced no dermal irritation. Test material T-6067 produced slight erythema reactions in one animal treated at 4 mg/kg, in two animals treated at 16 mg/kg, and in two animals treated at 40 mg/kg. Both fabric test materials, T-6068 and T-6069, produced slight to moderate erythema and slight edema and desquamation reactions. Pathology Individual animal pathology comments are presented in Table 4. There were no lesions observed in any of the animals. Page 15 contains a pathology report by the study pathologist. DISCUSSION The acute systemic absorption/toxicity and relative skin irritancy of T-6067, T-6068, and T-6069 were evaluated in male and female albino rabbits when administered as a single dermal application. Application of the these materials did not result in any effects on body weight gain or macroscopic findings at necropsy. All animals appeared normal throughout the study. No dermal irritation was observed as a result of the application of the control material, very slight irritation was seen in a few of the animals treated with T-6067, and slight to moderate irritation was seen in the animals treated with T-6068 or T-6069. SIGNATURE MV Steven M. Glaza Study Director Acute Toxicology & ________ Date Page 14 of 39 HWI 6329-152 REFERENCE 1. Draize, J. H., "Acute Dermal Toxicity (Single Exposure)," In: Appraisal of the Safety of Chemicals in Foods, Drugs and Cosmetics - Dermal Toxicity, Association of Food and Drug Officials of the U.S., pp. 54-56 (1959). Page 15 of 39 HWI 6329-152 PATHOLOGY REPORT There were six rabbits (three males and three females) each from six dose levels euthanized and necropsied at the termination of the study. The test material, dose level, day of death, and gross observations recorded for each animal are in the Individual Pathology Commervts that follow this report. At necropsy, there were no visible lesions in any of the animals. The liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from one male and one female in each group were collected, frozen, and sent to the Sponsor. After necropsy, the animals were discarded. Pathologist (6329-152.Ihm) 020895 Date CQ2S7C Animal Number F52725 F52711 F52719 Mean Page 16 of 39 HWI 6329-152 Table 1 Individual and Mean Body Weights (g) Male_________________ Random ization Dav Dav -8 1 29 _______________ Female Random Animal ization Number Dav -8 Dav 1 29 Group I (Control) - Distilled Water (0 mo/ko) 2,699 2,592 2,456 2,796 2,726 2,494 3,135 3,085 2,671 F52721 F52806 F52727 2,463 2,524 2,639 2,651 2,627 2,713 2,963 3,086 3,103 2,582 2,672 2,964 2,542 2,664 3,051 F52718 F52723 F52713 Mean 2,380 2,482 2,602 2,488 Group 2 - T-6067 (4 mq/kq) 2,450 2,778 2,527 2,795 3,181 2,980 F52715 F52809 F52738 2,363 2,498 2,489 2,585 2,985 2,450 2,377 2,511 2,619 2,502 2,752 2,909 2,946 2,869 F52737 F52801 F52724 Mean 2,631 2,611 2,634 2,625 Group 3 - T-6067 (16 mq/kq) 2,634 2,699 2,616 2,878 3,113 2,933 F52733 F52788 F52720 2,543 2,575* 2,560 2,650 2,975 2,559 2,629 2,429 2,627 2,562 3,017 2,994 3,028 3,013 F52802 F52730 F52731 Mean 2,543 2,408 2,339 2,430 Group 4 - T-6067 f40 mq/kq) 2,785 2,520 2,489 3,088 2,766 2,715 F52714 F52734 F52774 2,523 2,557 2,364* 2,598 2,856 2,481 2,578 2,660 2,556 2,598 2,893 2,839 2,930 2,887 * Day -7 body weight. G026 Animal Number F52729 F52803 F52712 Mean Page 17 of 39 HWI 6329-152 Table 1 (Continued) Individual and Mean Body Weights (g) Male_________________ Random ization Dav Dav -8 1 29 _______________ Femal e Random Animal ization Number Dav -8 Day 1 29 Group 5 - T-6068 n o cm x 10 cm Section) 2,431 2,442 2,631 2,545 2,596 2,645 2,893 2,920 2,886 F52716 F52787 F52728 2,652 2,494* 2,495 2,778 2,544 2,619 3,211 3,017 3,009 2,501 2,595 2,900 2,547 2,647 3,079 F52736 F52717 F52735 Mean Group 6 - T-6069 (10 cm x 10 cm Section) 2,681 2,405 2,570 2,745 2,538 2,606 2,996 3,025 2,875 F52739 F52732 F52726 2,442 2,477 2,591 2,552 2,630 2,965 2,503 2,480 2,525 2,768 2,591 2,957 2,930 3,110 2,999 * Day -7 body weight G 027^U Sex Mal e Female Male Female Male Female Page 18 of 39 HWI 6329-152 Table 2 Individual Clinical Signs Animal Number Observation 1-4 Hours . (Day 1) ____Day 2 through 29 Group 1 (Control) - Distilled Water (0 ma/kal F52725 Appeared normal / F52711 Appeared normal / / F52719 Appeared normal / / F52721 Appeared normal / / F52806 Appeared normal / / F52727 Appeared normal / / Group 2 -'T-6067 (4 mq/kq) F52718 Appeared normal / / F52723 Appeared normal / F52713 Appeared normal / / F52715 Appeared normal / / F52809 Appeared normal / / F52738 Appeared normal / / GrouD 3 - T-6067 (16 mq/kq) F52737 Appeared normal / / F52801 Appeared normal / / F52724 Appeared normal / / F52733 Appeared normal / / F52788 Appeared normal / / F52720 Appeared normal / / / Condition existed. 002S7J Sex Male Female Male Female Male Female Page 19 of 39 HWI 6329-152 Table 2 (Continued) Individual Clinical Signs Animal Number Observation 1-4 Hours (Dav 11 Dav 2 throuoh 29 Group 4 - T-6067 (40 ma/ko) F52802 Appeared normal / / F52730 Appeared normal / / F52731 Appeared normal / / F52714 Appeared normal / / F52734 Appeared normal / / F52774 Appeared normal J J GrouD 5 - T-6068'(10 cm X 10 cm Section) F52729 Appeared normal / / F52803 Appeared normal / / F52712 Appeared normal / / F52716 Appeared normal / / F52787 Appeared normal / / F52728 Appeared normal / / Group 6 - T-6069 (10 cm X 10 cm Section) F52736 Appeared normal / / F52717 Appeared normal / / F52735 Appeared normal / / F52739 Appeared normal / / F52732 Appeared normal / / F52726 Appeared normal / / Condition existed. X, O f Page 20 of 39 HWI 6329-152 Table 3 Individual Dermal Irritation Scores Group 1 (Control) - Distilled Water (0 mg/kg) Dermal Reaction Hales______ . Studv Dav 1 _2_ 4 _8_ _______ Females Studv Dav I _2_ 4 8_ Animal No. F52725 Animal No. F52721 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0000 0000 0000 0000 0000 0000 0000 0000 0000 0000 0000 0000 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52711 0000 0000 0000 0000 0000 0000 Animal No. F52806 0000 0000 0000 0000 0000 0000 Erythema Edema Atonia Desquamation Cori aceousness Fissuring Animal No. F52719 0000 0000 0000 0000 0000 0000 Animal No. F52727 0000 0000 0000 0000 0000 0000 Page 21 of 39 HWI 6329-152 Table 3 (Continued) Individual Dermal Irritation Scores Group 2 - T-6067 (4 mg/kg) Dermal Reaction Males_______ _ Study Dav _ L _2_ 4 _8_ Animal No. F52718 _______ Females Studv Dav 1 _2_ 4 8 Animal No. F52715 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0000 0000 0000 0000 000 0 0000 0 100 00 0 0 0000 0000 00 0 0 0000 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52723 0000 0 00 0 0000 0000 0000 0000 Animal No. F52809 0000 0000 0000 0000 0000 00 0 0 Erythema Edema Atonia Desquamation Cori aceousness Fissuring Animal No. F52713 0000 0000 0000 0000 0000 0000 Animal No. F52738 00 0 0 0000 0000 0000 0000 0000 coze^ Page 22 of 39 HWI 6329-152 Table 3 (Continued) Individual Dermal Irritation Scores Group 3 - T-6067 (16 mg/kg) Dermal Reaction ________Males_______ _ Study Day______ 1 _2_ 4 8_ _______ Females ______ Study Day 1 _2_ 4 8_ Aniimal No. F52737 Aniimal No. F52733 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0 100 0000 0000 0000 0000 00 0 0 0000 00 0 0 00 0 0 0000 0000 0000 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Aniimal No. F52801 0000 0000 0000 0000 0000 0000 Aniimal No. F52788 0000 0000 00 0 0 0000 0000 0000 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Aniimal No. F52724 0 100 0000 0000 0000 0000 0000 Aniimal No. F52720 0000 00 00 0000 0000 0000 0000 C02S7f Page 23 of 39 HWI 6329-152 Table 3 (Continued) Individual Dermal Irritation Scores Group 4 - T-6067 (40 mg/kg) Dermal Reaction Males______ _ Studv Dav 1 JL 4 JL _______ Females Studv Dav 1 _2_ 4 JL Animal No. F52802 Animal No. F52714 Erythema Edema Atonia Desquamation Coriaceousness Fi ssuring 0 100 0000 0000 0000 0000 0000 0000 0000 0000 0000 0000 0000 Erythema Edema Atoni a Desquamation Coriaceousness Fissuring Animal No. F52730 0000 0000 0000 0000 0000 0000 Animal No. F52734 0000 0000 0 00 0 0000 0000 0000 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52731 0000 0000 0000 0000 0000 0000 Animal No. F52774 0 10 0 0000 0000 0000 0000 0000 TOiTuCW 002679 Page 24 of 39 HWI 6329-152 Table 3 (Continued) Individual Dermal Irritation Scores Group 5 - T-6068 (10 cm x 10 cm Section) Dermal Reaction Males Study Dav 1 _2_ 4 8 Females Study Dav 1 _2_ 4 8 Animal No. F52729 Animal No. F52716 Erythema Edema Atonia Desquamation Coriaceousness Fissuring 0111 0 00 0 0000 0001 0000 0000 0000 0000 0000 0000 0000 0000 Erythema Edema Atona Desquamation Coriaceousness Fissuring Animal No. F52803 0 100 0000 0000 0000 0 00 0 0000 Animal No. F52787 0 110 0000 0000 0000 0000 0000 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animal No. F52712 0 110 0000 0000 0000 0000 0000 Animal No. F52728 0 12 1 0000 0000 000 1 0000 0000 0 0 2SJfy Page 25 of 39 HWI 6329-152 Table 3 (Continued) Individual Dermal Irritation Scores Group 6 - T-6069 (10 cm x 10 cm Section) Dermal Reaction Males Studv Day J L _2_ 4 8_ Females Study Dav 1 JL 4 8_ Animai No. F52736 Animai No. F52739 Erythema Edema Atoni a Desquamation Coriaceousness Fissuring 0 12 1 0000 000 0 0001 0000 0000 0 12 1 0110 00 0 0 000 1 00 0 0 000 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animai No. F52717 0110 0000 0000 0000 0000 0000 Animai No. F52732 0110 0110 0000 0000 0000 000 0 Erythema Edema Atonia Desquamation Coriaceousness Fissuring Animai No. F52735 0 12 1 0 110 0000 0001 0000 0000 Animai No. F52726 012 1 00 10 000 0 000 1 0000 00 0 0 CO 26S0 Animal Number F52725 F52711 F52719 F52721 F52806 F52727 F52718 F52723 F52713 F52715 F52809 F52738 F52737 F52801 F52724 F52733 F52788 F52720 Page 26 of 39 Table 4 Individual Pathology Comments ______Test Dav Sex Died Sacrificed Necropsy Observation Group 1 (Control! - Distilled Water (0 mg/kq) M- 29 No visible lesions. M- 29 No visible lesions. M- 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible 1esions. F- 29 No visible lesions. Group 2 -'T-6067 (4 mq/ko) M- 29 No visible lesions. M- 29 No visible lesions. M- 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible lesions. Group 3 - T-6067 f16 mq/ko) M- 29 No visible 1esions. M- 29 No visible lesions. M- 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible lesions. F- 29 No visible lesions. HWI 6329-152 - Not applicable. 09263/ Page 27 of 39 HWI 6329-152 Table 4 (Continued) Individual Pathology Comments Animal ______ Test Day_____ Number Sex Died Sacrificed NecroDSV Observation Group 4 - T-6067 (40 mq/kq) F52802 M - 29 No visible lesions. F52730 M - 29 No visible lesions. F52731 M - 29 No visible lesions. F52714 F - 29 No visible lesions. F52734 F - 29 No visible lesions. F52774 F - 29 No visible lesions. GrouD 5 - T-6068 'HO cm x 10 cm Section) F52729 M - 29 No visible lesions. F52803 M - 29 No visible lesions. F52712 M - 29 No visible lesions. F52716 F - 29 No visible lesions. F52787 F - 29 No visible lesions. F52728 F - 29 No visible lesions. Group 6 - T-6069 flO cm x 10 cm Section) F52736 M - 29 No visible lesions. F52717 M - 29 No visible lesions. F52735 M - 29 No visible lesions. F52739 F - 29 No visible lesions. F52732 F - 29 No visible lesions. F52726 F _ 29 No visible lesions. - Not applicable 00268 Page 28 of 39 APPENDIX A Protocol TP3016 HWI 6329-152 0026S HAZLETON WISCONSIN P O M O l M M B O X / ', .1 M A D I M I O \A/ i >:t /! (Ii i / 114 1. Page 29 of 39 CORNING i." Sponsor: 3M St. Paul, Minnesota PROTOCOL TP3016.AB ' Study Title: Single-Dose Dermal Absorption/Toxicity Study of T-6067, T-6068, and T-6069 in Rabbits Date: December 6, 1994 Performing Laboratory. Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: HWI 6329-152 ) P tl u n t 1 o O ij 1 <1 1 4 ' i 1E \ (f< y N (VI A L L) L L IV l K > V Ml : K IM s M A N F it \ V A : ) -> ; N o U ei vV I C026S Page 30 of 39 ) TP3016.AB Page 2 STUDY IDENTIFICATION Single-Dose Dermal Absorption/Toxicity Study of T-6067, T-6068, and T-6069 in Rabbits HWI No. 6329-152 Test Materials 1. T-6067 2. T-6068 3. T-6069 Sponsor 3M Toxicology Services 220-2E-02 3M Center St. Paul, MN 55144 Sponsor's Representative John L. Butenhoff, PhD ' 3M Toxicology Services 220-2E-02 3M Center I St. Paul, MN 55144 (612) 733-1962 Study Director Steven M. Glaza Hazleton Wisconsin, Inc. P.0. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Study Location Hazleton Wisconsin, Inc. Building No. 3 3802 Packers Avenue Madison, WI 53704 Proposed Study Timetable Experimental Start Date Experimental Termination Date Draft Report Date December 8, 1994 January 5, 1995 February 16, 1995 ; (t u W t Page 31 of 39 1. Study Single-Dose Dermal Absorption/Toxicity Study in Rabbits TP3016.AB Page 3 2. Purpose To assess the systemic absorption and toxicity and relative skin irritancy of test materials when applied to the skin of rabbits 3. Regulatory Compliance This study will be conducted in accordance with the following Good Laboratory Practice Regulations/Standards/Guidelines: [ ] Conduct as a Nonregulated Study [X] 21 CFR 58 (FDA) [ ] 40 CFR 160 (EPA-FIFRA) [ ] 40 CFR 792 (EPA-TSCA) [ ] C(81)30 (Final) (OECD) [ ] 59 Nohsan No. 3850 (Japanese MAFF) [ ] Notification No. 313 (Japanese MOHW) All procedures in this protocol are in compliance with the Animal Welfare Act Regulations. In the opinion of the Sponsor and study director, the study does not unnecessarily duplicate any previous work. 4. Qua!itv Assurance The protocol, study conduct, and the final report will be audited by the Quality Assurance Unit in accordance with Hazleton Wisconsin (HW1) Standard Operating Procedures (SOPs) and policies. 5. Test Materials A. Identi fication 1. T-6067 2. T-6068 3. T-6069 B. Physical Description 1. (To be documented in the raw data) 2. (To be documented in the raw data) 3. (To be documented in the raw data) C. Purity and Stability The Sponsor assumes responsibility for purity and stability determinations (including under test conditions). ) C-O26S0 Page 32 of 39 TP3016.AB Page 4 D. Storage Room temperature E. Reserve Samples Reserve sample(s) of each batch/lot. of test and control materials will be taken for this study. The test and control material reserve samples will be stored at HWI in a freezer set to maiintain a temperature of -20*C 10*C for 10 years per HWI SOP. The Sponsor will be contacted after 10 years for disposition in accordance with the appropriate regulatory Good Laboratory Practices. F. Retention Any unused test materials will be returned to the Sponsor after completion of the in-life phase of the study. G. Safety Precautions As required by HWI SOPs and policies 6. Control Material A. Identification Distilled water B. Physical Description Clear, colorless liquid C. Purity and Stability The purity and stability of this manufactured material is considered to be adequate for the purposes of this study. D. Storage Conditions Room temperature E. Reserve Samples See Section 5. E. Reserve Samples F. Retention Any remaining control material may be used for other testing and will not be discarded after issuance of the final report. G. Safety Precautions As required by HWI SOPs and policies C-026S^ Page 33 of 39 TP3016.AB Page 5 7. Experimental Design A. Animals (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/HRP, Inc. (3) Age at Initiation Adult (4) Weight at Initiation 2.0 to 3.0 kg (5) Number and Sex 18 males and 18 females (6) Identification Individual numbered ear tag (7) Husbandry (a) Housing Individually, in screen-bottom stainless steel cages (heavy gauge) (b) Food A measured amount of Laboratory Rabbit Diet HF #5326 (PMI Feeds, Inc.). The food is routinely analyzed by the manufacturer for nutritional components and environmental contaminants. (c) Water Ad libitum from an automatic system. Samples of the water are analyzed by HWI for total dissolved solids, hardness, and specified microbiological content and for selected elements, heavy metals, organophosphates, and chlorinated hydrocarbons. (d) Contaminants There are no known contaminants in the food or water that would interfere with this study. (e) Environment Environmental controls for the animal room will be set to maintain a temperature of 19*C to 23*C, a relative humidity of 50% 20%, and a 12-hour 1ight/12-hour dark cycle. cozssg Page 34 of 39 TP3016.AB Page 6 (f) Acclimation At least 7 days (8) Selection of Test Animals Based on health and body weight according to HWI SOPs. An adequate number of extra animals will be purchased so that no animal in obviously poor health is placed on test. The animals will be placed into study groups using a stratified body weight randomization program within nine days of study initiation. (9) Justification for Soecies Selection Historically, the New Zealand White albino rabbit has been the animal of choice because of the large amount of background information on this species. B. Dose Administration (1) Test Groups GrouD Test Material Dose Level Number of Animals . (mq/kq) Males Females o>cn -tktorv>*-* (Control) Distilled water T-6067 T-6067 T-6067 T-6068 T-6069 0* 4 16 *40 ** 3 3 3 3 3 3 3 3 3 3 3 3 * To be administered at a dose volume of 2.0 mL/kg ** To be administered as a 10.0-cm x 10.0-cm piece of test material (fabric) (2) Preparation of Exposure Area On the day before test material application, the back and, if necessary (to obtain unblemished skin), the flanks of each rabbit will be clipped free of hair with an electric clipper. The shaved area will constitute approximately 20% of the total body surface area. The treatment sites (intact skin) will be inspected for interfering lesions, irritation, or defects that would preclude the use of any of the animals. The animals will be clipped as needed throughout the study. Page 35 of 39 TP3016.AB Page 7 (3) Dose Administration All animals will receive a single administration of the respective test or control material. The day of treatment will be designated as Day 1. The respective doses for the animals in Groups I, 2, 3, and 4 will be based on the animal's body weight just before administration and spread onto the area of exposure in a thin and uniform layer. The Group 1, 2, 3, and 4 materials will be applied undiluted. The Group 5 and 6 materials will be applied as a 10.0-cm x 10.0-cm piece of the respective test material moistened with distilled water. The area of application (Groups 1-6) will be covered with a 10-cm x 10-cm gauze bandage secured with paper tape around all edges and overwrapped with Saran Wrap* and Elastoplast* tape to provide an occlusive dressing. The rabbits will be collared during the 24-hour application period. (4) Reason for Route of Administration The dermal route is a potential route of exposure in humans. (5) Removal of Test Material Approximately 24 hours after test or control material application the bandages and collars will be removed and the residual test material will be removed using water or an appropriate solvent, if necessary. Observation of Animals (1) Clinical Observations For clinical signs before test or control material administration and for clinical signs and mortality at approximately 1, 2.5, and 4 hours after test material administration (Day 1) and daily thereafter for clinical signs, and twice daily (a.m. and p.m.) for mortality for at least 28 days. Observations may be extended when directed by the study director. (2) Reading of Dermal Irritation Before test or control material administration the initial dermal irritation reading will be made and recorded as the Day 1 reading (Attachment 1). Additional dermal irritation readings will be made approximately 30 minutes after bandage removal (Day 2) and on Study Days 4 and 8. Individual dermal irritation records will be maintained for each animal. (3) Body Weights Just before test or control material application (Day 1), on Day 29, and at unscheduled death (when survival exceeds I day) 002690 Page 36 of 39 TP3016.AB Page 8 (4) Sample Collections (a) Frequency Before initiation (Day 1), approximately 24 hours post-dose (Day 2), Days 4,. 8, 15, 22, and at experimental termination (Day 29) (b) Number of Animals All (c) Method of Collection Blood samples (approximately 4 ml) will be collected from the marginal ear vein of either ear on Days 1, 2, 4, 8, 15, and 22. Approximately 20 mL of blood (actual volume to be documented in the raw data) will be obtained from the posterior vena cava of each animal sacrificed in a moribund condition or sacrificed at the time of necropsy (Day 29). The samples will be stored at room temperature and then centrifuged, and the separate serum and cellular fractions stored- in a freezer set to maintain -20*C 10*C. The separated serum and cellular fractions will be sent frozen to the Sponsor after experimental termination. Samples will be shipped to: James D. Johnson 3M E.E. & P.C. Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106 James D. Johnson or alternate will be notified by telephone at (612) 778-5294 prior to the shipment of the samples. D. Pathology (1) Unscheduled Sacrifices and Deaths Any animal dying during the study or sacrificed in a moribund condition will be subjected to an abbreviated gross necropsy examination and all abnormalities will be recorded. Animals in a moribund condition will be anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the vena cava, and exsanguinated. After necropsy, the animals will be discarded. C02S9/ Page 37 of 39 TP3016.AB Page 9 (2) Scheduled Sacrifice At termination of the experimental phase (Day 29), surviving animals will be anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the vena cava, exsanguinated, and subjected to an abbreviated gross necropsy examination. The animals will be necropsied in random order and all abnormalities will be recorded. (a) Sample Collection The sites of test and control material application will be washed with lukewarm tap water prior to the necropsy procedure. The whole liver, bile, an approximate 1-cm x 1-cm section of the dermal application site from all animals, and both kidneys from the first male and female necropsied in each group will be collected and immediately placed in a freezer set to maintain a temperature of -20*C 10*C. After necropsy, the animals will be discarded. The tissues.(1iver, bile, dermal application site, kidneys) will be sent frozen on dry ice to the Sponsor after experimental termination. The samples will be shipped to the person listed in Section j 7.C.(4).(c). The Sponsor is responsible for the retention and disposition of the samples. E. Statistical Analyses No statistical analyses are required. 