Document 6ba7QEdxZoDdqNvxevqnkdQEo

9M61-O200 - 373 Single-dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits In-Vivo Study Reference Number: HW I#6329-159 Study Number: AMDT-042095.1 Test Substance: FC-95 (T-6246) Name and Address of Sponsor: 3M SCD Division 3 6 7 Grove Street St. Paul, MN 5510 6 & EPA-OTS 000811785U 0001175U Name and Address of Testing Facility: 3M Environmental Technology & Services 935 Bush Avenue St. Paul, MN 5510 6 NO5 o Method Numbers and Revisions: AMDT-M-1-0, Thermal Extraction o f Fluoride by means o f a Modified rcrdj CD 1 cn Dohrmann D X 2000 Organic Halide Analyzer-Liver ~~rn~ AMDT-M-2-0, Fluoride Measurement by Means o f an Orion EA940 ro Expandable Ion Analyzer CO AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver AMDT-M-8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode AMDT-M-14-0, Thermal Extraction o f Fluoride by Means of a Modified Dohrmann D X 2000 Organic Halide Analyzer-Serum :a : "O rr --ip- oc Initiation Date: See attached protocol 000081 poojl_- 000001 Z3 ! 1.0 SUMMARY Rabbit serum and liver were analyzed for fluorine content at various times after rabbits were dosed intravenously with a single dose o f T-6246, a 0.12% solution of FC-95. Doses are expressed as FC-95 equivalents. The fluorine in serum was detected after a single dose of 0.12 mg/kg and after 0.60 mg/kg, but not after a 0.012 mg/kg dose at 28 days post dose. The method is not sensitive enough to detect and measure fluoride after the 0.012 mg/kg dose. The half-life o f perfluorooctanesulfonate anion was found to be one month or greater in serum and liver. The average liver fluoride ion concentration in Group 5 animals (0.60 mg/kg), at day 14 post dose was 1.48 ppm and for four other animals in the same group sacrificed at day 28 the concentration was 1.23 ppm. Thus, the decrease in concentration in liver from 14 days to 28 days post dose is slow; based on just these two points the half-life is greater than 1 month. 2.0 INTRODUCTION This study was designed to provide information as to whether the perfluorooctanesulfonate anion does go to the liver and other tissues when the material is administered in an intravenous dose, and to ascertain the change in concentration with time after dose in serum and liver. It is known from studies done previously with rats that the half-life o f perfluorooctanesulfonate anion is quite long (>1 month). It was expected that the half-life in rabbits would also be long. Perfluorooctanesulfonate anion is a likely biotransformation product o f several fluorochemicals that are to be tested for dermal absorption in other studies. The pharmacokinetics of the perfluorooctane sulfonate anion is very relevant to these dermal absorption studies. 3.0 TEST MATERIALS 3.1 Test, Control, and Reference Substances and Matrices 3.1.1 Analytical Reference Substance; FC-95, lot 161 or 171. They are equivalent. 3.1.2 Analytical Reference Matrix: Bovine liver and bovine serum 3.1.3 Analytical Control Substance: None 3.1.4 Analytical Control Matrix: Bovine liver and bovine serum 000082 0000022 3.2 Source of Materials: 3M ICP/PCP Division For FC-95. Bovine liver from grocery store, bovine serum from Sigma Chemical Company 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 Materials: Room temperature for FC-95. For biological samples, the storage is -2010 C. 3.6 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. 4.0 EXPERIMENTAL-Overview Serum and tissues from animals dosed as described (H W I#6329-159), were available for analysis for fluorine compounds. Since perfluorooctanesulfonate anion is not biotransformed, the analysis was accomplished with combustion and subsequent analysis for fluorine. The fluorine data are related directly to perfluorooctane sulfonate concentration. The fluorine analysis of serum collected at different time intervals after dosing provides data which can be interpreted pharmacokinetically. Additional analysis o f liver samples provides evidence that the perfluorooctane sulfonate anion is still present at 28 days. 5.0 EXPERIMENTAL - Methods 5.1 AMDT-M-1-0, Thermal Extraction o f Fluoride by Means of a Modified Dohrmann D X 2000 Organic Halide Analyzer-Liver 5.2 AMDT-M-2-0, Fluoride Measurement by Means o f an Orion EA 940 Expandable Ion Analyzer 5.3 AMDT-M-4-0, Extraction o f Fluorochemicals from Rabbit Liver 5.4 AMDT-M-8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 5.5 AMDT-M-14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer-Serum 000083 000003 3 6.0 DATA ANALYSIS Raw data are shown in Table I (Appendix 9.11). Doses are adjusted for the FC-95 equivalents. The administered dose was T-6246 which is a water solution o f 0.12% FC-95. Serum concentrations are obviously not different than the control concentrations for the 0.006 and 0.012 mg/kg doses. The concentration in serum is plotted for Group 4 and Group 5 (see pages 140-141). For Group 5 (0.60 mg/kg dose), if the average concentration is fit to a least square regression Y=ae'KT, r2 =0.95, a=1.62, k=-1.099 x 10'3, k=0.693 over t1/2, and t1/2=26 days. Similarly, ti/2=48 days for Group 4. Thus, if Perfluorooctanesulfonate ion is formed from biotransformation, a large portion of the amount formed will be present in the serum at 2 8 days. The average liver fluoride ion concentration in Group 5 animals, at day 14 post dose was 1.48 ppm and for four other animals in the same group sacrificed at day 28 the concentration was 1.23 ppm. Thus, the decrease in concentration in liver from 14 days to 28 days post dose is slow; based on just these two points the half-life is greater than 1 month. Overall, for all time periods the serum and liver levels are as expected for the intravenous dose. The ability to detect levels in liver after an intravenous dose o f 120 ug/kg suggests that when perfluorooctanesulfonate is the marker, dermal absorption studies will be quite sensitive. 6.1 Circumstances That May Affect the Quality of the Data: These values for concentration are from combustion analyses o f the biological material. The recoveries are based on spiking the biological material with known amounts o f FC-95. The fluorine is measured by selective ion electrode and the Orion meter is calibrated over a 5 point range. However, if there is a different response at different concentrations o f FC-95 other than the point used as a calibration check, there could be a bias at those concentrations even though the amount o f fluorine is being measured accurately. This can come from other variables in the method such as the efficiency o f combustion. Thus, the values reported for concentration are not absolute values. The values are certainly sufficient to make the point that the perfluorooctanesulfonate anion is present at 28 days in liver and serum, that the halflife is on the order o f a month or longer, and that the method can detect perfluorooctanesulfonate in serum after a dose somewhere between 0.012 and 0.12 mg/kg. This method allows sufficient sensitivity for dermal absorption studies where some o f the applied fluorochemical is biotransformed to perfluorooctane sulfonate anion. 000084 0(KM>044 7-0 CONCLUSION The half-life of perfluorooctanesulfonate in liver and serum after an intravenous dose is one month or greater in rabbits. The perfluorooctanesulfonate anion can be detected by the combustion method and subsequent fluoride analysis at 28 days after either a 0.12 or a 0.60 mg/kg intravenous dose. The 0.012 mg/kg intravenous dose does not provide sufficient fluoride ion for this method. The method would probably be able to detect a dose somewhere between 0.012 and 0.12 mg/kg. Analysis o f fluorine at 28 days in liver and semm provides a marker for dermal absorption for dermally applied fluorochemicals that are biotransformed to perfluorooctanesulfonate anion. 8.0 MAINTENANCE OF RAW DATA 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. 9.0 APPENDICES_______________________________________________________ 9.1 Protocol and Amendments 9.1.1 Protocol and Final Report: H W I#6329-159 "Single D ose Intravenous Pharmacokinetic Study of T-6246 in Rabbits" (Protocol type TP8084.PK for dosing o f animals, tissue collection, etc.) 9.1.2 Analytical protocol AMDT-042095.1 9.2 Signed Reports from Individual Scientists: None 9.3 Quality Assurance Unit Statement: See attached 9.4 Key Personnel Involved in the Study: See attached 9.5 Materials and Equipment: See methods 9.6 Solutions, Reagents, and Standards: See methods 9.7 Sample Preparation: See methods 9.8 Quality Control Practices: See methods 000085 000005 5 9.9 Test Methods: See Protocol AMDT-042095.1 9.10 Instrument 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; ug F in whole liver as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. 9.11.3 Summary and raw data; ppm F in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. 9.11.4 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. 000086 000006 9.1.1 Protocol and Final Report: HW I#6329-159 "Single Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits" (Protocol type TP8084.PK for dosing of animals, tissue collection, etc.) 000087 000007 H A ZLETO N WISCONSIN OX 75J5 707-7545 O cu Sponsor: 3M St. Paul, Minnesota a C O R N I N G Company FINAL REPORT Study Title: Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Author: Steven M. Glaza Study Completion Date: September 14, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Pro.iect Identification: HWI 5329-159 P h o n e 608-241 -4 4 7 i E X P R ESS -MAIL DELIVERY Page 1 of 38 000008 330 ! KINSMAN F_a X _ 6 0 8 - 2 4 1 - 7 2 2 7 B LV D . MADISON. W I 5 3 70 4 000088 Page 2 of 38 HWI 6329-159 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 Phase Date Reported to Date to Study Director Manaaement 03/22/95 04/10/95 04/27/95 07/05/95 09/12/95 03/22/95 04/10/95 04/27/95 07/10/95 09/12/95 Protocol Review Protocol Amendment Animal Observation Data/Report Review Report Rereview 03/22/95 04/10/95 04/27/95 07/10/95 09/12/95 03/10/95 05/10/95 05/10/95 08/10/95 10/10/95 Date 000089 000003 Page 3 of 38 STUDY IDENTIFICATION Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits HWI 6329-159 Test Material T-6246 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. 3301 Kinsman Boulevard Madison, WI 53704 Study Timetable Study Initiation Date Experimental (In-life) Start Date In-life End Date Experimental Termination Date Study Completion Date March 30, 1995 April 4, 1995 May 1, 1995 September 14, 1995 September 14, 1995 000090 000010 Page 4 of 38 Acute Toxicoloov Steven M. Glaza Study Director Manager KEY PERSONNEL HWI 6329-159 Qua!itv Assurance Sherry R. W. Petsel Manager Francis (Bud) W. McDonald Study Coordinator Patricia Padgham In-life Supervisor Laboratory Animal Medicine Cindy J. Cary, DVM Diplomate, ACLAM Supervisor Rose M. Bridge Report Supervisor Anatomical Patholoov Toxicoloqy SuDDort Kathy Myers Manager Calvin L. Horton Supervi sor Thomas E. Palmer, PhD Anatomical Pathologist Jack Serfort/ Deborah L. Pirkel Supervi sors Necropsy Anne Mosher Supervisor Pathology Data 000031 OOOOll Page 5 of 38 CONTENTS Quality Assurance Statement Study Identification Key Personnel Summary Objective Regulatory Compliance Test and Control Materials Test System Procedures Results Discussion Signature Pathology Report Tabl e 1 Individual Body Weights (g) 2 Individual Clinical Signs 3 Individual Pathology Comments 4 Individual Animal Tissue Weights and Bile Volumes Appendix A Protocol Deviations Protocol TP8084.PK Protocol Amendment No. 1 HWI 6329-159 Page 2 3 4 6 .8 8 8 9 10 13 13 14 15 16 18 21 22 25 26 27 37 000092 OOOC12 Page 6 of 38 SUMMARY HWI 6329-159 This study was done to assess the level of systemic exposure of T-6246 when administered by a single intravenous injection to rabbits. The study was conducted using four male and four female acclimated rabbits of the Hra:(NZW)SPF strain for each treatment group as follows: GrouD 1 (Control) 2 (Low) 3 (Medium) 4 (High) 5 (High) Test Material Sterile water T-6246 T-6246 T-6246 T-6246 Dose Level Number of Animals (mq/kq) Mai es Females 04 54 10 4 100 4* 500 4* 4 4 4 4 4 * One animal sacrificed after treatment due to a possible broken back and replaced with another male animal. The animals received a single intravenous injection of the test material at the indicated dose level into the marginal ear vein of the right ear. The dose volume was 0.5 mL/kg of body weight for Groups 1 through 4 and was 0.75 mL/kg for Group 5. Two animals/sex/dose level were sacrificed on Day 15 and the remaining animals (two animals/sex/dose level) were sacrificed on Day 28. Clinical observations were conducted predose and at approximately 0.5, 2, and 4 hours after intravenous injection. Additional clinical observations and twice a day mortality checks were conducted daily thereafter until the scheduled sacrifice interval (Day 15 or Day 28). Body weights were determined on Day -7 for randomization purposes, before test or control material administration (Day 1), and at the scheduled sacrifice interval (Day 15 or Day 28). A blood sample (approximately 4 mL) was collected from a marginal ear vein (left ear) of the animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. An approximate 4-mL blood sample was also collected on Days 15 and 22 for the animals scheduled for sacrifice on Day 28. In addition, at the time of the scheduled sacrifice (Day 15 or Day 28), approximately 20 mL of blood was obtained from each animal. All samples were centrifuged, separated into serum and cellular fractions, and sent to the Sponsor. On Day 15 or 28, the animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, and exsanguinated. An abbreviated gross necropsy examination was not done, however, tissues were collected. The whole liver, bile, and both kidneys from each animal were collected, weighed (volume only determined for bile), and sent frozen to the Sponsor. 