8. Report A final report including those items listed below will be submitted. Description of the test and control materials Description of the test system Procedures Dates of experimental initiation and termination Tabulation of mortality data by sex and dose level Description of any toxic effects/dermal irritation Tabulation of mean body weights by sex and dose level Gross pathology findings/gross pathology report ) Uuuuft G0269 Page 38 of 39 TP3016.AB Page 10 9. Location of Raw Data, Records, and Final Report Original data, or copies thereof, will be available at HWI to facilitate auditing the study during its progress and before acceptance of the final report. When the final report is completed, all original paper data, including those item listed below will be retained in the archives of HWI according to HWI SOP. Protocol and protocol amendments Dose preparation records In-life records Body weights Dose administration Observations Anatomical pathology records Sample collection records Shipping records Study correspondence Final report (original signed copy) The following supporting records will be retained at HWI but will not be archived with the study'data. Animal receipt/acclimation records ^ Water analysis records Animal room temperature and humidity records Refrigerator and freezer temperature records Instrument calibration and maintenance records ) UUUU'fll C 0269J Page 39 of 39 PROTOCOL APPROVAL TP3016.AB Page 11 John L. Butenhoff, PhD Sponsor's Representative 3M Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. 0 L A._j -A ) __________________ Refj'resentative , Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-152.protdskl) lZ-i'2--in Date Date v~L-'o-c\M Date ) 00269^ 9.1.2 Analytical protocol AMDT-011095.1 fX J Q O 'V r* C02S9^ 3M Environmental Laboratory_________ Protocol - Analytical Study Single-Dose Dermal Absorption/Toxicity Study of T-6067, T-6068 and T-6069 in Rabbits In-Vivo Study Reference Number: HWI#6329-152 Study Number: A M D T-011095.1 Test Substance: L -13492 (T-6067, T-6068 and T-6069) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55106 Name and Address of Testing Facility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 55106 Proposed Initiation Date: July 25, 1995 Proposed Completion Date: August 25, 1995 M ethod Numbers and Revisions: AM DT-M -1-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver A M D T-M -2-0, Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer AMDT- M -4-0, Extraction o f Fluorochemicals from Rabbit Liver AMDT- M -5-0, Analysis o f Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry AM DT-M -8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode AM DT-M -14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum Author: James D. Johnson roved Bv; /Jam es D. / Study Di /y/v/ Date z. V t John Butenhoff, PhD Sponsor Representative Date 1 TWTJTrTj y 00269 1.0 PURPOSE This study is designed to provide information as to whether L -13492 (T-6067, T-6068 and T-6069) is dermally absorbed. The analytical aspect o f this study is to determine total organic fluorine in the tissue and serum o f rabbits at various times post dose dermal application o f L-13492. 2.0 TEST MATERIALS 2.1 Test, Control, and Reference Substances and M atrices 2.1.1 Analytical Reference Substance: FC-95, lot 161 or 171. They are equivalent. 2.1.2 A nalytical Reference M atrix: Bovine liver and bovine serum 2.1.3 Analytical Control Substance: None 2.1.4 Analytical Control Matrix: Bovine liver and bovine serum 2.2 Source o f M aterials: 3M ICP/PCP Division (2.1.1), Grocery store (2.1.2, 2.1.4 liver), Sigma Chemical Company (2.1.2, 2.1.4 serum) 2.3 N um ber o f Test and Control Samples: Tissues and fluids from 30 test animals and 6 control animals. Tissues and fluids include liver, kidney, serum, cellular fraction, dermal application site and bile. Analysis o f these tissues will be at the discretion o f the Study Director. 2.4 Identification o f Test and Control Samples: The samples are identified using the HWI animal identification number which consists o f a letter and five digit number, plus the tissue identity and day identity (serum). 2.5 Purity and Strength of Reference Substance: To be determined by Sponsor. 2.6 Stability o f Reference Substance: To be determined by Sponsor. 2.7 Storage Conditions for Test M aterials: Room temperature (2.1.1), -20 10C (2.1.2, 2.1.4). Test and Control samples will be received according to A M D T -S-10-0. 2.8 Disposition o f Specimens: Biological tissues and fluids will be retained per GLP Regulation for the time period required for studies longer than 28 days. 2.9 Safety Precautions: Refer to appropriate MSDS. Wear appropriate laboratory attire. U se caution when handling knives for cutting the samples. 2 eo269r 3.0 EXPERIMENTAL - Overview The tissues from animals dosed as described (HW I#6329-152), are available for analysis for fluorine compounds. At the discretion o f the Study Director, a series o f analytical tests can be performed. The screening for fluoride in liver via combustion (See Methods--next Section) is the appropriate analysis to present definitive data for fluorine in the liver. Not all o f the tissues and fluid samples w ill be analyzed. When sufficient data has been collected to meet the objectives o f the study in the opinion o f the Study Director, analysis will cease. 4.0 EXPERIM ENTAL - Methods 4.1 Liver and Serum screening methods: (attached) 4.1.1 A M DT-M -1-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer-Liver 4.1.2 AM DT-M -2-0, Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer 4.1.3 AM DT-M -4-0, Extraction o f Fluorochemicals from Rabbit Liver 4.1.4 AM DT-M -5-0, Analysis o f Rabbit Liver Extract for Fluorochemicals using Electrospray Mass Spectrometry 4.1.5 AM D T-M -8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 4.1.6 AM DT-M -14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum 5.0 DATA ANALYSIS 5.1 Data Reporting: Data will be reported as a concentration (weight/weight) o f fluoride per tissue or fluid, or as FC-143 (electrospray mass spectrometry) per unit o f tissue or fluid. Statistics used, at the discretion o f the Study Director, may include regression analysis o f serum concentrations with time and averages and standard deviations o f concentrations for different dose groups. If necessary (in the opinion o f the Study Director), simple statistical tests such as Student's t test may be applied to determine statistical difference. ^t TvW Iw w C0269g 6.0 MAINTENANCE OF RAW DATA AND RECORDS 6.1 R aw Data and Records: Raw data, approved protocol, appropriate specimens, approved final report, and electronic data will be maintained in the AMDT archives. 7.0 REFERENCES 7.1 AM D T-S-10-0, Sample Tracking System 8JLAI-TACHMEKIS_________________________________________ 8.1 AM DT-M -1-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver 8.2 A M D T-M -2-0, Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer 8.3 AM DT-M -4-0, Extraction o f Fluorochemicals from Rabbit Liver 8.4 A M D T-M -5-0, Analysis o f Rabbit Liver Extract for Fluorochemicals Using Electrospray Mass Spectrometry 8.5 AM D T-M -8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 8.6 AM DT-M -14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer-Serum 3IVI Environmental Laboratory M ethod Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann DX2000 Organic Halide Analyzer - Liver M ethod Identification Number: AMDT-M-1 Revision Number: 0 Adoption Date: Revision Date: None Author: Rich Youngblom Approved by: Quality Assurance /z>- / - f r ~ Date Software: MS Word 5 .la Affected Documents: AMDT-M-2 Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer AMDT-EP-3 Routine Maintenance of a Modified Dohrmann DX2000 Organic Halide Analyzer 002700 1.0 SCOPE . APPLICABLE CO M PO UNDS. AND M ATRICES 1.1 Scope: This method is for the operation of a Dohrmann DX2000 when it is used to extract fluoride from various matrices. The fluoride is typically collected in TISAB solution for analysis with an ion selective electrode. 1.2 Applicable C om pounds: Fluorochemicals or other fluorinated compounds. 1.3 M atrices: Biological tissues, particularly liver. 2,0 K EY W O R D S______________________________________________________ 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, DX2000, fluorochemicals. 3.0 PR E C A U T IO N S___________________________________________________ 3.1 Glassware and exhaust gases can be extremely hot. 3.2 Glassware is fragile, broken glass may cause injuries. 3.3 Pressurized gases, proper compressed gas handling practices required. 3.4 Solvent based samples may flash, may need to allow them to dry down before starting run. 3.5 Potential biohazards due to the biological matrices. Use appropriate personal protective equipment. 4.0 SU PPLIE S AND M ATERIALS______________________________________ 4.1 Compressed Oxygen, Hydrocarbon free, regulated to 30 PSI. 4.2 Compressed Helium, High Purity Grade, regulated to 45 PSI. 4.3 Quartz glass sample boat with TeflonTM tubing, Dohrmann 890-097 or equivalent. 4.4 Quartz glass combustion tube, Reliance Glass G-9405-012 or equivalent. 4.5 Orion 940999 Total Ionic Strength Adjustment Buffer (TISAB II ) or equivalent. 4.6 Sample collection vials, HDPE. 4.7 Milli-QTM water 4.8 Polystyrene pipettes. 4.9 Activated Charcoal, E. Merck 2005 or equivalent. 4.10 Hamilton Syringe or equivalent. 4.11 Miscellaneous laboratory glassware 5.0 E Q U IP M E N T __________________________________________________________ 5.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer, modified for fluoride extraction. 5.2 IBM compatible 386 or 486 computer. 5.3 DX2000 software, version 1.00, modified for fluoride extraction. 5.4 Excel Spreadsheet, version 5.0 or greater 6.0 IN T E R FE R E N C E S____________________________________________________ 6.1 Sample size is limited to approximately 150 mg, depending on sample moisture content. This may vary from matrix to matrix. C0270/ 7.0 SAM PLE HANDLING 7.1 Samples are not to be handled with bare hands. Fluoride may leach from the skin to the sample. Use forceps or probe to transfer tissues. 7.2 Samples o f liver are cut from frozen liver and placed in a tared and labeled weigh boat. Use a clean scalpel and cutting board. The cutting board and scalpel should be cleaned with water, methanol, or methanol-water solution after each liver is cut. 8.0 CALIBRATIO N AND STANDARDIZATION 8.1 P reparation of C alibration Standards 8.1.1 The standards required for each project will need to be appropriate for that individual project. Refer to protocol for that project. 8.1.2 Typically 50-500 ppm FC-95 in methanol standards are used. 8.1.3 For rabbit liver studies, use beef liver as the matrix. Cut a piece of frozen beef liver (100 150 mg) and weigh it in a labeled and tared weigh boat. 8.2 C alibration - Overview The normal calibration is the fluoride curve (AMDT-M-2). However, if an optional spiked liver curve is required the procedure listed below is used. 8.2.1 A calibration curve for the DX2000 is generated by spiking samples with known standards and combusting them using the same methods and matrix type as the samples to be tested. 8.2.2 Typically, three replicates of each standard and five concentrations o f standards will be spiked. 8.2.3 Standard curve will be plotted as Mass Spiked F (ug) on the x-axis and Standard Mass Recovered F (ug) on the y-axis. Generate a regression curve and calculate the equation for the line and the r^ value. 8.2.4 Mass Spiked F (ug) = (Amount spiked in mL) x ( Cone, o f standard in ppm) x (0.6004)* *FC-95 is 60.04% F therefore 0.6004 is the factor used to convert FC-95 to F 8.2.5 Standard Mass Recovered F (ug) = (TISAB volume in mL) x (Orion reading in ppm) 8.3 C alibration - Procedure 8.3.1 S ta rt Up 8.3.1.1 Run 2 or more Clean Cycles when starting instrument each day. More clean cycles may be used if the previous samples contained high concentrations o f fluoride. 8.3.2 Blanks 8.3.2.1 Prepare sample using the same methods and type of matrix as the test sample. 8.3.2.2 For rabbit studies, use beef liver as the matrix. Prepare at least 3 samples o f beef liver (100 - 150 mg) for blanks. 8.3.2.3 Put sample in Dohrmann boat. Combust each sample as described in section 9.0 and analyze sample according to method AMDT-M-2 for the ion selective electrode analysis. C0270^ 8.3.2.4 For rabbit studies, the meter reading for a blank sample should be 0.03 ppm or lower before proceeding with the calibration. Bum samples until this limit is reached, or until in the judgement of the operator the reading is stable with respect to historical readings (previous 48 hours). 8.3.2.5 For non-rabbit studies, the blank readings should reach a predetermined ion concentration before proceeding with the calibration. 8.3.2.6 It may be necessary to mix approximately 50 mg of charcoal with the sample to aid combustion. 8.3.3 S tan d ard C urve 8.3.3.1 Weigh out at least 15 matrix samples (5 standards with 3 replicates each) in tared and labeled weigh boats. For rabbit studies, weigh 100-150 mg beef liver samples. Record weights in study data. Store the matrix samples on dry ice or ice packs to keep them frozen until used. 8.3.3.2 Place weighed beef liver sample in Dohrmarm sample boat. 8.3.3.3 Start with the lowest standard concentration. Using a Hamilton syringe, eject a fixed quantity of the standard on or in the matrix. For rabbit studies, use 4 uL o f standard and eject it on or in the beef liver. 8.3.3.4 At least 3 replicates should be used for the lowest standard concentration; more replicates may be used at the discretion of the analyst. 8.33 . 5 Combust the sample as described in section 9.3 and analyze according to AMDT-M-2. 8.3.3.6 Run all 15 standards. If one replicate is significantly different from the other two replicates, mn another sample for that standard. Indicate in data that the new replicate replaces the old replicate and that the new replicate will be used to calculate the regression curve. 8.3.3.7 When all standards have been run, calculate the r^. r^ must be at least 0.95. If it is not at least 0.95, consult with supervisor. 8.3.3.8 A new standard curve should be run when the combustion tube or sample matrix is changed. New standard curve may also be run at the discretion of the analyst. 8.4 Storage Conditions for Standards 8.4.1 Storage requirements for standards are dependent on the individual standards used. Typically, standards are stored at room temperature in plastic screw top bottles. 8.4.2 New FC-95 standards should be prepared at least once a month. 9.0 P R O C E D U R E S_________________________________ 9.1 Typical O perating Conditions: 9.1.1 Combustion tube temperature = 950C. 9.1.2 Oxygen and Helium flow = 50 cc/minute. 9.1.3 Vaporization/Drying time = 240 seconds. 9.1.4 Bake time = 300 seconds. 9.2 S tart Up Procedure: 9.2.1 If the program is not started, start the EOX program on the PC. 9.2.2 Open the SYSTEM SETUP window. 9.2.3 Put the furnace module and the cell in the READY mode. 9.2.4 Close the SYSTEM SETUP window. 0Q270jf 9.2.5 When the oven has reached the READY temperature, run the CLEAN BOAT program found in the CELL CHECK menu. 9.2.6 See AMDT-EP-3 for details o f the Dohrmann software. 9.3 Sample Extraction Procedure: 9.3.1 Open the SAMPLE HATCH and place the sample in the BOAT. It may be necessary to mix approximately 50 mg o f charcoal with the sample to aid combustion. If this is done, charcoal should also be mixed in while establishing the baseline and when generating the standard curve. 9.3.2 Close SAMPLE HATCH. 9.3.3 Add appropriate volume ofTISAB solution or 1:1 TISAB:Milli-QTM water mixture to a labeled sample collection vial. Typically 0.6 mL to 15 mL are used. For rabbit studies, use 1.0 or 2.0 mL of 1:1 TISAB:Milli-QTM water mixture. 9.3.4 Place the vial so that the tip of the COMBUSTION TUBE is in the TISAB at least 0.25 inches. Gases released during pyrolysis must bubble through the TISAB. 9.3.5 Run the EOX-SOLIDS program found in the RUN menu. 9.3.6 When the EOX program is finished, remove the collection vial from the combustion tube. 9.3.7 If undiluted TISAB was used to collect the sample, add an equal volume of Milli-QTM water to the TISAB to make 1:1 TISAB:Milli-QTM. 9.3.8 Rinse the end of the combustion tube with Milli-QTM water and wipe with a KIMWIPE to remove any TISAB remaining on the tube. 9.3.9 Open the sample hatch and remove any remaining ash from the boat. Ash can be removed with a cotton tipped applicator or vacuumed out. It may be necessary to scrap particles off the bottom with a spatula or other similar device. A drop of Milli-QTM water may be added to the boat to aid in the Clean Cycle. 9.3.10 Close the hatch. 9.3.11 Run the CLEAN BOAT program. 9.3.12 Sample is ready for analysis by ion selective electrode (AMDT-M-2). 9.4 Sample Calculations- 9.4.1 Use the standard curve to calculate the sample value. 9.4.2 Sample Mass Recovered F (ug) = (TISAB vol in mL) x (Orion reading in ppm - intercept! (Slope) 10.0 VALIDATION 10.1 Q uality Control 10.1.1 Daily S tart Up Check Samples: Once the standard curve is established, each day of analysis is started by analyzing QC samples. The QC samples are to be the same as the lowest concentration spiked samples used to generate the standard curve. Each concentration must be done in triplicate unless the first two replicates are within 20% of the standard curve, then a third replicate is not necessary. 10.2 Precision and Accuracy: See method development analysis and sample analysis in Fluoride Notebooks 2,3, and 5. Precision and accuracy varies when analyzing samples o f different matrices and different reference compounds. 10.3 O ther Validation Param eters: NA 5 027Olf 11.0 DATA ANALYSIS 11.1 Calculations 11.1.1 For the standard curve, use regression analysis in Excel, version 5.0 or greater. 11.1.2 To calculate the fluoride contraction in the sample, see method AMDT-M-2. 11.2 Analyzing the D ata 11.2.1 r2 must be at least 0.95 or greater. "Outliers" may be excluded if two of the three replicates are within 20% of each other and the outlier is greater than 200% o f the average o f those two or less than 50% o f the average of those two. Any such outliers should be pointed out in the data and noted in the Final Report along with the reason it was considered an outlier. 12.0 A T T A C H M E N T S_____________________________________________________ None 13.0 R EFER EN C ES________________________________________________________ 13.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer Operator's Manual (Manual 915349, revision B, December 1993) 13.2 AMDT-M-2 Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer 13.3 AMDT-EP-3 Routine Maintenance of a Modified Dohrmann DX2000 Organic Halide Analyzer 14.0 R E V ISIO N S__________________________________________________________ Revision Number Reason for Change Revision B ats___ CQ270 3M Environm ental Laboratory M ethod Fluoride M easurement by M eans of an Orion EA940 Expandable Ion Analyzer M ethod Identification Number: AMDT-M-2 Revision Number: 0 Adoption Date: Revision Date: None Author: Rich Youngblom Approved By: CL^ Grodti Leader /J --- / /-* // s Date Quality Assurance Date Software: MS Word 5.1a Affected Documents: AMDT-M-1 Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer 1 C 0 2 7 o 1.0 SCOPE . APPLICABLE CO M PO UND S. AND M ATRICES 1.1 SCOPE: This method is for the calibration and operation of an Orion EA940 Expandable Ion Analyzer. 1.2 A PPLICABLE COM POUNDS: Fluoride. 1.3 APPLICABLE M ATRICES: Liquid samples in an appropriate buffer solution. Preferred pH of 6.0. 2.0 K EY W O R D S_________________________ 2.1 Fluoride, fluorine, ion selective electrode 3.0 PR E C A U TIO N S______________________________________________________ 3.1 No hazards identified with this method. 4.0 SU PPLIE S AND M ATERIALS_______________________________________ 4.1 Orion 940999 Total Ionic Strength Adjustment Buffer II (TISABII) or equivalent. 4.2 Orion Model 900001 electrode filling solution (AgCl) or equivalent. 4.3 Orion 940907 100 ppm fluoride standard or equivalent. 4.4 Milli-QTM water or equivalent. 4.5 Magnetic stir bars. 4.6 Lab tissues. 4.7 Sample collection vials. 4.8 Plastic 100 mL volumetric flasks. 4.9 Polystyrene pipettes. 4.10 Miscellaneous laboratory glassware. 5.0 E Q U IPM E N T _________________________________________________________ 5.1 Orion Model EA940 Expandable Ion Analyzer or equivalent. 5.2 Orion Model 960900 Solid State Combination Fluoride electrode or equivalent. 5.3 Magnetic Stir Plate. 5.4 IBM compatible 386 or 486 computer (only needed if using Orion 3E software). 5.5 Orion RS232 interface cable (only needed if using Orion 3E software). 5.6 Microsoft Excel 5.0 (only needed if using Orion 3E software). 6.0 IN T E R FER EN C ES____________________________________________________ 6.1 It is recommended that the pH be at or near 6.0. A 1:1 mixture o f TISAB and sample/MilliQTM water will generally bring sample to pH o f 6.0. 6.2 Sample temperature may effect fluoride measurement. It is recommended that the sample be at room temperature as the standards were when the meter was calibrated. 6.3 The rate the samples are stirred at should be consistent with the rate the standards were stirred. 2 CQ270f 6.4 Air bubbles trapped under electrode can give erroneous readings. Make sure no air is trapped under electrode. 7.0 SAM PLE H ANDLING______________________________________________ __ 7.1 No special handling necessary. 8.0 C A LIBR A TIO N AND STA N D A R D IZA TIO N __________________________ 8.1 P reparation of C alibration Standards 8.1.1 Measure 50 mL o f TISAB II into 5 100 mL plastic volumetric flasks. 8.1.2 Label the flasks as 0.05, 0.1,0.5, 1.0, and 1.5 ppm F-, along with the date and your initials. 8.1.3 Pipette 0.05, 0.1, 0.5, 1.0, and 1.5 mL of 100 ppm fluoride standard into the appropriately labeled flasks. 8.1.4 Add approximately 30 mL o f Milli-QTM water to each flask. 8.1.5 Shake the flasks to mix the solutions. 8.1.6 Eliminate air bubbles from the flasks by tipping the flasks on their sides and rolling the air in the flasks over the air bubbles. 8.1.7 Bring the volume in the flasks up to the 100 mL mark with Milli-QTM water. 8.1.8 Invert and shake the flasks for the final mixing. 8.1.9 Record standards in Standards Log Book. 8.2 C alibration 8.2.1 If necessary, remove tape from electrode filling hole. 8.2.2 Invert probe to wet top seal. 8.2.3 Eject a few drops of filling solution from bottom o f electrode to wet lower seal. 8.2.4 Fill the electrode with filling solution. 8.2.5 The meter and the F- electrode are typically calibrated by direct measurement with no blank correction, using standards with concentrations o f 0.05, 0.1, 0.5, 1.0, and 1.5 ppm F-, following the manufacturer's instructions. 8.2.6 Record the slope in the appropriate log book. 8.2.7 Clean the electrode by rinsing with Milli-QTM water and wiping the sides down with lab tissues. 8.3 Storage Conditions for Standards 8.3.1 Calibration standards are stored at room temperature. 9.0 PR O C ED U R ES________________________________________________________ 9.1 C alibration and M easurem ent, Standard method: 9.1.1 The sample to be measured needs to be mixed with TISAB using the proportions recommended by the TISAB manufacturer. 9.1.2 Place a stir bar in the sample and place the sample on the stir plate. 9.1.3 Allow the sample to mix for a few seconds before inserting the electrode. When the electrode is inserted, make sure there are no air bubbles trapped under the electrode. 9.1.4 The sample should be the same temperature as the calibration standards and stirred at the same rate as the calibration standards. 9.1.5 When the readings have stabilized, record the reading in the appropriate log book. 3 9.2 C alibration And M easurem ent, Using O rion 3E Software: 9.2.1 C alibration: 9.2.1.1 Follow steps 8.2.1 to 8.2.4. 9.2.1.2 Press Function Key #8 (F8). 9.2.1.3 The computer screen will ask you to confirm the number of standards to be used, concentration o f the standards, and whether or not a blank is to be included in the calibration. Make any necessary changes to the information presented and click on CONTINUE. 9.2.1.4 Place the electrode in the first standard on the stir plate and click on CONTINUE. 9.2.1.5 Observe the readings on the graphic display on the computer. When the readings have stabilized, press ACCEPT READING. 9.2.1.6 Repeat step 9.2.1.4 and 9.2.1.5 for the remaining standards. 9.2.1.7 After the final standard, the computer will display the slope of the curve, as well as the intercept and correlation. Record the slope, intercept, and correlation in the appropriate log book and click on CONTINUE. The calibration data is automatically copied to C:\Orion\Data\Calib.txt. 9.2.2 D ata Spreadsheet: 9.2.2.1 Select either NEW or OPEN from the FILE menu to open a new or existing spreadsheet to store data in. 9.2.2.2 Record the name of the spreadsheet used in the appropriate log book. 9.2.3 Fluoride M easurem ent: 9.2.3.1 Follow steps 9.2.1 through 9.2.4 9.2.3.2 Enter the name o f the sample in the appropriate place on the screen. 9.2.3.3 Click on the NEW SAMPLE button ' 9.2.3.4 When the readings have stabilized, click on the RECORD button and write the result in the appropriate log book. 10.0 V A L ID A T IO N ________________________________________________________ 10.1 Q uality Control: 10.2 Precision and Accuracy 10.3 O ther Validation Param eters According to Reference 13.2, the range o f detection is 0.02 ppm fluoride up to a saturated solution of fluoride. 11.0 DATA ANALYSTS____________________________________________________ 11.1 Calculations None necessary. 11.2 Analyzing the Data None necessary. 12.0 A T T A C H M EN TS____________________________________________________ None 13.0 REFERENCES 4 W t o TTu x 0027 13.1 Orion Model EA940 Expandable Ion Analyzer Instruction Manual, Orion Research Incorporated, 1991. 13.2 Orion Model 960900 Solid State Combination Fluoride Electrode Instruction Manual, Orion Research Incorporated, 1991. 14.0 REVISIONS Revision Number Reason fpr hanpe Revision Date 002710 3M Environm ental Laboratory M ethod Extraction of Fluorochem icals from Rabbit Livers SOP Identification Num ber: AMDT-M-4 Revision Num ber: 0 A doption D ate: / o - ? t - <''C Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: roup Leader /0-31-lS Date Quality Assurance Date Software: MS Word, 6.0 Affected Documents: M-5, Analysis of Rabbit Extract for Fluorochemicals Using Electrospray Mass Spectroscopy. ooo m a 1 C 02718 1.0 SC O P E _____________________________________________________________ 1 .1 Scope: This method is for the extraction of fluorochemicals from rabbit livers. Ethyl acetate is used to extract fluorochemicals from the livers for analysis by electrospray mass spectroscopy. 1 .2 Applicable Compounds: Fluorochemicals or other fluorinated compounds. 1 .3 M atrices: Rabbit Livers. 2.0 K E Y W O R D S _________________________ _______________________________ 2 .1 Fluorochemicals, rabbit livers, electrospray mass spectrometer, fluorinated compounds, extraction. 3.0 PRECAUTIONS______________________________________ 3 .1 Use gloves when handling the rabbit livers, they may contain pathogens. 4 .0 S U P P L IE S A N D M A T E R IA L S_______________________________________ 4.1 Supplies 4 .1 .1 Syringe, capable of measuring 100 pL 4 .1 .2 Eppendorf type or disposable pipets 4 .1 .3 Gloves 4 .1 .4 Plastic grinding tubes 4 .1 .5 Plastic centrifuge tubes, 15 mL 4 .1 .6 Labels 4 .1 .7 Nitrogen 4 .1 .8 Timer 4 .1 .9 Filters, Titan nylon syringe filters, 0.2 pm. 4 .1 .1 0 Analytical pipets: glass volumetric pipets. 4 .1 .1 1 Disposable plastic 3 cc syringes. 4 .1 .1 2 Crimp cap autovials. 4.2 Reagents 4 .2 .1 Aqueous Ammonium Acetate (Aldrich), approx. 250 ppm: Prepare a 2500 ppm aqueous solution of ammonium acetate by adding 250 mg ammonium acetate to a 100 mL volumetric flask and dilute to volume with Milli-Q water. Dilute this solution 1:10 for a 250 ppm solution. 4 .2 .2 Sodium carbonate/Sodium Bicarbonate Buffer (J.T. Baker), (Na^COj/NaHCOj) 0.25 M: Weigh 26.5 g of sodium carbonate (N a ^ O j) and 21.0 g of sodium bicarbonate (NaHC03) into a 1 L volumetric flask and bring to volume with Milli-Q water. 4 .2 .3 Dilute acetonitrile solution, dilute acetonitrile 1:1 with Milli-Q water. 4 .2 .4 Ethyl Acetate 4 .2 .5 Methanol 4 .2 .6 Milli-Q water 4 .2 .7 1H,1H,2H,2H - periluorooctanesulfonic acid (Aldrich) 4 .2 .8 FC-95 (3M Specialty Chemical Division) 2 C0271JL/ 5J) EQUIPMENT___________________________ 5 .1 Ultra-Turrax T25 Grinder for grinding liver samples. 5 .2 Vortex mixer 5 .3 Centrifuge 5 .4 Shaker 5 .5 Analytical Evaporator 6JD IN T E R F E R E N C E S __________________ ___ 6 .1 There are no known interferences at this time. 7.0 S A M P L E H A N D L IN G ________________________________________________ 7 .1 The rabbit livers are received frozen, and must be kept frozen until the extraction is performed. 8.0 C A L IB R A T IO N A N D S T A N D A R D IZ A T IO N _______________________ 8 .1 Preparation of Internal Standards 8 .1 .1 Prepare an internal standard of approximately 12 ppm 1H,1H,2H,2Hperfluorooctanesulphonic acid to be added to each liver sample. 8 .1 .2 Weigh at least 0.1 g of lH,lH,2H,2H-perfluorooctanesulphonic acid into a 100 mL volumetric flask. Record the actual weight. 8 .1 .3 Bring it up to volume with methanol, this is the stock standard. 8 .1 .4 To a 250 mL volumetric flask, add 3 mLs of the stock standard and bring to volume with Milli-Q water. Calculate the actual concentration of the standard. actual mg perfluoroctane- sulphonic acid X 3 mL = 0.1 L 250 mL actual concentration, ppm 8 .2 Prepare FC-95 Anion Standards 8 .2 .1 Prepare FC-95 standards for the standard curve. 8 .2 .2 Weigh approximately 100 mg of FC-95 into a 100 mL volumetric flask. Record tire actual weight. 8 .2 .3 Bring up to volume with dilute acetonitrile. 8 .2 .4 Dilute the solution with dilute acetonitrile 1:10 for a solution of approximately 100 ppm. Dilute this solution 1:10 with dilute acetonitrile for a solution of approx. 10 ppm. 8 .2 .5 Use the 10 ppm solution to make working standards with values close to 5.0 ppm, 1.0 ppm and 500 ppb. 8 .3 Prepare Beef Liver Homogenate to Use for Standards 8 .3 .1 Weigh 40 g of Bovine liver into a 250 mL Nalgene bottle containing 200 mLs Milli-Q water. Grind to a homogenous solution. 8 .3 .2 Add 1 mL of the solution to a 15 mL centrifuge tube. Prepare a total of eight 1 mL aliquots of the solution in 15 mL centrifuge tubes. Be sure to re suspend solution by shaking it between aliquots. CQZ71J 8 .3 .3 Spike seven of the 1 mL aliquots with the following amounts of working standards in step 9.12 of the procedure. One 1 mL aliquot serves as the blank. Working Standard (Approximate Cone.) 500 ppb 500 ppb 500 ppb 500 ppb Ippm 5 ppm 5 ppm uL 100 200 300 400 500 2 300 Approximate final concentration of FC-95 in liver Blank 0.292 ppm 0.5&4 ppm 0.877 ppm 1.168 ppm .024 ppm 5.&48 ppm 8.772 ppm 8 .4 Calculate the actual value of the standards: uL of standard x concentration fin ppm! = final concentration (ppm) 171 mg liv e r '/1 ml homogenate of FC -95 in liver *Average weight of bovine liver in solution as determined by weighing 1 mL homogenates of 40 mg liver in 200 mL of Milli-Q water. The amount of FC-95 is reported as equivalents o f FC-95 potassium salt. 8.5 C alibration 8 .5 .1 Extract the spiked beef liver homogenate following 9.13 to 9.23 o f this method. Use these standards to establish your curve on the mass spectrometer. 8 .5 .2 Alternatively, a standard curve may be generated using ratios of responses o f the perfluorooctansulfonate anion and the internal standard anion versus concentration of the perfluorooctanesulfonate anion. 8.6 Storage Conditions for Standards 8 .6 .1 New standards are prepared with each analysis. Standards are stored in covered plastic centrifuge tubes until the analysis on the mass spectrometer is performed. 8.7 Storage Conditions for Standards 8 .7 .1 Beef liver homogenates may be frozen after preparation. 9.0 P R O C E D U R E S _____________________________________________________ 9 .1 Obtain frozen liver samples. In spent tissue, note that the liver has not been packaged with other tissues. 9 .2 Use a dissecting scalpel and cut off approximately 1 g of liver. 9 .3 Weigh the sample directly into a tared plastic grinding tube. 9 .4 Record the liver weight in the study note book. 9 .5 Put a label on the vial with the study number, weight, rabbit ED, date and analyst initials. T T m /u T ro 4 9 .6 Add 2.5 mLs water. 9 .7 Grind the sample. Put the grinder probe in the sample and grind for about 2 minutes, until the sample is a homogeneous solution with no large chunks. 9 .8 Rinse the probe off into the sample with 2.5 mLs water using a pipet. 9 .9 Take the grinder apart and clean it with methanol after each sample. Follow AMDT-EP-22. 9 .1 0 Cap the sample and vortex for 15 seconds. 9 .1 1 Pipet 1 mL into a 15 mL centrifuge tube. Label the centrifuge tube with the identical information as the grinding tube. (See AMDT-M-4 Worksheet for documenting the remaining steps.) 9 .1 2 Spike the beef liver homogenates with the appropriate amount of FC-95 standard as described in 8.3. 9 .1 3 Spike the samples and beef liver homogenates with 100 uL o f internal standard. 9 .1 4 Add 1 mL of the sodium carbonate/sodium bicarbonate buffer and 1 mL ammonium acetate. 9 .1 5 Using an analytical pipet, add 5 mL ethyl acetate. 9 .1 6 Cap the sample and vortex 20 to 30 seconds. 9 .1 7 Put them in the shaker for 20 min. 9 .1 8 Centrifuge for 20 to 25 minutes, until the layers are well separated. Set the power on the centrifuge to 25. 9 .1 9 Remove 4 mLs of the top organic layer to a fresh 15 mL centrifuge tube with a 5 mL graduated glass pipet. Transfer the label to the fresh tube. 9 .2 0 Blow the sample down on the analytical evaporator to near dryness with nitrogen, approximately 30 to 40 minutes. 9 .2 1 Bring the remaining sample up in 1 mL dilute acetonitrile with an analytical pipet. 9 .2 2 Vortex 15 seconds. 9 .2 3 Transfer the sample to a 3 mL syringe. Attach a 0.2 (im nylon mesh filter, and filter the sample into a fresh centrifuge tube or a autovial. Label the tube or vial with the study number and animal number. 9 .2 4 Cap and hold for analysis by electrospray mass spectroscopy. 9 .2 5 Complete AMDT-M-4 worksheet and attach to page of study notebook. 10.0 V A L ID A T IO N __________________________________ 10 .1 Quality Control - not applicable 10 .2 Precision and Accuracy- not applicable 1 0 .3 Other Validation Parameters- not applicable 11.0 D A T A A N A L Y S IS ______________________________ 11.1 None 12.0 A T T A C H M E N T S ________________________________ 1 2 .1 Worksheet AMDT-M-4 13.0 R E F E R E N C E S ___________________________________ 1 3 .1 AMDT-EP-22 Routine Maintenance of Ultra-Turrax T-25 14.0 R E V IS IO N S _____________ Revision Number Reason for Change Revision Date WWW 5 G0271JF W orksheet AMDT-M-4 Study # _ _ _ . _ _ 1 Sam ple N um ber set # B lank F iver F C -95 approx 0.5 ppm a c tu a l ppm #W _ 100 u l. 200 ul. 300 uL 400 uL _ _ _ _ _ _ _ F C -95 approx 1 ppm actual ppm #W _ - _ - 500 uL _ ,, _ * _ _ F C -9 5 approx. 5 ppm a c tu a l ppm #W - - D ate and Initials for Std. 200 uL 300 ul. _ _ _ _ _ _ .. 1 1 s tu d y n u m b e r v.'h e re th e o rig in a l w o r k s h e e t is lo c a te d a n d n la c e a c o n v .____ Il II L iv e r E x tra c tio n P ro c e s s :__________ _________________ D a te & I n itia ls ______ P in et 1 m L o f T.iver S o lu tio n P in e t 100 u L o f .1 2 n n m In te rn a l S ta n d a r d _____________ S td . # ____________ V ortex 15 see. P in e t 1 m i . n f 2 5 0 n n m A m m o n iu m A c e ta te ____________ S td # ____________ P inet 1 mT. o f 0.25 N a,C O ,/0 25M N aH C O , B uffer P inet 5 m l. o f Fthvl A cetate V ortex 20-30 sec Shake 20 m in C entrifune 20-25 m in R em ove a 4 m l. alinuot o f organic laver B low dow n to near drvness f< 0 25 m i l w ith NA dd 1 m- o f M A cetonitrile/H -O V ortex 1S cec ___ T b t ____________ Filter usine a 3cc B -D . syringe a iti] a 0 2 n m S R I f ilte r in to a 1 5 m l a u to s a m n lc vial____________ 6- 002719 3M Environm ental Laboratory M ethod Analysis of Rabbit Liver Extract for Fluorochem icals using Electrospray M ass Spectroscopy SOP Identification Number: AMDT-M-5 Revision Number: 0 Adoption Date: $-- Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: Software: MS Word, 6.0 Affected Documents: M-4, Extraction of Fluorochemicals from Rabbit Livers _ j_ C0271^ LO SCOPE________________________ ________________________ 1.1 Scope: This method is for the analysis of extracts of rabbit liver or other tissues or fluids for fluorochemicals using the electrospray mass spectrometer. The analysis is performed by single ion monitoring of FC-95 anion, M/Z= 499, the internal standard M/Z = 427, and other appropriate masses. 1.2 Applicable Compounds: Fluorochemicals or other fluorinated compounds. 1 .3 M atrices: Rabbit Livers (samples), Beef Liver (standards), other tissues and fluids. 2.0 K E Y W O R D S_________________________ ;_________________________________ 2 .1 Fluorochemicals, fluorinated compounds, electrospray mass spectroscopy, mass spectrometer, rabbit livers. 3.0 P R E C A U T IO N S _______________________________________________________ 3 .1 Use caution with the voltage cable for the probe. When the voltage cable is plugged into the probe DO NOT TOUCH THE PROBE, there is risk of electrical shock. 3 .2 Do not run the pump above it's capacity of 4000 psi. If pressure goes over 4000 psi stop and release pressure. The peak tubing may be plugged. Troubleshoot back to find the plug and replace the plugged tubing. See AMDT-EP-15 3 .3 Do not run the pump to dryness. 4 .0 SU P P L IE S AND M A T E R IA L S_____________________________________ 4.1 Supplies 4 .1 .1 Nitrogen gas regulated to 140 psi. 4 .1 .2 Fluofix column or equivalent. 4 .1 .3 100 uL or 250 uL flat tip syringe for sample injection. 4.2 Reagents 4 .2 .1 Dilute acetonitrile mobile phase, dilute acetonitrile 1:1 with Milli-Q water. 4 .2 .2 Milli-Q water, all water used in this method should be Milli-Q water. 5.0 E Q U IP M E N T _______________________________________________________ 5 .1 VG Trio 2000 Electrospray Mass Spectrometer or equivalent. 5 .2 ISCO Syringe Pump 5 .3 Spectraphysics AS300 Autosampler 5 .4 100 uL Assembly 5 .5 Autovials or capped centrifuge tubes. 6.0 IN T E R F E R E N C E S ______________________ 6 .1 There are no known interferences at this time. 7.0 SA M P L E H A N D L IN G ________________________________________________ 7 .1 Keep the extracted samples in capped 15 mL centrifuge tubes or in capped autovials until ready for analysis. 00271g 8.0 C A L IB R A T IO N AND S T A N D A R D IZ A T IO N _______________________ 8.1 Preparation of Calibration Standards 8 .1 .1 Seven beef liver standards and one blank beef liver are prepared during the extraction procedure. (See AMDT-M-4, section 8.0) 8.2 Calibration 8 .2 .1 Run the seven beef liver standards twice, starting with the lowest standard to obtain the standard curve. 8 .2 .2 Typically one standard is run after each 5 to 7 samples. Choose a standard in the same range of concentration as the samples. 8.3 Storage Conditions for Standards 8 .3 .1 Fresh standards are prepared with each analysis. Standards are stored in covered plastic centrifuge tubes until the analysis on the mass spectometer is performed. Samples and standards are NOT refrigerated. 8.4 Storage Conditions for Beef Liver Homogenates 8 .4 .1 Beef liver homogenates may be frozen after preparation. 9 .0 P R O C E D U R E ________________________________________________________ 9.1 Initial Set-up 9 .1 .1 Set software to "Operate on", Ion Mode ES'. 9 .1 .2 Record backing pressure in the instrument log. 9 .1 .3 Fill the solvent cylinder with mobile phase. 9 .1 .4 Set the pump to "Run". Set the flow to 1000 uL/min. Observes droplets coming out of the tip of the probe. The pressure should be at 1700 to 1800 psi. 9 .1 .5 Check the fused silica at the end of the probe. Use an eye piece to check for chips. The tip should be flat with no jagged edges. If any chips are found cut off the tip of the silica with a column cutter and pull the silica through to the appropriate length. 9 .1 .6 Check your nitrogen supply. Turn on the nitrogen. There should be no nitrogen leaking around the tip of the probe. A fine mist should be coming out of the tip. 9 .1 .7 Carefully guide the probe into the opening. Insert it until it won't go any further. Connect the voltage cable to the probe. 9 .1 .8 Go to the "Editor" page, and set Ionization Mode to ES', and the appropriate masses to 427 and 499. 9 .1 .9 If it is not in single ion mode go to "Option" and set SIR. 9 .1 . lOStart Acquisition. Assign a file name, MO-DAY-YR + letter. Record it in the log book. 9 .1 .1 1 Run the beef liver samples first, running each standard twice at the beginning of the run.. Run a QC check by running one standard after every 5 to 7 samples. 9.2 M anual Injection 9 .2 .1 Draw 150 uL of sample into a syringe. Inject the sample into the rheodyne injection port. Inject slowly. Record the sample ID in the log book. 9 .2 .2 Turn the valve to "On". 9 .2 .3 Wait two minutes, and inject the next sample. 9 .2 .4 Record the scan number for each sample in the logbook. c027/9 9 .3 Using the Autosampler 9 .3 .1 Set up sample tray A, B, or C. 9 .3 .2 Record the samples and their positions in the instrument log book. Up to 17 vials may be in each run. 9 .3 .3 Set-up the sampler: 9 .3 .3 .1 Push the sample button 9 .3 .3 .2 Set sample loop size = 100 uL 9 .3 .3 .3 Set inject/sample = 2 9 .3 .3 .4 Set Cycle time = 0 9 .3 .3 .5 Name the file: Livers 9 .3 .3 .6 Identify the tray used 9 .3 .3 .7 Add the samples to Queue by pressing "Enter' 9 .3 .3 .8 Press "Run" to start 10.0 V A L ID A T IO N __________________________________ ___________________ 10.1 Q uality Control 1 0 .1 . IR un a standard every 5 to 7 samples. If a significant change( 50%) in peak height occurs stop the run. Only the samples before the last acceptable standard will be used. The remaining samples will be reanalyzed. 10.2 Precision and Accuracy 1 0 .2 . lS ee Method Validation Report number AMDT-M-5.0.V1 10.3 O ther Validation Param eters 1 0 .4 Refer to Method Validation Report Number AMDT-M-5.0.V 1 11.0 D A T A A N A L Y SIS__________________________________________________ 11.1 11.2 Calculations Plot the standard curve, using the mean of the two values obtained for each standard. 1 1 .2 . IR ead peak heights or areas for the samples from the printout. Use linear regression to determine the sample concentrations. 1 1 .2 .2 Calculate the mg of FC-95 anion, or other fluorochemical in the total rabbit liver: mg FC-95 anion in the total rabbit liver = mg FC-95 anion from std. curve gms of liver used for analysis x Total mass of liver, gms 11.3 11.4 Make a results table and enter it in the study book. Print a chromatogram for each sample, with the peaks labeled with the sample or standard ED. Write the study number on the printout, initial, date, and put it in the study folder. Staple all chromatograms together and number pages. 4 12.0 ATTACH M ENTS None 13.0 REFERENCES 1 3 .1 AMDT-EP-17 14.0 REV ISIO NS Revision Number Reason for change Revision Date 3M Environmental Laboratory M ethod A nalysis of Fluoride Using the Skalar Segm ented Flow A nalyzer With Ion Selective Electrode Method Identification Number: AMDT-M-8 Adoption Date: Revision Number: 0 Revision Date: None Author: Deb Wright / Cynthia Weber Approved By: Software: IBM MS Word, 6.0 Affected Documents: AMDT-EP-26, Operation and Maintenance of the Skalar Segmented Flow Analyzer 1 1.0 SC O P E _______________________________________________ _________________ 1.1 This method is for the analysis for fluoride, thermally extracted from samples using the Dohrmann DX2000 (AMDT-M-1), and collected in TISAB for analysis with an Ion Selective Electrode (ISE). The analysis is performed using the Skalar Segmented Flow Analyzer with ISE. 1.2 Samples can be tissues, serum, biological material, or other materials extracted on the Dohrmann. 2.0 K E Y W O R D S___________________________ 2 .1 Skalar, segmented flow, fluoride. 3 .0 P R E C A U T IO N S ________________________ 3 .1 Follow standard laboratory safety practices. 4 .0 S U P P L IE S A N D M A T E R IA L S____________________________ _ 4.1 Supplies 4 .1 .1 Sample cups, 4 mL plastic cups with caps 4 .1 .2 Autopipets, oxford or equivalent with plastic tips 4 .1 .3 Polypropylene volumetric flasks, 100 mL 4 .1 .4 Cartridge components, refer to the Skalar Methods for components and part numbers. 4 .1 .5 Sample prefilters, Evergreen 4.2 Reagents 4 .2 .1 Brij 35, 30% S.F.A.S. Detergent 4 .2 .2 TISAB II buffer solution: Purchase TISAB II from Orion. To 1 liter of TISAB II add 2.5 mL or 100 ppm fluoride solution and 1 mL Brij. 4 .2 .3 Sampler rinsing solution: Dilute TISAB II 1:1 with Milli-Q water. 4 .2 .4 Nitric acid solution for decontamination, 1 N (lab grade): Slowly add 64 mLs concentrated nitric acid (H N 03) to 250 mLs of Milli-Q water. Bring the volume up to 1 L with Milli-Q water. 