000093 000013 Page 7 of 38 HWI 6329-159 Intravenous injection of T-6246 did not result in any test material-related changes in body weight gain. All animals appeared clinically normal throughout the study with the exception of one Group 4 and one Group 5 male animals that were sacrificed on Days 2 and 1, respectively, due to injury (possible broken backs). These animals were replaced and the replacement animals appeared clinically normal throughout the study. 000094 000C14 Page 8 of 38 HWI 6329-159 OBJECTIVE The objective of this study was to assess the level of systemic exposure to the test material, T-6246, when administered as a single intravenous injection to 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, with the exception that analysis of the test mixtures for concentration, homogeneity/solubility, and stability was not conducted. All procedures used in this study were 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 material was identified as T-6246 and described as a clear, colorless liquid. The control material was Sterile Water for Injection, USP (Abbott Laboratories, Lot No. 86-748-DM-02; Exp. April 1, 1996), and was described as a clear, colorless liquid. Purity and Stabi1ity The Sponsor assumes responsibility for test material purity and stability determinations (including under test conditions). Analysis of the test material mixtures for concentration, homogeneity/solubility, and stability was not conducted or requested by the Sponsor. The purity and stability of the control material were considered to be adequate for the purposes of this study. Storage and Retention The test material was stored at room temperature. The control material was stored refrigerated. Any unused test material was returned to the Sponsor after completion of all in-life testing according to Hazleton Wisconsin (HWI) Standard Operating Procedure (SOP). Any remaining vehicle may be used for other testing and will not be discarded after issuance of the final report. 000095 000015 Page 9 of 38 Safety Precautions HWI 6329-159 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 received from HRP, Inc., Kalamazoo, Michigan on March 8, 1995 and maintained at the Hazleton Wisconsin facility at 3301 Kinsman Boulevard, 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 Tibi turn 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 -7. 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. 000096 000016 Page 10 of 38 Study Design HWI 6329-159 Animals weighing from 2,523 to 3,276 g at initiation of treatment were placed into the following study groups: Grouo Dose Test/Control Dose Level Voi urne Number of Animals' Materi al (mq/kq) (mL/kq) Mal es Females 1 (Control) 2 (Low) 3 (Medium) 4 (High) 5 (High) Sterile water T-6246 T-6246 T-6246 T-6246 0 5 10 100 500 0.5 0.5 0.5 0.5 0.75 4 4 4 4* 4* 4 4 4 4 4 * One animal sacrificed after treatment due to a possible broken back and replaced with another male animal. a Two animals/sex/dose level were sacrificed on Day 15. The remaining animals (two animals/sex/dose level) were sacrificed on Day 28. 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 Dose Preparation and Administration The test material was diluted with sterile water to achieve a specific concentration for each dose level in Groups 2 to 5. An individual dose of each respective test solution or control was calculated for each animal based on its body weight on the day of treatment. The respective test solution was administered by intravenous injection into the marginal ear vein of the right ear over approximately 22 to 60 seconds. The prepared test solutions were stored at room temperature until administered. After administration, any remaining test solutions were discarded. Reason for Route of Administration Intravenous injection is an acceptable route to assess systemic exposure. 000097 000017 Page 11 of 38 Observations of Animals HWI 6329-159 Clinical observations were conducted predose and at approximately 0.5, 2, and 4 hours after intravenous injection. Additional clinical observations and twice a day mortality checks were conducted daily thereafter until the scheduled sacrifice interval (Day 15 or Day 28). Body weights were determined on Day -7 for randomization purposes and before test or control material administration (Day 1). Additional body weights were determined at the scheduled sacrifice interval (Day 15 or Day 28) or at unscheduled sacrifices (when survival exceeded 1 day). Sample Collections A blood sample (approximately 4 mL) was collected from the marginal ear vein (left ear) of the animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. An approximate 4-mL blood sample was also collected on Days 15 and 22 for the animals scheduled for sacrifice on Day 28. In addition, at the time of necropsy, approximately 20 mL of blood was obtained from the posterior vena cava of each animal. All samples were stored at room temperature, and then centrifuged, 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 The animals sacrificed on Days 1 and 2 due to apparent broken backs were necropsied in the same manner as the animals surviving to the scheduled sacrifices with the exception that they received an abbreviated gross necropsy examination and any abnormalities were recorded. On Day 15, the first two animals/sex assigned to each dose level (based on the group assignment randomization) were anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the posterior vena cava, and exsanguinated. An abbreviated gross necropsy examination was not done, however, tissues were collected. The whole liver, bile, and both kidneys from each animal were collected, weighed (volume only determined for bile), and immediately placed in a freezer set to maintain a temperature of -20C 10C. After tissue/bile collection, the animals were discarded. The remaining two animals/sex/dose level were anesthetized, bled, and exsanguinated on Day 28 in the same manner as the animals sacrificed on Day 15. 000098 000015 Page 12 of 38 Shipment of Tissues HWI 6329-159 After completion of the in-life phase, the blood samples (serum and cellular fractions), livers, bile, 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), along with their corresponding weights or volumes. The Sponsor is responsible for the retention and disposition of the samples. HWI does not accept any responsibility for the analysis of the 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. 000099 000019 Page 13 of 38 RESULTS HWI 6329-159 Body Weights Individual body weights are in Table 1. There was no meaningful effect on body weight gain during the study. Clinical Observations Individual clinical signs are in Table 2. All animals appeared normal throughout the study with the following exceptions: One Group 4 male (No. F54106) treated with T-6246 at 100 mg/kg appeared to have injured its back on Day 2 and was sacrificed, necropsied, and replaced with No. F54099. The replacement animal appeared normal throughout the study. One Group 5 male (No. F54076) treated with T-6246 at 500 mg/kg appeared to have injured its back on the day of treatment and was sacrificed, necropsied, and replaced with No. F54112. The replacement animal appeared normal throughout the study. Patholoov Individual animal pathology comments are presented in Table 3. Individual animal tissue weights and bile volumes are in Table 4. The necropsy of animal No. F54106 revealed the skeletal muscle surrounding the lumbar spinal cord was diffusely dark red. In animal No. F54076, the skeletal muscle surrounding both hind limbs and the sacral spinal cord was diffusely dark red. The remaining animals survived to their scheduled sacrifice and were not examined grossly, although tissues were saved. Page 15 contains a pathology report by the study pathologist. DISCUSSION The level of systemic exposure of T-6246 was evaluated in male and female albino rabbits when administered as a single intravenous injection at levels of 5, 10, 100, and 500 mg/kg. There were no test material-related effects in any of the animals following administration of this material. OOOIOO 000020 Steven M. Glaza Study Director Acute Toxicology Page 14 of 38 SIGNATURE Q Date HWI 6329-159 000101 O O O O iii Page 15 of 38 PATHOLOGY REPORT HWI 6329-159 There were two male rabbits sacrificed on Day 1 or Day 2 and necropsied. At necropsy, the skeletal muscle surrounding the lumbar spinal cord in Animal No. F54106 (100 mg/kg of body weight) was diffusely dark red. In Animal No. F54076 (500 mg/kg of body weight), the skeletal muscle surrounding both hind limbs and the sacral spinal cord was diffusely dark red. The findings in these animals are indicative of injury of undetermined etiology to the caudal region of the back. The liver, bile, and both kidneys from these animals were collected as required by protocol. After necropsy, the animals were di scarded. Pathologi st (6329-159.lhm) 051295 Date 0 0 J.0 2 000022 Page 16 of 38 Table 1 Individual Body Weights (g) HWI 6329-159 Random- Animal ization Initial Terminal Sex Number (Day -7) (Dav 11 Dav 15 Dav 28 GrouD 1 (Control! - Sterile Water for In.iection (0 mq/kql Mal e F54105 F54075 F54081 F54107 2,603 2,531 2,373 2,544 2,761 2,539 2,526 2,731 2,933 2,703 - - - 2,874 3,056 Female F54104 F54086 F54074 F54085 2,665 2,526 2,540 2,553 2,890 2,632 2,703 2,667 3,130 2,775 - - _ 2,990 3,014 Mal e Female F54083 F54100 F54077 F54094 F54091 F54108 F54102 F54097 Group 2 - T-6246 (5 mq/kql 2,381 2,675 2,459 2,466 2,598 2,869 2,592 2,601 2,538 3,030 - - 2,717 2,412 2,690 2,439 2,785 2,622 2,839 2,533 2,996 2,790 - - . - 2,859 2,916 . _ 3,290 2,928 Mal e Female F54071 F54093 F54095 F54101 F54110 F54096 F54090 F54078 Group 3 - T- 6246 (10 mq/kql 2,487 2,652 2,662 2,622 2,523 2,776 2,772 2,642 2,709 2,993 - - 2,719 2,772 2,542 2,710 2,802 2,914 2,740 2,842 2,959 3,022 - - . - 3,126 3,162 _ - 3,022 3,244 - Not required. 000103 0000^3 Sex Mai e Female Page 17 of 38 Table 1 (Continued) Individual Body Weights (g) HWI 6329-159 Animal Number Random ization (Day -7) Initial (Day 1) Terminal Day 15 Dav 28 GrouD 4 - T- 6246 (100 ma/kol F54088 F541063 F54089 F54087 F54099 F54080 F54092 F54109 F54114 2,494 2,677 2,597 2,578 2,360 2,498 2,400 2,592 2,776 2,582 2,833 2,662 2,665 2,764 2,622 2,540 2,657 2,933 2,771 . 2,803 2 - - 3,004 2,835 2,667 - - _ - 2,911 3,060 - _ - 2,979 3,296 Male Female GrouD 5 - T- 6246 (500 ma/kal F54076b F54113 F54082 F54111 F54112 2,655 2,667 2,734 2,547 2,996 2,678 2,753 2,843 2,784 3,276 _ 2,896 - - 3,530 F54098 F54084 F54079 F54116 2,565 2,510 2,468 2,442 2,679 2,600 2,526 2,611 2,856 2,821 - - _ 3,216 3,228 - - 2,959 3,002 Not required. a Animal No. F54106 was originally selected by the randomization program for use in the study and was treated. This animal was sacrificed on Day 2 due to a broken back and was replaced with No. F54099. ( ) Number in superscripted parentheses indicates the day the body weight was taken. b Animal No. F54076 was originally selected by the randomization program for use in the study and was treated. This animal was sacrificed on Day 1 due to a broken back and was replaced with No. F54112. 000104 000024 Page 18 of 38 HWI 6329-159 Table 2 Individual Clinical Signs Animal Sex Number Observation Hour Dax. 0.5 2 4 Dav 2 3 to 15 16 to 28 Male GrouD 1 (Control) - Sterile Water for In.iection (0 mq/kq) F54105 Appeared normal //// / F54075 Appeared normal //// / * * F54081 Appeared normal //// / / Female F54107 F54104 F54086 Appeared normal Appeared normal Appeared normal //// //// /// / / / / * * F54074 Appeared normal //// / / F54085 Appeared normal //// / / Mai e F54083 F54100 GrouD 2 - T- 6246 5 mq/kfll Appeared normal // Appeared normal /// / / / / * * F54077 Appeared normal /// / / Female F54094 F54091 F54108 Appeared normal Appeared normal Appeared normal //// //// //// / / / / * * F54102 F54097 Appeared normal Appeared normal //// //// / / / / * Animal sacrificed on Day 15. / Condition existed. 000105 00005 Page 19 of 38 Table 2 (Continued) Individual Clinical Signs HWI 6329-159 Animal Sex Number Observation Hour (Day 1) __________ Day 0.5 2 4 2 3 to 15 16 to 28 GrouD 3 - T -6246 (10 mq/kq) Mai e Female F54071 Appeared normal F54093 Appeared normal F54095 Appeared normal F54101 Appeared normal F54110 'Appeared normal F54096 Appeared normal /// / //// //// //// /// / //// / / / / / / * * / / * * F54090 F54078 Appeared normal Appeared normal //// /// / / / / / Mai e F54088 F54106 Group 4 - T- 6246 (100 mq/kq) Appeared normal //// Appeared normal /// - Broken back /- - - Moribund sacrifice - - -/ / * F54089 Appeared normal //// / / Female F54087 Appeared normal F54099a Appeared normal F54080 Appeared normal F54092 Appeared normal F54109 Appeared normal F54114 Appeared normal /// / //// //// //// //// //// / / / / / / / * * * / / * Animal sacrificed on Day 15. / Condition existed. - Condition not evident. a Replacement animal for Animal No. F54106. 000106 OOOO^C Sex Mai e Female Page 20 of 38 Table 2 (Continued) Individual Clinical Signs HWI 6329-159 Animal Number Observation Hour (Day 11 __________ Day 0.5 2 4 2 3 to 15 16 to 28 GrouD 5 - T-6246 (500 mq/kq). F54076 Appeared normal /// Broken back t- - Moribund sacrifice - - t F54113 Appeared normal //// / * F54082 Appeared normal // / / F54111 'Appeared normal F541123 Appeared normal F54098 Appeared normal F54084 Appeared normal //// //// //// //// / / / / / * * * F54079 Appeared normal /// / / F54116 Appeared normal //// / * Animal sacrificed on Day 15. / Condition existed. - Condition not evident. f Condition evident at time of 12-hour bleeding interval, a Replacement animal for Animal No. F54076. 000107 0000^7 Page 21 of 38 Table 3 Individual Pathology Comments HWI 6329-159 Animal ______Test Day_____ Number Sex Died Sacrificed Necropsy Observation Group 4 - T-6246 (100 mq/kg) F54106 M - 2 The skeletal muscle surrounding the lumbar spinal cord is diffusely dark red. F54076 M Group 5 - T-6246 1500 mq/kgl - 1 The skeletal muscle surrounding both hind limbs and sacral spinal cord is diffusely dark red. 000108 000028 Page 22 of 38 Table 4 Individual Animal Tissue Weights and Bile Volumes HWI 6329-159 Animal Sacrifice Weiaht (q) Bile Sex Number Dav Liver Kidnevs Volume (mL) Grouo 1 (Control) - Sterile Water for In.iection _(0 mq/kq) Male F54105 15 86.35 14.92 0.6 F54075 15 60.25 15.97 0.1 F54081 28 80.597 14.764 1.2 F54107 28 80.649 15.620 0.4 Female F54104 15 98.40 16.03 0.7 F54086 15 77.30 15.23 0.7 F54074 28 82.895 18.793 1.6 F54085 28 80.950 14.925 1.3 Mal e Female F54083 F54100 F54077 F54094 F54091 F54108 F54102 F54097 Group 2 - T-6246 (5 mo/ko) 15 73.15 14.87 15 77.90 17.62 28 75.699 17.864 28 78.689 17.213 15 84.90 14.92 15 79.90 15.40 28 101.494 18.308 28 73.621 16.214 0.7 0.7 0.3 0.8 1.6 0.4 1.7 1.4 000109 000029 Page 23 of 38 HWI 6329-159 Table 4 (Continued) Individual Animal Tissue Weights and Bile Volumes Animal Sacrifice Weioht (q) Bile Sex Number Dav Liver Kidnevs Volume i GrouD 3 - T-6246 flO mo/kal Mai e Female F54071 F54093 F54095 F54101 F54110 F54096 F54090 F54078 15 15 28 28 15 15 28 28 77.10 81.40 85.504 85.668 83.68 78.10 76.220 83.460 15.88 15.50 15.561 16.120 15.79 18.70 15.878 18.130 0.5 0.8 1.2 1.0 1.4 1.1 2.1 1.0 Mai e Femal e GrouD 4 - T--6246 (100 ma/kal F54088 15 84.26 16.98 F54106 2 82.301 16.550 F54089 28 71.796 16.434 F54087 28 83.710 18.591 F540993 15 82.143 16.038 F54080 15 75.42 18.32 F54092 15 78.36 16.68 F54109 28 73.958 16.915 F54114 28 80.966 19.404 0.4 2.0 1.0 0.6 0.5 1.2 * 1.6 2.0 a Replacement animal for Animal No. F54106. * This animal had no gallbladder, so no bile was collected. 000110 000030 Page 24 of 38 HWI 6329-159 Table 4 (Continued) Individual Animal Tissue Weights and Bile Volumes Animal Sacrifice Weiqht (q) Bile Sex Number Day Liver Kidnevs Volume i GrouD 5 - T-6246 (500 ma/kal Male F54076 1 78.058 14.695 2.0 F54113 15 91.52 16.57 1.0 F54082 28 88.965 19.431 . 1.6 F54111 28 83.823 19.405 1.5 F54112a 15 97.824 21.069 1.4 Female F54098 15 78.95 14.72 1.2 F54084 15 66.42 13.18 0.8 F54079 28 83.117 14.785 1.8 F54116 28 71.962 15.793 2.0 a Replacement animal for Animal No. F54076. O O O lll 000071 Page 25 of 38 APPENDIX A Protocol Deviations Protocol TP8084.PK Protocol Amendment No. 1 HWI 6329-159 000112 0000-32 Page 26 of 38 Protocol Deviations HWI 6329-159 Protocol___________ _ Page 6, 7. Experimental Design, C. Dosing Procedures, (1) Dosing Route. Intravenous injection into the marginal ear vein of the right ear over approximately 30 to 60 seconds. ________ Actual Procedure The rate of injection for one Group 5 female (No. F54098) was 22 seconds. Also, the time for dose administration for one male animal (Group 4, No. F54087) was incorrectly recorded thus the exact duration of dosing can not be determined. These deviations are not considered to have had an adverse effect on the outcome of the study. 000113 000033 Cl 2 . HAZLETON WISCONSIN O S T O f f i c e B O X 7 5 <1 S a d i s o n . w i n i ; / ;..if . Page 27 of 38 a C O R N IN G Company Sponsor: 3M . St. Paul, Minnesota PROTOCOL TP8084.EXT Study Title: Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Date: March 30, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: HWI 6329-159 P h o n e 5 0 S 2 4 i 4 ' i E X P R E S S MAIL OeilVfS'i 000114 000034 ijl KI NSMAN Bl VO Fax 6 0 8-74 [-7 2 2 t M A D I S O N . Wl 5370 4 Page 28 of 38 STUDY IDENTIFICATION TP8084.EXT Page 2 Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits HWI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Draft Report Date 6329-159 T-6246 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 John L. Butenhoff, PhD 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 (612) 733-1962 Steven M. Glaza Hazleton Wisconsin, Inc. P.O. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, WI 53704 April 4, 1995 May 1, 1995 June 12, 1995 000115 000035 Page 29 of 38 TP8084.EXT Page 3 1. Study Single-Dose Intravenous Pharmacokinetic Study in Rabbits 2. Purpose To assess the level of systemic exposure when the test material is administered as a single intravenous injection to rabbits 3. Regulatory Compliance This study will be conducted in accordance with the following Good Laboratory Practice Regulations/Standards/Guidelines with the exception that analysis of the test material mixtures for concentration, solubility, homogeneity, and stability will not be conducted: [ ] 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) [ 3 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. Quality Assurance The protocol, study conduct, and the final report will be audited by the Quality Assurance Unit in accordance with Hazleton Wisconsin (HWI) Standard Operating Procedures (SOPs) and policies. 5. Test Material A. Identification T-6246 B. Physical Description (To be documented in the raw data) C. Purity and Stability The Sponsor assumes responsibility for purity and stability determinations (including under test conditions). Samples of test material/vehicle mixture(s) for concentration, solubility, homogeneity, and stability analyses will be taken before administration if requested by the Sponsor. These samples (if taken) will be sent to the Sponsor after experimental termination. D. Storage Room temperature 000116 000036 Page 30 of 38 TP8084.EXT Page 4 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 maintain a temperature of -20C 10C 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 material 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 Sterile water for injection B. Physical Description Clear, colorless liquid C. Purity and Stability The purity and stability of this USP grade material is considered adequate for the purposes of this study. D. Storage Refrigerated 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 7. Experimental Design A. Animals (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/HRP, Inc. 000117 000037 Page 31 of 38 TP8084.EXT Page 5 (3) Aoe at Initiation Adult (4) Weight at Initiation 2.5 to 3.5 kg (5) Number and Sex 20 males and 20 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, specified microbiological content, 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) Envi ronment Environmental controls for the animal room will be set to maintain a temperature of 19C to 23C, a relative humidity of 50% +20%, and a 12-hour 1ight/I2-hour dark cycle. (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. 000118 000038 Page 32 of 38 TP8084.EXT Page 6 (9) 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. B. Dose Administration (1) Test Groups GrouD Test/Control Materi al Dose Level Number of Animalsb (mg/kg)a Males Females 1 (Control) Sterile water 0 4 4 2 (Low) T-6246 5 4 4 3 (Medium) T-6246 10 44 4 (High) T-6246 100 4 4 5 (High) T-6246 500 44 a The dose volume will be 0.5 mL/kg of body weight, b Two animals/sex/dose level will be sacrificed on Day 15. The remaining animals (two animals/sex/dose level) will be sacrificed on Day 28. C. Dosing Procedures (1) Dosing Route Intravenous injection into the marginal ear vein of the right ear over approximately 30 to 60 seconds. (2) Reason for Dosing Route Intravenous injection is an acceptable route to assess systemic exposure. (3) Dosing Duration Single dose (4) Dose Preparation The day of treatment will be designated as Day 1. The Group 1 animals will be treated with sterile water at a dose volume of 0.5 mL/kg. The test material will be diluted with sterile water to achieve a specific concentration for each dose level in Groups 2-5. Individual doses will be calculated based on the animal's body weight taken just before test material administration. The prepared test mixtures will be stored at room temperature until administration. J 000119 :0 0 0 0 3 Page 33 of 38 TP8084.EXT Page 7 D. Observation of Animals (1) Clinical Observations The animals will be observed for clinical signs of toxicity before test or control material administration, at approximately 0.5, 2.0, 4.0 hours post-injection (Day 1), and daily thereafter for clinical signs and twice daily (a.m. and p.m.) for mortality until the scheduled sacrifice interval (Day 15 or Day 28). Observations may be extended when directed by the Study Director. (2) Body Weights For randomization, before test or control material injection (Day 1), at the scheduled sacrifice interval (Day 15 or Day 28), and at unscheduled death and sacrifices (when survival exceeds 1 day) (3) Sample Collections (a) Frequency 4-, 8-, 12-, 24-, and 48-hours post-injection, on Days 8, 15, 22, and at the scheduled sacrifice interval (Day 15 or Day 28) (b) Method of Collection/Number of Animals Blood samples (approximately 4 mL) will be collected from the marginal ear vein (left ear) of all animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. Additional samples will be collected on Days 15 and 22 for the animals scheduled for sacrifice on Day 28. Approximately 20 mL of blood (actual volume to be documented in the raw data) will be obtained from the posterior vena cava at the time of the scheduled sacrifice (Day 15 or Day 28). Approximately 20 mL of blood will be collected from moribund animals during the study, also, if possible. 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 a temperature of -20C 10C. The separated serum and cellular fractions will be sent frozen on dry ice to the Sponsor after experimental termination. The Sponsor is responsible for the retention and disposition of the samples. 000120 COOOmO TP8084.EXT Page 8 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 his alternate will be notified by telephone at (612) 778-5294 prior to the shipment of the samples. Termination (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. Tissues, as described in section 7.E. (3) Sample Collection, will be collected from any animal dying during the study or sacrificed in a moribund condition. After necropsy, the animals will be discarded. (2) Scheduled Sacrifices On Day 15, the first two animals/sex assigned to each dose level (based on the group assignment randomization) will be anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the vena cava, and exsanguinated. The remaining two animals/sex/dose level will be anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the vena cava, and exsanguinated on Day 28. An abbreviated gross necropsy examination will not be done, however, tissues (as described in section 7.E. (3) Sample Collection) will be collected. (3) Sample Collection The whole liver, bile, and both kidneys from each animal will be collected, weighed (volume only determined for bile), and immediately placed in a freezer set to maintain a temperature of -20C 10C. After sample collection, the animals will be discarded. The samples (liver, bile, and kidneys) will be sent frozen on dry ice to the Sponsor after experimental termination. The samples and their corresponding weights or volumes will be shipped to the person listed in Section 7.D.(3).(b). The Sponsor is responsible for the retention and disposition of the samples. 000121 Page 35 of 38 F . Statistical Analyses No statistical analyses are required. TP8084.EXT Page 9 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 Description of any toxic effects Gross pathology findings (if applicable) Gross pathology report (if applicable and requested by the Study Director) Individual animal tissue weights and bile volumes 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-1ife records Body weights Dose administration Observations Sample collection records Shipping records Pathology 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 .) 