4.3 Standards 4 .3 .1 Stock solution, 100 ppm F: purchased from Orion. 4 .3 .2 Intermediate standard, 10 ppm: Dilute 10 mLs of stock solution to 100 mLs with Milli-Q water. Use polypropylene volumetric flasks. 4 .3 .3 Working standard: Make up the following working standards by adding the _volumes of intermediate or stock standard indicated on the table, using Oxford or pumpmate pipets, to 50 mLs of TISAB and diluting to 100 mLs ___________________ with Milli-Q water.___________________________________________________ Working Standard mLs of Stock Standard mLs of Intermediate Standard 0.015 ppm - 0.15 0.03 ppm - 0.3 6.06 ppm - 0.6 0.09 ppm - 0.9 0.12 ppm - 1.2 0.15 ppm - 1.5 0.3 ppm 0.3 - 0.6 ppm 0.6 - 2 ^ ..E Q UIPMENT___________________________________ _ 5 .1 Skalar Segmented Flow Auto Analyzer Sanspiu` System equipped with ISE 6.Q. INTERFERENCES___ __________________________ _ 6 .1 High concentrations of alkalinity, chloride, phosphate, sulfate or iron can cause interferences. LQ .SAMPLE HANDLING_____________________________ _ 7 .1 Samples should be stored in polyethylene bottles. Samples should be analyzed within 30 days. M .CALIBRATION. AND-STANPARDIM.TIQN______________ _ _ 8.1 Preparation of Calibration Standards 8 .1 .1 Prepare calibration standards as in section 4.3. 8.2 Calibration 8 .2 .1 The standards are analyzed at the beginning of the run. 8.3 Storage Conditions for Standards 8 .3 .1 Standards are stored in capped polypropylene volumetric flasks. New standards are prepared at a minimum of every six months, or as necessary. ^ j L E R O C E P U R E ___________________________________________________________ 9.1 Start Up Procedure 9 .1 .1 Clamp down the pumpdecks, air bars and sampler-pump tubing. 9 .1 .2 Put the fluoride electrodes in the electrode chamber. 9 .1 .3 Turn on the power of the sampler, pumps, offset potentiometer and heating bath. 9 .1 .4 Put the reagent-lines in the appropriate bottles. 9 .1 .5 Turn on the interface, computer, display and printer. M ake su re you tu rn on the interface before the com puter. 9 .1 .6 Let the system stabilize for approximately 30 minutes. 9.2 Starting a Run 9 .2 .1 Create a sample table by selecting FILES, TABLE, and CREATE, type in the name of the file, and press ENTER. 9 .2 .2 Print the sample table, inserted in the system table by pushing ESC, PRINT, GROUP 1. This will print the entire run. 9 .2 .3 Dial the sampler settings to the appropriate number o f samples, number of seconds for sample wash, and number of seconds for the sample. 9 .2 .4 Fill the sample tray with the standards, samples, washes and drifts. IW and FW/RUNOUT cups on the sampler do not need to be filled. 9 .2 .5 Set the baseline. 0027; 9 .2 .5 .1 Select GRAPHICS, REAL TIME. If you cannot get real-time, you may be in the Data Handling Panel. Switch to the Analysis Panel by selecting CONTROL PANEL and pushing F7. 9 . 2 . 5 . 2Use the small screwdriver for the offset potentiometer to set the base line. Adjust the baseline until it is approximately 3/4 inch from the bottom of the screen. 9 . 2 . 5 . 3Check the highest standard and adjust the gain, if necessary, with the interface screw #3. 9 .2 .6 Go to CONTROL PANEL, and to analysis panel. Deselect the analysis that will not be run. (Select or deselect analysis by pressing ENTER.) Press Tab to return to the Analysis Panel. 9 .2 .7 Press the spacebar to bring up the local menu. 9 .2 .8 Select START to start the analysis. 9 .2 .9 Type your ID (initials), the sample table which you created under 9.2.1 (or press ENTER for choices), choose running with or without the system table and select START ANALYSIS. 9 .2 .1 0 After starting the software, start the sampler. Make sure that the sampler is set to the right number of samples and that the sample/wash/air times are OK. 9 .2 .1 1 Select GRAPHICS, REAL TIME to view the progress of the analysis. 9 .3 Loading and Printing the Data-File 9 .3 .1 Go to CONTROL PANEL, press the spacebar to bring up the local menu and select LOAD. Select AUTOCALCULATION and enter the filename (or highlight the file to be printed and press ENTER). 9 .3 .2 To view the calibration curve, go to GRAPHICS, CALIBRATION CURVE. 9 .3 .3 To print the high level curve, push PRINT SCREEN. 9 .3 .4 To print the low level screen, push ESC to get out of graphics. Select SETTINGS. Change the max y value to approximately 900. Go to CAL CURVE and press ESC, andEnter. Press PRINT SCREEN. 9 .3 .5 Return to SETTINGS and change the max value back to 4095, go to EDIT, press ENTER and PRINT SCREEN to print sample peaks. 9 .3 .6 To print the results go to CONTROL PANEL, SPACEBAR, OUTPUT, OUTPUT. Select PRINTER for the Epson or PRN for the Laser. 9 .4 Shutdown 9 .4 .1 Put all the reagent-lines in Milli-Q water. 9 .4 .2 Let the system rinse for approximately 30 minutes. 9 .4 .3 After the system has rinsed completely, turn off the sampler, pump and offset potentiometer. Turn off the heating bath on weekends. Leave liquid in the lines. 9 .4 .4 Take the electrode out and soak in 100 ppm F overnight. 9 .4 .5 Release the pump-decks, air bars and sampler pump-tubing. 9 .4 .6 Select FILES, press ALT F and select QUIT to exit the program. 9 .4 .7 On Friday, turn off the computer, display and interface for the weekend. 10.0 V A L ID A T IO N _______________________________________________________ 10.1 Quality Control 1 0 .1 . IRun a standard (mid to high concentration) every 10 samples. If a significant change in peak height occurs, only the samples before the last acceptable standard will be used. The remaining samples will be reanalyzed. C0272* 4 10.2 P recision an d A ccuracy 10. ?. lS ee Method Validation Report number AMDT-M-8.0.VI 1 0 .3 Other Validation Parameters 1 0 .4 Refer to Method Validation Report Number AMDT-M-8.0. VI 11.0 D A T A A N A L Y S IS________________________________________________ _ _ 11.1 Calculations 1 1 .1 . IT he standard curve is plotted by the Skalar software. 1 1 .1 .2 All calculations are done by the Skalar software, r2 should be 0.995 or better. 1 1 .2 Prepare spreadsheets to summarize data. Include sample volume, weights used etc. 1 1 .3 Write the study number on the printouts, initial, date the printout, and bind together with all package documents and place in the study folder. Make a copy of the summary sheet and tape into the study notebook. Back up all data and spreadsheets onto study disk and backup disks. 1 1 .4 Electronic Data 1 1 .4 .1GLP studies: Electronic data is copied onto the Study floppy disk for each study, and also data is copied onto a floppy disk that is stored in the lab. 1 1 .4 .2 Other studies: All data is copied onto a floppy disk that is stored in the lab. 12.0 A T T A C H M E N T S _____________________________________________________ None 13.0 R E F E R E N C E S ____________ ;___________________________________________ 13.1 13.2 13.3 AMDT-M-1, Thermal Extraction of Fluoride by Means of a Modified Dohrmann DX2000 Organic Halide Analyzer-Liver Skalar Methods, #335, Skalar Methods Manual AMDT-EP-26, Operation and Maintenance of the Skalar Segmented Flow Analyzer 14.0 R E V ISIO N S Revision Number Reason for change Revision Dais__ 5 027; 3M Environm ental Laboratory M ethod Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer - Serum M ethod Identification Num ber: AMDT-M-14 Revision N um ber: 0 Adoption Date: / o - 3 c" Revision Date: None Author: Rich Youngblom Approved by: roup Leader Date Quality Assurance Date Software: MS Word 5.1a Affected Documents: AMDT-M-2 Fluoride Measurement by Means o f an Orion EA940 Expandable Ion Analyzer AMDT-EP-3 Routine Maintenance o f a Modified Dohrmann DX2000 Organic Halide Analyzer 1.0 SCOPE . APPLICABLE COM POUNDS. AND M ATRICES 1.1 Scope: This method is for the operation o f a Dohrmann DX2000 when it is used to extract fluoride from various matrices. The fluoride is typically collected in TISAB solution for analysis with an ion selective electrode. 1.2 A pplicable C om pounds: Fluorochemicals or other fluorinated compounds. 1.3 M atrices: Biological fluids, particularly serum. 2.0 K E Y W O R D S__________________________________________________________ 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, DX2000, fluorochemicals. 3.0 P R E C A U T IO N S_______________________________________________________ 3.1 Glassware and exhaust gases can be extremely hot. 3.2 Glassware is fragile, broken glass may cause injuries. 3.3 Pressurized gases, proper compressed gas handling practices required. 3.4 Solvent based samples may flash, may need to allow them to dry down before starting run. 3.5 Potential biohazards due to the biological matrices. Use appropriate personal protective equipment. 4,0 SUPPLIES AND M ATERIALS 4.1 Compressed Oxygen, Hydrocarbon free, regulated to 30 PSI. 4.2 Compressed Helium, High Purity Grade, regulated to 45 PSI. 4.3 Quartz glass sample boat with TeflonTM tubing, Dohrmann 890-097 or equivalent. 4.4 Quartz glass combustion tube, Reliance Glass G-9405-012 or equivalent. 4.5 Orion 940999 Total Ionic Strength Adjustment Buffer (TISAB I I ) or equivalent. 4.6 Sample collection vials, HDPE. 4.7 Milli-QTM water 4.8 Polystyrene pipettes. 4.9 Activated Charcoal, E. Merck 2005 or equivalent. 4.10 Hamilton Syringe or equivalent. 4.11 Miscellaneous laboratory glassware 5.0 EQ UIPM ENT 5.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer, modified for fluoride extraction. 5.2 IBM compatible 386 or 486 computer. 5.3 DX2000 software, version 1.00, modified for fluoride extraction. 5.4 Excel Spreadsheet, version 5.0 or greater 6.0 IN T E R F E R E N C E S 6.1 Sample size is limited to approximately 100 jil. This may vary from matrix to matrix. 7.0 S A M P L E H A N D L IN G 7.1 Samples are to be handled with plastic pipettes. A new pipette is to be used for each sample. 8.0 C A L IB R A T IO N AND ST A N D A R D IZ A T IO N ___________________________ 8.1 Preparation of Calibration Standards 8.1.1 The standards required for each project will need to be appropriate for that individual project. Refer to protocol for that project. 8.1.2 Typically 50-500 ppm FC-95 in methanol standards are used. 8.1.3 For rabbit serum studies, use beef serum as the matrix. 8.2 Calibration - Overview The normal calibration is the fluoride curve (AMDT-M-2). However, if an optional spiked serum curve is required the procedure listed below is used. 8.2.1 A calibration curve for the DX2000 is generated by spiking samples with known standards and combusting them using the same methods and matrix type as the samples to be tested. 8.2.2 Typically, three replicates of each standard and five concentrations o f standards will be spiked. 8.2.3 Standard curve will be plotted as Mass Spiked F (ug) on the x-axis and Standard Mass Recovered F (ug) on the y-axis. Generate a regression curve and calculate the equation for the line and the r^ value. 8.2.4 Mass Spiked F (ug) = (Amount spiked in mL),x ( Cone, o f standard in ppm) x (0.6004)* *FC-95 is 60.04% F therefore 0.6004 is the factor used to convert FC-95 to F 8.2.5 Standard Mass Recovered F (ug) = (TISAB volume in mL) x (Orion reading in ppm) 8.3 Calibration - Procedure 8.3.1 Start Up 8.3.1.1 Run 2 or more Clean Cycles when starting instrument each day. More clean cycles may be used if the previous samples contained high concentrations o f fluoride. 8.3.2 Blanks 8.3.2.1 Prepare sample using the same methods and type o f matrix as the test sample. 8.3.2.2 For rabbit studies, use beef serum as the matrix. 8.3.2.3 Put serum blank in Dohrmann boat. Combust sample as described in section 9.0 and analyze sample according to method AMDT-M-2 for the ion selective electrode analysis. 8.3.2.4 For rabbit studies, the meter reading for a blank sample should be 0.03 ppm or lower before proceeding with the calibration. Bum samples until this limit is reached, or until in the judgement o f the operator the reading is stable with respect to historical readings (previous 48 hours). 8.3.2.5 For non-rabbit studies, the blank readings should reach a predetermined ion concentration before proceeding with the calibration. 8.3.2.6 It may be necessary to mix approximately 50 mg o f charcoal with the sample to aid combustion. C02' 8.3.3 Standard Curve 8.3.3.1 If beef serum is frozen, thaw at least enough to complete the standard curve analysis for the day (=30 mL). 8.3.3.2 Pipette lOOfiL of beef serum into Dohrmann sample boat. 8.3.3.3 Start with the lowest standard concentration. Using a Hamilton syringe, eject a fixed quantity of the standard on or in the matrix. For rabbit studies, use 4 uL o f standard and eject it on or in the beef serum. 8.3.3.4 At least 3 replicates should be used for the lowest standard concentration; more replicates may be used at the discretion o f the analyst. 8.3.3.5 Combust the sample as described in section 9.3 and analyze according to AMDT-M-2. 8.3.3.6 Run all 15 standards. If one replicate is significantly different from the other two replicates, run another sample for that standard. Indicate in data that the new replicate replaces the old replicate and that the new replicate will be used to calculate the regression curve. 5.3.3.7 When all standards have been run, calculate the r^. r^ must be at least 0.95. If it is not at least 0.95, consult with supervisor. 8.3.3.8 A new standard curve should be run when the combustion tube or sample matrix is changed. New standard curve may also be run at the discretion o f the analyst. 8.4 Storage Conditions for Standards 8.4.1 Storage requirements for standards are dependent on the individual standards used. Typically, standards are stored at room temperature in plastic screw top bottles. 8.4.2 New FC-95 standards should be prepared at least once a month. 9.0 P R O C E D U R E S____________________ __________________________________ 9.1 Typical Operating Conditions: 9.1.1 Combustion tube temperature = 950C. 9.1.2 Oxygen and Helium flow = 50 cc/minute. 9.1.3 Vaporization/Drying time = 240 seconds. 9.1.4 Bake time = 300 seconds. 9.2 Start Up Procedure: 9.2.1 If the program is not started, start the EOX program on the PC. 9.2.2 Open the SYSTEM SETUP window. 9.2.3 Put the furnace module and the cell in the READY mode. 9.2.4 Close the SYSTEM SETUP window. 9.2.5 When the oven has reached the READY temperature, run the CLEAN BOAT program found in the CELL CHECK menu. 9.2.6 See AMDT-EP-3 for details o f the Dohrmann software. 9.3 Sample Extraction Procedure: 9.3.1 Open the SAMPLE HATCH and pipette lOOpL o f sample into the BOAT. It may be necessary to mix approximately 50 mg of charcoal with the sample to aid combustion. If this is done, charcoal should also be mixed in while establishing the baseline and when generating the standard curve. 9.3.2 Close SAMPLE HATCH. 0273 4 4 9.3.3 Add appropriate volume of TISAB solution or 1:1 TISAB.Milli-QTM water mixture to a labeled sample collection vial. Typically 0.6 mL to 15 mL are used. For rabbit studies, use 1.0 or 2.0 mL o f 1:1 TISAB:Milli-QTM water mixture. 9.3.4 Place the vial so that the tip of the COMBUSTION TUBE is in the TISAB at least 0.25 inches. Gases released during pyrolysis must bubble through the TISAB. 9.3.5 Run the EOX-WATER program found in the RUN menu. 9.3.6 When the EOX program is finished, remove the collection vial from the combustion tube. 9.3.7 If undiluted TISAB was used to collect the sample, add an equal volume o f Milli-QTM water to the TISAB to make 1:1 TISAB:Milli-QTM. 9.3.8 Rinse the end o f the combustion tube with Milli-QTM water and wipe with a KIMWEPE to remove any TISAB remaining on the tube. 9.3.9 Open the sample hatch and remove any remaining ash from the boat. Ash can be removed with a cotton tipped applicator and/or vacuumed out. It may be necessary to scrap particles off the bottom with a spatula or other similar device. A drop o f Milli-QTM water may be added to the boat to aid in the Clean Cycle. 9.3.10 Close the hatch. 9.3.11 Run the CLEAN BOAT program. 9.3.12 Sample is ready for analysis by ion selective electrode (AMDT-M-2). 9.4 Sample Calculations 9.4.1 Use the standard curve to calculate the sample value. 9.4.2 Sample Mass Recovered F (ug) = (TISAB vol in mL) x /Orion reading in ppm - intercept^ (Slope) 10.0 V A L ID A T IO N ____________________ 10.1 Q uality Control 10.1.1 Daily S ta rt Up Check Samples: Once the standard curve is established, each day o f analysis is started by analyzing QC samples. The QC samples are to be the same as the lowest concentration spiked samples used to generate the standard curve. Each concentration must be done in triplicate unless the first two replicates are within 20% o f the standard curve, then a third replicate is not necessary. 10.2 Precision and Accuracy: See method development analysis and sample analysis in Fluoride Notebooks 2,3, and 5. Precision and accuracy varies when analyzing samples o f different matrices and different reference compounds. 10.3 O th er Validation Param eters: NA 11.0 D A TA AN ALYSIS___________________________________________ 11.1 C alculations 11.1.1 For the standard curve, use regression analysis in Excel, version 5.0 or greater. 11.1.2 To calculate the fluoride contraction in the sample, see method AMDT-M-2. C0272 11.2 Analyzing the Data 11.2.1 r2 must be at least 0.95 or greater. "Outliers" may be excluded if two o f the three replicates are within 20% o f each other and the outlier is greater than 200% o f the average o f those two or less than 50% of the average of those two. Any such outliers should be pointed out in the data and noted in the Final Report along with the reason it was considered an outlier. 12.0 A T T A C H M E N T S______________________________________________________ None 13,( REFERENCE S_____________________________________________ 13.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer Operator's Manual (Manual 915349, revision B, December 1993) 13.2 AMDT-M-2 Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer 13.3 AMDT-EP-3 Routine Maintenance of a Modified Dohrmann DX2000 Organic Halide Analyzer 14.0 R E V IS IO N S_______________ ;___________________________________________ Revision Number Reason for Change Revision Date C 0Z 73y 9.1J Amendment to analytical protocol AMDT-011095.1 00273J Attachment I GLP Study Protocol Amendment Study Number: A M DT-011095.1 Study Title: Single Dose Dermal Absoption/Toxicity o f T-60676, T-6068 and T-6069 in Rabbits Study Director: James D. Johnson Amendment Date: November 10, 1995 Amendment Number: 1 This amendment modifies the following portion o f the protocol: AM DT-M -14-0 specifies using bovine serum for blanks and QC check samples in the thermal extraction. However, rabbit senim from the control animals (AM DT-110394.1) was used was used because the bovine serum blanks were higher than the samples. Approved by: Date 00273 9.3 Q uality Assurance Unit Statem ent Attachment D GLP Study Quality Assurance Statement ypflgfijd & Sigfo Piaster ' v \ Xopfeatt QAUKte* 1 Study Title: Single-dose Dermal A bsorption/Toxicity Study of T-6067, T-6068 and T-6069 in R abbits Study N um ber AMDT-011095.1 Name of Auditor: Kari Rambo This study has been inspected by the Quality Assurance Unit as indicated in the following table. The findings were reported to the study director and management. Inspection Dates From Is ______ Phase______________ 11-15-95 11-20-95 Final Report Date Inspection Reported to Management Study Director 11-20-95 11-20-95 / QAU Auditor //-2c j r Date 9.4 K ey Personnel Involved in the Study C -Q 2 7 3 jr \7x7Tj^*9m 3M Environm ental Laboratory Key Personnel Thermal extraction followed by analysis using Orion ion analyzer: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Analysis o f liver extracts using electrospray mass spectrometry: Jim Johnson Dave Christenson Thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Docum entation and Reporting: Jim Johnson Rich Youngblom Q uality Assurance Unit: Gale Van Buskirk Cynthia Weber Kari Rambo C0272J 9.11 Data C027 9.11.1 Summary and raw data; ug F* in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. 00274 Summary of Combustion Data - Liver AMDT-011095.1, HWI 6329-152 As Referenced in Final Report section 6.0 DATA ANALYSIS Total ug Fluoride in Whole Liver Mean per Dose Group* pg Std. Dev. Control Group 12.5 + 2.1 4.0 mg/kg dose (T6067) 12.5 + 1.6 16 mg/kg dose (T6067) 14.7 + 4.0 40 mg/kg dose (T6067) 16.5 + 5.2 Fabric exposure (T6068)** 15.7 + 4.2 Fabric exposure (T6069)** 12.8 + 3.7 Calculated as the mean of triplicate samples from each of three male and three female rabbits. ^Administered as a 10 cm x 10 cm piece of test material (fabric). 00274* RPT152L.XLS L13492 AB ID liver blank-1 liver spk-1 liver spk-2 liver spk-3 liver spk-4 liver spk-5 liver spk-6 liver blank-2 F52711-1 F52711-2 F52711-3 F52719-1 F52719-2 F52719-3 Liver blk 1 Liver blk 2 Liver spk 1 Liver spk 2 F52725-1 F52725-2 F52725-3 F52713-1 F52713-2 F52713-3 F52718-1 F52718-2 F52718-3 F52723-1 F52723-2 F52723-3 F52721-1 F52721-2 F52721-3 F52727-1 F52727-2 F52727-3 F52806-1 F52806-2 F52806-3 F52715-1 F52715-2 F52715-3 F52738-1 F52738-2 F52738-3 % rcvry 97% 85% 85% 107% 97% 95% 97% 98% Actual ppm Fin liver (W/W) 0.213 1.10 0.872 0.883 2.63 2.00 2.55 0.159 0.167 0.144 0.149 0.166 0.157 0.205 0.250 0.169 1.19 1.37 0.228 0.231 0.230 0.172 0.152 0.170 0.161 0.156 0.233 0.175 0.230 0.155 0.218 0.191 0.142 0.194 0.206 0.160 0.158 0.155 0.166 0.226 0.255 0.211 0.178 0.169 0.166 Average ppm Fin liver (WAV) 0.153 0.176 0.230 0.164 0.183 0.187 0.184 0.187 0.160 0.230 0.171 Liver burned (grams) 0.125 0.134 0.147 0.147 0.123 0.147 0.112 0.140 0.134 0.141 0.149 0.137 0.124 0.133 0.129 0.142 0.124 0.109 0.126 0.116 0.124 0.154 0.166 0.163 0.128 0.135 0.126 0.171 0.150 0.156 0.101 0.116 0.154 0.153 0.136 0.134 0.134 0.137 0.125 0.127 0.133 0.122 0.134 0.126 0.131 Whole liver weight (grams) 66.1 66.1 66.1 76.5 76.5 76.5 69.5 69.5 69.5 71.4 71.4 71.4 57.7 57.7 57.7 72.6 72.6 72.6 62.9 62.9 62.9 69.2 69.2 69.2 69.8 69.8 69.8 62.1 62.1 62.1 64.6 64.6 64.6 Total Fin whole liver (pg) 10.1 13.5 16.0 11.7 10.6 13.6 11.6 12.9 11.1 14.3 11.0 Dosage (mg/kg) 0.0 0.0 0.0 4.0 4.0 4.0 0.0 0.0 0.0 4.0 4.0 Page 1 002743, RPT152L.XLS L13492 AB ID F52809-1 F52809-2 F52809-3 F52724-1 F52724-2 F52724-3 F52737-1 F52737-2 F52737-3 liver bink-1 liverspk-1 liverspk-2 liverspk-3 F52801-1 F52801-2 F52801-3 F52720-1 F52720-2 F52720-3 F52733-1 F52733-2 F52733-3 F52788-1 F52788-2 F52788-3 Liver blk 1 Liver blk 2 Liver spk 1 Liver spk 2 F52730-1 F52730-2 F52730-3 F52731-1 F52731-2 F52731-3 F52802-1 F52802-2 F52802-3 F52714-2 F52714-1 F52714-3 F52734-1 F52734-2 F52734-3 F52774-1 F52774-2 F52774-3 % rcvry 89% 86% 91% 111% 108% Actual ppm Fin liver (W/W) 0.432 0.159 0.168 0.187 0.235 0.192 0.163 0.145 0.155 0.344 1.10 0.973 0.924 0.372 0.278 0.161 0.252 0.206 0.225 0.224 0.182 0.208 0.179 0.154 0.165 0.760 0.301 1.06 1.02 0.339 0.237 0.380 0.163 0.202 0.160 0.286 0.542 0.244 0.247 0.454 0.281 0.223 0.297 0.183 0.287 0.215 0.174 Average ppm Fin liver (W/W) 0.253 0.204 0.154 0.271 0.228 0.205 0.166 0.318 0.175 0.357 0.327 0.234 0.226 Liver burned (grams) 0.138 0.128 0.112 0.119 0.139 0.151 0.129 0.147 0.142 0.134 0.123 0.134 0.149 0.112 0.119 0.148 0.128 0.133 0.130 0.123 0.139 0.139 0.143 0.146 0.121 0.140 0.116 0.160 0.160 0.123 0.113 0.159 0.158 0.108 0.158 0.150 0.110 0.122 0.154 0.152 0.151 0.111 0.146 0.143 0.138 0.122 0.137 Whole liver weight (grams) 55.4 55.4 55.4 82.0 82.0 82.0 62.5 62.5 62.5 76.7 76.7 76.7 68.7 68.7 68.7 67.7 67.7 67.7 68.0 68.0 68.0 57.9 57.9 57.9 59.0 59.0 59.0 71.9 71.9 71.9 45.5 45.5 45.5 58.1 58.1 58.1 71.1 71.1 71.1 Total Fin whole liver (pg) 14.0 16.8 9.6 20.7 15.7 13.9 11.3 18.4 10.3 25.7 14.9 13.6 16.0 Dosage (mg/kg) 4.0 16.0 16.0 16.0 16.0 16.0 16.0 40.0 40.0 40.0 40.0 40.0 40.0 Page 2 RPT152L.XLS L13492 AB ID F52712-1 F52712-2 F52712-3 F52729-1 F52729-2 F52729-3 F52803-1 F52803-2 F52803-3 F52716-1 F52716-2 F52716-3 F52728-1 F52728-2 F52728-3 F52787-1 F52787-2 F52787-3 Liver blk 1 Liver blk 2 Liver stnd 1 Liver stnd 2 52717-1 52717-2 52717-3 52735-1 52735-2 52735-3 52736-1 52736-2 52736-3 52726-1 52726-2 52726-3 52732-1 52732-2 52732-3 52739-1 52739-2 52739-3 Liver spk-3 Liver spk-4 Liver spk-5 Liver spk-6 Liver spk-7 Liver spk-8 Liver spk-9 % rcvry 87% 88% 0.69 0.8222 0.8295 1.039 0.769 0.8718 0.9319 Actual ppm Fin liver (W/W) 0.442 0.336 0.210 0.291 0.185 0.184 0.294 0.193 0.148 0.179 0.165 0.177 0.192 0.166 0.215 0.177 0.242 0.190 0.255 0.250 1.14 1.13 0.360 0.202 0.265 0.206 0.203 0.191 0.158 0.168 0.155 0.135 0.143 0.173 0.162 0.148 0.163 0.313 0.127 0.133 0.923 0.888 1.11 1.23 2.29 1.82 2.35 Average ppm Fin liver (W/W) 0.330 0.220 0.212 0.173 0.191 0.203 0.276 0.200 0.160 0.150 0.158 0.191 ' Liver burned (grams) 0.115 0.134 0.111 0.127 0.135 0.132 0.118 0.115 0.140 0.117 0.137 0.121 0.125 0.126 0.106 0.141 0.110 0.128 0.147 0.122 0.115 0.118 0.119 0.121 0.122 0.137 0.129 0.111 0.160 0.117 0.145 0.152 0.165 0.113 0.156 0.130 0.113 0.127 0.162 0.160 0.113 0.140 0.113 0.128 0.102 0.145 0.120 Whole liver weight (grams) 72.0 72.0 72.0 63.1 63.1 63.1 71.1 71.1 71.1 67.3 67,3 67.3 79.9 79.9 79.9 71.2 71.2 71.2 73.0 73.0 73.0 53.9 53.9 53.9 72.9 72.9 72.9 74.4 74.4 74.4 66.1 66.1 66.1 64.9 64.9 64.9 Total Fin whole liver (pg) 23.7 13.9 15.1 11.7 15.3 14.5 20.1 10.8 11.7 11.2 10.4 12.4 Dosage (mg/kg) Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Fabric Page 3 C0274$ 9.11.2 Summaiy and raw data; analysis of liver extracts using electrospray mass spectrometry. This data, although supportive, in the opinion o f the Study Director is not required to reach the conclusion stated in Final Report Section 6.0, and therefore is not discussed in detail. C -Q 27^r HWI# 6329-152 A Study: Protocol Numbor: Tost Material: Matrix: R Squared Value: Response Factor Amount: Analyst: Date Analyzed: Method: Instrument: LABBASE FILE Single Dose Dermal Absorption TP3016.AB T-6067, T-6068. and T-6069 In Rabbits (L13492) Liver Screening N/A DLC 4/25/95 Fisons VG 2000 Electrospray MS 042595D A-l y w CctteWiAS pe.CC-5 4-I A -4 /? " /<'5 / 1 5 F O' Lui aC MOT C ir c -e .W W .v P u. -Vo i-orUooid efr tJO CtwSWi4V e. 'oc.cn Ai L AA L4S 'AQvo vfccEcrriiW Uvsr.i't-. A" `&LC- lVt5/<l5 Group Do m Group 1: Distilled Water 2.0 mL/kg Group 2: 4 mg/kg T-6067 Group 3: 16 mg/kg T-6067 Group 4: 40 mg/kg T-6067 Group 5: Fabric T-6068 Sam ple# F52725 F52711 Ion Count Area * Extracted wt g Dilution factor N/D N/D Concentration pg/g ** Total mass of liver 9 Total amount of FC-95 per liver mg N/D N/D N/D N/D F52715 F52718 F52723 N/D N/D N/D N/D N/D N/D N/D N/D N/D F52724 F52637 F52788 F52733 N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D F52774 F52731 F52734 F52714 F52802 F52728 F52729 F52716 F52712 N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/D N/0 N/D N/D N/D Group 6: Fabric T-6068 F52739 F52736 F52726 N/D N/D N/D N/D N/D N/D N/D N/D N/D \\l(6iy>` SIR of M 412 + 413 ** The concentration was calculated by using the standard curve and multiplying the result by 4/5. The 4/5 factor Is the result of a miscalculation In applying formula 8.4 In Method AMDT-M-4-0. 137 mg of liver was used In this calculation rather than 171 mg. The concentrations In the standard curve are therefore 5/4 larger than they should be. By multiplying the calculated concentration in the standard curve by 4/5, the correct result is obtained. CO2 7 4 0 ^ HWI # 6329-151 & 152; CMPD L13492 j 042595D Sm (Mn, 2x3); Sm (Mn, 2x3) 100 i% SIR of 4 Channels ES427.00 6.92e4 Area 2982 O v -.r t Scan 3000 C0274g 9.11.3 Summary and raw data; ppm F' in serum as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. C-02750 mJ ' I Z81*0 RE: 6329-152 SERUM SAMPLES AMDT 11095.1 Date of Analysis: 8/22, 8/23, and 8/28/95 Analyst: DDW The samples are burned in the Dohrman at 950 C using 0.10 mL of the serum. The gas is collected in 2.0 mL of 1:1 TISAB/Milli-Q water. The samples are then analyzed on a Skalar Segmented Flow Analyzer using the Ion Specific Electrode (ISE) Method. TISAB buffer is added to each sample as it proceeds through the system. The sample then goes through a heated mixing coil before the potential between the ion selective electrode and the reference electrode is measured. The signal is amplified and related to the fluoride concentration. The instrument was calibrated in the ranges of 0.015 - 0.15 ppm and 0 . 1 5 - 1 . 5 0 ppm fluoride. The standard curve for the high range was plotted using the inverse logarithm option. The standard curve for the low range is linear. All standards and samples were then calculated by the Skalar software using these curves. All results below 0.0001 ppm appear on the raw data as #.####. A quality control standard was analyzed every 10 samples to check for accuracy and drift. Raw data is taken from the appropriate calibrated range of the Skalar printout and summarized on an Excel spreadsheet. The final results are adjusted for the collection volume and any subsequent dilutions. CQ275J. SUMMARY OF 6329-152 SERUM SAMPLES AMDT 11095.1 U tjiJ *14*1*15 GROUP 1 Dose Level : 0 mg/kg FJ2711-1 0.84 0.42 0.48 0.45 F52719-1 0.68 0.34 0.73 ND F52725-1 0.66 0.31 0.96* 0.78 F5272I-1 0.51 ND 0.84* 0.63 F52727-1 0.48 0.60 0.82* 0.53 F52806-1 0.53 0.62 0.92 0.49 GROUP 2 Dose Level : 4 mg/kg F52713-1 1.36 0.49 0.64 0.31 F52718 1.16 0.42 0.99 0.33 F52723-1 0.93 0.52 1.25 0.32 F52715-1 1.00 0.54 0.69 0.49 F52738-1 1.17 0.43 1.40 0.38 F52809-1 1.49 0.61 0.72 0.33 GROUP 3 Dose Level : 16 mg/kg F52724-1 1.21 0.52 0.64 0.74 F52737-1 1.17 0.48 0.64 0.72 F52801-1 1.05 0.92 1.20 1.29 F52720-2 0.73 0.92 0.92 F52733-1 1.11 0.64 0.70 0.54 F52788-1 0.32 0.74 0.50 ND GROUP 4 Dose Level : 40 mg/kg F52730-1 1.04 0.59 0.39 F52731-1 0.95 0.03 ' 0.39 F52802-1 1.11 0.68 0.59 F52714-1 0.87 0.85 0.93 F52734-1 1.07 0.69 1.35 F52774-1 1.23 0.72 0.92 F52712-1 1.28 0.95 0.69 F52729-1 1.08* 0.62 0.67 GROUP 5 F52803-1 1.31 0.63 0.92 Dose Level : 10 x 10 fabric F52716-1 1.38 0.65 0.77 F52728-1 1.44 0.77 0.68 F52787-1 1.18 0.68 0.76 GROUP 6 Dose Level : 10 x 10 fabric F52717-1 F52735-1 F52736-1 F52726-1 F52732-1 F52739-1 1.71 1.48 1.16 0.94 1.33 0.90 0.62 0.74 0.57 0.79* 0.69 0.58 1.03 0.67 0.60* 0.54 0.42 0.33 * Average of duplicate analysis taken for final result. DDW BEH COPifAVAIUlijjj; 152S-SUM.XLS C 0 2 7 5 jL t Page 1 SERUM CURVE 2 7-27-95 CARLTON s u U_ HuCT u 27.9 -IS 2 _ ,/W b T J l O ^ S . 1 Sample ID Spk 34-1 Spk 34-2 Spk 34-3 Spk 40-1 Spk 40-2 Spk 40-3 Spk 62-1 Spk 62-2 Spk 62-3 Spk 100-1 Spk 100-2 Spk 100-3 Spk 124-1 Spk 124-2 Spk 124-3 Spk 250-1 Spk 250-2 Spk 250-3 Spk 500-1 Spk 500-2 Spk 500-3 Skalar Result (ppm) DLTISAB mL FC 95 Cone final voi Solution FC 95 Soln (mL) Spiked (ppm) Mass Spiked (ug F-) Average Mass Recovered ("g F-) % Recovery 0.07 2.0 0.004 34.00 0.06 2.0 0.004 34.00 0.08 0.12 151% 0.06 2.0 0.004 34.00 0.06 2.0 0.004 40.00 0.06 2.0 0.004 40.00 0.10 0.13 132% 0.07 2.0 0.004 40.00 0.08 2.0 0.004 62.00 0.08 2.0 0.004 62.00 0.15 0.17 114% 0.09 2.0 0.004 62.00 0.10 2.0 0.004 100.0 0.11 2.0 0.004 100.0 0.24 0.22 93% 0.12 2.0 0.004 100.0 0.12 2.0 0.004 124.0 0.14 2.0 0.004 124.0 0.30 0.28 94% 0.15 2.0 . 0.004 124.0 0.33 2.0 0.004 250.0 0.23 2.0 0.004 250.0 0.60 0.52 87% 0.23 2.0 0.004 250.0 0.47 2.0 0.004 500.0 0.41 2.0 0.004 '500.0 1.20 0.92 77% 0.50 2.0 0.004 500.0 SERUM CURVE 1 (CARLTON) 7-27-95 MASS SPIKED (ug) SERCRV2C.RPT 0 0 2 7 5 ,5 Paga 1 3 SERUM CURVE 2 7-27-95 CARLTON Sample ID Spk 34-1 Spk 34-2 Spk 34-3 Spk 40-1 Spk 40-2 Spk 40-3 Spk 62-1 Spk 62-2 Spk 62-3 Spk 100-1 Spk 100-2 Spk 100-3 Spk 124-1 Spk 124-2 Spk 124-3 Spk 250-1 Spk 250-2 Spk 250-3 Spk 500-1 Spk 500-2 Spk 500-3 Skalar Result (ppm) DLTISAB mL FC 95 Cone final voi Solution FC 95 Soin (mL) Spiked (ppm) Mass Spiked (us F-) Mass % Recovered Recovery (ug F-) 0.07 2.0 0.004 34.00 0.08 0.14 171% STANDARD DEVIATION : 0.1787 0.06 2.0 0.004 34.00 0.08 0.11 138% % RSD : 11.8165 0.06 2.0 0.004 34.00 0.08 0.12 145% 0.06 2.0 0.004 40.00 0.10 0.12 123% STANDARD DEVIATION : 0.1072 0.06 2.0 0.004 40.00 0.10 0.12 128% % RSD : 8.1286 0.07 2.0 0.004 40.00 0.10 0.14 144% 0.08 2.0 0.004 62.00 0.15 0.16 110% STANDARD DEVIATION : 0.1072 0.08 2.0 0.004 62.00 0.15 0.16 106% % RSD : 9.4074 0.09 2.0 0.004 62.00 0.15 0.19 126% 0.10 2.0 0.004 100.0 0.24 0.20 85% STANDARD DEVIATION : 0.0666 0.11 2.0 0.004 100.0 0.24 0.23 96% % RSD : 7.1794 0.12 2.0 0.004 100.0 0.24 0.23 98% 0.12 2.0 0.004 124.0 0.30 0.25 84% STANDARD DEVIATION : 0.0975 0.14 2.0 0.004 124.0 0.30 0.29 97% % RSD : 10.3277 0.15 2.0 0.004 124.0 0.30 0.31 103% 0.33 2.0 0.004 250.0 0.60 0.66 109% STANDARD DEVIATION : 0.1942 0.23 2.0 0.004 250.0 0.60 0.45 76% % RSD : 22.3705 0.23 2.0 0.004 250.0 0.60 0.45 76% 0.47 2.0 0.004 500.0 1.20 0.93 77% STANDARD DEVIATION : 0.0795 0.41 2.0 0.004 500.0 1.20 0.82 68% % RSD : 10.3961 0.50 2.0 0.004 500.0 1.20 ' 1.01 84% SERUM CURVE 1 (CARLTON) 7/27/95 y = 0.7212X + 0.063 R2= 0.967 SERCRV2C.SUM e0275ff Page 1 _ va fs tz o o 152S-A.XLS 1995-08-22 13:41 OutPut of : 950822B1 Operator : DDW Date of the Analysis : 1995-08-22 09:04 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950822B1 Sample Sample Skalar Standard (PP*p ) Skalar Result % DLTISAB Qty Samp! Recovery' final voi ( m L ) (ntl) , , 1 Tracer 1.50 2 Drift 1.50 3 Wash 4 Standard 1 0.015 5 Standard 2 0.03 6 Standard 3 0.06 7 Standard 4 0.09 8 Standard 5 0.12 9 Standard 6 0.15 10 Standard 7 0.30 11 Standard 8 0.60 12 Standard 9 1.20 13 Standard 10 1.50 14 Drift 1.50 15 Wash 16 SERUM BLK 1 17 SERUM BLK 2 18 SPK 62-1 19 SPK 62-2 20 SPK 62-3 21 SPK 250-1 22 SPK 250-2 23 SPK 250-3 24 BLK 25 BLK 26 Drift 1.50 27 Wash 28 F52713-1 29 F52718-29 30 F52723-1 1.46 1.47 ND 0.015 0.03 0.06 0.09 0.12 0.15 0.28 0.62 1.23 1.47 1.48 ND 0.07 0.04 0.07 0.11 0.12 0.12 0.27 0.27 0.09 0.07 1.46 ND 0.07 0.06 0.05 98% 98% 103% 94% 105% 100% 98% 101% 93% 103% 103% 98% 99% 97% 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 1 Actual mi FC95 Solution Cone Mass Mass % ^ 5 ReH*red Hocovcry >.O>aVa C'wa oo 1.48 0.82 1.46 0.004 62 00 0.15 2.18 0.004 62.00 0.15 2.49 0.004 62.00 0.15 2.49 0.004 250.0 0.60 5.44 0.004 250.0 0.60 5.34 0.004 250.0 0.60 1.71 1.35 0.15 98% 0.22 146% 0.25 167% 0.25 42% 0.54 91% 0.53 89% 1.36 1.16 0.93 Sample M 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 Sample IP F52715-1 F52738-1 F52809-1 F52724-1 F52737-1 F52801-1 F52729-1 Drift Wash SPK62-1 SPK62-2 SPK 250-1 SPK 250-2 BLK BLKC SPK62-1 SPK62-2 SPK 250-1 SPK 250-2 Drift Wash SPK 250-3 SPK 250-4 BLK BLK F52733-1 F52788-1 F52730-1 F52731-1 F52802-1 F52714-1 Drift Wash F52734-1 F52774-1 F52712-1 Skalar Standard ippiB) , Skalar Resoli fcpm> 152S-A.XLS % PI TISAB Qty Sampt Reewexy finalvoi (m U ^ ....z. Actual HJLFC95 Cone Mass Mass % SSIr sw , wovered Rjwowwy i F) 0.05 2.0 0.06 2.0 0.07 2.0 0.06 2.0 0.06 2.0 0.05 2.0 0.04 2.0 1.50 1.46 98% ND 0.07 2.0 0.12 2.0 0.17 2.0 0.26 2.0 0.06 2.0 0.04 2.0 0.08 2.0 0.08 2.0 0.20 20 0.22 2.0 1.50 1.48 99% ND 0.24 2.0 0.24 2.0 0.05 2.0 0.04 2,0 0.06 2.0 0.02 2.0 0.05 2.0 0.05 2.0 0.06 2.0 0.04 2.0 1.50 1.46 97% ND 0.05 2.0 0.06 2.0 0.06 2.0 0.10 1.00 0.10 1.17 0.10 1.49 0.10 1.21 0.10 1.17 0.10 1.05 0.10 0.81 0.10 1.43 0.004 62.00 0.15 0.10 2.42 0.004 62.00 0.15 0.10 3.32 0.004 250.0 0.60 0.10 5.14 0.004 250.0 0.60 0.10 1.16 0.10 0.88 0.10 1.59 0.004 62.00 0.15 0.10 1.66 0.004 62.00 0.15 0.10 4.06 0.004 250.0 0.60 0.10 4.38 0.004 250.0 0.60 0.10 4.86 0.004 250.0 0.60 0.10 4.76 0.004 250.0 0.60 0.10 1.03 0.10 0.72 0.10 1.11 0.10 0.32 0.10 1.04 0.10 0.95 0.10 1.11 0.10 0.87 0.10 1.07 0.10 1.23 0.10 1.28 0.14 96% 0.24 163% 0.33 55% 0.51 86% 0.16 107% 0.17 111% 0.41 68% 0.44 73% 0.49 81% 0.48 79% Page 2 BEST COPY AVMUBLE Sample 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 Sample Skalar Standard tppra) F52729-1 SPK 62-1 SPK 62-2 SPK 62-3 SPK 250-1 BLK F52803-1 Drift Wash F52716-1 F52728-1 F52787-1 F52717-1 F52735-1 F52736-1 F52726-1 F52732-1 F52739-1 SPK 62-1 Drift Wash SPK 62-2 SPK 250-1 SPK 250-2 Drift Wash 1.50 1.50 1.50 Skalar ftesmli (gpOi) 152S-A.XLS % DI TISAB Qtv Sanrni Hecovmy final vq irctlA i l 0.07 2.0 0.08 2.0 0.10 2 0 0.14 2.0 0.26 2.0 0.17 2.0 0.07 2.0 1.48 98% ND 0.07 2.0 0.07 2.0 0.06 2.0 0.09 2.0 0.07 2.0 0.06 2.0 0.05 2.0 0.07 2.0 0.05 2.0 0.08 2.0 1.50 100% ND 0.13 2.0 0.34 2.0 0.36 2.0 1.51 101% ND 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 3 BEST COPYm\l ABU fi Actual mU FC 95 Cone Mass Mass p p m * SoM on PC P i Soli* ''S t 'vjwyjY/TZ&yMWttslwsii Recovered M m E lk 1.35 1.57 0.004 62.00 0.15 1.96 0.004 62.00 0.15 2.71 0.004 62.00 0.15 5.10 0.004 250.0 0.60 3.40 1.31 0.16 0.20 0.27 0.51 % 105% 132% 182% 85% 1.38 1.44 1.18 1.71 1.48 1.16 0.94 1.33 0.90 1.68 0.004 62.00 0.15 0.17 113% 2.66 0.004 62.00 0 15 6.72 0.004 250.0 0.60 7.28 0.004 250.0 0.60 0.27 178% 0.67 112% 0.73 121% 152S-B.XLS 1995-08-23 11:10 OutPut of : 950822C1 Operator : DDW Date of the Analysis : 1995-08-22 13:41 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950822C1 impie # VAV fY fW /Y S///tV ySi Sample ID Skalar Skalar % DJ.TI5AB Qty Sampi Standard R<sH Recovery final voi (mL> (ppm) j J H f c d , , <W . -J 1 Tracer 1.50 2 Drift 1.50 3 Wash 4 Standard 1 0.015 5 Standard 2 0.03 6 Standard 3 0.06 7 Standard 4 0.09 8 Standard 5 0.12 9 Standard 6 0.15 10 Standard 7 0.30 11 Standard 8 0.60 12 Standard 9 1.20 13 Standard 10 1.50 14 Drift 1.50 15 Wash 16 SERUM BLK 1 17 SERUM BLK 2 18 SPK62-1 19 SPK 62-2 20 SPK 62-3 21 SPK 250-1 22 SPK 250-2 23 SPK 250-3 24 BLK 25 BLK 26 Drift 1.50 27 Wash 28 F52711-1 29 F52719-1 30 F52725-1 1.48 1.49 ND 0.015 0.03 0.06 0.09 0.12 0.15 0.28 0.61 1.24 1.46 1.49 ND 0.05 0.05 0.12 0.09 0.11 0.19 0.30 0.28 0.08 0.06 1.45 ND 0.04 0.03 0.03 98% 99% 103% 94% 106% 98% 99% 101% 94% 102% 103% 97% 99% 97% 4 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 1 es^so o Acftial mkFC93 Cooc Mass Mass % pn tS TS --O 1.06 0.99 2.48 0.004 62.00 0.15 1.77 0.004 62.00 0.15 2.24 0.004 62.00 0.15 3.74 0.004 250.0 0.60 5.98 0.004 250.0 0.60 5.50 0.004 250.0 0.60 1.65 1.11 0.25 167% 0.18 119% 0.22 150% 0.37 62% 0.60 100% 0.55 92% 0.84 0.68 0.66 V- Sample Sample / ~,W',ZI7DZ'~ 152S-B.XLS Skalar Skalar % DI TISAB Standard insali Pecovery finalvol t- {PP10) / t 31 F52721-1 0.03 2.0 0.10 32 F52727-1 0.02 2.0 0.10 33 F52806-1 0.03 2.0 0.10 34 F52711-2 0.02 2.0 0.10 35 F52719-2 0.02 2.0 0.10 36 F52725-2 0.02 2.0 0.10 37 F52721-2 ND 2.0 0.10 38 Drift 1.50 1.49 99% 39 Wash ND 40 SPK62-1 0.11 2.0 0.10 41 SPK250-1 0.31 2.0 0.10 42 BLK 0.06 2.0 0.10 43 BLK 0.05 2.0 0.10 44 SPK62-1 0.07 2.0 0.10 45 SPK62-2 0.09 2.0 0.10 46 SPK62-3 0.09 2.0 0.10 47 SPK62-4 0.10 2.0 0.10 48 SPK250-1 0.18 2.0 0.10 49 SPK250-2 0.29 2.0 0.10 50 Drift 1.50 1.45 97% 51 Wash ND 52 SPK250-3 0.27 2.0 0.10 53 BLK 0.06 2.0 0.10 54 BLK 0.04 2.0 0.10 55 F52727-2 0.03 2.0 0.10 56 F52806-2 0.03 2.0 0.10 57 F52713-2 0.02 2.0 0.10 58 F52718-2 0.02 2.0 0.10 59 F52723-2 0.03 2.0 0.10 60 F52715-2 0.03 2.0 0.10 61 F52738-2 0.02 2.0 0.10 62 Drift 1.50 1.53 102% 63 Wash ND 64 F52809-2 0.03 2.0 0.10 65 F52724-2 0.03 2.0 0.10 66 F52737-2 0.02 2.0 0.10 002759 Page 2 0.51 0.48 0.53 0.42 0.34 0.31 ND 2.22 0.004 62.00 0.15 6.26 0.004 250.0 0.60 1.16 0.92 1.34 0.004 62.00 0.15 1.88 0.004 62.00 0.15 1.76 0.004 62.00 0.15 2.09 0.004 62.00 0.15 3.58 0.004 250.0 0.60 5.88 0.004 250.0 0.60 5.46 0.004 250.0 0.60 1.11 0.73 0.60 0.62 0.49 0.42 0.52 0.54 0.43 0.22 149% 0.63 104% 0.13 90% 0.19 126% 0.18 118% 0 21 141% 0.36 60% 0.59 98% 0.55 91% 0.61 0.52 0.48 m GO CO -o JS- Sample u 67 68 69 70 71 72 Sample SPK 62-1 SPK 62-2 SPK 250-1 SPK 250-2 Drift Wash Skalar Standard (ppm) 152S-B.XLS Skalar Resoli ,% m m m Q V 'S m p b - <fcpm> //m.'/Z 0.05 2.0 0.07 2.0 0.14 2.0 0.24 2.0 1.50 1.51 101% ND 0.10 0.10 0.10 0.10 r> ro O mo Page 3 Actual mtFCfSj Cone Mass Mass % Solution FC?5SoMt fteooveroti Hwovety tmmstiiW&m gaa&l Ci 1.03 0.004 62.00 0.15 1.39 0.004 62.00 0.15 2.88 0.004 250.0 0.60 4.70 0.004 250.0 0.60 0.10 69% 0.14 94% 0.29 48% 0.47 78% BEST COPY AVAIlABLE 152S-C.XLS 1995-08-23 11:00 OutPut of : 950823A1 Operator : DDW Date of the Analysis : 1995-08-23 06:55 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950823A1 Sample Sample Skalar Standard Skalar % DI.TISAB Qtj Sampl Result Recovery final voi (ro L ) (ppm) / / < / ,///././////'/i//Y////. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 o N 27 28 29 & 30 Tracer Drift Wash Standard 1 Standard 2 Standard 3 Standard 4 Standard 5 Standard 6 Standard 7 Standard 8 Standard 9 Standard 10 Drift Wash SERUM BLK SERUM BLK SPK 62-1 SPK 62-2 SPK 62-3 SPK 250-1 SPK 250-2 SPK 250-3 SPK 250-4 SPK 250-5 Drift Wash SPK 250-6 BLK BLK 1.50 1.50 0.015 0.03 0.06 0.09 0.12 0.15 0.30 0.60 1.20 1.50 1.50 1.50 1.45 1.47 ND 0.018 0.03 0.06 0.09 0.12 0.15 0.28 0.62 1.23 1.47 1.51 ND 0.06 0.05 0.08 0.10 0.10 0.14 0.21 0.23 0.23 0.26 1.51 ND 0.39 0.07 0.03 97% 98% 117% 89% 101% 102% 99% 100% 93% 103% 103% 98% 101% 100% * 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 1 A<s#al mi.FC^S Cow Mass Mass SoJwtwn FC95Sotn Spited jfe . MM ito -.'i % BEST COPY AVAIIABLE 1.15 0.94 1.51 0.004 62.00 0.15 2.01 0.004 62.00 0.15 2.02 0.004 62.00 0.15 2.74 0.004 250.0 0.60 4.16 0.004 250.0 0.60 4.50 0.004 250.0 0.60 4.54 0.004 250.0 0.60 5.18 0.004 250.0 0.60 0.15 101% 0.20 135% 0.20 136% 0.27 46% 0.42 69% 0.45 75% 0.45 76% 0.52 86% 7.74 0.004 250.0 0.60 1.30 0.64 0.77 129% j i-0 o Sample # 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 r > 61 62 63 0 64 & 65 f%v D*1 66 Sample IP F52801-2 F52720-2 F52733-2 F52788-2 F52730-2 F52731-2 F52802-2 Drift Wash F52714-2 F52734-2 F52774-2 SPK62-1 SPK 250-1 SPK 250-2 BLK BLK SPK 62-1 SPK 62-2 Drift Wash SPK 62-3 SPK 250-1 SPK 250-2 SPK 250-3 SPK 250-4 BLK BLK F52712-2 F52729-2 F52803-2 Drift Wash F52716-2 F52728-2 F52787-2 Skalar Standard (ppm> 1.50 1.50 1.50 Skalar Reseli fce*5 152S-C.XLS % DITJSAB QtySampl Rcwvety final voi / Mtl- 'V, 4 I I I Z *f/f m m wm . 0.05 2.0 0.04 2.0 0.03 2.0 0.04 2.0 0.03 2.0 0.00 2.0 0.03 2.0 1.51 100% ND 0.04 2.0 0.03 2.0 0.04 2.0 0.10 2.0 0.15 2.0 0.17 2.0 0.05 2.0 0,05 2.0 0.05 2.0 0.08 2.0 1.53 102% ND 0.12 2.0 0.13 2.0 0.22 2.0 0.29 2.0 0.25 2.0 0.07 2.0 0.06 2.0 0.05 2.0 0.03 2.0 0.03 2.0 1.52 101% ND 0.03 2.0 0.04 2.0 0.03 2.0 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 2 0.92 0.73 0.64 0.74 0.59 0.03 0.68 0.85 0.69 0.72 1.92 2.92 3.46 1.03 1.03 1.03 1.64 2.46 2.55 4.36 5.82 5.06 1.30 1.13 0.95 0.62 0.63 0.65 0.77 0.68 I tab FC 95 Com % - Solmion FC95 Solo Sp>kcd Rwiwwt Rocovcfj 0.004 0.004 0.004 62.00 250.0 250.0 0.15 0.60 0.60 0.004 0.004 62.00 62.00 0.15 0.15 0.004 0.004 0.004 0.004 0.004 62.00 250.0 250.0 250.0 250.0 0.15 0.60 0.60 0.60 0.60 0.19 129% 0.29 49% 0.35 58% 0.10 69% 0.16 110% 0.25 165% 026 43% 0.44 73% 0.58 97% 0.51 84% Sample * 67 68 69 70 71 72 73 74 Sample w / ID F52717-2 SPK 62-1 SPK 62-2 SPK 250-1 SPK 250-2 SPK 250-3 Drift Wash Scalar Standard ppm) 1.50 Skalar ResvU 0.03 0.05 0.09 0.10 0.24 0.18 1.54 ND 152S-C.XLS % Dl TISABQtySampl #8W |; W *4fk', VMif. A m ai mitPC95 Cone Mass Mass % FC 95 Sol Spiked Recovered , K cw w i WutiW*4 $ Y y g j 102% 2.0 2.0 2.0 2.0 2.0 2.0 0.10 0.62 0.10 0.97 0.004 62.00 0.15 0.10 65% 0.10 1.83 0.004 62.00 0.15 0.18 123% 0.10 2.06 0.004 250.0 0.60 0.21 34% 0.10 4.76 0.004 250.0 0.60 0.48 79% 0.10 3.64 0.004 250.0 0.60 0.36 61% BEST COPY AVAUAELE V 3L Z 00 Page 3 152S-D.XLS 1995-08-23 15:22 OutPul of : 950823B1 Operator : DDW Date of the Analysis : 1995-08-23 10:50 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950823B1 Sample # Sample ID Skalar Skalar Standaid Result - (ppl* , ,, )?p?r) , % DITISAB QtySampl Recovery final vol t mR) // / , ,,,, /// 1 Tracer 1.50 1.48 98% 2 Drift 1.50 1.48 99% 3 Wash ND 4 Standard 1 0015 0.016 106% 5 Standard 2 0.03 0.03 95% 6 Standard 3 0.06 0.06 101% 7 Standard 4 0.09 0.09 102% 8 Standard 5 0.12 0.12 98% 9 Standard 6 0.15 0.15 100% 10 Standard 7 0.30 0.28 92% 11 Standard 8 0.60 0.62 104% 12 Standard 9 1.20 1.23 102% 13 Standard 10 1.50 1.47 98% 14 Drift 1.50 1.49 99% 15 Wash ND 16 SERUM BLK 0.05 2.0 17 SERUM BLK 0.05 2.0 18 SERUM BLK 0.03 2.0 19 SPK62-1 0.05 2.0 20 SPK 62-2 0.10 2.0 21 SPK 62-3 0.11 2.0 22 SPK 250-1 0.10 2.0 23 SPK 250-2 0.23 2.0 24 SPK 250-3 0.21 2.0 25 SPK 124-1 0.20 2.0 26 Drift 1.50 1.47 98% 27 Wash ND 28 SPK 124-2 0.21 2.0 29 BLK 0.13 2.0 30 BLK 0.06 2.0 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 1 G027G4 S3 1.04 0.93 0.67 1.03 0.004 62.00 0.15 2.0S 0.004 62.00 0.15 2.13 0.004 62.00 0.15 1.94 0.004 250.0 0.60 4.66 0.004 250.0 0.60 4.16 0.004 250.0 0.60 4.08 0.004 124.0 0.30 0.10 69% 0.21 140% 0.21 143% 0.19 32% 0.47 78% 0.42 69% 0.41 137% 4.12 0.004 124.0 0.30 2.53 1.17 0.41 138% pn Sample # 31 32 33 34 35 36 :37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 Sample II> F52726-2 F52735-2 F52732-2 F52736-2 F52739-2 F52711-4 F52721-4 Drift Wash F52719-4 F52727-4 F52725-4 F52723-4 SPK 124-1 BLK BLK SPK 62-1 SPK 62-2 SPK 62-3 Drift Wash SPK 250-1 SPK 250-2 SPK 250-3 BLK BLK BLK F52726-2 F52732-2 F52739-2 F52735-2 Drift Wash F52736-2 F52711-4 BLK Skalar Standard (ppm) 1.50 1.50 1.50 152S-D.XLS Skalar % DH15AB Qty SampJ Result RcWKty final vot tppm) w m m m m fn&) 0.04 2.0 0.04 2.0 0.03 2.0 0.03 2.0 0.03 2.0 0.03 2.0 0.02 2.0 1.50 100% ND 0.26 2.0 0.04 2.0 0.03 2.0 0.06 2.0 0.17 2.0 0.04 2.0 0.04 2.0 0.06 2.0 0.08 2.0 0.10 2.0 1.48 99% ND 0.17 2.0 0.25 2.0 0.29 2.0 0.11 2.0 0.08 2.0 0.02 2.0 0.04 2.0 ND 2.0 ND 2.0 0.02 2.0 1.45 96% ND 0.03 2.0 ND 2.0 O il 2.0 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 2 Aciual m L FC 95 