000122 or Page 36 of 38 TP8084.EXT Page 10 PROTOCOL APPROVAL L .J U C Z A a M ,7 John L. Butenhoff, PhD Sponsor's Representative 3M Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. Representative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-159.protdsk2) Date Date 3 -?o-73~ Date 000123 00004; HAZLETON WISCONSIN POST OFFICE BOX 7545 M A D I S O N , Wl 53 707 7545 Page 37 of 38 a C O R N IN G Company PROTOCOL TP8084.EXT Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits HWI 6329-159 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 Contractor Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, WI 53704 Study Director Steven M. Glaza Amendment No. 1 This amendment modifies the following portions of the protocol: Effective April 4, 1995 1. Page 6, 7, Experimental Design: B. Dose Administration; (1) Test Groups. The test material mixture for Group 5 could not be prepared at the concentration needed to utilize a dose volume of 0.5 mL/kg. Modify footnote "a" in this section with the following underlined addition: a The dose volume will be 0.5 mL/kg of body weight for Groups 1-4 and 0.75 mL/kg for Group 5. ; 000124 00004 P h on e 6 0 3 - 2 4 1 4 4 7 1 EXPRESS-MAIL DELIVERY 330 KINSMAN BLVD F a x 6 0.8 - 2 4 1 - 7.2 2.7 MADISON. Wl 53704 Page 38 of 38 Amendment No. HWI 6329-159 Page 2 Effective April 5, 1995 2. Page 6, 7. Experimental Design; B. Dose Administration; U ) Test Group?;. Animal No. F54076 (Group 5 male) and Animal No. F54106 (Group 4 male) were sacrificed on Days 1 and 2, respectively, due to injuries (apparent broken backs). Add the following paragraph to this section: Due to the sacrifice on Day 1 of one Group 5 male (Animal No. F54076) and on Day 2 of one Group 4 male (Animal No. F54106) because of injuries (apparent broken backs), replacement animals will be treated at the same dose levels in the same manner as for the initial animals in the study. The observations (clinical observations, body weights and sample collections) and the termination of the animals (unscheduled sacrifices and deaths, scheduled sacrifices, and sample collection) will be conducted in the same manner as for the other animals in the study terminating on Day 15. PROTOCOL AMENDMENT APPROVAL 2. John L. Butenhoff, PhD Sponsor's Representative 3M Toxicology Service Medical Department Date gz. z - Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. _________ Date Representative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-159.Ami.dsk3) Date 00012S 00004 9.1.2 Analytical protocol AMDT-042095.1 000126 0()004 3M Environmental Laboratory Protocol - Analytical Study Single-dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits In-Vivo Study Reference Number: H W I#6329-159 Study Number: AMDT-042095.1 Test Substance: FC-95 (T-6246) Name and Address of Sponsor: 3M SCD Division 367 Grove Street St. Paul, MN 55 1 0 6 Name and Address of Testing Facility: 3M Environmental Technology and Services 935 Bush Avenue St. Paul, MN 5 5 1 0 6 Proposed Initiation Date: July 25, 1995 Proposed Completion Date: August 25, 1995 Method Numbers and Revisions: AMDT-M-1-0, Thermal Extraction o f Fluoride by Means of a Modified Dohrmann D X 2000 Organic Halide Analyzer-Liver AMDT-M-2-0, Fluoride Measurement by Means o f an Orion E A 940 Expandable Ion Analyzer AMDT-M-4-0, Extraction o f Fluorochemicals from Rabbit Liver AMDT-M-8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode AMDT-M-14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer-Serum Author: James D. Johnson iproved. Tames ihnson Study frector /m/fs Date John Butenhoff, PhD Sponsor Date 000127 000047 i 1.0 PURPOSE This study is designed to provide information as to whether the perfluorooctanesulfonate anion does go to the rabbit liver and other tissues when the material is administered in an intravenous dose, and to ascertain the change in concentration with time after dose in semm and liver. 2.0 TEST MATERIALS 2.1 Test, Control, and Reference Substances and Matrices 2.1.1 Analytical Reference Substance: FC-95, lot 161 or 171. They are equivalent. 2.1.2 Analytical Reference Matrix: 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 of Materials: 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 Num ber of Test and Control Samples: Liver and serum from 32 test animals and 8 control animals. Two animals were replaced at 24 hours (animal F 54076 was replaced with animal F 54112, and animal F 54106 was replaced with animal F 54099). Other biological tissues (kidney, bile, cellular fraction) will be available for analysis if deemed appropriate by the Study Director. 2.4 Identification of Test and Control Samples: The samples are identified using the F1WI 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 of Reference Substance: To be determined by Sponsor. 2.7 Storage Conditions for Test Materials: Room temperature (2.1.1), -20 10C (2.1.2, 2.1.4). Test and Control samples will be received according to AMDT-S-10-0. 000128 C0C04S 2 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. Use caution when handling knives for cutting the samples. 3.0 EXPERIM ENT AL - Overview________________________________________ The tissues from animals dosed as described (H W I#6329-159), 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. For these analyses, bovine liver appears to be a good approximation to rabbit liver for matrix spikes and recovery determinations. Since perfluorooctanesulfonate anion is not biotransformed, fluorine content will be an accurate estimate o f the concentration o f the compound. 4.0 EXPERIM ENTAL - Methods__________________________________________ 4.1 Liver and Serum screening methods: (attached) 4.1.1 AMDT-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 AMDT-M-2-0, Fluoride Measurement by Means o f an Orion E A 940 Expandable Ion Analyzer 4.1.3 AMDT-M-4-0, Extraction o f Fluorochemicals from Rabbit Liver 4.1.4 AMDT-M-8-0, Analysis o f Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 4.1.5 AMDT-M-14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer-Serum 5.0 DATA ANALYSIS____________________________________________________ 5.1 Data Reporting: Data will be reported as a concentration (weight/weight) of fluoride per tissue or fluid, or as FC-95 per 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 of concentrations for 000129 0000-1 0 3 different dose groups. If necessary, simple statistical tests such as Student's t test may be applied to determine statistical difference. Half-life o f disappearance from serum will be determined from the linear regression o f the averages of the concentrations at different time points for the post distribution part o f the serum concentration versus time curve. 6.0 M AINTENANCE OF RAW DATA AND RECORDS_________________ __ 6.1 R aw D ata 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 AMDT-S-10-0, Sample Tracking System 8.0 ATTACHM ENTS___________________ 8.1 AMDT-M-1-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer-Liver 8.2 AMDT-M-2-0, Fluoride Measurement by Means o f an Orion EA 940 Expandable Ion Analyzer 8.3 AMDT-M-4-0, Extraction of Fluorochemicals from Rabbit Liver 8.4 AMDT-M-8-0, Analysis of Fluoride Using the Skalar Segmented Flow Analyzer with Ion Selective Electrode 8.5 AMDT-M-14-0, Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer-Serum 000130 CCOOSO 4 3M Environmental Laboratory Method Thermal Extraction of Fluoride by Means o f a Modified Dohrmann D X 2000 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 /a- /-fi" Date Software: MS Word 5.1a Affected Documents: AMDT-M-2 Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer AMDT-EP-3 Routine Maintenance of a Modified Dohrmann DX2000 Organic Halide Analyzer oooiai 1 000051 1.0 SCOPE . APPLICABLE COMPOUNDS. AND MATRICES 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 Compounds: Fluorochemicals or other fluorinated compounds. 1.3 Matrices: Biological tissues, particularly liver. 2.0 KEYW ORDS___________________________________________ _ _ _ _ 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann, halide, DX2000, fluorochemicals. 3.0 PRECAUTIONS____________________________________________________ 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 MATERIALS_____________________________________ 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 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 INTERFERENCES______________________________________________ ____ 6.1 Sample size is limited to approximately 150 mg, depending on sample moisture content. This may vary from matrix to matrix. 000132 2 0000 7.0 SAMPLE 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 of 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 CALIBRATION AND STANDARDIZATION_______________________ __ 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 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 Calibration - 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 of 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, of 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 of 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 of 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. 000133 3 0000 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 Standard Curve 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 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 of 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. 8.3.3.5 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. 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 PROCEDURES________________________________ 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. 000134 OOOO 3 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 of 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 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. 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 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 fOrion reading in ppm - intercept! (Slope) 10.0 VALIDATION 10.1 Quality 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 of different matrices and different reference compounds. 10.3 O ther Validation Parameters: NA 000135 5 OOOO" 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 Data 11.2.1 r^ 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% of the average of 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 ATTACHM ENTS__________________________________________________ None 13.0 REFERENCES 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 REVISIONS Revision Number Reason for Change Revision Date 000136 6 0000 j 3M Environmental Laboratory Method Fluoride Measurement by Means of an Orion EA 940 Expandable Ion Analyzer Method Identification Number: AMDT-M-2 Revision Number: 0 Adoption Date: Revision Date: None Author: Rich Youngblom Approved By: Grodt/ Leader / } /A/?s Date At 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 000137 1 00005 1.0 SCOPE . APPLICABLE COMPOUNDS. AND MATRICES 1.1 SCOPE: This method is for the calibration and operation of an Orion EA940 Expandable Ion Analyzer. 1.2 APPLICABLE COMPOUNDS: Fluoride. 1.3 APPLICABLE MATRICES: Liquid samples in an appropriate buffer solution. Preferred pH of 6.0. 2.0 KEYW ORDS_________________________________________________ 2.1 Fluoride, fluorine, ion selective electrode 3.0 PRECAUTIONS_____________ 3.1 No hazards identified with this method. 4.0 SUPPLIES AND MATERIALS 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 EQUIPM ENT_________________ _____________________________ 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 INTERFERENCES______________________ ___________________ 6.1 It is recommended that the pH be at or near 6.0. A 1:1 mixture of TISAB and sample/MilliQTM water will generally bring sample to pH of 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. 000138 OOOO.' 2 6.4 Air bubbles trapped under electrode can give erroneous readings. Make sure no air is trapped under electrode. 7.0 SAM PLE HANDLING______________________________________ 7.1 No special handling necessary. 8.0 CALIBRATION AND STANDARDIZATION_______________________ __ 8.1 Preparation of Calibration Standards 8.1.1 Measure 50 mL of 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 of 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 Calibration 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 of 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 of 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 PROCEDURES______________________________________________________ 9.1 C alibration and Measurement, 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. 000139 3 0000 9.2 Calibration And Measurement, Using Orion 3E Software: 9.2.1 Calibration: 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 of 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 Data 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 Measurement: 9.2.3.1 Follow steps 9.2.1 through 9.2.4 9.2.3.2 Enter the name of 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 VALIDATION_____________________________________________________ 10.1 Quality Control: 10.2 Precision and Accuracy 10.3 O ther Validation Parameters According to.Reference 13.2, the range of 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 ATTACHM ENTS_______________________________________________ __ None 13.0 REFERENCES____________________________________________________ 000140 4 0(100 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 for Change Revision Date 000141 5 00000 3M Environm ental Laboratory M ethod Extraction of Fluorochemicals from Rabbit Livers SOP Identification Number: AMDT-M-4 Revision Number: 0 Adoption Date: - r? /- v s; Revision Date: None Author: Dave Christenson/Cynthia Weber Approved By: J'uroup Leader / ' ,, iy ---------- - Quality Assurance Date ________________ ________________________ Date o 1 k Software: MS Word, 6.0 Affected Documents: M-5, Analysis of Rabbit Extract for Fluorochemicals Using Electrospray Mass Spectroscopy. 000142 OOOOO i 1.0 SC O PE___________________________________________________________ 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 Matrices: Rabbit Livers. 2.0 K EY W O R D S_____________________________________________________ 2 .1 Fluorochemicals, rabbit livers, electrospray mass spectrometer, fluorinated compounds, extraction. 3 .0 PR E C A U T IO N S____________________________________________ 3 .1 Use gloves when handling the rabbit livers, they may contain pathogens. 4 .0 SU PPLIES AND M ATERIALS_____________________________________ 4.1 Supplies 4 .1 . 1 Syringe, capable of measuring 100 )iL 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 jim. 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^Oj/NaHCC^) 0.25 M: Weigh 26.5 g of sodium carbonate (Na2C 0 3) and 21.0 g of sodium bicarbonate (N aH C03) 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 - perfluorooctanesulfonic acid (Aldrich) 4 . 2 . 8 FC-95 (3M Specialty Chemical Division) 000143 0000G3 2 5.0 EQUIPMENT____________________________________ 5 .1 Ultra-Tunrax T25 Grinder for grinding liver samples. 5 .2 Vortex mixer 5 .3 Centrifuge 5 .4 Shaker 5 .5 Analytical Evaporator 6.0 IN T E R FE R E N C E S_____________________ 6 .1 There are no known interferences at this time. 7.0 SA M PL E HANDLING ______________________________________________ 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 AND ST 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 the 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. 000144 00006 3 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 lppm 5 ppm 5 ppm uL - 100 200 300 400 500 200 300 Approximate final concentration of FC-95 in liver Blank 0.292 ppm 0.584 ppm 0.877 ppm 1.168 ppm 2.924 ppm 5.848 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 liver*/ 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 of FC-95 potassium salt. 8 .5 Calibration 8 . 