Com? Mass Mas % 0.83 0.74 0.69 0.57 0.58 0.53 0.47 5.10 0.70 0.60 1.25 3.30 0.004 124.0 0.30 0.89 0.84 1.14 0.004 62.00 0.15 1.67 0.004 62.00 0.15 2.06 0.004 62.00 0.15 0.33 111% O il 76% 0.17 112% 0.21 139% 3.42 0.004 250.0 0.60 0.34 57% 4.96 0.004 250.0 0.60 0.50 83% 5.88 0.004 250.0 0.60 0.59 98% 2.23 1.57 0.48 0.75 ND ND 0.43 0.57 ND 2.25 BEST COPY m u m Sample * 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 Sample IP Skalar Standard (ppm) Skalar Result (ppm) 152S-D.XLS % DfTISAB QtySampj Recovery final voi ' ' ' C v M Z ' BLK 0.18 2.0 BLK 0.04 2.0 BLK 0.02 2.0 SPK 62-1 0.08 2.0 SPK 62-2 0.08 2.0 SPK 250-1 0.17 2.0 SPK 250-2 0.18 2.0 Drift 1.50 1.49 100% Wash ND SPK 250-3 0.21 2.0 SPK 124-1 0.17 2.0 SPK 124-2 0.16 2.0 BLK 0.08 2.0 BLK 0.07 2.0 F5?711-4 0.04 2.0 F52719-4 0.04 2.0 SPK 124-1 0.13 2.0 Drift 1.50 1.47 98% Wash ND 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 C 02766 Page 3 ; X ml-F+C< 9'5" ^1C/ s .u m / ir , ' , / ^ Mass' *, *' - Mas % < 3.64 0.79 0.42 1.58 0.004 62.00 0.15 1.65 0.004 62.00 0.15 3.48 0.004 250.0 0.60 3.56 0.004 250.0 0.60 4.10 0.004 250.0 0.60 3.32 0.004 124.0 0.30 3.20 0.004 124.0 0.30 1.68 1.38 0.90 0.73 2.62 0.004 124.0 0.30 0.16 106% 0.17 111% 0.35 58% 0.36 59% 0.41 68% 0.33 111% 0.32 107% 0.26 88% 152S-E.XLS 1995-08-28 15:03 OutPut o f : 9S0828A1 Operator : DDW Date of the Analysis : 1995-08-28 07:32 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950828A1 Sample # Sample ID Skalar Skalar Standard Resoli (PP*) . (PPro) % Dl TISAB Q y Saippl Recovery' final *ol (mL> ,mp 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 o 27 o 28 w 29 30 G\ Tracer Drift Wash Standard 1 Standard 2 Standard 3 Standard 4 Standard 5 Standard 6 Standard 7 Standard 8 Standard 9 Standard 10 Drift Wash SERUM BLANK SERUM BLANK SPK 62-1 SPK 62-2 SPK 250-1 SPK 250-2 SPK 124-1 SPK 124-2 BLANK F52725-4 Drift Wash F52721-4 F52727-4 F52806-4 1.50 1.50 0.015 0.03 0.06 0.09 0.12 0.15 0.30 0.60 1.20 1.50 1.50 1.50 1.45 1.47 ND 0.014 0.03 0.06 0.09 0.12 0.15 0.28 0.62 1.23 1.47 1.49 ND 0.06 0.06 0.10 0.09 0.20 0.21 0.19 0.18 0.06 0.07 1.46 ND 0.06 0.05 0.05 97% 98% 91% 105% 102% 99% 99% 100% 93% 103% 103% 98% 99% 97% 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 1 Actual mL FC 95 BEST COPY AVAILABLE 1.15 1.15 1.92 0.004 62.00 0.15 1.73 0.004 62.00 0.15 3.90 0.004 250.0 0.60 4.28 0.004 250.0 0.60 3.84 0.004 1240 0.30 3.52 0.004 124.0 0.30 1.20 1.32 0.19 129% 0.17 116% 0.39 65% 0.43 71% 0.38 129% 0.35 118% 1.21 0.93 0.92 f J: t & 3LZ00 Sample ., u 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 : 1 64 ; 1 66 Sample . ID 152S-E.XLS Skalar Skalar % DLTJSAB Qty Satnpl Standard Result Recovery final voi i n & ) fl), ,<PW*>..,iVA4v//f////}VAVMSA F52713-4 0.03 F52718-4 0.05 F52723-4 0.04 F52715-4 0.03 F52738-4 0.07 SPK 62-1 0.05 SPK 62-2 0.12 Drift 1.50 1.48 99% Wash ND SPK 124-1 0.15 BLANK-1 0.05 BLANK-2 0.04 SPK 62-1 0.08 SPK 62-2 0.08 SPK 250-1 0.30 SPK 250-2 0.26 BLK 0.06 F52809-4 0.04 F52724-4 0.03 Drift 1.50 1.47 98% Wash ND F52737-4 0.03 F52801-4 0.06 F52720-4 0.05 F52733-4 0.04 F52788-4 0.02 F52730-4 0.02 F52731-4 0.02 F52802-4 0.03 SPK 62-1 0.05 SPK 62-2 0.10 Drift 1.50 1.49 99% Wash ND SPK 250-1 0.18 SPK 250-2 0.24 SPK 124-1 0.16 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 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 2 Actual L F C 9 5 Cone W * * Solution FC PSSoin aSajnoJe ' s # e d Mass Mass ^Recovered % 0.64 0.99 0.73 0.69 1.40 0.93 0.004 62.00 0.15 2.30 0.004 62.00 0.15 009 63% 0.23 155% 2.93 0.004 124.0 0.30 1.04 0.75 1.58 0.004 62.00 0.15 1.69 0.004 62.00 0.15 6.04 0.004 250.0 0.60 5.14 0.004 250.0 0.60 1.21 0.72 0.64 0.29 98% 0.16 106% 0.17 114% 0.60 101% 0.51 86% BEST COPY AVARABLE 0.64 1.20 0.92 0.70 0.50 0.39 0.39 0.59 0.95 0.004 62.00 0.15 2.05 0.004 62.00 0.15 0.10 64% 0.20 138% 3.52 0.004 250.0 0.60 4.70 0.004 250.0 0.60 3.20 0.004 124.0 0.30 0.35 59% 0.47 78% 0.32 107% Sample if 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 152S-E.XLS Sample " - ig SkaMr Standard . <Ppm) Skalar Kesnft Ob*) % Di'TfSAB Qty Sampl Actual Cene Mass Recovery inalvei t m b ) ppmF: ,, Soletto FC 9$ Soin , Spiked , ^ M M ^ B k j (bS Mass % Recovered Reeww i ^^ BLK 0.08 2.0 BLK 0.06 2.0 F52714-4 0.05 2.0 F52734-4 0.07 2.0 F52774-4 0.05 2.0 F52712-4 0.03 2.0 F52729-4 0.03 2.0 Drift 1.50 1.49 9 9 % Wash ND F52803-4 0.05 2.0 F52716-4 0.04 2.0 F52728-4 0.03 2.0 F52787-4 0.04 2.0 SPK62-1 0.07 2.0 SPK62-2 0.11 2.0 SPK 124-1 0.09 2.0 SPK 124-2 0.15 2.0 Drift 1.50 1.51 100% Wash ND 0.10 1.62 0.10 1.25 0.10 0.93 0.10 1.35 0 10 0.92 0.10 0.69 0.10 0.67 0.10 0.92 0.10 0.77 0.10 0.68 0.10 0.76 0.10 1.31 0.004 62.00 0.15 0.10 2.20 0.004 62.00 0.15 0.10 1.81 0.004 124.0 0.30 0.10 2.97 0.004 124.0 0.30 0.13 88% 0.22 148% 0.18 61% 0.30 100% BEST COPY AVAIlABLE 002769 Page 3 ~o 152S-F.XLS 1995-08-29 06:38 OutPut of : 950828B1 Operator : DDW Date of the Analysis : 1995-08-28 11:47 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950828B1 Sample Sample JO Skalar Standard ( m 'l. Skater Resell ,<wm> % DI.TlSABQtySampl Recovery final voi ( m L ) , ftnL) -- Actual Cose PPm F- Solution FC 95 Soto , S * nii-/! '' t % Recovered Recowry, o o w ovl 1 1 1 t c Po 1 Tracer 1.50 1.46 98% 2 Drift 1.50 1.47 98% 3 Wash ND 4 Standard 1 0.015 0.011 74% 5 Standard 2 003 0.04 131% 6 Standard 3 0.06 0.06 97% 7 Standard 4 0.09 0.09 96% 8 Standard 5 0.12 0.12 99% 9 Standard 6 0.15 0.15 101% 10 Standard 7 0.30 0.28 93% 11 Standard 8 0.60 0.62 103% 12 Standard 9 1.20 1.23 102% 13 Standard 10 1.50 1.47 98% 14 Drift 1.50 1.50 100% 15 Wash ND 16 SERUM BLK 1 0.04 2.0 17 SERUM BLK 2 0.03 2.0 18 SPK 62-1 0.11 2.0 19 SPK 62-2 0.09 2.0 20 SPK 124-1 0.29 2.0 21 SPK 124-2 0.13 2.0 22 BLK 0.06 2.0 23 BLK 0.04 2.0 24 F52717-4 0.05 2.0 25 F52735-4 0.03 2.0 26 Drift 1.50 1.46 97% 27 Wash ND 28 F52736-4 0.03 2.0 29 F52726-4 0.03 2.0 30 F52732-4 0.02 2.0 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 1 0.84 0.69 2.16 0.004 62.00 0.15 1.89 0.004 62.00 0.15 5.86 0.004 124.0 0.30 2.55 0.004 124.0 0.30 1.20 0.71 1.03 0.67 0.64 0.54 0.42 0.22 145% 0.19 127% 0.59 197% 0.26 86% GO Sample 8 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 6 64 65 vv! 66 Vi Sample H ?, Simla r Standard Skalar Result 152S-F.XLS % Pl;m 4B Q ty5aw pI Recovery final yol F52739-4 F52711-8 F52719-8 BLK SPK 62-1 SPK 62-2 SPK 124-1 Drift Wash SPK 124-2 BLK BLK F52725-8 F52721-8 F52727F52806-8 F52713-8 F52718-8 F52723-8 Drift Wash F52715-8 F52738-8 F52809-8 SPK 62-1 SPK 62-2 SPK 124-1 SPK 124-2 BLK BLK SPK 62-1 Drift Wash SPK 62-2 SPK 124-1 SPK 124-2 1.50 1.50 1.50 0.02 0.02 ND 0.03 0.05 0.06 0.08 1.46 97% ND 0.17 0.11 0.05 0.04 0.03 0.03 0.02 0.02 0.02 0.02 1.46 98% ND 0.02 0.02 0.02 0.05 0.06 0.10 0.12 0.04 0.02 0.08 1.46 97% ND 0.08 0.14 0.14 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 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 Page 2 Actual ml-FCSS:" ,C ^i, ife ; tym " ,% VW?* ,S#On FC9$ Soi,;,,Spiksd , R ^ r ! S w a y 0.33 0.45 ND 0.60 1.09 0.004 62.00 0.15 1.17 0.004 62.00 0.15 1.58 0.004 124.0 0.30 3.48 0.004 124.0 0.30 2.15 0.93 0.78 0.63 0.53 0.49 0.31 0.33 0.32 O il 73% 0.12 79% 0.16 53% 0.35 117% BEST COPY AVMlABli 0.49 0.38 0.33 0.90 0.004 62.00 0.15 1.30 0.004 62.00 0.15 1.98 0.004 124.0 0.30 2.44 0.004 124.0 0.30 0.72 0.39 1.50 0.004 62.00 0.15 0.09 61% 0.13 87% 0.20 67% 0.24 82% 0.15 101% 1.60 0.004 62.00 0.15 2.79 0.004 124.0 0.30 2.85 0.004 124.0 0.30 0.16 107% 0.28 94% 0.29 96% Skalar Skalar Sample Sample Standard lesoli ' '/ ,,TIP" ,, > (PPm), ,,$P0 67 BLK 0.05 68 BLK 0.04 69 F52724-8 0.04 70 F52737-8 0.04 71 F52801-8 0.06 72 F52720-8 0.05 73 F52733-8 0.03 74 Drift 1.50 1.47 75 Wash ND 76 F52788-8 ND 77 SPK 62-1 0.06 78 SPK 62-2 0.07 79 SPK 124-1 0.11 80 Drift 1.50 1.47 81 Wash ND 152S-F.XLS % PLTJSAB Qty Sampl y y^' Actual roi, FC95 Orne * ? FC^ " S * 2.0 2.0 2.0 2.0 2.0 2.0 2.0 98% 0.10 0.95 0.10 0.76 0.10 0.74 0.10 0.72 0.10 1.29 0.10 0.92 0.10 0.54 2.0 2.0 2.0 2.0 98% 0.10 ND 0.10 1.12 0.004 62.00 0.10 1.31 0.004 62.00 0.10 2.15 0.004 124.0 Mass Mass % ;W W ,j ,Recovered lisce* 0.15 0.11 75% 0.15 0.13 88% 0.30 0.21 72% CQ2772 Page 3 1995-0&-22 13:41 OutPut of : 950822B1 Software : version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-22 09:04 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950822B1 A W \ io<?s. \ HuJZ (* 3 zH -i5 7 - ( W "2- OaJ2.4ow. Fluoride 1.5 Calibration order = Inverse Logarithm Slope : s = #./#### [ - - C1 1 Result = 10*- s J X = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al aO = -0.00000 0.00072 -1.20078 Fluoride L Calibration order = 2 C o rre la tio n : r = 0.99917 Result = a2 * xa + al * x + aO a2 = al aO = -0.00000 0.00025 0.00551 Sampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 mm. Diluter needle Height dilution Factor dilution Volume Resample Dilution runs :80 :10 :2 . 5 :1 :1 mm ml. User file : Reproces : No TXT C02773 1995-08.-22 13:41 OutPut of : 950822B1 Fluoride 1.5 Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 3 Debubbled Yes 0 No No 4095 0 Off si sTandard Ignore s2 sTandard Ignore s3 sTandard Ignore s4 sTandard Ignore s5 sTandard Ignore s6 sTandard 0.150 s7 sTandard 0.300 s8 sTandard 0.600 s9 sTandard 1.200 slO sTandard 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : output : ##.### Fluoride L Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 0 Debubbled No 0 No No 4095 0 Off C02774 ^ 1995-08-22 13:41 Output of : 950822B1 Si BTandard 0.015 s2 sTandard 0.030 S3 sTandard 0.060 s4 sTandard 0.090 s5 sTandard 0.120 s6 sTandard 0.150 s7 sTandard Ignore s8 sTandard Ignore s9 sTandard Ignore slO sTandard Ignore Order : 2 Dimension : PPM start Value 500 DU trigger Limit 1800 Sec Peak shape Pointed stArt ignore 60 Sec eNd ignore 120 Sec Measure window 75 Filter No Regeneration No formula : c4:==c3 output : #.#### CQ2775 ^ 1995-08-22 13:41 OutPut of : 950822B1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time wt iw Initial Wash 3 0.063 1t Tracer 3 1.464 2d Drift 3 1.468 3w Wash 3 0.063 4 si Standard 1 3 0.067 5 s2 Standard 2 3 0.073 6 s3 Standard 3 3 0.091 7 s4 Standard 4 3 0.108 8 s5 Standard 5 3 0.127 9 s6 Standard 6 3 0.157 10 s7 Standard 7 3 0.278 11 s8 Standard 8 3 0.618 12 s9 Standard 9 3 1.232 13 SlO Standard 10 3 1.466 14 d Drift 3 1.480 15 w Wash 3 0.063 16 u SERUM BLK 1 3 0.098 17 u SERUM BLK 2 3 0.079 18 u SPK 62-1 3 0.097 19 u SPK 62-2 3 0.121 20 u SPK 62-3 3 0.133 21 u SPK 250-1 3 0.133 22 u SPK 250-2 3 0.272 23 u SPK 250-3 3 0.267 24 u BLK 3 0.105 25 u BLK 3 0.094 26 d Drift 3 1.457 27 w Wash 3 0.063 28 u F52713-1 3 0.094 29 u F52718-29 3 0.088 30 u F52723-1 3 0.082 31 u F52715-1 3 0.084 32 u F52738-1 3 0.089 33 u 34 u F52809-1 3 0.098 F52724-1 3 0.090 35 u F52737-1 3 0.089 36 u F52801-1 3 0.085 37 u F52729-1 3 0.079 38 d Drift 3 1.464 39 w Wash 3 0.063 40 u SPK 62-1 3 0.096 41 u SPK 62-2 3 0.131 42 u SPK 250-1 3 0.166 43 u SPK 250-2 3 0.257 44 u BLK 3 0.088 45 u BLK C 3 0.081 46 u SPK 62-1 3 0.101 47 . u SPK 62-2 3 0.103 48 u SPK 250-1 3 0.203 49 u SPK 250-2 3 0.219 50 . d Drift 3 1.479 51 w 52 u 53 u Wash 3 0.063 SPK 250-3 3 0.243 SPK 250-4 3 0.238 65 4 0.0055 210 4 0.6754 384 4 0.6770 626 4 0.0055 731 4 0.0154 909 4 0.0282 1085 4 0.0629 1257 4 0.0902 1435 4 0.1171 1611 4 0.1512 1786 4 0.2529 1960 4 0.4152 2136 4 0.6046 2310 4 0.6763 2486 4 0.6810 2727 4 0.0055 2837 4 0.0738 3007 4 0.0410 3185 4 0.0730 3363 4 0.1088 3539 4 0.1246 3713 4 0.1246 3888 4 0.2488 4063 4 0.2453 4237 4 0.0856 4416 ,, 4 0.0673 4588 4 0.6732 4823 4 0.0055 4938 4 0.0681 5113 4 0.0582 5289 4 0.0467 5461 4 0.0502 5635 4 0.0584 5813 4 0.0743 5991 4 0.0604 6163 4 0.0587 6340 4 0.0527 6515 4 0.0405 6690 4 0.6754 6929 4 0.0055 7042 4 0.0715 7229 4 0.1212 7392 4 0.1612 7566 4 0.2380 7741 4 0.0579 7917 4 0.0440 8092 4 0.0794 8267 4 0.0829 8441 4 0.1957 8617 4 0.2097 8792 4 0.6807 9031 4 0.0055 9142 4 0.2278 9318 4 0.2241 o o 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 1 of 2 C 0 2 7 7 6 av 1995-08-22 13:41 OutPut of : 950822B1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 u 72 u 73 u 74 d 75 -w 76 u 77 u 78 u 79 u 80 u 81 u 82 u 83 u 84 u 85 u 86 d 87 w 88 u 89 u 90 u 91 d 92 w wt rw BLK 3 0.085 BLK 3 0.077 F52733-1 3 0.087 F52788-1 3 0.068 F52730-1 3 0.085 F52731-1 3 0.083 F52802-1 3 0.087 F52714-1 3 0.081 Drift 3 1.462 Wash 3 0.063 F52734-1 3 0.086 F52774-1 3 0.090 F52712-1 3 0.092 F52729-1 3 0.094 SPK 62-1 3 0.100 SPK 62-2 3 0.114 SPK 62-3 3 0.143 SPK 250-1 3 0.255 BLK 3 0.170 F52803-1 3 0.093 Drift 3 1.475 Wash 3 0.063 F52716-1 3 0.095 F52728-1 3 0.097 F52787-1 3 0.089 F52717-1 3 0.105 F52735-1 3 0.098 F52736-1 3 0.088 F52726-1 3 0.083 F52732-1 3 0.093 F52739-1 3 0.081 SPK 62-1 3 0.104 Drift 3 1.503 Wash 3 0.063 SPK 62-2 3 0.140 SPK 250-1 3 0.336 SPK 250-2 3 0.364 Drift 3 1.508 Wash 3 0.063 RunOut Wash 3 0.063 9490 4 0.0517 9668 4 0.0362 9842 4 0.0554 10017 4 0.0161 10193 4 0.0520 10367 4 0.0477 10545 4 0.0557 10719 4 0.0435 10893 4 0.6748 11126 4 0.0055 11240 4 0.0537 11417 4 0.0617 11596 4 0.0641 11785 4 0.0676 11945 4 0.0785 12120 4 0.0981 12294 4 0.1356 12469 4 0.2371 12645 4 0.1650 12820 4 0.0654 12995 4 0.6793 13230 4 0.0055 13345 4 0.0691 13521 4 0.0720 13695 4 0.0592 13871 . 4 0.0853 14045 4 0.0738 14221 4 0.0579 14396 4 0.0472 14571 4 0.0666 14745 4 0.0450 14920 4 0.0839 15097 4 0.6888 15338 4 0.0055 15450 4 0.1328 15621 4 0.2886 15800 4 0.3039 15972 4 0.6906 16213 4 0.0055 16447 4 0.0055 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 002777 21 Calibration curve of 950822B1 : Fluoride L u o ) e_i ) u a o u o o Order : 2 r : 0.99917 Calibration curve of 950822B1 : Fluoride 1.5 Measured Order : Inverse Logarithm s 6378 4095 | 002779 Raw data of 95082281 : Fluoride 1.5 Esc=Exit ! Fl=Help C rtl-P = E d it peaks ! C 0 2 7 3 1 31 3*- 002732 CQ2753 V 1995-08-22 17:22 OutPut of : 950822C1 Software : version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-22 13:41 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950822C1 Sv %W\W-S ~r w o ^ S -l yluJ-E (e3 2 P l->s x S t C**aaM. "2- Cc Fluoride 1.5 Calibration order = Inverse Logarithm Slope : a = *.99999 [x -cl -1 -----sJ x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al = aO = -0.00000 0.00071 -1.19998 Fluoride L Calibration order = 2 C o rre la tio n : r = 0.99910 Result = a2 * x a + al * x + aO a2 = al <= aO = Sampler 0.00000 0.00023 0.00817 Type Number Sample Time Wash Time Air Time Take up sPecial needle Height : SA1000 :1 : 50 sec. : 120 sec. : 1 sec. : Single : None : 70 mm. Diluter needle Height dilution Factor dilution Volume Resample Dilution runs :80 :10 :2.5 :1 :1 mm ml. User file Reproces No TXT 1995-08-22 17:22 Output of : 950822C1 Fluoride 1. Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 3 Debubbled Yes 0 No No 4095 0 Off si sTandard Ignore s2 sTandard Ignore s3 sTandard Ignore s4 sTandard Ignore s5 sTandard Ignore s6 sTandard 0.150 s7 sTandard 0.300 s8 sTandard 0.600 s9 sTandard 1.200 slO sTandard 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : output : ##.### Fluoride L Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 0 Debubbled No 0 No No 4095 0 Off 1995-08-22 17:22 Output of : 950822C1 si sTandard 0.015 s2 sTandard 0.030 s3 sTandard 0.060 s4 sTandard 0.090 s5 sTandard 0.120 s6 sTandard 0.150 s7 sTandard Ignore s8 sTandard Ignore s9 sTandard Ignore slO sTandard Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 Filter : No Regeneration : No formula : c4:=c3 output : #.#### 1995-08-22 17:22 OutPut of : 950822C1 Fluoride 1.5 PPM Fluoride L y ' PPM Pos Typ Ident Ch Result F Time Ch Result F Time wt iw Initial Wash 3 0.063 1t Tracer 3 1.476 2d Drift 3 1.488 3w Wash 3 0.063 4 si Standard 1 3 0.066 5 s2 Standard 2 3 0.073 6 s3 Standard 3 3 0.092 7 s4 Standard 4 3 0.108 8 s5 Standard 5 3 0.129 9 s6 Standard 6 3 0.156 10 s7 Standard 7 3 0.281 11 s 8 Standard 8 3 0.614 12 s9 Standard 9 3 1.238 13 slO Standard 10 3 1.462 14 d Drift 3 1.488 15 w Wash 3 0.063 16 u SERUM BLK 1 3 0.086 17 u SERUM BLK 2 3 0.084 18 u SPK 62-1 3 0.134 19 u SPK 62-2 3 0.108 20 u SPK 62-3 3 0.124 21 . u SPK 250-1 3 0.187 22 u SPK 250-2 3 0.299 23 u SPK 250-3 3 0.275 24 . u BLK 3 0.104 25 u BLK 3 0.087 26 d Drift 3 1.449 27 w Wash 3 0.063 28 u F52711-1 3 0.080 29 u F52719-1 3 0.076 30 u F52725-1 3 0.075 31 u F52721-1 3 0.071 32 u F52727-1 3 0.071 33 u F52806-1 3 0.072 34 u F52711-2 3 0.069 35 u F52719-2 3 0.067 36 u F52725-2 3 0.066 37 u F52721-2 3 0.063 38 d Drift 3 1.489 39 w Wash 3 0.063 40 u SPK 62-1 3 0.124 41 u SPK 250-1 3 0.313 42 u BLK 3 0.089 43 u BLK 3 0.082 44 u SPK 62-1 3 0.094 45 u SPK 62-2 3 0.112 46 u SPK 62-3 3 0.108 47 u SPK 62-4 3 0.119 48 u SPK 250-1 3 0.179 49 u SPK 250-2 3 0.294 50 d Drift 3 1.454 51 w 52 u 53 u Wash 3 0.063 SPK 250-3 3 0.273 BLK 3 0.087 65 4 0.0082 210 4 0.9301 386 4 0.9386 619 4 0.0082 734 4 0.0154 911 4 0.0282 1086 4 0.0636 1261 4 0.0885 1435 4 0.1183 1611 4 0.1510 1785 4 0.2692 1959 4 0.4788 2134 4 0.7912 2308 4 0.9213 2483 4 0.9386 2723 4 0.0082 2831 4 0.0528 3006 4 0.0493 3184 4 0.1240 3360 4 0.0885 3533 4 0.1120 3710 4 0.1854 3884 4 0.2835 4060 4 0.2644 4231 4 0.0823 4408 4 0.0556 4584 ' 4 0.9125 4789 4 0.0082 4935 4 0.0418 5111 4 0.0340 5283 4 0.0328 5459 4 0.0257 5633 4 0.0239 5811 4 0.0264 5977 4 0.0209 6155 4 0.0172 6335 4 0.0154 6509 4 0.0088 6685 4 0.9390 6926 4 0.0082 7033 4 0.1112 7213 4 0.2938 7384 4 0.0581 7558 4 0.0460 7736 4 0.0671 7911 4 0.0940 8084 4 0.0880 8261 4 0.1047 8434 4 0.1767 8611 4 0.2791 8785 4 0.9157 9025 4 0.0082 9136 4 0.2630 9309 4 0.0556 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 1 of 2 37 C-027S7 1995-08-22 17:22 OutPut of : 950822C1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 d 72 w wt rw BLK 3 0.077 F52727-2 3 0.074 F52806-2 3 0.074 F52713-2 3 0.071 F52718-2 3 0.069 F52723-2 3 0.072 F52715-2 3 0.072 F52738-2 3 0.069 Drift 3 1.526 Wash 3 0.063 F52809-2 3 0.074 F52724-2 3 0.072 F52737-2 3 0.071 SPJC 62-1 3 0.085 SPK 62-2 3 0.096 SPK 250-1 3 0.150 SPK 250-2 3 0.235 Drift 3 1.512 Wash 3 0.063 RunOut Wash 3 0.063 9482 9658 9838 10004 10184 10351 10536 10711 10886 11127 11234 11413 11587 11761 11936 12112 12286 12462 12703 12937 4 0.0367 4 0.0300 4 0.0312 4 0.0245 4 0.0209 4 0.0261 4 0.0271 4 0.0216 4 0.9653 4 0.0082 4 0.0305 4 0.0261 4 0.0241 4 0.0514 4 0.0697 4 0.1442 4 0.2310 4 0.9553 4 0.0082 4 0.0082 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2Page 2 of C-Q27S8 >* Calibration curve of 950822C1 : Fluoride L 0 Order : 2 Measured r : 0.99910 900 C027S9 Calibration curve of 950822C1 : Fluoride 1.5 Measured Order : Inverse Logarithn 6271 4095 C027S ho C02791 ^ 1995-08-23 11:00 OutPut of : 950823A1 Software : version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-23 06:55 Analysis Pile Name : C:\SKALAR\DATA\HWIDATA\SERUM\950823A1 O-u) tJ -s H 0 9 S .I C .'S X ^ - ? Fluoride 1.5 Calibration order = Inverse Logarithm S lo p e : s = #.##### [x - cl -j -----sJ x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al = aO = -0.00000 0.00076 -1.19531 Fluoride L Calibration order = 2 C o rrela tio n : r = 0.99897 Result = a2 * xa + al * x + aO a2 = al = aO * -0.00000 0.00028 0.00600 Sampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 mm. Diluter needle Height : 80 dilution Factor : 10 dilution Volume : 2.5 Resample :1 Dilution runs : 1 mm ml. User file : Reproces : No . TXT 002792 1995-08-23 11:00 OutPut of : 950823A1 Fluoride 1.5 Path number 3 Signal type : Debubbled Decolor : Yes system Number : 0 diLute : No Resample : No dil Threshold : 4095 diG output :0 Window event : Off si sTandard Ignore s 2 sTandard Ignore S3 sTandard Ignore s4 sTandard Ignore s 5 sTandard Ignore s6 sTandard 0.150 s7 sTandard 0.300 s8 sTandard 0.600 s9 sTandard 1.200 slO sTandard 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : output : ##.### Fluoride L Path number : 0 Signal type : Debubbled Decolor : No system Number : 0 diLute : No Resample : No dil Threshold : 4095 diG output :0 Window event : Off C 02793 43 1995-08-23 11:00 OutPut of : 950823A1 si sTandard 0.015 s2 sTandard 0.030 S3 sTandard 0.060 s4 sTandard 0.090 s5 sTandard 0.120 s6 sTandard 0.150 s7 sTandard Ignore s8 sTandard Ignore s9 sTandard Ignore slO sTandard Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 ft Filter : No Regeneration : No formula : c4 :=c3 output : #. *#** 002794w 1995-08-23 11:00 OutPut of : 950823A1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time wt iw Initial Wash 3 0.064 1t Tracer 3 1.452 2d Drift 3 1.469 3w Wash 3 0.064 4 si Standard 1 3 0.069 5 s2 Standard 2 3 0.073 6 S3 Standard 3 3 0.090 7 s4 Standard 4 3 0.109 8 s5 Standard 5 3 0.129 9 s6 Standard 6 3 0.157 10 s7 Standard 7 3 0.278 11 s8 Standard 8 3 0.618 12 s9 Standard 9 3 1.233 13 slO Standard 10 3 1.466 14 d Drift 3 1.510 15 w Wash 3 0.064 16 u SERUM BLK 3 0.088 17 u SERUM BLK 3 0.082 18 u SPK 62-1 3 0.098 19 u SPK 62-2 3 0.115 20 u SPK 62-3 3 0.115 21 u SPK 250-1 3 0.144 22 u SPK 250-2 3 0.208 23 u SPK 250-3 3 0.225 24 u SPK 250-4 3 0.227 25 u SPK 250-5 3 0.259 26 d Drift 3 1.505 27 w Wash 3 0.064 28 u SPK 250-6 3 0.387 29 u BLK 3 0.092 30 u BLK 3 0.075 31 u F52801-2 3 0.082 32 u F52720-2 3 0.077 33 u F52733-2 3 0.075 34 u F52788-2 3 0.077 35 u F52730-2 3 0.074 36 u F52731-2 3 0.062 37 u F52802-2 3 0.076 38 d Drift 3 1.506 39 w Wash 3 0.064 40 u F52714-2 3 0.080 41 u F52734-2 3 0.076 42 u F52774-2 3 0.077 43 u SPK 62-1 3 0.112 44 u SPK 250-1 3 0.153 45 u SPK 250-2 3 0.173 46 u BLK 3 0.085 47 u BLK 3 0.085 48 u SPK 62-1 3 0.085 49 u SPK 62-2 3 0.102 50 d Drift 3 1.529 51 w Wash 3 0.064 52 u SPK 62-3 3 0.132 53 u SPK 250-1 3 0.136 65 209 384 615 732 910 1086 1260 1436 1610 1786 1960 2134 2310 2485 2724 2833 3010 3186 3361 3533 3711 3887 4063 4237 4412 4587 4819 4937 5112 5284 5462 5636 5812 5986 6162 6338 6513 6688 6930 7037 7215 7391 7565 7740 7915 8089 8264 8438 8616 8790 9025 9140 9314 4 0.0060 4 0.5049 4 0.5067 4 0.0060 4 0.0175 4 0.0266 4 0.0605 4 0.0914 4 0.1186 4 0.1503 4 0.2419 4 0.3698 4 0.4794 4 0.5064 4 0.5109 4 0.0060 4 0.0576 4 0.0468 4 0.0753 4 0.1004 4 0.1012 4 0.1371 4 0.1954 4 0.2081 4 0.2094 4 0.2308 4 0.5104 4 0.0060 4 0.2952 4 0.0651 4 0.0321 4 0.0458 4 0.0365 4 0.0321 4 0.0371 4 0.0294 4 0.0015 4 0.0341 4 0.5106 4 0.0060 4 0.0425 4 0.0346 4 0.0362 4 0.0961 4 0.1460 4 0.1657 4 0.0517 4 0.0517 4 0.0517 4 0.0821 4 0.5129 4 0.0060 4 0.1230 4 0.1277 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 1 of 2 HS 002795 1995-08-23 11:00 OutPut of : 950823A1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 u 72 u 73 d 74 w wt rw SPK 250-2 3 0.218 SPK 250-3 3 0.291 SPK 250-4 3 0.253 BLK 3 0.092 BLK 3 0.087 F52712-2 3 0.083 F52729-2 3 0.075 F52803-2 3 0.075 Drift 3 1.522 Wash 3 0.064 F52716-2 3 0.075 F52728-2 3 0.078 F52787-2 3 0.076 F52717-2 3 0.075 SPK 62-1 3 0.083 SPK 62-2 3 0.109 SPK 250-1 3 0.117 SPK 250-2 3 0.238 SPK 250-3 3 0.182 Drift 3 1.537 Wash 3 0.064 Runout Wash 3 0.064 9490 9666 9839 10014 10189 10365 10535 10715 10891 11131 11238 11413 11592 11764 11941 12117 12292 12468 12645 12817 13048 13292 4 0.2034 4 0.2496 4 0.2269 4 0.0651 4 0.0563 4 0.0477 4 0.0310 4 0.0316 4 0.5121 4 0.0060 4 0.0324 4 0.0384 4 0.0341 4 0.0310 4 0.0487 4 0.0914 4 0.1030 4 0.2170 4 0.1746 4 0.5137 4 0.0060 4 0.0060 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 0 -0 2 7 9 6 Hu C02797 Calibration curve of 950823A1 : Fluoride 1.5 0 Measured Order : Inverse Logarithm s : 6330 4095 0 0 2 7 9 9 <1 002800 S* 0-025:01 1995-08-23 15:23 OutPut of ; 950823B1 Software : version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-23 10:50 Analysis Pile Name : C:\SKALAR\DATA\HWIDATA\SERUM\950823B1 HOiS \ H O I 3 2-* - SZ_ Sc/* 1/ -- irc i. Fluoride 1.5 Calibration order = Inverse Logarithm Slope : s = #. r x - cl i ------ Result = 1 0 L s * x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al = aO = -0.00000 0.00070 -1.18413 Fluoride L Calibration order = 2 C o rre la tio n : r = 0.99924 Result = a2 * xa + al * x + aO a2 = al aO = -0.00000 0.00027 0.00378 Sampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 mm. Diluter needle Height dilution Factor dilution Volume Resample Dilution runs :80 :10 :2.5 :1 :1 mm ml. User file : Reproces : No TXT CQ2EG2 52. 1995-08-23 15:23 OutPut of : 950823B1 Flubride 1. Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event Debubbled Yes 0 No No 4095 0 Off si sTandard Ignore s2 sTandard Ignore S3 sTandard Ignore s4 sTandard Ignore ss5o sTandard sTandard Igno0r.e150 s7 sTandard 0.300 s8 sTandard 0.600 s9 sTandard 1.200 slO sTandard 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula output : ##.### Fluoride L Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 0 Debubbled No 0 No No 4095 0 Off C02303 S3 1995-08-23 15:23 OutPut of : 950823B1 si sTandard 0.015 s2 sTandard 0.030 s3 sTandard 0.060 s4 sTandard 0.090 s5 sTandard 0.120 s6 sTandard 0.150 s7 sTandard Ignore s8 sTandard Ignore s9 sTandard Ignore slO sTandard Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : c4 :=c3 output : #. #### 1995-08-23 15:23 OutPut of : 950823B1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time wt iw Initial Wash 3 0.065 1t Tracer 3 1.476 2d Drift 3 1.484 3w Wash 3 0.065 4 si Standard 1 3 0.070 5 s2 Standard 2 3 0.076 6 S3 Standard 3 3 0.092 7 s4 Standard 4 3 0.111 8 s5 Standard 5 3 0.129 9 s6 Standard 6 3 0.157 10 s7 Standard 7 3 0.277 11 s8 Standard 8 3 0.621 12 s9 Standard 9 3 1.225 13 slO Standard 10 3 1.470 14 d Drift 3 1.489 15 w Wash 3 0.065 16 u SERUM BLK 3 0.087 17 u SERUM BLK 3 0.084 18 u SERUM BLK 3 0.078 19 u SPK 62-1 3 0.087 20 u SPK 62-2 3 0.119 21 u SPK 62-3 3 0.121 22 u SPK 250-1 3 0.114 23 u SPK 250-2 3 0.233 24 u SPK 250-3 3 0.208 25 u SPK 124-1 3 0.204 26 d Drift 3 1.468 27 vr Wash 3 0.065 28 u SPK 124-2 3 0.206 29 u BLK 3 0.136 30 u BLK 3 0.090 31 u F52726-2 3 0.082 32 u F52735-2 3 0.080 33 u F52732-2 3 0.079 34 u F52736-2 3 0.076 35 u F52739-2 3 0.076 36 u F52711-4 3 0.075 37 u F52721-4 3 0.074 38 d Drift 3 1.504 39 w Wash 3 0.065 40 u F52719-4 3 0.255 41 u F52727-4 3 0.079 42 u F52725-4 3 0.077 43 u F52723-4 3 0.093 44 u SPK 124-1 3 0.165 45 u BLK 3 0.083 46 u BLK 3 0.082 47 u SPK 62-1 3 0.090 48 u SPK 62-2 3 0.105 49 u SPK 62-3 3 0.118 50 d Drift 3 1.483 51 w 52 u 53 u Wash 3 0.065 SPK 250-1 3 0.171 SPK 250-2 3 0.248 65 212 386 627 730 911 1087 1263 1439 1614 1786 1960 2136 2312 2487 2727 2840 3012 3189 3363 3538 3714 3888 4065 4239 4415 4589 ' 4825 4940 5116 5288 5464 5638 5817 5994 6163 6336 6503 6691 6933 7055 7217 7393 7566 7742 7917 8092 8267 8438 8618 8792 9029 9142 9317 4 0.0038 4 0.5182 4 0.5190 4 0.0038 4 0.0159 4 0.0284 4 0.0604 4 0.0921 4 0.1175 4 0.1507 4 0.2461 4 0.3798 4 0.4895 4 0.5177 4 0.5196 4 0.0038 4 0.0519 4 0.0467 4 0.0334 4 0.0514 4 0.1042 4 0.1066 4 0.0971 4 0.2171 4 0.1981 4 0.1951 4 0.5174 4 0.0038 4 0.1966 4 0.1266 4 0.0583 4 0.0413 4 0.0368 4 0.0347 4 0.0287 4 0.0289 4 0.0263 4 0.0236 4 0.5211 4 0.0038 4 0.2321 4 0.0350 4 0.0302 4 0.0624 4 0.1594 4 0.0444 4 0.0418 4 0.0568 4 0.0837 4 0.1032 4 0.5190 4 0.0038 4 0.1649 4 0.2273 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 1 of 2 oJ- S CiO C02S05 1995-08-23 15:23 OutPut of : 950823B1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 u 72 u 73 u 74 d 75 w 76 u 77 u 78 u 79 u 80 u 81 u 82 u 83 u 84 d 85 w wt rw SPK 250-3 3 0.294 BLK 3 0.124 BLK 3 0.102 BLK 3 0.074 F52726-2 3 0.080 F52732-2 3 0.070 F52739-2 3 0.068 F52735-2 3 0.073 Drift 3 1.447 Wash 3 0.065 F52736-2 3 0.076 F52711-4 3 0.063 BLK 3 0.125 BLK 3 0.182 BLK 3 0.081 BLK 3 0.073 SPK 62-1 3 0.102 SPK 62-2 3 0.105 SPK 250-1 3 0.174 SPK 250-2 3 0.178 Drift 3 1.494 Wash 3 0.065 SPK 250-3 3 0.205 SPK 124-1 3 0.166 SPK 124-2 3 0.160 BLK 3 0.105 BLK 3 0.096 F52711-4 3 0.084 F52719-4 3 0.079 SPK 124-1 3 0.139 Drift 3 1.471 Wash 3 0.065 RunOut Wash 3 0.065 9494 9669 9841 10017 10193 10367 10543 10719 10891 11082 11245 11433 11594 11769 11942 12123 12289 12469 12645 12819 12993 13222 13343 13519 13693 13869 14044 ' 14214 14396 14567 14743 14966 15218 4 0.2558 4 0.1115 4 0.0787 4 0.0239 4 0.0373 4 0.0140 4 0.0100 4 0.0215 4 0.5152 4 0.0038 4 0.0287 4 *.*### 4 0.1127 4 0.1761 4 0.0397 4 0.0212 4 0.0792 4 0.0827 4 0.1685 4 0.1717 4 0.5200 4 0.0038 4 0.1953 4 0.1599 4 0.1539 4 0.0839 4 0.0688 4 0.0449 4 0.0366 4 0.1311 4 0.5177 4 0.0038 4 0.0038 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 v/T T T T W U 002S07 C o n c e n tra t i o n C02S08 C02S09 002810 C02S1 61 Raw data o f 950823B1 : F lu o rid e 1 .5 Esc=Exit Fl=Help ! C rtl-P = E d it peaks ! 002812 Software : version 6.1 cl990,93 Operator : DDW Date of the Analysis : 1995-08-28 07:32 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950828A1 OCW 1 A nfcT )\09 S 'l C.-32-^ Sui.f'V' Fluoride 1.5 Calibration order = Inverse Logarithm Slope : a = #.##### [x - cl -| -----s -I x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a2 = al = aO = -0.00000 0.00075 -1.16846 Fluoride L Calibration order = 2 C o rre la tio n : r = 0.99897 Result = a2 * xa + al * x + aO a2 = al = aO * -0.00000 0.00032 0.00531 Sampler Type Number Sample Time Wash Time Air Time Take up special needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 mm. Diluter needle Height dilution Factor dilution Volume Resample Dilution runs :80 :10 :2 . 5 :1 :1 mm ml. User file : Reproces : No - . TXT 002813 3 1995-08-28 15:03 OutPut of : 950828A1 Fluoride 1. 5 Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 3 Debubbled Yes 0 No No 4095 0 Off si sTandard Ignore s2 sTandard Ignore s3 sTandard Ignore s4 sTandard Ignore s5 sTandard Ignore s6 sTandard 0.150 s7 sTandard 0.300 s8 sTandard 0.600 s9 sTandard 1.200 slO sTandard 1.500 Order : Inverse Logarithm Dimension : PPM start Value 500 DU trigger Limit 1800 Sec Peak shape Pointed stArt ignore :60 Sec eNd ignore :120 Sec Measure window :75 % Filter :No Regeneration :No formula : output : ##.### Fluoride L Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event Debubbled No 0 No No 4095 0 Off C02S14 1995-08-28 15:03 Output of : 950828A1 si sTandard 0.015 s2 sTandard 0.030 S3 sTandard 0.060 s4 sTandard 0.090 s5 sTandard 0.120 s6 sTandard 0.150 S7 sTandard Ignore s8 sTandard Ignore S9 sTandard Ignore slO sTandard Ignore Order : 2 Dimension : PPM start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Filter : No Regeneration : No formula : c4 :=c3 output : #. #### 002815 1995-08-28 15:03 OutPut of : 950828A1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time oooooooooooooooooooooooooooooooooooooooooooooooooooooo wt iw Initial Wash 3 0.068 65 4 0.0053 1t Tracer 3 1.453 209 4 0.2787 2d Drift 3 1.471 385 4 0.2740 3w Wash 3 0.068 587 4 0.0053 4 si Standard 1 3 0.071 733 4 0.0136 5 s2 Standard 2 3 0.078 901 4 0.0315 6 s3 Standard 3 3 0.092 1085 4 0.0609 7 S4 Standard 4 3 0.109 1260 4 0.0895 8 s5 Standard 5 3 0.129 1436 4 0.1188 9 s6 Standard 6 3 0.156 1611 4 0.1507 10 s7 Standard 7 3 0.279 1786 4 0.2370 11 s8 Standard 8 3 0.616 1960 4 0.3222 12 s9 Standard 9 3 1.233 2136 4 0.3177 13 slO Standard 10 3 1.467 2312 4 0.2751 . 14 d Drift 3 1.485 2488 4 0.2699 15 w Wash 3 0.068 2687 4 0.0053 16 u SERUM BLANK 3 0.091 2836 4 0.0577 17 u SERUM BLANK 3 0.091 3012 4 0.0577 18 u SPK 62-1 3 0.113 3188 4 0.0961 19 u SPK 62-2 3 0.107 . 3362 4 0.0867 20 u SPK 250-1 3 0.195 3538 4 0.1857 21 u SPK 250-2 3 0.214 3713 4 0.1995 22 u SPK 124-1 3 0.192 3887 4 0.1833 23 u SPK 124-2 3 0.176 4063 4 0.1697 24 u BLANK 3 0.092 4237 4 0.0598 25 u F52725-4 3 0.095 4413 4 0.0659 26 d Drift 3 1.462 4589 ' 4 0.2764 27 w Wash 3 0.068 4830 4 0.0053 28 u F52721-4 3 0.092 4940 4 0.0606 29 u F52727-4 3 0.085 5113 4 0.0467 30 u F52806-4 3 0.085 5285 4 0.0458 31 u F52713-4 3 0.079 5464 4 0.0321 32 u F52718-4 3 0.087 5640 4 0.0497 33 u F52723-4 3 0.081 5812 4 0.0367 34 u F52715-4 3 0.080 5986 4 0.0345 35 u F52738-4 3 0.097 6164 4 0.0700 36 u SPK 62-1 3 0.085 6334 4 0.0467 37 u SPK 62-2 3 0.126 6514 4 0.1152 38 d Drift 3 1.482 6690 4 0.2708 39 w Wash 3 0.068 6932 4 0.0053 40 u SPK 124-1 3 0.152 7039 4 0.1466 41 u BLANK-1 3 0.088 7213 4 0.0518 42 u BLANK-2 3 0.081 . 7391 4 0.0376 43 u SPK 62-1 3 0.102 7565 4 0.0791 44 u SPK 62-2 3 0.106 7740 4 0.0847 45 u SPK 250-1 3 0.302 7915 4 0.2478 46 u SPK 250-2 3 0.257 8091 4 0.2256 47 u BLK 3 0.092 8263 4 0.0604 48 u F52809-4 3 0.080 8435 4 0.0361 49 u F52724-4 3 0.079 8614 4 0.0321 50 d Drift 3 1.469 8791 4 0.2746 51 w Wash 3 0.068 9031 4 0.0053 52 u F52737-4 3 0.079 9138 4 0.0321 53 u F52801-4 3 0.092 9315 4 0.0601 1 1 I 1 1 1 ' I 1 I 1995-08-28 15:03 OutPut of : 950828A1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Tine Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 u 72 u 73 u 74 d 75 w 76 u 77 u 78 'u 79 u 80 u 81 u 82 u 83 u 84 d 85 w wt rw F52720-4 3 0.085 F52733-4 3 0.080 F52788-4 3 0.075 F52730-4 3 0.073 F52731-4 3 0.073 F52802-4 3 0.077 SPK 62-1 3 0.086 SPK 62-2 3 0.117 Drift 3 1.492 Wash 3 0.068 SPK 250-1 3 0.176 SPK 250-2 3 0.235 SPK 124-1 3 0.160 BLK 3 0.103 BLK 3 0.093 F52714-4 3 0.085 F52734-4 3 0.096 F52774-4 3 0.085 F52712-4 3 0.080 F52729-4 3 0.079 Drift 3 1.488 Wash 3 0.068 F52803-4 3 0.085 F52716-4 3 0.081 F52728-4 3 0.079 F52787-4 3 0.081 SPK 62-1 3 0.095 SPK 62-2 3 0.122 SPK 124-1 3 0.109 SPK 124-2 3 0.154 Drift 3 1.506 Wash 3 0.068 RunOut Wash 3 0.068 9484 4 0.0458 9666 4 0.0351 9840 4 0.0249 10016 4 0.0196 10192 4 0.0193 10366 4 0.0293 10536 4 0.0476 10715 4 0.1024 10892 4 0.2679 11096 4 0.0053 11241 4 0.1694 11415 4 0.2128 11591 4 0.1543 11769 4 0.0811 11941 4 0.0624 12113 4 0.0467 12291 4 0.0674 12461 4 0.0461 12641 4 0.0345 12817 4 0.0336 12993 4 0.2690 13224 4 0.0053 13336 4 0.0461 13516 4 0.0385 13688 4 0.0342 13866 4 0.0382 14042 ' 4 0.0653 14218 4 0.1101 14391 4 0.0903 14570 4 0.1483 14744 4 0.2632 14981 4 0.0053 15219 4 0.0053 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 C02817 0.2314781 Calibration curve of 950828Al Fluoride L Concentrt ion 0.0053119 002818 Calibration curve of 95082881 ' Fluoride 1.5 0 Measured Order : Inverse Logarithm 4095 6G51 C02S19 092820 7 002821 T Raw data of 950828ft1 ' Fluoride 1.5 Esc=Exit ! Fl=Help ! C rtl-P = E d it peaks ! Printout. of Sample Table : N2946- Inserted in System Table of 0 iw 1t 2d 3w 4 si 5 s2 6 s3 7 s4 8 s5 9 s6 10 s7 141 s~8n initial Wash l ^sTracer l sift l S 3. Standard l^l Standard 2 Standard 3 Standard 4 Standard 5 \l 1 Standard 6 1 Standard 7 1 S14t-a-n--dJard r8t 14 1.0000 1.000 1.000 / 1.000 ( 1.000 k 1.000 S' 1.000 ^M.ooo 1>QOO 1.0 o a 1.000 14.0A0A0A group 1 1995-08-29 06:38 OutPut of : 950828B1 Software : version 6.1 cl990,93 Operator : DOW Date of the Analysis : 1995-08-28 11:47 Analysis Pile Name : C:\SKALAR\DATA\HWIDATA\SERUM\950828B1 T* Fluoride 1.5 Calibration order = Inverse Logarithm Slope : a = .** r x - cl I I ------ I Result = 10L s J x = corrected value of the sample cl - corrected value of the concentration 1 s = Slope of the electrode a2 = al = aO = -0.00000 0.00071 -1.18575 Fluoride L Calibration order = 2 C o rre la tio n : r = 0.99313 Result = a2 * xa + al * x + aO a2 al = aO = 0.00000 0.00020 0.00438 Sampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 mm. Diluter needle Height dilution Factor dilution Volume Resample Dilution runs :80 :10 :2.5 :1 :1 mm ml. User file : Reproces : No . TXT 2. t 1995-08-29 06:38 Output of : 950828B1 Fluoride 1.5 Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event Debubbled Yes 0 No No 4095 0 Off si sTandard : Ignore s2 sTandard : Ignore S3 sTandard : Ignore s4 sTandard : Ignore s5 sTandard : Ignore s6 sTandard : 0.150 s7 sTandard : 0.300 s8 sTandard : 0.600 s9 sTandard : 1.200 slO sTandard : 1.500 Order : Inverse Logarithm Dimension : PPM start Value 500 DU trigger Limit 1800 Sec Peak shape Pointed stArt ignore 60 Sec eNd ignore 120 Sec Measure window 75 % Filter No Regeneration No formula : output : ##.### Fluoride L Path number Signal type Decolor system Number diLute Resample dil Threshold diG output Window event 0 Debubbled No 0 No No 4095 0 Off 1995-08-29 06:38 OutPut of : 950828B1 si sTandard 0.015 s2 sTandard 0.030 s3 sTandard 0.060 s4 sTandard 0.090 s5 sTandard 0.120 s6 sTandard 0.150 s7 sTandard Ignore s8 sTandard Ignore s9 sTandard Ignore slO sTandard Ignore Order : 2 Dimension : PPH start Value : 500 DU trigger Limit : 1800 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec Measure window : 75 % Pilter : No Regeneration : No formula : c4:=c3 output : #.#### C92526 it 1995-08-29 06:38 OutPut of : 950828B1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time wt iw Initial Wash 3 0.065 1t Tracer 3 1.464 2d Drift 3 1.474 3w Wash 3 0.065 4 si Standard 1 3 0.069 5 s2 Standard 2 3 0.085 6 S3 Standard 3 3 0.095 7 s4 Standard 4 3 0.113 8 s5 Standard 5 3 0.133 9 s6 Standard 6 3 0.156 10 s7 Standard 7 3 0.280 11 s8 Standard 8 3 0.618 12 s9 Standard 9 3 1.228 13 slO Standard 10 3 1.470 14 d Drift 3 1.498 15 w Wash 3 0.065 16 u SERUM BLK 1 3 0.086 17 u SERUM BLK 2 3 0.082 18 u SPK 62-1 3 0.127 19 u SPK 62-2 3 0.118 20 u SPK 124-1 3 0.293 21 'u SPK 124-2 3 0.139 22 u BLK 3 0.097 23 u BLK 3 0.082 24 ' u F52717-4 3 0.092 25 u F52735-4 3 0.081 26 d Drift 3 1.455 27 w Wash 3 0.065 28 u F52736-4 3 0.080 29 u F52726-4 3 0.078 30 u F52732-4 3 0.074 31 u F52739-4 3 0.072 32 u F52711-8 3 0.075 33 u F52719-8 3 0.070 34 u BLK 3 0.079 35 u SPK 62-1 3 0.093 36 u SPK 62-2 3 0.096 37 u SPK 124-1 3 0.108 38 d Drift 3 1.460 39 w Wash 3 0.065 40 u SPK 124-2 3 0.174 41 u BLK 3 0.126 42 u BLK 3 0.089 43 u F52725-8 3 0.084 44 u F52721-8 3 0.080 45 u F52727- 3 0.077 46 u F52806-8 3 0.076 47 u F52713-8 3 0.071 48 u F52718-8 3 0.072 49 u F52723-8 3 0.072 50 d Drift 3 1.463 51 w Wash 3 0.065 52 u F52715-8 3 0.076 53 u F52738-8 3 0.073 65 212 387 616 727 909 1087 1263 1439 1615 1787 1963 2137 2313 2487 2686 2832 3011 3189 : 3363 3540 3714 3892 4064 4242 4414 4588 4815 4936 5116 5288 5463 5635 5812 5992 6165 6341 6515 6689 6904 7041 7215 7389 7566 7739 7914 8092 8266 8439 8613 8789 9028 9133 9313 4 0.0044 4 1.5923 4 1.6075 4 0.0044 4 0.0111 4 0.0394 4 0.0580 4 0.0861 4 0.1182 4 0.1522 4 0.3134 4 0.6720 4 1.2898 4 1.6007 4 1.6438 4 0.0044 4 0.0420 4 0.0343 4 0.1082 4 0.0946 4 0.3291 4 0.1277 4 0.0602 4 0.0353 4 0.0516 4 0.0334 4 1.5790 4 0.0044 4 0.0318 4 0.0269 4 0.0210 4 0.0163 4 0.0227 4 0.0123 4 0.0298 4 0.0545 4 0.0585 4 0.0791 4 1.5857 4 0.0044 4 0.1784 4 0.1074 4 0.0467 4 0.0390 4 0.0316 4 0.0267 4 0.0247 4 0.0157 4 0.0163 4 0.0161 4 1.5898 4 0.0044 4 0.0243 4 0.0191 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 1 of 2 e c r s ; -7 -n 1995-08-29 06:38 OutPut of : 950828B1 Fluoride 1.5 Fluoride L PPM PPM Pos Typ Ident Ch Result F Time Ch Result F Time 54 u 55 u 56 u 57 u 58 u 59 u 60 u 61 u 62 d 63 w 64 u 65 u 66 u 67 u 68 u 69 u 70 u 71 u 72 u 73 u 74 d 75 'w 76 u 77 u 78 u 79 u 80 d 81 w wt rw F52809-8 SPK 62-1 SPK 62-2 SPK 124-1 SPK 124-2 BLK BLK SPK 62-1 Drift Wash SPK 62-2 SPK 124-1 SPK 124-2 BLK BLK F52724-8 F52737-8 F52801-8 F52720-8 F52733-8 Drift Wash F52788-8 SPK 62-1 SPK 62-2 SPK 124-1 Drift Wash Runout Wash 3 0.072 3 0.088 3 0.100 3 0.121 3 0.136 3 0.083 3 0.074 3 0.106 3 1.460 3 0.065 3 0.109 3 0.147 3 0.149 3. 0.089 3 0.084 3 0.083 3 0.083 3 0.099 3 0.088 3 0.078 3 1.465 3 0.065 3 0.071 3 0.094 3 0.100 3 0.126 3 1.468 3 0.065 3 0.065 9488 4 0.0165 9662 4 0.0451 9838 4 0.0648 10014 4 0.0991 10190 4 0.1220 10364 4 0.0362 10538 4 0.0197 10714 4 0.0752 10888 4 1.5857 11129 4 0.0044 11240 4 0.0799 11414 4 0.1393 11592 4 0.1425 11765 4 0.0477 11941 4 0.0378 12114 4 0.0369 12290 4 0.0362 12466 4 0.0643 12640 4 0.0460 -12814 4 0.0269 12988 4 1.5940 13219 4 0.0044 13336 4 0.0154 13514 4 0.0558 13692 4 0.0653 13866 4 0.1074 14039 ' 4 1.5982 14280 4 0.0044 14514 4 0.0044 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 C0Z.SZ8 -* Calibration curve of 950828B1 : Fluoride L Order : 2 r : 0.99313 002829 -l Calibration curve of 950828B1 : Fluoride 1.5 C02830 So 002831 002S32 002833 93 I 002834 9.11.4 Summary and raw data; ppm F' in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. 002835 HW I6329-152 AMDT 011095.1 Dohrmann Serum Analysis Analysis Dates: 08/02/95 - 08/18/95 A ll serum samples were thermally extracted by a modified Dohrmann D X 2000 Organic Halide Analyzer and collected in a 1:1 milli Q water and TISAB solution. The samples were measured on an Orion EA940 expandable ion analyzer. The Dohrmann was calibrated using 34ppm, 40ppm, 62ppm, lOOppm, 124ppm, 250ppm, and 500ppm FC-95 standards. The Orion was calibrated by direct measurement with no blank correction using 0.05ppm, O.lppm, 0.5ppm, l.Oppm and 1.5ppm F standards. The slope, intercept, and correlation were recorded in die appropriate logbook. A summary table is included, showing the ppm F ' in each sample (see page 2). An initial calibration curve with standard deviation, %RSD, R2 value and equation of the line is on pages 3 - 5 . Pages 6 - 1 9 show the excel spreadsheet that was generated when the samples were being analyzed. The Dohrmann FC-95 initial calibration curve was not used to generate the data in the spreadsheet. Any Orion reading below 0.05 is below the Orion calibration and should be considered an estimate. The FC-95 initial calibration curve was spiked into bovine serum. However, due to problems with blanks having higher readings than the samples, the blank samples and QC check samples after 08/11/95 were taken from study # HWI 6329-123 rabbit # F52423, F52346, and F52330. This would affect all the day 4, day 8 samples, and F52726, F52732, F52739, F52736 o f the day 2 samples. This study was discontinued after day 8 because nothing was found in the first 3 days of sampling. ***G l*f S 002836 L3492 Group 1 Dosage: 0 mg/kg HWI 6329-152 Fluoride concentration in rabbit serum (ppm F-) Sample F52711 F52719 F52725 F52721 F52727 F52806 Day 1 0.593 0.505 0.531 0.443 0.408 0.472 Day 2 0.381 0.337 0.296 0.257 0.526 0.594 Day 4 0.594 0.518 0.691 0.606 0.496 0.459 Day 8 0.484 0.328 0.600 0.556 0.461 0.462 Group 2 Dosage: 4 mg/kg Sample F52713 F52718 F52723 F52715 F52738 F52809 Day 1 0.994 0.810 0.730 0.749 0.942 1.12 Day 2 0.400 0.402 0.460 0.410 0.382 0.384 Day 4 0.371 0.590 0.414 0.377 0.827 0.391 Day 8 0.341 0.349 0.346 0.344 0.331 0.303 Group 3 Dosage: 16 mg/kg Sample F52724 F52737 F52801 F52720 F52733 F52788 Day 1 0.866 0.870 0.761 0.644 0.775 0.337 Day 2 0.369 0.371 0.609 0.464 0.451 0.511 Day 4 0.373 0.827 0.718 0.531 0.436 0.383 Day 8 0.578 0.563 0.856 0.788 0.491 0.259 Group 4 Dosage: 40 mg/kg Sample F52730 F52731 F52802 F52714 F52734 F52774 Day 1 0.785 0.772 0.854 0.71 0.772 0.924 Day 2 0.472 0.372 0.482 0.515 0.434 0.434 Day 4 0.348 0.298 0.464 0.580 0.892 0.570 Group 5 Sample Dosage: 10 X 10 fabric F52712 F52729 F52803 F52716 F52728 F52787 Day 1 0.934 0.686 0.948 0.964 1.06 0.890 Day 2 0.649 0.457 0.448 0.419 0.549 0.536 Day 4 0.436 0.452 0.474 0.411 0.452 0.443 Group 6 Sample Dosage: 10 X 10 fabric F52717 F52735 F52736 F52726 F52732 F52739 Day 1 1.27 1.09 0.894 0.725 1.08 0.675 Day 2 0.510 0.472 0.493 0.653 0.337 0.352 Day 4 0.987 0.638 0.503 0.511 0.425 0.328 002837 Sample ID blank-1 blank-2 blank-3 blank-4 blank-5 blank-6 blank-7 blank-8 blank-9 blank-10 blank-11 blank-12 blank-13 Sample <>_Date 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 7/26/95 Actual reading 0.04392 0.03695 0.03742 0.04193 0.03201 0.03174 0.02625 0.03425 0.04265 0.03228 0.03017 0.03311 0.02693 Sample TISAB m L FC95 Qty final vol spiked jmLjong) (mL| 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 0.1 2.0 one. FC9 solution MjjgmK 34ppm-1 34ppm-2 34ppm-3 7/26/95 0.05378 7/26/95 0.04430 7/26/95 0.04601 0.1 0.1 0.1 2.0 2.0 2.0 0.004 34 0.004 34 0.004 ' 34 40ppm-1 40ppm-2 40ppm-3 7/26/95 0.04739 7/26/95 0.04881 7/26/95 0.05178 0.1 0.1 0.1 2.0 2.0 2.0 0.004 0.004 0.004 40 40 40 62ppm-1 62ppm-2 62ppm-3 7/26/95 0.06585 7/26/95 0.06287 7/26/95 0.06969 0.1 0.1 0.1 2.0 2.0 2.0 0.004 0.004 0.004 62 62 62 100ppm-1 100ppm-2 100ppm-3 7/26/95 0.08098 7/26/95 0.08998 7/26/95 0.1047 0.1 0.1 0.1 2.0 2.0 2.0 0.004 0.004 0.004 100 100 100 124ppm-1 124ppm-2 124ppm-3 7/26/95 0.09999 7/26/95 0.1159 7/26/95 0.1219 0.1 0.1 0.1 2.0 2.0 2.0 0.004 0.004 0.004 124 124 124 STUDY# 6329-152 SERUM % Actual recovery ppm F- Jkig/ug^jr^amgte 0.8784 0.7390 0.7484 0.8386 0.6402 0.6348 0.5250 0.6850 0.8530 0.6456 0.6034 0.6622 0.5386 Mass spiked Mass recovered 0.0878 0.0739 0.0748 0.0839 0.0640 0.0635 0.0525 0.0685 0.0853 0.0646 0.0603 0.0662 0.0539 132% 109% 113% 1.0756 0.0817 0.8860 0.0817 0.9202 0.0817 0.1076 STDEV: 0.0886 %RSD: 0.0920 A V E R A G E : 0.010105 11 0.0961 99% 102% 108% 0.9478 0.0961 0.9762 0.0961 1.0356 0.0961 0.0948 STDEV: 0.0976 %RSD: 0.1036 A V ER A G E : 0.00448 4.5 0.0987 88% 1.3170 0.1489 0.1317 STDEV: 0.006838 84 % 1.2574 0.1489 0.1257 %RSD: 5.2 94% 1.3938 0.1489 0.1394 AVER A G E: 0.1323 67% 1.6196 0.2402 0.1620 STDEV: 0.023949 75 % 1.7996 0.2402 0.1800 %RSD: 13 87% 2.0940 0.2402 0.2094 AVER A G E: 0.1838 67% 1.9998 0.2978 0.2000 STDEV: 0.022645 78 % 2.3180 0.2978 0.2318 %RSD: 10 82% 2.4380 0.2978 0.2438 AVERAG E: 0.2252 STUDY# 6329-152 SERUM Sample ID Sample Date Actual reading (ppm F-) Sample TISAB mLFC95 one. FC9 % Actual Qty final vol spiked solution recovery ppm F- (mL or g) (mL) (ppm) (ug/ug) in sample Mass spiked (ug F-) Mass recovered (ug F-) 250ppm-1 7/26/95 0.1947 0.1 2.0 250ppm-2 7/26/95 0.302 0.1 2.0 250ppm-3 7/26/95 0.1981 0.1 2.0 250ppm-4 7/26/95 0.1972 0.1 2.0 0.004 0.004 0.004 0.004 250 65% 250 101% 250 66% 250 66% 3.8940 6.0400 3.9620 3.9440 0.6004 0.6004 0.6004 0.6004 0.3894 0.6040 0.3962 0.3944 STDEV: %RSD: AVERAGE: 0.105373 24 0.4460 500ppm-1 7/26/95 0.3486 0.1 2.0 500ppm-2 7/26/95 0.4286 0.1 2.0 500ppm-3 7/26/95 0.3745 0.1 2.0 500ppm-4 7/26/95 0.3566 0.1 2.0 0.004 0.004 0.004 0.004 500 58% 500 71% 500 62% 500 59% 6.9720 8.5720 7.4900 7.1320 1.2008 1.2008 1.2008 1.2008 0.6972 0.8572 0.7490 0.7132 STDEV: %RSD: AVERAGE: 0.072032 9.6 0.7542 G028G CDI -i SERUM CURVE 2 CARLTON Oo K 09 > O \ Sample ID BLANK BLANK 62PPM-1 62PPM-2 62PPM-3 250PPM-1 250PPM-2 250PPM-3 BLANK BUNK BUNK BUNK F52713DAY1 F52718DAY1 F52723DAY1 F52715DAY1 F52738DAY1 F52809DAY1 F52724DAY1 F52737DAY1 F52801DAY1 F52720DAY1 62ppm-1 62ppm-2 250ppm-1 250ppm-2 BUNK BUNK SERUM BUN K SERUM BUN K SERUM BUN K SERUM BUN K SERUM B U N K SERUM B U N K Actual Sample TISAB mL FC95 Cone. FC95 reading Qty final vol spiked (ppm F-) (mLorg) (mL) solution (ppm) 0.0512 0.1 2.0 0.0314 0.1 2.0 0.0557 0.1 2.0 0.004 62 0.0867 0.0995 0.1 0.1 2.0 0.004 2.0 0.004 62 62 0.0982 0.1 2.0 0.004 250 0.240 0.1 2.0 0.004 250 0.235 0.1 2.0 0.004 250 0.164 0.0878 0.1 0.1 2.0 2.0 0.0631 0.1 2.0 0.0404 0.1 2.0 0.0497 0.1 2.0 0.0405 0.1 2.0 0.0365 0.1 2.0 0.0374 0.1 2.0 0.0471 0.1 2.0 0.0561 0.0433 0.1 0.1 2.0 2.0 0.0435 0.1 2.0 0.0381 0.1 2.0 0.0322 0.1 2.0 0.0501 0.1 2.0 0.004 62 0.0838 0.1 2.0 0.004 62 0.134 0.1 2.0 0.004 250 0.233 0.1 2.0 0.004 250 0.123 0.1 2.0 0.0323 0.1 2.0 0.0478 0.1 2.0 0.0524 0.1 2.0 0.0709 0.1 2.0 0.0612 0.1 2.0 0.0542 0.1 2.0 0.0542 0.1 2.0 002841 s- * t e 5 ? x - i 5 z 5 eeuM % Actual Mass Mass recovery ppm F- spiked recovered 75% 116% 134% 33% 80% 78% 67% 113% 45% 78% 1.024 0.628 1.11 1.73 1.99 1.96 4.80 4.70 3.27 1.76 1.26 0.807 0.994 0.810 0.730 0.749 0.942 1.12 0.866 0.870 0.761 0.644 1.00 1.68 2.68 4.67 2.46 0.646 0.956 1.05 1.42 1.22 1.08 1.08 0.149 0.149 0.149 0.600 0.600 0.600 0.149 0.149 0.600 0.600 0.102 0.0628 0.111 0.173 0.199 0.196 0.480 0.470 0.327 0.176 0.126 0.0807 0.0994 0.0810 0.0730 0.0749 0.0942 0.112 0.0866 0.0870 0.0761 0.0644 0.100 0.168 0.268 0.467 0.246 0.0646 0.0956 0.105 0.142 0.122 0.108 0.108 Sample ID SERUM BUNK SERUM B U N K SERUM BUNK SERUM BUN K SERUM B U N K SERUM BUNK SERUM BUNK SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 BUNK BUNK BUNK BUNK BUNK BUNK F52733-DAY1 F52788-DAY1 F52730-DAY1 F52731-DAY1 F52802-DAY1 F52714-DAY1 F52734-DAY1 F52774-DAY1 F52712-DAY1 F52729-DAY1 62-PPM-1 62-PPM-2 62-PPM-3 250-PPM-1 BUNK Actual Sample TISAB mL FC95 reading Qty final vol spiked (PPm F-) (mLorg) (mL) 0.0636 0.1 2.0 0.0540 0.1 2.0 0.0526 0.1 2.0 0.0585 0.1 2.0 0.0464 0.1 2.0 0.0369 0.1 2.0 0.0318 0.1 2.0 0.0604 0.1 2.0 0.004 0.0610 0.1 2.0 0.004 0.177 0.184 0.1 0.1 2.0 0.004 2.0 0.004 0.211 0.1 2.0 0.004 0.204 0.1 2.0 0.004 0.170 0.1 2.0 0.0644 0.1 2.0 0.0476 0.0413 0.1 0.1 2.0 2.0 0.0346 0.1 2.0 0.0274 0.1 2.0 0.0388 0.1 2.0 0.0168 0.1 2.0 0.0392 0.1 2.0 0.0386 0.1 2.0 0.0427 0.1 2.0 0.0355 0.1 2.0 0.0386 0.1 2.0 0.0462 0.1 2.0 0.0467 0.1 2.0 0.0343 0.1 2.0 0.0604 0.1 2.0 0.004 0.0768 0.1 2.0 0.004 0.104 0.1 2.0 0.004 0.226 0.1 2.0 0.004 0.0983 0.1 2.0 Z& 8Z09 -U STLibV (<>323-102 3 6 .'