5 . 1 Extract the spiked beef liver homogenate following 9.13 to 9.23 of 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 of 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. 2JLERQ.CEPURES_____________________________________ 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 ID, date and analyst initials. 000145 COCO 4 9.6 9.7 9. 8 9.9 9 .1 0 9.11 9.12 9.13 9 .1 4 9 .1 5 9 .1 6 9 .1 7 9 .1 8 9 .1 9 9 .2 0 9 .2 1 9 .2 2 9 .2 3 9.24 9 .2 5 Add 2.5 mLs water. 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. Rinse the probe off into the sample with 2.5 mLs water using a pipet. Take the grinder apart and clean it with methanol after each sample. Follow AMDT-EP-22. Cap the sample and vortex for 15 seconds. 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.) Spike the beef liver homogenates with the appropriate amount of FC-95 standard as described in 8.3. Spike the samples and beef liver homogenates with 100 uL of internal standard. Add 1 mL of the sodium carbonate/sodium bicarbonate buffer and 1 mL ammonium acetate. Using an analytical pipet, add 5 mL ethyl acetate. Cap the sample and vortex 20 to 30 seconds. Put them in the shaker for 20 min. Centrifuge for 20 to 25 minutes, until the layers are well separated. Set the power on the centrifuge to 25. 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. Blow the sample down on the analytical evaporator to near dryness with nitrogen, approximately 30 to 40 minutes. Bring the remaining sample up in 1 mL dilute acetonitrile with an analytical pipet. Vortex 15 seconds. Transfer the sample to a 3 mL syringe. Attach a 0.2 |m 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. Cap and hold for analysis by electrospray mass spectroscopy. Complete AMDT-M-4 worksheet and attach to page of study notebook. 10.0 V A L ID A T IO N _________________________________ 1 0 .1 Quality Control - not applicable 1 0 .2 Precision and Accuracy- not applicable 1 0 .3 Other Validation Parameters- not applicable 1 1.0 D A TA A N ALYSIS_____________________________ 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 1 4.0 R E V ISIO N S_____________ Revision Number Reason for Change Revision Date 000146 00006C 5 W orksheet AMDT-M-4 Study # - _ - _ - 1 Sam ple FC-95 N um ber approx 0.5 ppm actual ppm set # #W B la n k L iv e r ____ - ________ lO C u L . 2 0 0 nL ________3 0 0 uL ________ 4 0 0 uL _ _ _ _ _ _ _ _ _ - _ _ F C -95 approx 1 ppm actual ppm #W - _ _ 500 uL _ _ - _ _ _ _ _ F C -95 approx. 5 ppm actual ppm #W > _ 200 uL .100 uL _ . _ _ _ _ _ _ D ate and In itials for Std. _ ' s tu d y n u m b e r v /h e r e th e o r ig in a l w o r k s h e e t is lo c a te d a nri n la c e a c o n v .____ ___________________ r ____________ r _____ i i L iv e r E x tr a c tio n P m c e s v __________ __________________ P a t e & In itia ls__ P in et 1 m L n f L iver Solution P in e t 1 0 0 u L n f __ 1 2 n n m Intern al S ta n d a rd _____________ S td . # ____________ V nrtex. 1 5 sen P in e t 1 mT. 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 m L o f 0 .2 5 N a ,C O ,/0.25M N aH C O , Buffer Pinet. 5 mT. o f F.thvl A cetate Vnrtex 2 0 - 3 0 sec Shake 2 0 m in. C entrifiipe 20-2.5 m in R e m o v e a 4 m l . a liq u o t o f o r g a n ic la v e r __________________ B lo w d ow n to near d rvness (< 0 2 5 m L) w ith N , A dd 1 n r n f 1 1 Aeetonitrile/T-LO _ Vnrtex 1 5 sec T N # ... . F i l t e r u s in g a S e e B - D s y r in g e w ith a 0 2 n m S R I f ilt e r in to a 1 5 m l . a u tn s a m n le v ia l._______________________________ 000147 000067 6 3M Environmental Laboratory M ethod Analysis of Fluoride Using the Skalar Segm ented Flow Analyzer 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 Row Analyzer 000148 00006S i 1,Q SCOPE______________________________________ 1 .1 This method is for the analysis for fluoride, thermally extracted from samples using the Dohrmann DX2000 (AMDT-M-l), 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 SU PPL IE S AND M ATERIALS__________________________________ ___ 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 0.06 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 - 000149 2 0000(9 1.2 ppm 1.5 ppm 1.2 1.5 - 5,9 EQUIPMENT________________________________________ 5 .1 Skalar Segmented Flow Auto Analyzer Sanspiui System equipped with ISE 6 .0 IN T E R FE R E N C E S_______________________________________________ 6 .1 High concentrations of alkalinity, chloride, phosphate, sulfate or iron can cause interferences. 7 .0 SA M PLE H A N D LIN G _______________________________________i . 7 .1 Samples should be stored in polyethylene bottles. Samples should be analyzed within 30 days. 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 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. 2.,Q PROCEDURE_________________________________________________________ 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 Tum on the interface, computer, display and printer. Make sure you turn on the interface before the computer. 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 of 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. 000150 3 9 .2 .5 . I 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 . 2 Use 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 . 3 Check 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, and Enter. 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, tum off the sampler, pump and offset potentiometer. Tum 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 _____________________________________________________ 1 0 .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. 000151 000071 1 0 .2 Precision and Accuracy 10. ? . lSee Method Validation Report number AMDT-M-8.0.V1 1 0 .3 Other Validation Parameters 1 0 .4 Refer to Method Validation Report Number AMDT-M-8.0. V 1 11.0 D ATA ANALYSIS___________________________________________ _____ 11.1 1 1 .2 11.3 Calculations 1 1 .1 . IThe 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. Prepare spreadsheets to summarize data. Include sample volume, weights used etc. 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. 1 2 .0 A TT A C H M EN TS___________________________________________________ None 13.0 R E FE R E N C E S_________ :____________________________________________ 1 3 .1 1 3 .2 1 3 .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 1 4 .0 R E V ISIO N S__________________________:__________________ ___________ Revision Number Reason for change Revision Date 000152 5 0000 3M Environm ental Laboratory Method Thermal Extraction o f Fluoride by Means o f a Modified Dohrmann D X 2000 Organic Halide Analyzer - Serum Method Identification Number: AMDT-M-14 Revision Number: 0 Adoption Date: j o ~ Revision Date: None Author Rich Youngblom Approved by: V r <=\ J ^r o u p Leader y /0/ 3 / ? s Date Quality Assurance r~ Date Software: MS Word 5.1a Affected Documents: AMDT-M-2 Fluoride Measurement by Means of an Orion EA940 Expandable Ion Analyzer AMDT-EP-3 Routine Maintenance of a Modified Dohrmann DX2000 Organic Halide Analyzer 000153 0000 1.0 SCOPE . APPLICABLE COMPOUNDS. AND MATRICES 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 Compounds: Fluorochemicals or other fluorinated compounds. 1.3 Matrices: Biological fluids, particularly serum. 2.0 K EYW O RD S_______________________________________________________ 2.1 Fluoride, fluorine, extraction, pyrolysis, ionization, ion selective electrode, Dohrmann,. halide, DX2000, fluorochemicals. 3.0 PRECAUTIONS____________________________________________________ 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 MATERIALS_____________________________________ 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 EN T_____________________________________________________ 5.1 Rosemount Dohrmann DX2000 Organic Halide Analyzer, modified for flupride 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 INTERFERENCES________________________________________________ 6.1 Sample size is limited to approximately 100 (il. This may vary from matrix to matrix. 000154 00007 2 7.0 SAMPLE HANDLING 7.1 Samples are to be handled with plastic pipettes. A new pipette is to be used for each sample. 8.0 CALIBRATION AND STANDARDIZATION 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 of 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, of 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 of 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 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 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. 000155 00007 3 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 IOOjuL 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 of 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 of 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. 8.3.3.7 When all standards have been run, calculate the 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 PROCEDURES 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 of the Dohrmann software. 9.3 Sample Extraction Procedure: 9.3.1 Open the SAMPLE HATCH and pipette lOOpL of 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. 000156 4 0000 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 raL 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-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 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 and/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 Quality Control 10.1.1 Daily Start 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 of different matrices and different reference compounds. 10.3 O ther Validation Parameters: NA 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. 000157 5 oooo 11.2 Analyzing the Data 11.2.1 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% of the average of 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 ATTACHM ENTS______________________________________________ _ None 13.0 REFERENCES_______________________________________________ __ 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 REVISIONS Revision Number Reason for Change Revision Date 000158 9.3 Quality Assurance Unit Statement 000159 Attachment D GLP Study Quality Assurance Statement Original to: Study birestor Copies to; QAtlFiles Study Tide: Single-dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Study Number: AMDT-042095.1 Name of Auditor: KariRambo 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 To ______ Phase_____________ 11-13-95 11-13-95 Final Report Date Inspection Reported to Management Study Director 11-13-95 11-13-95 /QAU Auditor Date 000180 000o s o 1 9.4 Key Personnel Involved in the Study 000161 000081 3M Environmental Laboratory Key Personnel Thermal extraction followed by analysis using Orion ion analyzer: Jim Johnson * Deb Wright 1 Rich Youngblom Deann Plummer Thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode: Jim Johnson Deb Wright Rich Youngblom Deann Plummer Documentation and Reporting: Jim Johnson Rich Youngblom Quality Assurance Unit: Gale Van Buskirk Cynthia Weber Kari Rambo 000162 00008 9.11 Data 000163 000083 9.11.1 Summary and raw data; ug F' in whole liver as determined by thermal extraction followed by analysis using Orion ion analyzer. 000164 000084 Summary of Combustion Data - Liver AMDT-042095.1, HWI 6329-159 As Referenced in Final Report section 6.0 D ATA A N A L Y S IS Total jig Fluoride in Whole Liver Mean per Dose Group per Day Sacrifised* (ig Std. Dev. Control Group, Day 15** 27.1 3.1 Control Group, Day 28 38.7 16.3 100 mg/kg dose (T6246), Day 15 37.4 7.9 (0.12 mg/kg)*** 100 mg/kg dose (T6246), Day 28 29.6 1.3 (0.12 mg/kg)*** 500 mg/kg dose (T6246), Day 15 123.1 14.0 (0.60 mg/kg)*** 500 mg/kg dose (T6246), Day 28 100.5 5.9 (0.60 mg/kg)*** *CalcuIated as a mean triplicate samples from each of two male and two female rabbits. **Only two livers analyzed from this group. ***T est material is a 0.12% solution of FC-95, actual dose in parenthesis. 000165 000085 FC95 ID liver blank-1 liver blank-2 liver blank-3 F54075-1 F54075-2 F54075-3 F54081-1 F54081-2 F54081-3 F54105-1 F54105-2 F54105-3 F54107-1 F54107-2 F54107-3 F54074-1 F54074-2 F54074-3 F54085-1 F54085-2 F540853 Liver Blk-1 Liver Blk-2 Liver Spk-1 Liver Spk-2 Liver Spk-3 Liver Spk-4 Liver Spk-5 blank 1 blank 2 spike-1 spike-2 spike-3 spike-4 spike-5 spike-6 blk 1 blk 2 spk 1 spk 2 spk 3 spk 4 F54082-1 F54082-2 F54082-3 F54111-1 F54111-2 F54111-3 Actual Average ppm F- ppm F% in liver in liver rcvry (W/W) (W/W) 0.776 0.500 0.480 0.401 0.523 0.412 0.313 0.308 0.448 0.387 0.404 0.267 0.431 0.339 0.319 1.34 0.542 0.780 0.459 0.350 0.265 0.336 0.393 0.312 0.272 0.407 0.637 0.437 0.185 40% 0.553. 45% 0.469 40% 0.389 39% 0.418 54% 0.660 0.528 0.334 91% 1.26 106% 1.11 118% 1.15 97% 1.04 136% 1.65 96% 1.06 0.530 0.264 119% 1.15 113% 1.14 106% 0.979 103% 1.11 1.22 1.25 1.23 1.21 1.21 1.11 1.17 1.17 Whole Total F- in liver liver whole burned weight liver (grams) (grams) 0.113 (ug) 0.117 0.131 0.131 60.3 0.120 60.3 24.8 0.132 60.3 0.125 80.6 0.116 80.6 31.2 0.108 80.6 0.151 86.4 0.105 86.4 29.3 0.117 86.4 0.107 80.6 0.135 80.6 62.9 0.129 80.6 0.123 82.9 0.144 82.9 27.9 0.111 82.9 0.149 81.0 0.149 81.0 33.0 0.101 81.0 0.097 0.148 0.110 0.145 0.156 0.142 0.125 0.097 0.148 0.110 0.145 0.156 0.142 0.125 0.137 0.153 0.161 0.157 0.150 0.164 0.141 0.160 89.0 0.163 89.0 109.4 0.156 89.0 0.121 83.8 0.115 83.8 97.7 0.136 83.8 Dosage (mg/kg) 0.0 0.0 0.0 0.0 0.0 0.0 500 500 Page 1 000166 RPT159L.XLS 00086 FC95 Actual Average ppm F- ppm F- ID % in liver in liver rcvry (W/W) (W/W) F54079-1 1.17 F54079-2 1.20 1.18 F54079-3 1.16 F54116-1 1.36 F54116-2 1.33 1.35 F54116-3 1.36 F54112-1 1.44 F54112-2 1.55 1.46 F54112-3 1.40 F54076-1 1.69 F54076-2 1.81 1.76 F54076-3 1.78 F54113-1 1.24 F54113-2 1.33 1.33 F54113-3 1.42 F54084-1 1.58 F54084-2 1.73 1.66 F54084-3 1.68 F54098-1 1.46 F54098-2 1.39 1.49 F54098-3 1.61 F54087-1 0.400 F54087-2 0.346 0.376 F54087-3 0.382 F54088-1 1.33 F54088-2 0.485 0.765 F54088-3 0.482 LIVER BLK 1 0.259 LIVER BLK 2 0.132 LIVER SPK 1 89% 1.05 LIVER SPK 2 111% 1.30 LIVER BLANK 0.243 LIVER SPIKE 3 111% 1.27 liver blank 0.355 F54089-1 0.398 F54089-2 0.394 0.394 F54089-3 0.391 F54099-1 0.502 F54099-2 0.504 0.483 F54099-3 0.444 F54114-1 0.337 F54114-2 0.372 0.359 F54114-3 0.367 F54109-1 0.383 F54109-2 0.387 0.400 F54109-3 0.431 F54080-1 0.460 F54080-2 0.475 0.356 F54080-3 0.132 Whole Total F- in liver liver whole burned weight liver Dosage (grams) (grams) (ug) (mg/kg) 0.157 83.1 0.142 83.1 97.9 500 0.141 83.1 0.154 72.0 0.140 72.0 97.1 500 0.157 72.0 0.134 97.8 0.158 97.8 142.9 500 0.146 97.8 0.139 78.1 0.124 78.1 137.4 500 0.114 78.1 0.145 91.5 0.123 91.5 121.7 500 0.117 91.5 0.134 66.4 0.140 66.4 110.4 500 0.120 66.4 0.135 79.0 0.147 79.0 117.3 500 0.125 79.0 0.130 83.7 0.131 83.7 31.5 100 0.135 83.7 0.134 84.3 0.148 84.3 64.4 100 0.134 84.3 0.164 0.158 0.128 0.129 0.140 0.133 0.111 0.153 71.8 0.157 71.8 28.3 100 0.147 71.8 0.139 82.1 0.140 82.1 39.7 100 0.158 82.1 0.157 81.0 0.136 81.0 29.0 100 0.155 81.0 0.133 74.0 0.134 74.0 29.6 100 0.154 74.0 0.132 75.4 0.132 75.4 26.8 100 0.103 75.4 Page 2 000167 RPT159L.XLS COO0S7 FC95 ID F54092-1 F54092-2 F54092-3 F54106-1 F54106-2 F54106-3 Blank Liver spike 1 Liver spike 2 Liver spike 3 Liver spike 4 Liver spike 5 Liver spike 6 liver blank-1 liver blank-2 liver blank-3 liver spike 63-1 liver spike 63-2 liver spike 63-3 liver spike 63-4 F54088-1 F54088-2 F54088-3 liver blank-4 liver spike 63-1 liver spike 63-2 liver spike 63-3 liver spike 63-4 Actual Average ppm F- ppm F% in liver in liver rcvry (W/W) (W/W) 0.537 0.463 0.478 0.433 0.639 0.665 0.634 0.598 0.181 82% 1.09 81% 0.755 85% 0.897 76% 1.47 66% 1.36 70% 1.40 0.549 0.454 0.278 103% 1.09 96% 1.11 159% 1.69 101% 1.41 0.485 0.655 0.544 0.491 0.292 117% 1.30 89% 0.972 89% 0.922 90% 0.910 Whole Total F- in liver liver whole burned weight liver (grams) (grams) 0.117 78.4 (ug) 0.145 78.4 0.138 78.4 37.4 0.134 82.3 0.125 82.3 0.121 82.3 52.2 0.133 0.113 0.158 0.144 0.156 0.147 0.152 0.147 00..111303 0.143 0.132 0.143 0.108 0.145 84.3 0.123 84.3 45.8 0.132 84.3 0.123 0.136 0.139 0.147 0.149 Dosage (mg/kg) 100 100 100 RPT159L.XLS Page 3 000168 ooooss 9.11.2 Summary and raw data; ug F' in whole liver as determined by thermal extraction followed by analysis using Skalar segmented flow analyzer with ion selective electrode. 000169 0000 RE: 6329-159 LIUER SAMPLES RMDT 42095.1 Date o f Analysis: 5-23 and 5-24-95 Rnalyst: DDLU The samples are burned in the Dohrman at 950 C using between 0.1 and 0.2 grams of the liuer. The gas is collected in 2.0 mL o f 1:1 TISRB/Milli-Q w ater. The samples are then analyzed on a Skalar Segmented Flow Analyzer using the Ion Specific Electrode (ISE) Method. TISRB b u f f e r 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 selectiue electrode and the reference electrode is measured. The signal is amplified and related to the fluoride concentration. The in strum en t was calibrated in the ranges o f 0.015 - 0.15 ppm and 0.15 - 1.50 ppm fluoride. The standard curue fo r the high range was plotte d using the inuerse logarithm option. The standard curue f o r the low range is linear. Rll standards and samples were then calculated by the Skalar software using these curues. Rll results below 0.0001 ppm appear on the raw data as # . # # # # . R quality control standard was analyzed euery 10 samples to check f o r accuracy and drift. Raw data is taken from the appropriate calibrated range of the Skalar prin to u t and summarized on an Excel spreadsheet. The final results are adjusted fo r the collection uolume and any subsequent dilutions. 000170 ooooyo GROUP 1 Dose Level : 0 c ''4i F54075-1 F54075-2 F54075-3 F54081-1 F54081-2 F54081-3 F54105-1 F54105-2 F 54105-3 F54107-1 F 54107-2 F54107-3 F54074-1 F54074-2 F54074-3 F54085-1 F54085-2 F54085-3 u ) ^1-2.1\=vs SUMMARY of 6329-159 LIVER SAMPLES AMDT 42095.1 0.03 0.03 0.02 0.02 0.02 0.02 0.02 0.02 0.02 0.45 0.05 0.03 0.02 0.02 0.02 0.02 0.02 0.06 SjHHBpjBil 2.0 0.1311 0.50 60.2500 30.06 2.0 0.1197 0.55 0.52 60.2500 33.32 27.4 2.0 0.1318 0.31 60.2500 18.93 2.0 0.1246 0.32 80.597 25.87 2.0 0.1161 0.36 0.33 80.597 29.30 26.8 2.0 0.1075 0.31 80.597 25.34 2.0 0.1507 0.25 86.35 21.54 2.0 0.1053 0.46 0.36 86.35 39.53 31.2 2.0 0.1173 0.38 86.35 32.39 2.0 0.1069 8.38 80.649 675.67 2.0 0.1351 0.75 3.19 80.649 60.53 257 2.0 0.1286 0.43 80.649 34.37 2.0 0.1227 0.37 82.895 31.08 2.0 0.1441 0.25 0.34 82.895 21.05 28.3 2.0 0.1110 0.40 82.895 32.86 2.0 0.1485 0.23 80.95 18.97 2.0 0.1490 0.30 1.81 80.95 23 .9 0 45.8 2.0 0 .1005 1.17 80.95 94.40 GROUP4 Dose Level : 100 m g/kg F54087-1 F54087-2 F54087-3 F54088-1 F54088-2 F54088-3 F54092-1 F54092-2 F54092-3 F 54106-1 F 54106-2 F 54106-3 F54088-1 F54088-2 F54088-3 F54089-1 F54089-2 F54089-3 F54099-1 F54099-2 F54099-3 F54114-1 F 54114-2 F 54114-3 F54109-1 F 54109-2 F 54109-3 0.04 0.03 0.03 0.11 0.05 0.05 0.06 0.05 0.04 0.06 0.06 0.04 0.05 0.05 0.04 0.03 0.03 0.03 0.04 0.04 0.05 0.03 0.03 0.03 0.03 0.03 0.03 000171 2.0 0.1301 0.59 2.0 0.1314 0.47 2.0 0.1351 0.51 2.0 0.1339 1.70 2.0 0.1476 0.66 2.0 0.1344 0.68 2.0 0.1169 0.96 2.0 0.1453 0.69 2.0 0.1381 0.60 2.0 0.1336 0.85 2.0 0.1249 0.90 2.0 0.1213 0.71 2.0 0.1448 0.72 2.0 0.1228 0.89 2.0 0.1317 0.63 2.0 0.1532 0.42 2.0 0.1568 0.41 2.0 0.1474 0.39 2.0 0.1386 0.56 2.0 0.1403 0.53 2.0 0.1575 0.59 2.0 0.1573 0.39 2.0 0.1359 0.38 2.0 0.1552 0.40 2.0 0.1326 0.38 2.0 0.1339 0.38 2.0 0.1536 0.45 HWI-159L.SUM 0.52 1.01 0.75 0.82 0.75 0.41 0.56 0.38 40.30 83.71 83.71 83.71 84.26 84.26 84.26 78.36 78.36 78.36 82.301 82.301 82.301 84.26 84.26 84.26 71.796 71.796 71.796 82.143 82.143 82.143 80.966 80.966 80.966 73.958 73.958 73.958 49.42 39.24 42.51 143.47 55.26 57.30 75.61 54.15 46.75 70.35 74.33 58.76 60.63 75.07 53.49 29.81 29.12 28.15 46.35 43.21 48.29 31.19 31.10 32.14 28.45 28.17 33.13 43.7 85.3 58.8 67.8 63.1 29 45.95 31.5 29.9 Page 1 000031 SUMMARY of 6329-159 LIVER SAMPLES AMDT 42095.1 W l Saiapte J " Resali' - m sm dSlBiki F54080-1 0.03 F54080-2 0.03 F54080-3 ND 2.0 2.0 2.0 0.1322 0.1323 0.1028 0.46 0.50 ND 0.32 75.42 75.42 75.42 34.91 37.74 ND 24.2 GROUP 5 Dose Level : 500 m g/kg F54082-1 F54082-2 F54082-3 F54111-1 F54111-2 F54111-3 F54079-1 F54079-2 F54079-3 F54116-1 F 54116-2 F 54116-3 F54112-1 F 54112-2 F 54112-3 F54076-1 F54076-2 F54076-3 F54113-1 F54113-2 F54113-3 F54084-1 F54084-2 F54084-3 F54098-1 F54098-2 F54098-3 0.12 0.12 0.11 0.09 0.08 0.10 0.11 0.11 0.10 0.13 0.12 0.13 0.12 0.15 0.13 0.15 0.15 0.13 0.12 0.11 0.11 0.14 0.16 0.13 0.13 0.14 0.13 2.0 0.1595 1.44 2.0 0 .1626 1.48 2.0 0 .1558 1.47 2.0 0 .1 2 0 7 1.41 2.0 0 .1 1 4 7 1.34 2.0 0 .1360 1.50 2.0 0.1571 1.46 2.0 0 .1420 1.52 2.0 0 .1 4 0 6 1.46 2.0 0.1 5 4 1 1.71 2.0 0.1403 1.68 2.0 0 .1 5 7 0 1.68 2.0 0 .1 3 4 0 1.82 2.0 0.1583 1.84 2.0 0 .1458 1.79 2.0 0.1385 2.22 2.0 0.1243 2.43 2.0 0.1136 2.35 2.0 0.1449 1.65 2.0 0 .1227 1.85 2.0 0.1171 1.94 2.0 0.1338 2.09 2.0 0.1402 2.24 2.0 0.1200 2.22 2.0 0 .1 3 4 6 1.93 2.0 0.1469 1.88 2.0 0.1253 2.12 1.46 1.42 1.48 1.69 1.82 2.33 1.81 2.18 1.98 88.965 88.965 88.965 83.823 83.823 83.823 83.117 83.117 83.117 71.962 71.962 71.962 97.824 97.824 97.824 78.058 78.058 78.058 91.52 91.52 91.52 66.42 66.42 66.42 78.95 78.95 78.95 128.40 131.75 130.42 118.48 112.69 125.36 121.58 126.67 121.07 122.82 121.15 120.73 177.98 180.45 175.25 173.59 189.65 183.60 151.08 169.17 177.26 138.90 148.76 147.23 152.03 148.33 167.23 130 119 123 122 178 182 166 145 156 000172 HWI-159LSUM Page 2 CPOO2 HWI-159L.XLS 1995-06-13 10:27 OutPut of : 950523A1 Operator : DDW Date of the Analysis : 1995-05-23 10:18 Analysis File Name : C:\SKALAR\DATA\HWIDATA\LIVERS\950523A1 000173 1 Tracer 1.50 1.47 98% 2 Drift 1.50 1.48 98% 3 Wash 0.00 4 Standard 1 0.015 0.02 125% 5 Standard 2 0.03 0.03 85% 6 Standard 3 0.06 0.06 100% 7 Standard 4 0.09 0.09 102% 8 Standard 5 0.12 0.12 99% 9 Standard 6 0.15 0.15 103% 10 Standard 7 0.30 0.29 95% 11 Standard 8 0.60 0.61 102% 12 Standard 9 1.20 1.24 103% 13 Standard 10 1.50 1.46 97% 14 Drift 1.50 1.53 102% 15 Wash 0.00 16 BLK 1 0.05 2.0 0.1332 0.79 17 BLK 2 0.04 2.0 0.1172 0.60 18 BLK 3 0.04 2.0 0.1313 0,60 19 F54075-1 0.03 2.0 0.1311 0.50 60.2500 30.06 20 F54075-2 0.03 2.0 0.1197 0.55 60.2500 33.32 21 F54075-3 0.02 2.0 0.1318 0.31 60.2500 18.93 22 F54081-1 0.02 2.0 0.1246 0.32 80.597 25.87 23 F54081-2 0.02 2.0 0.1161 0.36 80.597 29.30 24 F54081-3 0.02 2.0 0.1075 0.31 80.597 25.34 25 F54105-1 0.02 2.0 0.1507 0.25 86.35 21.54 26 Drift 1.50 1.53 102% 27 Wash 0.00 28 F54105-2 0.02 2.0 0.1053 0.46 86.35 39.53 29 F54105-3 0.02 2.0 0.1173 0.38 86.35 32.39 30 F54107-1 0.45 2.0 0.1069 8.38 80.649 675.67 Page 1 000003 HWI-159L.XLS 31 F54107-2 32 F54107-3 33 F54074-1 34 F54074-2 35 F54074-3 36 F54085-1 37 F54085-2 38 Drift 1.50 39 Wash 40 F54085-3 41 BLK 1 42 BLK2 43 SPK 1 44 SPK 2 45 SPK 3 46 SPK 4 000174 47 SPK 5 48 SPK 6 49 BLK 1 50 Drift 1.50 51 Wash 52 BLK 2 53 SPK 1 54 SPK 2 55 SPK 3 56 SPK 4 57 F54082-1 58 F54082-2 59 F54082-3 60 F54111-1 61 F54111-2 62 Drift 1.50 63 Wash ** 64 F54111-3 65 F54079-1 m 66 F54079-2 0.05 0.03 0.02 0.02 0.02 0.02 0.02 1.52 0.00 0.06 0.02 0.03 0.15 0.10 0.11 0.09 0.12 0.09 0.04 1.53 0.00 0.02 0.11 0.10 0.10 0.11 0.12 0.12 0.11 0.09 0.08 1.51 0.00 0.10 0.11 0.11 101% 102% 101% 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 0.1351 0.75 80.649 60.53 0.1286 0.43 80.649 34.37 0.1227 0.37 82.895 31.08 0.1441 0.25 82.895 21.05 0.111 0.40 82.895 32.86 0.1485 0.23 80.95 18.97 0.149 0.30 80.95 23.90 0.1005 0.0971 0.1479 0.1096 0.1446 0.1556 0.1416 0.1246 0.1369 0.153 1.17 0.50 0.39 2.67 1.36 1.39 1.21 1.97 1.32 0.58 80.95 94.40 0.004 0.004 0.004 0.004 0.004 0.004 63.00 63.00 63.00 63.00 63.00 63.00 0.1609 0.27 0.1568 1.37 0.004 0.1502 1.35 0.004 0.1636 1.17 0.004 0.1412 1.51 0.004 0.1595 1 .4 4 88.965 1 2 8 . 4 0 0.1626 1 .4 8 88.965 1 3 1 . 7 5 0.1558 1 .4 7 88.965 1 3 0 . 4 2 0.1207 1 .4 1 83.823 1 1 8 . 4 8 0.1147 1 .3 4 83.823 1 1 2 . 6 9 63.00 63.00 63.00 63.00 0.136 0.1571 0.142 1.50 1.46 1.52 Page 2 83.823 83.117 83.117 125.36 121.58 126.67 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.29 0.20 0.22 0.17 0.25 0.18 194% 130% 143% 114% 162% 119% 0.22 0.20 0.19 0.21 142% 134% 126% 141% Stator Stator S m pte s' SiUjpf s in to n i ^ ' D /f$ ' 67 F54079-3 68 F54116-1 69 F54116-2 70 F54116-3 71 F54112-1 72 F54112-2 73 F54112-3 74 Drift 75 Wash 76 F54076-1 77 F54076-2 78 F54076-3 79 F54113-1 80 F54113-2 81 F54113-3 82 F54084-1 83 F54084-2 84 F54084-3 85 F54098-1 86 Drift 87 Wash 88 F54098-2 89 F54098-3 90 F54087-1 91 F54087-2 92 F54087-3 93 F54088-1 94 F54088-2 95 F54088-3 96 Drift 97 Wash 1.50 1.50 1.50 0.10 0.13 0.12 0.13 0.12 0.15 0.13 1.48 0.00 0.15 0.15 0.13 0.12 0.11 0.11 0.14 0.16 0.13 0.13 1.49 0.00 0.14 0.13 0.04 0.03 0.03 0.11 0.05 0.05 1.49 0.00 HWI-159L.XLS % , PtTO A f* QiySantpl Achat! , Totol Tcfa V, i,FC95 ; {0 , % , , T ts s ^ W tfp e r Uss^b 2 . 0 0.1406 1 . 4 6 83.117 1 2 1 . 0 7 2 . 0 0.1541 1 .7 1 71.962 1 2 2 . 8 2 2 . 0 0.1403 1 . 6 8 71.962 1 2 1 . 1 5 2.0 0.157 1 . 6 8 71.962 1 2 0 . 7 3 2.0 0.134 1 . 8 2 97.824 1 7 7 . 9 8 2 . 0 0.1583 1 . 8 4 97.824 1 8 0 . 4 5 2 . 0 0.1458 1 .7 9 97.824. 1 7 5 . 2 5 99% 2 . 0 0.1385 2 . 2 2 78.058 1 7 3 . 5 9 2.0 0.1243 2.43 78.058 1 8 9 . 6 5 2 . 0 0.1136 2 . 3 5 78.058 1 8 3 . 6 0 2.0 0.1449 1.65 91.52 151.08 2.0 0.1227 1.85 91.52 169.17 2.0 0.1171 1.94 91.52 177.26 2.0 0.1338 2.09 66.42 138.90 2.0 0.1402 2.24 66.42 148.76 2.0 0.12 2.22 66.42 147.23 2.0 0.1346 1.93 78.95 152.03 99% 2.0 0.1469 1.88 78.95 148.33 2.0 0.1253 2.12 78.95 167.23 2.0 0.1301 0.59 83.71 49.42 2.0 0.1314 0.47 83.71 39.24 2.0 0.1351 0.51 83.71 42.51 2.0 0.1339 1.70 84.26 143.47 2.0 0.1476 0.66 84.26 55.26 2.0 0.1344 0.68 84.26 57.30 99% S4.TOOO L'GOOOO Page 3 HWI159L2.XLT 1995-06-13 10:27 OutPut of : 950524A1 Operator : DDW Date of the Analysis : 1995-05-24 07:50 Analysis File Name : C:\SKALAR\DATA\HWIDATA\LIVERS\950524A1 ,, Skalar Skalar ' % , DTJTSAB QtySampI Actual Total 000176 1 Tracer 1.50 1.45 97% 2 Drift 1.50 1.47 98% 3 Wash 0.00 4 Standard 1 0 . 0 1 5 0.02 107% 5 Standard 2 0.03 0.03 94% 6 Standard 3 0 . 0 6 0.06 101% 7 Standard 4 0.09 0.09 103% 8 Standard 5 0 . 1 2 0.12 97% 9 Standard 6 0.15 0.16 103% 10 Standard 7 0.30 0.28 94% 11 Standard 8 0 . 6 0 0.61 102% 12 Standard 9 1.20 1.24 103% 13 Standard 10 1 .5 0 1.46 97% 14 Drift 1.50 1.50 100% 15 Wash 0.00 16 F54092-1 0.06 2.0 0.1169 0.96 78.36 75.61 17 F54092-2 0.05 2.0 0.1453 0.69 78.36 54.15 18 F54092-3 0.04 2.0 0.1381 0.60 78.36 46.75 19 F54106-1 0.06 2.0 0.1336 0.85 82.301 70.35 20 F54106-2 0.06 2.0 0.1249 0.90 82.301 74.33 21 F54106-3 0.04 2.0 0.1213 0.71 82.301 58.76 22 BLK 0.01 2.0 0.1326 0.18 23 SPK 1 0.08 2.0 0.1133 1.34 0.004 63.00 24 SPK 2 0.08 2.0 0.1577 0.99 0.004 63.00 25 SPK 3 0.09 2 . 