.u#1 Cone. FC95 % Actual solution recovery ppmF- (ppm) (ug/ug) in sample 1.27 1.08 1.05 1.17 0.928 0.737 0.636 62 81% 1.21 62 82% 1.22 250 59% 3.54 250 61% 3.67 250 70% 4.22 250 68% 4.08 3.41 1.29 0.952 0.825 0.692 0.547 0.775 0.337 0.785 0.772 0.854 0.710 0.772 0.924 0.934 0.686 62 81% 1.21 62 103% 1.54 62 139% 2.07 250 75% 4.53 1.97 Mass spiked (ug F-) 0.149 0.149 0.600 0.600 0.600 0.600 0.149 0.149 0.149 0.600 Mass recovered (ug F-) 0.127 0.108 0.105 0.117 0.0928 0.0737 0.0636 0.121 0.122 0.354 0.367 0.422 0.408 0.341 0.129 0.0952 0.0825 0.0692 0.0547 0.0775 0.0337 0.0785 0.0772 0.0854 0.0710 0.0772 0.0924 0.0934 0.0686 0.121 0.154 0.207 0.453 0.197 Sample ID F52803-DAY1 F52716-DAY1 F52728-DAY1 F52787-DAY1 F52717-DAY1 F52735-DAY1' F52736-DAY1 F52726-DAY1 F52732-DAY1 F52739-DAY1 62-PPM-1 62-PPM-2 250-PPM-1 250-PPM-2 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 BLANK Actual Sample TISAB mL FC95 reading Qty final vol spiked (ppm F-) (mLorg) (mL) 0.0474 0.1 2.0 0.0482 0.0529 0.0445 0.0636 0.0547 0.0447 0.0362 0.0538 0.0338 0.0636 0.105 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 0.286 0.272 0.0766 0.0844 0.0673 0.0606 0.0588 0.0621 0.0616 0.0571 0.0469 0.0586 0.0422 0.0358 0.0363 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 2.0 0.004 2.0 0.004 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 0.104 0.0708 0.0881 0.162 0.1 0.1 0.1 0.1 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 0.267 0.245 0.140 0.1 0.1 0.1 2.0 0.004 2.0 0.004 2.0 t)TUOy it (p 3 z^- 152. S--IAM Cone. FC95 % Actual solution recovery ppm F- (ppm) (ug/ug) insample 0.948 0.964 1.06 0.890 1.27 1.09 0.894 0.725 1.08 0.675 62 85% 1.27 62 141% 2.09 250 95% 5.73 250 91% 5.44 1.53 1.69 1.35 1.21 1.18 1.24 1.23 1.14 0.937 1.17 0.844 0.716 0.725 62 139% 2.07 62 95% 1.42 62 118% 1.76 250 54% 3.24 250 89% 5.34 250 81% 4.89 2.81 Mass spiked (ug F-) 0.149 0.149 0.600 0.600 0.149 0.149 0.149 0.600 0.600 0.600 Mass recovered (ugF-) 0.0948 0.0964 0.106 0.0890 0.127 0.109 0.0894 0.0725 0.108 0.0675 0.127 0.209 0.573 0.544 0.153 0.169 0.135 0.121 0.118 0.124 0.123 0.114 0.0937 0.117 0.0844 0.0716 0.0725 0.207 0.142 0.176 0.324 0.534 0.489 0.281 Sample ID BLANK BLANK F52711-DAY1 F52719-DAY1 F52725-DAY1 F52721-DAY1 F52727-DAY1 F52806-DAY 1 F52711-DAY2 F52719-DAY2 F52725-DAY2 F52721-DAY2 SPIKE 62-1 SPIKE 250-1 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 62-4 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 BUNK BUNK BUNK BUNK Actual Sample TISAB mL FC95 reading Qty final vol spiked 0.0564 0.0406 0.0296 0.0253 0.0266 0.0222 0.0204 0.0236 0.0191 0.0169 0.0148 0.0128 0.0922 0.281 0.0668 0.0515 0.0787 0.0452 0.0484 0.0490 0.0526 0.0381 0.0407 0.0528 0.0742 0.0688 0.0842 0.149 0.258 0.241 0.137 0.0685 0.0492 0.0394 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 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 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 002844 -0 s ru y * u>v- is z SeuM Cone. FC95 % Actual solution recovery ppm F- (ppm) (ug/ug) in sample 1.13 0.812 0.593 0.505 0.531 0.443 0.408 0.472 0.381 0.337 0.296 0.257 62 124% 1.84 250 94% 5.62 1.34 1.03 1.57 0.904 0.967 0.979 1.05 0.761 0.814 62 71% 1.06 62 100% 1.48 62 92% 1.38 62 113% 1.68 250 49% 2.97 250 86% 5.15 250 80% 4.82 2.73 1.37 0.984 0.787 Mass spiked (ug F-) 0.149 0.600 0.149 0.149 0.149 0.149 0.600 0.600 0.600 Mass recovered (ug F-) 0.113 0.0812 0.0593 0.0505 0.0531 0.0443 0.0408 0.0472 0.0381 0.0337 0.0296 0.0257 0.184 0.562 0.134 0.103 0.157 0.0904 0.0967 0.0979 0.105 0.0761 0.0814 0.106 0.148 0.138 0.168 0.297 0.515 0.482 0.273 0.137 0.0984 0.0787 Sample 10 BLANK BLANK F52727 DAY2 F52806 DAY2 F52713 DAY2 F52718DAY2 F52723 DAY2 F52715 DAY2 F52738 DAY2 F52809 DAY2 F52724 DAY2 F52737 DAY2 SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250-5 SPIKE 250-6 BLANK BLANK BLANK Actual Sample TISAB mL FC95 reading Qty final vol spiked (PPm F-) (mL org) (mL) 0.0407 0.1 2.0 0.0310 0.1 2.0 0.0263 0.1 2.0 0.0297 0.1 2.0 0.0200 0.1 2.0 0.0201 0.1 2.0 0.0230 0.1 2.0 0.0205 0.1 2.0 0.0191 0.1 2.0 0.0192 0.1 2.0 0.0185 0.1 2.0 0.0186 0.1 2.0 0.0389 0.0546 0.118 0.1 0.1 0.1 2.0 0.004 2.0 0.004 2.0 0.004 0.223 0.1 2.0 0.004 0.103 0.1 2.0 0.0522 0.1 2.0 0.0474 0.1 2.0 0.0420 0.1 2.0 0.0330 0.1 2.0 0.0299 0.1 2.0 0.0517 0.1 2.0 0.004 0.0747 0.1 2.0 0.004 0.0763 0.1 2.0 0.004 0.112 0.1 2.0 0.004 0.179 0.1 2.0 0.004 0.197 0.1 2.0 0.004 0.194 0.1 2.0 0.004 0.223 0.1 2.0 0.004 0.330 0.1 2.0 0.004 0.102 0.1 2.0 0.0411 0.1 2.0 0.0185 0.1 2.0 C02845 D `o T U ttf Cone. FC95 % Actual solution recovery ppm F- (ppm) (ug/ug) insample 0.814 0.619 0.526 0.594 0.400 0.402 62 52% 62 73% 250 ' 39 % 250 74% 0.460 0.410 0.382 0.384 0.369 0.371 0.777 1.09 2.37 4.45 2.06 1.04 0.948 0.840 0.659 0.599 62 69% 1.03 62 100% 1.49 62 102% 1.53 250 37 % 2.23 250 60% 3.57 250 66% 3.94 250 65% 3.89 250 74% 4.46 250 110% 6.60 2.04 0.822 0.370 Mass spiked (ug F-) 0.149 0.149 0.600 0.600 0.149 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.600 Mass recovered (ugF-) 0.0814 0.0619 0.0526 0.0594 0.0400 0.0402 0.0460 0.0410 0.0382 0.0384 0.0369 0.0371 0.0777 0.109 0.237 0.445 0.206 0.104 0.0948 0.0840 0.0659 0.0599 0.103 0.149 0.153 0.223 0.357 0.394 0.389 0.446 0.660 0.204 0.0822 0.0370 152. 56^ -U ^ Sample ID F52801-DAY2 F52720-DAY2 F52733-DAY2 F52788-DAY2 F52730-DAY2 F52731-DAY2 F52802-DAY2 F52714-DAY2 F52734-DAY2 F52774-DAY2 SPIKE 62-1 SPIKE 250-1 SPIKE 250-2 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 BLANK BLANK BLANK BLANK BLANK Actual Sample TISAB mL FC95 reading Qty final vol spiked (ppm F-) (mL or gl (mL) 0.0304 0.1 2.0 0.0232 0.0225 0.1 0.1 2.0 2.0 0.0255 0.0236 0.0186 0.0241 0.0258 0.0217 0.0217 0.0673 0.115 0.215 0.0910 0.0568 0.0451 0.0438 0.0467 0.0435 0.0436 0.0335 0.0352 0.0395 0.0631 0.0926 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 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 0.0998 0.198 0.1 0.1 2.0 0.004 2.0 0.004 0.256 0.217 0.1 0.1 2.0 0.004 2.0 0.004 0.180 0.0994 0.0783 0.0525 0.0465 0.1 0.1 0.1 0.1 0.1 2.0 2.0 2.0 2.0 2.0 S* uoy # ^ 32^ - lS2 s Cone. FC95 solution (ppm) 62 250 250 62 62 62 250 250 250 250 % recovery (ug/ug) 90% 38% 72% 53% 85% 124% 33% 66% 85% 72% Actual ppm Fin sample 0.609 0.464 0.451 0.511 0.472 0.372 0.482 0.515 0.434 0.434 1.35 2.30 4.31 1.82 1.14 0.903 0.877 0.934 0.870 0.872 0.670 0.704 0.790 1.26 1.85 2.00 3.96 5.13 4.34 3.60 1.99 1.57 1.05 0.930 Mass spiked (u9 F-) 0.149 0.600 0.600 0.149 0.149 0.149 0.600 0.600 0.600 0.600 Mass recovered (ug F-) 0.0609 0.0464 0.0451 0.0511 0.0472 0.0372 0.0482 0.0515 0.0434 0.0434 0.135 0.230 0.431 0.182 0.114 0.0903 0.0877 0.0934 0.0870 0.0872 0.0670 0.0704 0.0790 0.126 0.185 0.200 0.396 0.513 0.434 0.360 0.199 0.157 0.105 0.0930 002847 Sample ID BLANK BLANK F52712 DAY2 F52729 DAY2 F52803 DAY2 F52716 DAY2 F52728 DAY2 F52787 DAY2 F52717 DAY2 SPIKE 62-1 SPIKE 62-2 SPIKE250-1 SPIKE250-2 SPIKE250-3 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 62-4 SPIKE 62-5 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250-5 SPIKE 250-6 SPIKE 124-1 SPIKE 124-2 Actual Sample TISAB mLFC95 reading Qty final vol spiked (ppmF-) (mLorg) (mL) _____ 0.0417 0.0385 0.0325 0.0229 0.0224 0.1 0.1 0.1 0.1 0.1 2.0 2.0 2.0 2.0 2.0 0.0209 0.0275 0.0268 0.0255 0.0356 0.0697 0.0820 0.221 0.182 0.114 0.0836 0.0620 0.0521 0.0426 0.0375 0.0341 0.0471 0.0592 0.0481 0.0574 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 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 0.0469 0.0871 0.167 0.184 0.187 0.162 0.196 0.171 0.0473 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 S T u b y -*{3 2 ^ - 152. Cone. FC95 % Actual solution recovery ppm F- (Ppm) Jug/uglinsampje 0.834 0.770 0.649 0.457 0.448 0.419 0.549 0.536 0.510 62 48% 0.712 62 94% 1.39 250 27% 1.64 250 74% 4.41 250 60 % 3.63 2.28 1.67 1.24 1.04 0.852 0.751 0.682 62 63% 0.943 62 79% 1.18 62 65% 0.963 62 77% 1.15 62 63% 0.939 250 29% 1.74 250 56% 3.33 250 61% 3.69 250 62% 3.73 250 54% 3.24 250 65% 3.91 124 115% 3.42 124 32% 0.946 Mass spiked (ug F-) 0.149 0.149 0.600 0.600 0.600 0.149 0.149 0.149 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.600 0.298 0.298 Mass recovered top-) 0.0834 0.0770 0.0649 0.0457 0.0448 0.0419 0.0549 0.0536 0.0510 0.0712 0.139 0.164 0.441 0.363 0.228 0.167 0.124 0.104 0.0852 0.0751 0.0682 0.0943 0.118 0.0963 0.115 0.0939 0.174 0.333 0.369 0.373 0.324 0.391 0.342 0.0946 O O w 00 00 OJ Sample ID SPIKE 124-3 SPIKE 124-5 SPIKE 124-6 SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52726 DAY 2*** F52735 DAY 2" * F52732 DAY 2*** F52736 DAY 2*** F52739 DAY 2*** F52711 DAY 4 "* F52721 DAY 4*** F52719 DAY 4*** F52727 DAY 4*** F52725 DAY 4*** SPIKE 62-1 SPIKE 124-1 BLANK BLANK 62PPM-1 Actual Sample TISAB mL FC95 reading Qty final vol spiked (ppm F-) (mL or g) (mL) 0.0133 0.161 0.155 0.143 0.0822 0.0677 0.0383 0.0366 0.0272 0.0431 0.0855 0.0877 0.0769 0.217 0.186 0.179 0.182 0.0966 0.0395 0.0264 0.0225 0.0208 0.0179 0.0177 0.0164 0.0146 0.0171 0.0168 0.0170 0.0407 0.130 0.0311 0.0291 0.0407 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 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 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 0.004 2.0 0.004 2.0 2.0 2.0 0.004 srubV * io2>iS\-152. See .m Cone FC95 % Actual solution recovery ppm F- (ppm) (ug/ug) insample 124 9 % 0.265 124 108% 3.21 124 104% 3.09 2.85 1.64 1.35 0.767 0.732 0.545 62 58 % 0.863 62 115% 1.71 62 118% 250 26 % 250 7 2 % 250 ' 62 % 1.75 1.54 4.35 3.72 124 120% 3.58 124 122% 3.64 1.93 0.790 0.529 0.450 0.416 0.357 0.354 0.328 0.293 0.342 0.335 0.341 62 55 % 0.813 124 88 % 2.61 0.623 0.582 62 55 % 0.814 Mass Mass spiked recovered (ug F-) (ug F-) 0.298 0.298 0.298 0.149 0.149 0.149 0.600 0.600 0.600 0.298 0.298 0.149 0.298 0.149 0.0265 0.321 0.309 0.285 0.164 0.135 0.0767 0.0732 0.0545 0.0863 0.171 0.175 0.154 0.435 0.372 0.358 0.364 0.193 0.0790 0.0529 0.0450 0.0416 0.0357 0.0354 0.0328 0.0293 0.0342 0.0335 0.0341 0.0813 0.261 0.0623 0.0582 0.0814 otoo mBu 00 * * Sample ID 62PPM-2 62PPM-3 250PPM -1 250PPM -2 250PPM -3 BLANK BUNK BUNK F52726 DAY2 F52732 DAY2 F52739 DAY2 F52735 DAY2 F52736 DAY2 F52711 DAY2 BUNK BUNK BUNK BUNK SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 124-1 SPIKE 124-2 BUNK BUNK F52711 DAY4 F52719 DAY4 SPIKE 62-1 SPIKE 124-1 BUNK BUNK BUNK Actual Sample TISAB mL FC95 Cone. FC95 reading Qty final vol spiked solution (ppm F-) (mLorg) (mL) (PPm) 0.0613 0.1 2.0 0.004 62 0.0798 0.1 2.0 0.004 62 0.141 0.1 2.0 0.004 250 0.223 0.1 2.0 0.004 250 0.267 0.1 2.0 0.004 250 0.0865 0.1 2.0 0.0582 0.1 2.0 0.0213 0.1 2.0 0.0326 0.1 2.0 0.0169 0.1 2.0 0.0176 0.1 2.0 0.0236 0.1 2.0 0.0246 0.1 2.0 0.0064 0.1 2.0 0.0888 0.1 2.0 0.150 0.1 2.0 0.0251 0.1 2.0 0.0175 0.1 2.0 0.0556 0.1 2.0 0.004 62 0.0570 0.1 2.0 0.004 62 0.133 0.1 2.0 0.004 250 0.138 0.1 2.0 0.004 250 0.159 0.1 2.0 0.004 250 0.122 0.1 2.0 0.004 124 0.121 0.1 2.0 0.004 124 0.0568 0.1 2.0 0.0457 0.1 2.0 0.0297 0.1 2.0 0.0259 0.1 2.0 0.0563 0.1 2.0 0.004 62 0.125 0.1 2.0 0.004 124 0.0673 0.1 2.0 0.0302 0.1 2.0 0.0334 0.1 2.0 eruM 152. aeri ,u^\ % Actual recovery ppm F- 82% 107% 47% 74% 89% 75% 77% 44% 46% 53% 82% 81% 76% 84% 1.23 1.60 2.81 4.45 5.35 1.73 1.16 0.426 0.653 0.337 0.352 0.472 0.493 0.128 1.78 3.00 0.501 0.350 1.11 1.14 2.66 2.76 3.18 2.43 2.42 1.14 0.913 0.594 0.518 1.13 2.51 1.35 0.603 0.668 Mass spiked 0.149 0.149 0.600 0.600 0.600 0.149 0.149 0.600 0.600 0.600 0.298 0.298 0.149 0.298 Mass recovered (ugF-) 0.123 0.160 0.281 0.445 0.535 0.173 0.116 0.0426 0.0653 0.0337 0.0352 0.0472 0.0493 0.0128 0.178 0.300 0.0501 0.0350 0.111 0.114 0.266 0.276 0.318 0.243 0.242 0.114 0.0913 0.0594 0.0518 0.113 0.251 0.135 0.0603 0.0668 I Cn o U Sample ID SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250-5 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52725 DAY 4 F52721 DAY 4 F52727 DAY 4 F52806 DAY 4 F52713 DAY 4 F52718 DAY 4 F52723 DAY 4 F52715 DAY 4 F52738 DAY 4 SPIKE 62-1 SPIKE 62-2 SPIKE 124-1 SERUM BLANK SERUM BLANK SERUM BLANK SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 Actual Sample TISAB mL FC95 reading Qty final vol spiked (ppm F-) (mL or g) (mL) 0.0317 0.0656 0.0567 0.0942 0.171 0.180 0.158 0.172 0.153 0.129 0.0708 0.0293 0.0346 0.0303 0.0248 0.0230 0.0185 0.0295 0.0207 0.0189 0.0414 0.0244 0.0784 0.126 0.0334 0.0290 0.0239 0.0414 0.0534 0.0564 0.127 0.223 0.172 0.264 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 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 Sruby * *32^-152 Sexi 1 Cone. FC95 solution (ppm) 62 62 62 250 250 250 250 250 124 124 62 62 124 62 62 62 250 250 250 250 % recovery (ug/ug) 43% 88% 76% 31% 57% 60% 53% 57% 103% 87% 33% 105% 85% 56% 72% 76% 42% 74% 57% 88% Actual ppm Fin sample 0.635 1.31 1.13 1.88 3.42 3.60 3.16 3.44 3.06 2.58 1.42 0.585 0.691 0.606 0.496 0.459 0.371 0.590 0.414 0.377 0.827 0.487 1.57 2.52 0.668 0.580 0.478 0.827 1.07 1.13 2.54 4.45 3.45 5.29 Mass spiked (ug F-) 0.149 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.298 0.298 0.149 0.149 0.298 0.149 0.149 0.149 0.600 0.600 0.600 0.600 Mass recovered (ug F-) 0.0635 0.131 0.113 0.188 0.342 0.360 0.316 0.344 0.306 0.258 0.142 0.0585 0.0691 0.0606 0.0496 0.0459 0.0371 0.0590 0.0414 0.0377 0.0827 0.0487 0.157 0.252 0.0668 0.0580 0.0478 0.0827 0.107 0.113 0.254 0.445 0.345 0.529 OON V J 00 cn i-* Sample ID SPIKE 250-5 blank blank F52809 DAY 4 F52724 DAY 4 F52737 DAY 4 F52801 DAY 4 F52720 DAY 4 F52733 DAY 4 F52788 DAY 4 F52730 DAY 4 F52731 DAY 4 F52802 DAY4 SPIKE 62-1 SPIKE 62-2 SPIKE 250-1 SPIKE 250-2 SPIKE 250-3 SPIKE 250-4 SPIKE 250 -5 SPIKE 250-6 SPIKE 250-7 SPIKE 250-8 SPIKE 124-1 BLANK BLANK BLANK BLANK BLANK BUNK F52714 DAY 4 F52734 DAY4 F52774 DAY 4 F52712 DAY 4 Actual Sample TISAB mL FC95 Cone. FC95 reading Qty final vol spiked solution (ppm F-) (mL org) (mL) (PPm) 0.230 0.1 2.0 0.004 250 0.177 0.1 2.0 0.0331 0.1 2.0 0.0196 0.1 2.0 0.0187 0.1 2.0 0.0164 0.1 2.0 0.0359 0.1 2.0 0.0265 0.1 2.0 0.0218 0.1 2.0 0.0192 0.1 2.0 0.0174 0.1 2.0 0.0149 0.1 2.0 0.0232 0.1 2.0 0.0325 0.1 2.0 0.004 62 0.0722 0.1 2.0 0.004 62 0.130 0.1 2.0 0.004 250 0.123 0.1 2.0 0.004 250 0.199 0.1 2.0 0.004 250 0.0964 0.1 2.0 0.004 250 0.119 0.1 2.0 0.004 250 0.183 0.1 2.0 0.004 250 0.138 0.1 2.0 0.004 250 0.192 0.1 2.0 0.004 250 0.114 0.1 2.0 0.004 124 0.0825 0.1 2.0 0.0818 0.1 2.0 0.0578 0.1 2.0 0.0562 0.1 2.0 0.0495 0.1 2.0 0.0379 0.1 2.0 0.0290 0.1 2.0 0.0446 0.1 2.0 0.0285 0.1 2.0 0.0218 0.1 2.0 % Actual recovery ppm F^^ug/ug^Jnsan^ 77% 4.60 3.54 0.663 0.391 44% 97% 43% 41% 66% 32% 40% 61% 46% 64% 76% 0.373 0.328 0.718 0.531 0.436 0.383 0.348 0.298 0.464 0.649 1.44 2.59 2.47 3.98 1.93 2.38 3.66 2.76 3.83 2.27 1.65 1.64 1.16 1.12 0.990 0.758 0.580 0.892 0.570 0.436 Mass spiked (ug F-) 0.600 0.149 0.149 0.600 0.600 0.600 0.600 0.600 0.600 0.600 0.600 0.298 Mass recovered (up F-) 0.460 0.354 0.0663 0.0391 0.0373 0.0328 0.0718 0.0531 0.0436 0.0383 0.0348 0.0298 0.0464 0.0649 0.144 0.259 0.247 0.398 0.193 0.238 0.366 0.276 0.383 0.227 0.165 0.164 0.116 0.112 0.0990 0.0758 0.0580 0.0892 0.0570 0.0436 Actual Sample TISAB mL FC95 Sample reading Qty final vol spiked ID (PPm F-) (mL or g) (mL) F52729 DAY 4 0.0226 0.1 2.0 F52803 DAY 4 0.0237 0.1 2.0 F52716 DAY4 0.0206 0.1 2.0 F52728 DAY 4 0.0191 0.1 2.0 F52787 DAY 4 0.0222 0.1 2.0 SPIKE 62-1 0.0307 0.1 2.0 0.004 SPIKE 62-2 0.0409 0.1 2.0 0.004 SPIKE 62-3 0.0710 0.1 2.0 0.004 SPIKE 124-1 0.0571 0.1 2.0 0.004 SPIKE 124-2 0.109 0.1 2.0 0.004 SERUM BLANK 0.0397 0.1 2.0 SERUM BLANK 0.0379 0.1 2.0 SERUM BLANK 0.0379 0.1 2.0 SERUM BLANK 0.0306 0.1 2.0 SERUM BLANK 0.0277 0.1 2.0 SPIKE 62-1 0.0898 0.1 2.0 0.004 SPIKE 62-2 0.0913 0.1 2.0 0.004 SPIKE 62-3 0.0769 0.1 2.0 0.004 SPIKE 124-1 0.113 0.1 2.0 0.004 SPIKE 124-2* 0.0326 0.1 2.0 SPIKE 124-2* 0.270 0.1 2.0 SPIKE 124-4 0.121 0.1 2.0 0.004 blank 0.0485 0.1 2.0 blank 0.0299 0.1 2.0 F52717 DAY4 0.0493 0.1 2.0 F52735 DAY4 0.0319 0.1 2.0 F52736 DAY4 0.0251 0.1 2.0 F52726 DAY4 0.0256 0.1 2.0 F52732 DAY4 0.0213 0.1 2.0 F52739 DAY4 0.0164 0.1 2.0 F52711 DAY 8 0.0242 0.1 2.0 O F52719 DAY 8 0.0164 0.1 2.0 O SPIKE 62-1 0.0332 0.1 2.0 0.004 W 00 SPIKE 62-2 0.0500 0.1 2.0 0.004 C/T io STUfcV * U.373 - \^TL SE.V.UM ne. FC95 olution (ppm) 62 62 62 124 124 62 62 62 124 124 62 62 % recovery (ug/ug) 41% 55% 95% 38% 73% 121% 123% 103% 76% 81% 45% 67% Actual ppm Fin sample 0.452 0.474 0.411 0.381 0.443 0.615 0.817 1.42 1.14 2.19 0.794 0.758 0.758 0.612 0.554 1.80 1.83 1.54 2.25 0.652 5.40 2.41 0.970 0.597 0.987 0.638 0.503 0.511 0.425 0.328 0.484 0.328 0.663 0.999 Mass spiked (ugF-) 0.149 0.149 0.149 0.298 0.298 0.149 0.149 0.149 0.298 0.298 0.149 0.149 Mass recovered <ug F-) 0.0452 0.0474 0.0411 0.0381 0.0443 0.0615 0.0817 0.142 0.114 0.219 0.0794 0.0758 0.0758 0.0612 0.0554 0.180 0.183 0.154 0.225 0.0652 0.540 0.241 0.0970 0.0597 0.0987 0.0638 0.0503 0.0511 0.0425 0.0328 0.0484 0.0328 0.0663 0.100 Sample SPIKE 62-3 SPIKE 62-4 SPIKE 62-5 SPIKE 62-6 SPIKE 62-7 SPIKE 62-8 SPIKE 62-9 SPIKE 124-1 SPIKE 124-2 BLANK BLANK F52725 DAY 8 F52721 DAY 8 F52727 DAY 8 F52806 DAY 8 F52713 DAY 8 F52718 DAY 8 F52723 DAY 8 F52715 DAY 8 F52738 DAY 8 F52809 DAY 8 SPIKE 62-1 SPIKE 62-2 SPIKE 62-3 SPIKE 62-4 SPIKE 124-1 SPIKE 124-2 SPIKE 124-3 BLANK BUNK SERUM BUNK SERUM BUNK SERUM BUN K SPIKE 62-1 Actual Sample TISAB mL FC95 reading Qty final vol spiked 0.0498 0.0392 0.126 0.0354 0.0421 0.0508 0.0537 0.0710 0.144 0.0847 0.0392 0.0300 0.0278 0.0230 0.0231 0.0170 0.0175 0.0173 0.0172 0.0165 0.0152 0.0245 0.0353 0.0414 0.0579 0.0925 0.0865 0.1048 0.0868 0.0414 0.0310 0.0284 0.0195 0.0558 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 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 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 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 0.004 c STUfcV * ^ 3- ^ _ i5 X S E JIM nc. FC95 solution (ppm) 62 62 62 62 62 62 62 124 124 % recovery (ug/ug) 67% 53% 169% 48% 57% 68% 72% 48% 96% v 62 33% 62 47% 62 56% 62 78% 124 62% 124 58% 124 70% 62 75% Actual ppm Fin sample 0.995 0.784 2.52 0.709 0.842 1.02 1.07 1.42 2.87 1.69 0.784 0.600 0.556 0.461 0.462 0.341 0.349 0.346 0.344 0.331 0.303 0.490 0.706 0.828 1.16 1.85 1.73 2.10 1.74 0.828 0.620 0.568 0.390 1.12 Mass spiked (ug F-) 0.149 0.149 0.149 0.149 0.149 0.149 0.149 0.298 0.298 0.149 0.149 0.149 0.149 0.298 0.298 0.298 0.149 Mass recovered (ugF-) 0.100 0.0784 0.252 0.0709 0.0842 0.102 0.107 0.142 0.287 0.169 0.0784 0.0600 0.0556 0.0461 0.0462 0.0341 0.0349 0.0346 0.0344 0.0331 0.0303 0.0490 0.0706 0.0828 0.116 0.185 0.173 0.210 0.174 0.0828 0.0620 0.0568 0.0390 0.112 <o UaG>N * k>325l - ISTZ. SEU. i Sample ID SPIKE 62-2 SPIKE 62-3 SPIKE 124-1 SPIKE 124-2 SPIKE 124-3 SPIKE 124-4 BLANK BLANK BLANK F52724 DAY8 F52737 DAY8 F52801 DAY8 F52720 DAY8 F52733 DAY8 F52788 DAY8 spike 62-1 spike 62-2 spike 124-1 Actual Sample TISAB mL FC95 Cone. FC95 reading Qty final vol spiked solution (PPm F-) (mL org) (ml) (ppm) 0.0619 0.0604 0.0933 0.0551 0.109 0.109 0.0837 0.0367 0.0305 0.0289 0.0282 0.0428 0.0394 0.0245 0.0129 0.0430 0.0518 0.104 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 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 0.004 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 2.0 0.004 2.0 0.004 62 62 124 124 124 124 62 62 124 % recovery (ug/ug) 83% 81% 63% 37% 73% 73% 58% 70% 70% Actual Ppm Fin sample 1.24 1.21 1.87 1.10 2.19 2.17 1.67 0.734 0.610 0.578 0.563 0.856 0.788 0.491 0.259 0.860 1.04 2.08 Mass spiked (ug F-) 0.149 0.149 0.298 0.298 0.298 0.298 0.149 0.149 0.298 Mass recovered (ug F-) 0.124 0.121 0.187 0.110 0.219 0.217 0.167 0.0734 0.0610 0.0578 0.0563 0.0856 0.0788 0.0491 0.0259 0.0860 0.104 0.208 * Forgot to spike blank serum with FC-95 therefore, another replicate was analyzed. DP2 (08/17/95) ** Instrument malfunction, the boat did not go all the way into the oven, another replicate was analyzed. DP2 (08/21/95) *** Samples reanalyzed on following day, due to QC not meeting requirements. DP2 (08/21/95) 00 1 tf* * >