0 0.1441 1 .1 9 0.004 63.00 26 Drift 1.50 1.45 97% 27 Wash 0.00 28 SPK 4 0.15 2.0 0.1556 1.89 0.004 126.00 29 SPK 5 0.13 2.0 0.1474 1.72 0.004 126.00 30 SPK 6 0.14 2.0 0.1518 1.82 0.004 126.00 Page 1 0.15 0.15 0.15 0.15 0.16 0.17 100% 104% 113% 0.30 0.30 0.30 0.29 0.25 0.28 97% 84% 91% 000036 . s % & - , Skalar f . / '& , ' ' p '^'^Iprafn) ''"' (nptft) ' "" HWI159L2.XLT v ',I>fcTJS4B QtySamp " Attuai, ToM ~ Total f y , R&F95, o m ,, , Msss 'M iss ,, Y* ' {mW * j ' " ' gitns)' ;Tjssr WL' ; psrii5M 'i$tfop''. I g l f e Lfe- ' ' istffF-3: xsT* cw'v 31 LIVER BLK-1 CCMO LIVER BLK-2 33 LIVER BLK-3 34 SPK 63-1 35 SPK 63-2 36 SPK 63-3 37 SPK 63-4 38 Drift 1.50 39 Wash 40 F54088-1 41 F54088-2 42 F54088-3 43 BLK 4 44 SPK 63-1 45 SPK 63-2 46 SPK 63-3 c 47 SPK 63-4 48 BLK-1 O O h* 49 50 51 BLK-2 Drift Wash 1.50 52 SPK-1 53 SPK-2 54 BLK 3 55 SPK-3 56 SPK-4 57 F54089-1 58 F54089-2 59 F54089-3 60 F54099-1 61 F54099-2 62 Drift 1.50 63 Wash 64 F54099-3 65 F54114-1 66 F54114-2 0.05 2.0 0.1467 0.70 0.03 2.0 0.1328 0.52 0.02 2.0 0.1101 0.29 0.09 2.0 0.1425 1.25 0.004 63.00 0.09 2.0 0.1316 1.36 0.004 63.00 0.15 2.0 0.1428 2.03 0.004 63.00 0.09 2.0 0.1079 1.73 0.004 63.00 1.49 100% 0.00 0.05 2.0 0.1448 0.72 84.26 60.63 0.05 2.0 0.1228 0.89 84.26 75.07 0.04 2.0 0.1317 0.63 84.26 53.49 0.02 2.0 0.1227 0.35 0.12 2.0 0.1358 1.71 0.004 63.00 0.08 2.0 0.1393 1.22 0.004 63.00 0.08 2.0 0.1468 1.15 0.004 63.00 0.08 2.0 0.1494 1.14 0.004 63.00 0.02 2.0 0.1643 0.24 0.00 2.0 0.1578 0.06 1.49 99% 0.00 0.08 2.0 0.1283 1.32 0.004 63.00 0.10 2.0 0.1292 1.49 0.004 63.00 0.02 2.0 0.1396 0.24 0.10 2.0 0.1325 1.48 0.004 63.00 0.02 2.0 0.1109 0.30 0.004 63.00 0.03 2.0 0.1532 0.42 71.796 29.81 0.03 2.0 0.1568 0.41 71.796 29.12 0.03 2.0 0.1474 0.39 71.796 28.15 0.04 2.0 0.1386 0.56 82.143 46.35 0.04 2.0 0.1403 0.53 82.143 43.21 1.49 99% 0.00 0.05 2.0 0.1575 0.59 82.143 48.29 0.03 2.0 0.1573 0.39 80.966 31.19 0.03 2.0 0.1359 0.38 80.966 31.10 0.15 0.18 0.15 0.18 0.15 0.29 0.15 0.19 118% 119% 192% 123% 0.15 0.23 0.15 0.17 0.15 0.17 0.15 0.17 154% 112% 112% 112% 0.15 0.17 0.15 0.19 0.15 0.20 0.15 0.03 112% 127% 129% 22% GOOO Page 2 ,,9 a s u * Skater, Skater % , D i m *w m?Mm* m & 67 F54114-3 68 F54109-1 69 F54109-2 70 F54109-3 71 F54080-1 72 F54080-2 73 F54080-3 74 Drift 75 Wash 1.50 0.03 0.03 0.03 0.03 0.03 0.03 0.00 1.47 98% 0.00 2.0 2.0 2.0 2.0 2.0 2.0 2.0 HWI159L2.XLT 0.1552 0.40 80.966 32.14 0.1326 0.38 73.958 28.45 0,1339 0.38 73.958 28.17 0.1536 0.45 73.958 33.13 0.1322 0.46 75.42 34.91 0.1323 0.50 75.42 37.74 0.1028 0.00 75.42 , 0.00 oO >1 00 ,M9>s? ,, Mass , % 4 2 o o ( 05 Page 3 995-05-23 15:25 OutPut of : 950523A1 oftware : v e r s i o n 6. 1 c l 9 9 0 ,93 perator : DDW ate of the Analysis : 1995-05-23 10:18 nalysis File Name : C:\SKALAR\DATA\HWIDATA\LIVERS\9 5052 3A1 luoride 1.5 alibration order = Inverse Logarithm 'lope : s = #.##### r X - Cl -| esult = 10 L s x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode 2 = -0.00000 1 = 0.00065 0 = -1.22333 luoride L alibration order = 2 Correlation : r = 0.99847 esult = a2 * x 2 + al * x + aO 2 = - 0.00000 1 = 0.00024 0 = -0.00181 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec 120 sec 1 sec Single None : 70 m m . iluter needle Height dilution Factor dilution Volume Resample Dilution runs : 80 : 10 : 2.5 :1 :1 mm ml. User file : Reproces : No . TXT \ is 000179 000099 995-05-23 15:26 OutPut of : 950523A1 luoride 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 sT andard 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 stA rt ignore : 60 Sec eNd ignore : 120 Sec M e a s u r e wi n d o w : 75 % Filter : No Regeneration : No formula : output : ##.### luoride 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 000180 000100 995-05-23 15:26 OutPut of : 950523A1 si sTandard 0.015 s2 sTa nd ard 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 Measur e w i n d o w : 75 % Filter : No Regeneration : No formula : c4:=c3 output : #.#### 000181 000101 995-05-23 15:26 OutPut of : 950523A1 Flu or ide 1 .5 Fluoride L PPM PPM os Typ Ident Ch Result F Time h iw Initial Wash 3 0.060 t Tracer 3 1.472 d Drift 3 1.476 w Wash 3 0.060 si S t a n d a r d 1 3 0.063 s2 S t a n d a r d 2 3 0.071 s3 S t a n d a r d 3 3 0.088 s4 S t a n d a rd 4 3 0.108 s5 S t a n d a r d 5 3 0.127 s6 S t a n d a r d 6 3 0.154 0 s7 Standard 7 3 0.285 s8 Standard 8 3 0.611 2 s9 Standard 9 3 1.238 3 SlO Standard 10 3 1.461 4d Drift 3 1.532 0w Wash 3 0.060 6u BLK 1 3 0.084 7u BLK 2 3 0.076 3u BLK 3 3 0.077 9u F54075-1 3 0.074 Du F54075-2 3 0.075 1u 2u F54075-3 3 0.069 F54081-1 3 0.069 3u F 54081-2 3 0.069 4u F54081-3 3 0.067 5u F54105-1 3 0.068 5n/ d w Drift j 1.526 Wash 3 0.060 3u F54105-2 3 0.070 9u F 54105-3 3 0.070 ju i_ u 7u F54107-1 F54107-2 F54107-3 3 0.866 3 0.083 3 0.072 3u F54074-1 3 0.070 4u F54074-2 3 0.068 ju F54074-3 3 0.070 3u iu F54085-1 o 0.068 F54085-2 3 0.070 3d Drift 3 1.515 3w Wash 3 0.060 3u F54085-3 3 0.087 Lu 1u 3u BLK 1 3 0.071 BLK 2 SPK 1 3-Jt 0.072 0.151 4u SPK 2 3 0.112 3u SPK 3 3 0.119 3u SPK 4 3 0.104 ru SPK 5 3 0.130 )u )u )d SPK 6 3 0.106 BLK 1 0 0.080 D r i f t 3 1 .533 w Wash 3 0.060 u BLK 2 3 0.069 u SPK 1 3 0.119 65 213 389 627 740 916 1090 1264 1442 1616 1792 1966 2140 2314 2490 2723 2839 3014 3191 3366 3541 3717 3891 4065 4238 4415 4591 4832 4941 5115 5291 5465 5640 5820 5992 616 6 6342 6514 6692 6925 7044 7216 7393 7583 7744 7918 8095 8271 8445 8619 8793 9030 9141 9319 Ch Result F Time 4 #.#### 4 0.5569 4 0.5576 4 #.#### 4 0.0188 4 0.0255 4 0.0599 4 0.0917 4 0.1189 4 0.1501 4 0.2516 4 0.3836 4 0.5188 4 0.5552 4 0.5664 4 #.#### 4 0.0527 4 0.0354 4 0.0393 4 0.0327 4 0.0331 4 0.0207 4 0.0200 4 0.0211 4 0.0169 4 0.0188 4 0.5654 4 #.#### 4 0.0241 4 0.0220 4 0.4478 4 0.0507 4 0.0274 4 0.0230 4 0.0183 4 0.0220 4 0.0174 4 0.0220 4 0.5638 4 #.#### 4 0.0586 4 0.0244 4 0.0285 4 0.1464 4 0.0984 4 0.1080 4 0.0860 4 0.1228 4 0.0901 4 0.0446 4 0.5666 4 #.#### 4 0.0218 1 0.1076 0 0 0 0 0 0 0 0 0 0 c 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 .V.tAo. c* 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00012 Page 1 of 2 000102 995-05-23 15:26 OutPut of': 950523A1 Fluoride 1.5 Fluoride L PPM PPM OS Typ Ident Ch Result F Time -,1t u 5u 5u 7u 3u 9u 3u 1u 2d 3w 4u ju 5u 7u 3u 9u 0u 1u 1u 3u 4d 5w 3u 7u 3u 9u 0u 1u 2u 3u a. u 5u d 7w 3u )u 9u Lu 2u Ju 4u 5 1J -5 d 7 V7 r rw SPK 2 3 0.114 SPK 3 3 0.110 SPK 4 3 0.118 F54082-1 3 0.124 F54082-2 3 0.128 F54082-3 3 0.124 F54111-1 3 0.103 F 54111 -2 3 0.098 Drift 3 1.512 Wash 3 0.060 F54111-3 3 0.114 F54079-1 3 0.124 F54079-2 3 0.119 F54079-3 3 0.115 F54.116-1 3 0.138 F54116-2 3 0.127 F54116-3 3 0.138 F54112-1 3 0.130 F 54112-2 3 0.150 F 54112-3 3 0.137 Drift 3 1.483 Wash 3 0.060 F54076-1 F54076-2 3n 0.154 0.151 F54076-3 3 0.139 F 54113-1 3 0.128 F54113-2 3 0.123 F54113-3 3 0.123 F 54084-1 3 0.145 F54084-2 3 0.157 F54084-3 3 0.139 F54098-1 3 0.136 Drift 3 1.490 Wash 3 0.060 F54098-2 3 0.143 F54098-3 3 0.139 F54087-1 3 0.077 F54037-2 J 0.073 F54087-3 F54088-1 F54088-2 F 54038-3 3 0.075 2 0.123 3 0.032 q 0.081 Drift 0 1.489 Wash 3 0.060 RunOut Wash 3 0.060 9495 9670 9845 10020 10196 10370 10546 10720 10394 11133 11245 11420 11595 11770 11944 12118 12296 12470 12645 12821 12995 13234 13347 13520 13697 13373 14047 14221 14397 14571 14747 14921 15095 15327 15447 15621 15796 15969 16142 16322 16496 16672 16846 17079 17321 Ch Result F Time 4 0.1015 4 0.0956 4 0.1065 4 0.1151 4 0.1204 4 0.1142 4 0.0353 4 0.0771 4 0.5633 4 n .#### 4 0.1017 4 0.1149 4 0.1082 4 0.1024 4 0.1315 4 0.1131 4 0.1317 4 0.1219 4 0.1460 4 0.1306 4 0.5587 4 #.#### 4 0.1501 4 0.1462 4 0.1336 4 0.1196 4 0.1134 4 0.1134 4 0.1399 4 0.1530 4 0.1330 4 0.1296 4 0.5593 4 #.#### 4 0.1380 4 0.1327 4 0.0384 4 0.0308 4 0.0343 4 0.1140 4 0.0484 4 0.0457 4 0.5596 4 #.#### 4 #.#### 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 2 of 2 000183 000101 .37TMH : I l` .1 I. N v : , i It fc i " ...k l . i I k i t i i ! - G.uOieOi t: 3"; Urcir fleasu reu BEST COPY AVAILABLE i i !! !: i G .00847 -----------. SOU 000184 000101 1 ~-~r;C! ddd'Vp dd"`^ d ;ddd7 " dr . - t.- - , .__ -____r- _ -- -- ..: - ---_: - ____________ i; -! i :i : lir:- i i.l i t i;i~a 5.: ..a 05S:?G40 %0 ' ?. .ri'.- r -:d n-.g- L x g ^ r i ih n BEST COPY AVAILABLE S -3i -- 000185 0 0 0 1 0 .7 K JOSS 1 Rau data o f SbUb.ARftl : rluoririe i.b O'. ;: W"" :"'c'i ! n A ! f : I : *\ a e* A MM! M M Mi i3-.Mh-r A * i\ i A:AJM! \Vi \ ' J 1i !!; :: i i\ J \l V A!i: ; A; ! : ! u, n M, / ! Is ', n t* ,0-^0 A\ M ;M ;>. ra{-A a /! 2 1 Ai 8 i\ a . j 27 ; iMa 5000 a it ; M -r ia A) i Ur i i-r-h d i i uaaki BEST COPY AVAILABLE 0001S6 00010f bGSb i#' au da ta c f 3 bOS"3fil : K l u e r i de i .b 3 cSn -- cd ' "'i j --: . " a - . - - - - - - - ______ s 5 i 4? ! i -fi V 0 IbHij \ 10 -:r: T iyh n *;, -'. i ': i i : ! i ! \ >\ ^ U u U U W ^ / \ / ' ! [ I '! [ ] '. j :. : i. -V '_J i j :-J !,-i \j ''-i ' \ y ' y' ; i. : : b< i -- u ^ :."-a >f;8-- 3 . f : - ; < l t r l - H a l p : : i - p - t d i i: p a a k a i T COPY AVAILABLE 000187 000107 Hay data at 35u S23h 1 Hluoride 1.5 1J j~K1!1-.;i'f! i c - t x 11 I F l - K a i p s C r t i - r - f c i i i t p e a k s 1 BEST COPY AVAILABLE 000188 00010S GOIOOO 68T000 ,-C.-Vr.~ F 318VHVV AdOO lS3fl -W *# l Ird-d-t '^ ? SWL ft\ h U U \ v HV ! * b \ sb ,U ^ tV V i 0 t *4> \ i\ Ml ; M r . i\ ; Mu : ! I\ :\ , !U U :; W U 4 , N U : ! ; M M j : M ;' i ! : ] M Ui ! ; - i! ;; i : !I ' ! i ' ! V ; f v f i: !: 4 if 15 b \ y ; ^ 'S* * \y ^ : 3.? K-:v-- S - cl i_ :" . , \V `V Ei i l >v m US- 3b ^ rrr^ f^ e r^---3=r:Cr---PzCnr a a a r e a g c S' l ^ p i J O - q j l'^Z cS Q S ; s tij.eo ney i- SSr 995-05-24 12:48 OutPut of : 950524A1 AHT-OHS .\ oftware : version 5.1 cl990,93 perator : DDW fc32.^-\S H BEST COPY AVAILABLE ate of the Analysis : 1995-05-24 07:50 .nalysis File Name : C:\SKALAR\DATA\HWIDATA\LIVERS\950524A1 'luoride 1.5 'alibration order = Inverse Logarithm Hope ; s = #.##### r X - Cl ! esult = 10 L1 ---s--- J! x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode .2 = .1 = .0 = 0.00000 0.00065 1.22861 luoride L a libration order = 2 'orrelation : r = 0.99392 esult = a2 * x 2 + al * x + 3 0 2= 1= 0= 0.00000 0.00021 0.00134 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height : SA1000 :1 : 50 sec. : 120 sec. : 1 sec. : Single : None 70 m m . needle Height dilution Factor dilution Volume Resample Dilution runs 80 mm 10 2 .5 ml 1 1 User file : Reproces : No . TXT 000190 000110 995-05-24 12:43 OutPut of : 950524A1 luoride 1.5 Path number : 3 Signal type : Debubbled Decolor : Yes system Number : 0 diLute : No Resample : No di.1 T h r e sh ol d : 4095 diG output :0 Window event : Off BEST COPY AVAILABLE si sTandard Ignore s2 sTandard Ignore s3 sTandard Ignore s4 sTandard Ignore s5 sT and ard Ignore s6 sTandard 0.150 s i sTandard 0.300 s8 sT andard 0.600 s9 sTandard 1.200 slO sTandard 1.500 Order : Inverse Logarithm Dimension : PPM start Value : 500 DU trigger Limit : 1S00 Sec Peak shape : Pointed stArt ignore : 60 Sec eNd ignore : 120 Sec M e as ur e w i n d o w : 75 % Filter : No Regeneration : No formula : outout : ##.### luoride 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 000191 000111 995-05-24 12:48 OutPut of : 950524A1 s i sTandard 0 015 s2 sTandard 0 030 S3 sTandard 0 060 s4 sTandard 0 090 s5 sTandard 0 . 120 3D sTandard 0 . 150 s7 sTanda rd Ignor e s3 sTandard Ignor e s9 sTandard Ignor e slO sTandard Ignor e Order 2 Dimension PPM start Value 500 DU trigger Limit 1800 Sec Peak shape Poin ted stArt ignore 60 Sec eNd ignore 120 Sec M e a s u r e w ind ow 75 % Filter No Regeneration No formula c4 :=c3 output #.#### 000192 000112 7 995-05-24 12:48 O u t P u t o f : 9 50 524A.1 Fluoride 1.5 Fluoride L PPM PPM w t o I- W W P O k CO ~ -J O ', e n >{>- l o I O I - 1 O ec< C O - J O i e n i p O J W I - 1 O U ) ( I ) v l O i U i t y p Ident Ch Result F Time Ch Result F Time iw Initial Wa s h 3 0.059 t Tracer 3 1.452 d Drift 3 1.469 W Wash 3 0.059 si S t a n d a r d 1 3 0.067 s2 S t a n d a r d 2 3 0.073 s3 S t a n d a r d 3 3 0.090 s4 Standard 4 3 0.110 35 S t a n d a r d 5 3 0.127 s6 S t a n d a r d 6 3"> 0.155 s7 S t a n d a r d 7 3 0.283 s3 Sta n d a rd 8 3 0.614 s9 S t a n d a r d 9 3 1.237 slO Standard 10 3 1.462 d Drift 3 1.495 w Wash 3 0 . 059 u F54092-1 3 0 . 088 u F54092-2 3 0 . 084 u F54092-3 3 0.079 u F 5 4 1 0 5 - 1 '3-i 0.088 u F54106-2 3 0 . 088 u F54106-3 3 0 . 080 u BLK 3 0.065 u SPK 1 3 0.099 u SPK 2 3 0 . 101 u SPK 3 3 0 . 105 d Drift 3 1 . 454 w W a s h 3o 0.059 u SPK 4 3 0.152 u SPK 5 3 0.135 u SPK 6 3 0.144 u LIVER BEK-1 3 0.085 u LIVER BLK-2 3 0.076 u LIVER 3LK-3 3 0.067 u SPK 63-1 3 0.108 u SPK 63-2 3 0.108 u S P K 63-3 3'*> 0.150 u SPK 63-4 3 0.110 d Drift 3 1.494 w Wash 3 0.059 u F54088-1 3 0.085 u F54088-2 3 0 . 087 u F54088-3 3 0 . 030 u BLK 4 3 0 . 069 u SPK 63-1 3 0 . 127 u SPK 63-2 3 0.105 u SPK 63-3 o 0.105 u SPK 63-4 *3 0 . 105 u BLK-1 3 0 . 069 u BLK-2 3 0.062 d Drift 1.487 w W a s h wi 0.059 u S P K - 1 3 0 . 10 5 u SPK-2 3 0 . 112 65 211 387 626 736 912 1086 1262 1438 1612 1788 1964 2138 2312 2488 2725 2838 3014 3190 3367 3541 3713 3883 4063 4241 4415 4589 4830 4939 5113 5291 546 3 5639 5813 5992 6168 6342 6516 6692 6923 7040 7218 7390 7566 7742 7916 8092 8268 8442 8614 8791 9029 9143 9317 4 # . #### 4 0.8505 4 0.8584 4 #.#### 4 0.0160 4 0.0282 4 0.0603 4 0.0931 4 0.1164 4 0.1510 4 0.2712 4 0.4713 4 0.7522 4 0.8550 4 0.8710 4 #.#### 4 0.0564 4 0.0502 4 0.0412 4 0.0571 4 0.0564 4 0.0433 4 0.0118 4 0.0760 4 0.0784 4 0.0854 4 0.8511 4 #.#### 4 0.1473 4 0.1269 4 0.1380 4 0.0515 4 0.0346 4 0.0162 4 0.0889 4 0.0898 4 0.1450 4 0.0931 4 0.8707 4 #.#### 4 0.0521 4 0.0547 4 0.0418 4 0.0217 4 0.1162 4 0.0847 4 0.0847 4 0.0849 4 0.0198 4 0.0049 4 0.8670 4 #.#### 4 0.0847 4 0.0960 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 60 1 9 3 W P O l O , --J I e n Page 1 of 2 000113 Ul 4- WIOH G U) ID ^ i. bJ K) H O 'vDOJ --J Ui 995-05-24 12:48 O u t p u t o f `: 9 5 0 5 2 4 A 1 Fluoride 1.5 Fluoride L PPM PPM Typ Ident Ch Result F Time u BLK 3 3 0.067 9491 u 3PK-3 3 0.114 9669 u SPK-4 3 0.067 9839 u F54089-1 3 0.074 10015 u F 54089-2 3 0.074 10193 u F54089-3 3 0.073 10368 u F54099-1 3 0.078 10544 u F54099-2 3 0.077 10718 d Drift 3 1.487 10893 w Wash 3 0.059 11107 u F54099-3 3 0.082 11244 u F54114-1 3 0.074 11416 u F54114-2 3 0.072 11594 u F54114-3 3 0.074 11768 u F54109-1 3 0.071 11940 u F54109-2 3 0.071 12115 u F54109-3 3 0.076 12296 u F54080-1 3 0.074 12469 u u F54080-2 F54080-3 OO3 0.075 12645 0.059 A 12818 d Drift 3 1.468 12993 w Wash 3 0.059 13185 rw RunOut Wash 3 0.059 13468 Ch Result F Time 4 0.0166 4 0.0979 4 0.0166 4 0.0318 4 0.0318 4 0.0289 4 0.0391 4 0.0369 4 0.8670 4 #.#### 4 0.0463 4 0.0303 4 0.0261 4 0.0308 4 0.0255 4 0.0255 4 0.0344 4 0.0306 4 0.0331 4 #.#### 4 0.8578 4 #.#### 4 #.#### 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 000194 000114 C a l i b r a t i o n curvo o f 0:>S2dAi : H u o r i d a L M is w iiiig g g m r . iO/^OOO 1 j E s m s s s s r -0 QQC094l j ! 0 fi Js I- y 5: wt 1 ,T 4 --0.00i33u Oi f----------:---------- :---------- i Crd-ar : 2 -----------1---------- ]----------1-----------f----------i---------- ; Measured 303 r : 0.33832 BEST COPY AVAILABLE 000195 000113 Calibration curva of 85 S24A: ' lue rida i .5 ESilirfm i ,3b?/bll 1 n rir' r'-^SfP'W iiJ S , - ^ - ,r .- - .Jh--Tr_Ta-a--^'..--!s:aa-rh.^^ J r ~ * "'---------- -------- ~ < --* - `--T_ J-- m e . 1 I Q :=! *`1 s r 5 g j .0 5 9 0 8 2 3 M ! 0 ..-4"' Haastireci Order In u a rss Logarithm 287 4G95 BIST COPYAVAILABLE 000196 000116 i *i >;~ Haw o a t i of 'H-urioa i a ;: tV R"!t ff' ns*fj, fi> * il ^i! J fl NiMH f ' M i ;; 1 A f\ i \ i \ M. J l* ,*. n IS 1 `fH :, , ,:, n % f ts / \ -.j M M :n l I /i & :i J: >1 :!1!!! ', . fI ; I ina ; h i - n a i i ) ! I.--:'t i ~ r :- b i 'l :. t p i i s i - BEST COPY AVAILABLE 000197 0 0 0 1 1 '? *} *85 ------------------------- Hau d a t a c-f 3 0 0 5 2 * 1 : l u c r i -s \ .s fi j W S W * * On"l l f f l l *2*420 ' ' S ' 2 * & 6P Ji j ; r--: < 2! - - if* i ; i: : i ? \i u i m -..J -J '-i V \_> \ *80 i 1? h; ^ [ MtS A1*' <1 J; h i \ i \ i V<--i-M:u ;\\.J;\! i\wf i If i iU\Jhi.u!MI:j>aw*T-,_ I iris V2 Hi --i wJ i f i\ *00 'ic.=ti<i i S J- i . - H a l o i-t'i 1- r - c . i l i o s a k s MST COPYAVAILABLE 000198 000115 >e. GTT0 0 0 66T000 319VIIVAV Ad031S38 bt- ? ; z w * '* HP3-d-I-i3 i -l wi i 3c't>'d a : : :>; ;i ; i k V! !Vi )* ) c fi r\ n. ---. i` },$ ' & \li' ;U\\ * & t * i* '3* B"H J B - f u S' l s p i ' - i o n i : ;Hp3S03 .40 e i= p - t f h 560p 9.11.3 Summary and raw data; ppm F" in serum as determined by thermal extraction followed by analysis using Orion ion analyzer. 000200 0001<i0 HWI 6329-159 AMDT 042095.1 Dohrmann Serum Analysis Analysis Dates: 04/28/95 - 05/10/95 All 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 E A 940 expandable ion analyzer. The Dohrmann was calibrated using 32ppm, 63 ppm, 126ppm, and 253ppm 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 the appropriate logbook. A summary table is included, showing the ppm F' in each sample (see page 2). The value o f "ND " has been entered for any sample with an Orion reading o f below 0.05. Pages 3 - 1 0 show the excel spreadsheet that was generated when the samples were analyzed. Pages 1 1 - 1 8 show the same spreadsheet with "N D " inserted where the Orion reading is below 0.05. 000201 Page 1 o f 18 000121 FC-95 PK osage: 0 mg/kg HWI 6329-159 Fluoride concentration in rabbit serum (ppm F-) Sample 4 hour 8 hour 12 hour 24 hour 48 hour F54074 ND ND ND ND F54075 ND ND ND ND ND F54081 ND ND ND ND F54085 ND ND ND ND F54086 ND ND ND ND F54104 ND ND ND ND F54105 ND ND ND ND ND F54107 ND ND ND ND Dosage: 5 mg/kg Sample F54077 F54083 F54091 F54094 F54097 F54100 F54102 F54108 4 hour ND ND ND ND ND ND ND ND 8 hour ND ND ND ND ND ND ND ND 12 hour 24 hour ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND Dosage: 10 mg/kg Sample F54071 F54078 F54090 F54093 F54095 F54096 F54101 F54110 4 hour ND ND ND ND ND ND ND ND 8 hour ND ND ND ND ND ND ND 1.5 12 hour 24 hour ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND Dosage: 100 mg/kg Sample F54080 F54087 F54088 F54089 F54092 F54099 F54106 F54109 F54114 4 hour ND ND ND ND ND ND ND ND ND 8 hour ND ND ND ND ND ND ND ND ND 12 hour 15 hour 24 hour 48 hours ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 1.5 ND 1.7 ND ND ND ND ND ND ND ND ND ND ND Day 8 ND ND ND ND ND ND ND ND ND Day 22 Day 28 ND ND ND ND ND ND ND ND ND Dosage: 500 mg/kg Sample F54076 F54079 F54082 F54084 F54098 F54111 F54112 F54113 F54116 4 hour ND 2.3 1.1 1.3 ND 1.9 ND 1.6 2.0 8 hour ND ND 1.3 ND 1.0 ND 1.2 ND 3.4 12 hour 24 hour 48 hours Day 8 ND ND 1.2 1.2 1.4 ND 1.1 1.3 1.5 ND 1.2 1.8 1.5 ND 1.3 1.3 1.3 ND 1.3 1.5 1.3 1.3 1.0 1.3 1.2 ND 1.3 1.4 1.6 ND 1.3 1.7 1.3 360 hour 1.3 ND 1.4 1.1 1.3 ND 1.3 1.4 Day 22 ND ND ND ND Day 28 ND ND ND ND 000202 00012 6329159S.XLS S T U D Y # 6329-159 384 (D serum blank-1 serum blank-2 serum spike 63-1 serum spike 63-2 serum spike 94.5-1 serum spike 94.5-2 serum spike 126-1 serum spike 126-2 serum spike 126-3 serum spike 126-4 F54081-4 F54075-4 F54107-4 F54105-4 F54074-4 Blank 1 Blank 2 Blank 3 Blank 4 Blank 5 Blank 6 Serum Spk-1 Serum Spk-2 Serum Spk-3 Serum Spk-4 Serum Spk-5 Serum Spk-6 Serum Spk-7 Serum Spk-8 Serum Spk-9 Serum Spk-10 F54085-4 F54086-4 F54104-4 F54077-4 F54083-4 F54094-4 F54100-4 F54091-4 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54097-4 F54102-4 F54108-4 F54071-4 mLof FC95 solutio spiked 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 Cone. Actual Quantity Dl: FC95 Meter sample T IS A B % Actual solution reading (mL or final revry ppm F- [ppm) (ppm F-) (grams) vot [mL) (ug/ug> in sample 0.036 0.10 2.6 0.72 0.020 0.10 2.0 0.40 63 0.070 0.10 2.0 9 3 % 1.4 63 0.070 0.10 2.0 9 2 % 1.4 94.5 94.5 126 126 0.063 0.046 0.15 0.099 0.10 0.10 0.10 0.10 2.0 56% 2.0 4 1 % 2.0 101% 2.0 65% 1.3 0.92 3.1 2.0 126 0.15 0.10 126 0.13 0.10 2.0 9 8 % 2.0 8 5 % 3.0 2.6 0.040 0.032 0.10 0.10 2.0 2.0 0.80 0.65 0.024 0.10 2.0 0.48 0.025 0.10 2.0 0.51 0.034 0.71 0.34 0.10 0.10 0.10 2.0 2.0 2.0 0.67 14 6.9 0.36 0.084 0.10 0.10 2.0 2.0 7.2 1.7 0.038 0.10 2.0 0.75 0.039 0.10 63 0.055 0.10 2.0 2.0 7 3 % 0.79 1.1 63 0.068 0.10 63 0.073 0.10 2.0 9 0 % 2.0 9 6 % 1.4 1.5 126 0.10 0.10 2.0 6 7 % 2.0 126 0.11 0.10 126 0.11 0.10 126 0.11 0.10 126 0.098 0.10 126 0.12 0.10 126 0.10 0.10 2.0 7 4 % 2.0 7 3 % 2.0 7 2 % 2.0 6 5 % 2.0 7 7 % 2.0 6 8 % 2.2 2.2 2.2 2.0 2.3 2.1 0.045 0.027 0.025 0.10 0.10 0.10 2.0 2.0 2.0 0.90 0.54 0.49 0.021 0.10 2.0 0.42 0.020 0.021 0.020 0.10 0.10 0.10 2.0 2.0 2.0 0.40 0.42 0.41 0.019 0.10 0.029 0.10 0.024 0.10 63 0.071 0.10 63 0.072 0.10 2.0 2.0 2.0 2.0 9 4 % 2.0 9 5 % 0.38 0.59 0.48 1.4 1.4 0.022 0.10 2.0 0.45 0.020 0.10 2.0 0.41 0.016 0.017 0.10 0.10 2.0 2.0 0.32 0.34 Mass Mass spiked revrd [ug F-) (ug F-) 0.072 0.15 0.15 0.23 0.23 0.30 0.30 0.30 0.30 0.040 0.14 0.14 0.13 0.092 0.31 0.20 0.30 0.26 0.080 0.065 0.048 0.051 0.067 1.4 0.69 0.72 0.17 0.075 0.079 0.15 0.11 0.15 0.14 0.15 0.15 0.30 0.20 0.30 0.22 0.30 0.22 0.30 0.30 0.30 0.30 0.22 0.20 0.23 0.21 0.090 0.054 0.049 0.042 0.040 0.042 0.041 0.038 0.15 0.059 0.048 0.14 0.15 0.14 0.045 0.041 0.032 0.034 000203 000123 6329159S.XLS S T U D Y # 6329-159 384 ID F54093-4 F54095-4 F54101-4 F54078-4 F54090-4 F54096-4 F54 1 10-4 F54087-4 F54088-4 F54089-4 F54106-4 F54080-4 F54092-4 F5 4 109-4 F54114-4 F54076-4 F54082-4 F54111-4 F54 113-4 F54079-4 F54084-4 F54098-4 F54 1 16-4 F54075-8 F54081-8 F54105-8 F54107-8 F54074-8 F54085-8 F54086-8 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54104-8 F54077-8 F54083-8 F54094-8 F 5 4 100-8 F54091-8 F54097- 8 F54102- 8 F54108- 8 F54071-8 F54093 - 8 F54095 - 8 F54101- 8 F54078- 8 mL of FC95 solutio spiked 0.004 0.004 Core. Actual Quantity Dl: FC95 Meter sample T fSA B solution reading (mL or final (ppm) (ppm F-) (grams) vol (mL) 0.020 0.016 0.015 0.021 0.020 0.017 0.020 0.025 0.028 0.020 0.032 0.025 0.019 0.021 0.039 0.041 0.057 0.096 0.082 0.12 0.067 0.046 0.10 0.025 0.020 0.021 0.015 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 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 0.015 0.10 2.0 0.021 0.10 2.0 0.031 0.10 2.0 0.029 0.10 2.0 0.028 0.10 2.0 63 0.080 0.10 2.0 63 0.069 0.10 2.0 0.031 0.10 2.0 0.023 0.10 2.0 0.018 0.10 2.0 0.025 0.017 0.014 0.014 0.013 0.013 0.015 0.019 0.026 0.016 0.021 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 % revry (ug/ug) 106% 91% Actual ppm Fm sample 0.41 0.32 0.29 0.41 0.39 0.34 0.39 0.50 0.55 0.41 0.63 0.51 0.38 0.42 0.78 0.81 1.1 1.9 1.6 2.3 1.3 0.93 2.0 0.49 0.40 0.43 0.30 0.30 0.43 0.62 0.58 0.56 1.6 1.4 0.62 0.46 0.35 0.51 0.35 0.28 0.28 0.26 0.25 0.30 0.37 0.51 0.32 0.42 Mass spiked (ug F-) 0.15 0.15 Mass revrd tug F-) 0.041 0.032 0.029 0.041 0.039 0.034 0.039 0.050 0.055 0.041 0.063 0.051 0.038 0.042 0.078 0.081 0.11 0.19 0.16 0.23 0.13 0.093 0.20 0.049 0.040 0.043 0.030 0.030 0.043 0.062 0.058 0.056 0.16 0.14 0.062 0.046 0.035 0.051 0.035 0.028 0.028 0.026 0.025 0.030 0.037 0.051 0.032 0.042 000204 0 0 0 1 -4 6329159S.XLS S T U D Y # 6329-159 384 ID F54090- 8 F54096- 8 F 5 4 1 10- 8 F54087- 8 F54088- 8 F54089- 8 F54080- 8 F54106- 8 F54092- 8 F54109- 8 F54114- 8 F5407S- 8 F54082- 8 F54111-8 F54113- 8 F54079- 8 F54084- 8 F54098- 8 F 5 4 1 16- 8 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54075-12 F54081-12 F54105-12 F54107-12 F54074-12 F54085-1 2 F5408S-12 F54086-12 F54077-12 F54083-1 2 F54094-12 F54100-12 F54091 -12 F64097-12 F54102-12 F54108-12 FS4077-12 F54071-1 2 FS4093-12 F54095-12 F54101 -12 F54 1 10-12 F54096-1 2 F54090-12 F54078-1 2 mL of FC95 solutio spiked 0.004 0.004 Conc- Actual Quantity Dl: FC95 Meter sample T ISA B % solution reading (mL or final rcvry (ppm) (ppm F-) (grams) yoi (mL) (ug/ug) 0.021 0.10 2.0 0.017 0.10 2.0 0.075 0.022 0.10 0.10 2.0 2.0 0.033 0.10 2.0 0.031 0.032 0.043 0.037 0.024 0.034 0.10 0.10 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 2.0 2.0 0.045 0.10 2.0 0.063 0.10 2.0 0.044 0.048 0.042 0.045 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 0.050 0.10 2.0 0.17 0.10 2.0 0.012 0.10 2.0 0.018 0.10 63 0.066 0.10 63 0.070 0.10 2.0 2.0 87% 2.0 9 3 % 0.028 0.10 2.0 0.021 0.10 2.0 0.026 0.10 2.0 0.019 0.017 0.10 0.10 2.0 2.0 0.018 0.10 2.0 0.020 0.10 2.0 0.025 0.10 2.0 0.038 0.032 0.035 0.022 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 0.029 0.10 2.0 0.025 0.10 2.0 0.022 0.10 2.0 0.014 0.046 0.10 0.10 2.0 2.0 0.021 0.10 2.0 0.016 0.10 2.0 0.016 0.10 2.0 0.020 0.10 2.0 0.016 0.10 2.0 0.012 0.10 2.0 0.017 0.014 0.10 0.10 2.0 2.0 Actual ppm Fin sample 0.42 0.34 1.5 0.44 0.66 0.62 0.64 0.86 0.74 0.49 0.68 0.89 1.3 0.88 0.95 0.84 0.89 1.0 3.4 0.25 0.36 1.3 1.4 0.56 0.42 0.53 0.39 0.35 0.37 0.41 0.50 0.76 0.64 0.71 0.44 0.58 0.49 0.44 0.29 0.92 0.41 0.33 0.32 0.40 0.31 0.25 0.34 0.28 Mass Mass spiked rcvrd (ug F-) (ug F-) 0.042 0.034 0.15 0.044 0.066 0.062 0.064 0.086 0.074 0.049 0.068 0.15 0.15 0.089 0.13 0.088 0.10 0.084 0.089 0.10 0.34 0.025 0.036 0.13 0.14 0.056 0.042 0.053 0.039 0.035 0.037 0.041 0.050 0.076 0.064 0.071 0.044 0.058 0.049 0.044 0.029 0.092 0.041 0.033 0.032 0.040 0.031 0.025 0.034 0.028 000205 0001 6329159S.XLS S T U D Y # 6329-159 384 ID F54088-12 F54106-12 F54089-12 F54087-12 F54080-1 2 F54092-12 F54109-12 F 5 4 1 14-12 F54076-12 F 5 4 1 13-12 F54082-12 F54111-12 F54098-12 F54084-12 F54079-12 F54116-12 Serum Blk-1 Serum Blk-2 Serum Spk-1 Serum Spk-2 Serum Spk-3 Serum Spk-4 Serum Spk-5 Serum Spk-6 Serum Spk-7 Serum Spk-8 F54105-24 F54075-24 F54081-24 F54107-24 F54104-24 F54086-24 F54074-24 Blank 1 Blank 2 spike 1 spike 2 spike 3 spike 4 spike 5 spike 6 Blank 3 Blank 4 F54085-24 F54083-24 F54100-24 F54077-24 F54094-24 F54091-24 mL of FC95 solutto spiked 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 Cone. FC95 solution [ppm) 63 63 63 63 63 63 63 63 63 63 63 63 63 63 Actual Quantity I: Meter sample TJSAB % reading (mL or final revry (ppm F-) (grams) vot (mL) (ug/ug) 0.025 0.10 2.0 0.027 0.10 2.0 0.019 0.10 2.0 0.019 0.10 2.0 0.018 0.10 2.0 0.017 0.10 2.0 0.021 0.10 2.0 0.026 0.10 2.0 0.027 0.10 2.0 0.022 0.10 2.0 0.034 0.10 2.0 0.046 0.10 2.0 0.042 0.10 2.0 0.044 0.10 2.0 0.043 0.10 2.0 0.047 0.10 2.0 0.033 0.10 2.0 0.030 0.10 2.0 0.034 0.10 2.0 4 4 % 0.040 0.10 2.0 5 2 % 0.050 0.10 2.0 6 6 % 0.054 0.10 2.0 7 2 % 0.083 0.10 2.0 110% 0.053 0.10 2.0 7 1 % 0.049 0.10 2.0 6 5 % 0.055 0.10 2.0 7 3 % 0.041 0.10 2.0 0.022 0.10 2.0 0.017 0.10 2.0 0.015 0.10 2.0 0.018 0.10 2.0 0.016 0.10 2.0 0.013 0.10 2.0 0.031 0.10 2.0 0.022 0.10 2.0 0.033 0.10 2.0 4 3 % 0.044 0.10 2.0 5 8 % 0.047 0.10 2.0 6 3 % 0.047 0.10 2.0 6 2 % 0.052 0.10 2.0 6 8 % 0.051 0.10 2.0 6 7 % 0.036 0.10 2.0 0.020 0.10 2.0 0.021 0.10 2.0 0.019 0.10 2.0 0.016 0.10 2.0 0.017 0.10 2.0 0.015 0.10 2.0 0.016 0.10 2.0 000206 Actual ppm Fin sample 0.51 0.54 0.38 0.37 0.36 0.33 0.41 0.52 0.55 0.44 0.69 0.91 0.84 0.87 0.85 0.94 0.66 0.60 0.67 0.79 0.99 1.1 1.7 1.1 0.99 1.1 0.81 0.44 0.35 0.30 0.37 0.33 0.27 0.61 0.43 0.65 0.88 0.95 0.94 1.0 1.0 0.72 0.39 0.41 0.38 0.32 0.33 0.31 0.31 Mass spiked lug F-) 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Mass revrd (ug F-> 0.051' 0.054 0.038 0.037 0.036 0.033 0.041 0.052 0.055 0.044 0.069 0.091 0.084 0.087 0.085 0.094 0.066 0.060 0.067 0.079 0.10 0.11 0.17 0.11 0.10 0.11 0.081 0.044 0.035 0.030 0.037 0.033 0.027 0.061 0.043 0.065 0.088 0.095 0.094 0.10 0.10 0.072 0.039 0.041 0.038 0.032 0.033 0.031 0.031 , 0001^ AoTOll (cOo10o) TcAoiconoI Ooi) cATni cooo> ocNT0AoJ1) C0oO> Tc4n| --co4 COoO) TA1i --O4) r(o0 AO0ncn0 icoo ov*cA(0nni N<0> TAOc0con0I a rcoo T01I --* _4 Kco> c*nn OMQ I-(O0- noAi -co* I(O0 To4Ii --O*) T(J1| C*O4 cnn 00 A00 00 40*0. cTnI c0o0 00 c4Tn*I -j C7AO* AO0MTcn0l 00 TcAnI --Co* 00 ATcnI --O*) NA) vTcAocnoiI KA> 0TaAo0Ii MtA* cTcAo0on0l IAO ATail _--4 KA> cTaAorooil KAl TaAil --k wa 323 (aAi 23 w NA) "(DA IO T7(A5C* CQA 233 er io (OA 233 cr -* TaAil A TaAli oco MA CTaAOroOil Tail 090 aTil 0v0i MA NA> IAO AOTa0co0il TaAil o0) rAo rAo 0T0aOA00il TaAOV9lii MA rA TaAOcooil oTaAcao>il TaAil o NA> rAo rAo aTAil O rAo TcnJ CaOi Tail CcoO TvaAoili rAo rAo rAo Tail cvoi rAo naAi oro rAo TaAil O00 rAo P 0co0 ffc oo oo op o4 c0o3 c0o3 iS=: eI aO 3, oa, 0 Ju2i Soi w oMe omu nO3 3 5' en p 3 ooooooopoooooppooooooooopppoooooooooooooooooooooo K s>cn co cbn b ccno b-t* CO cbCnTI cbn cn bvvij b 03 co b Ov)i b Oco) bo co b 0o3 0b0 cn bcn co b c__oA b03 vi bVi CO 0b3 03 0b3 00 b 03 a cbn co 0b3 0b3 *>. b 0co3 bccnn b03 -p b0IO0 bvi bIO 00 bccoo b 03 b_k b cn 03 b O) b -p* bb p* co b b co blo cn b_k b co b b a b IO co b cn bIO o b co b > ,3In::&C5-sO^:::ai^:: 2--. o pooo .a --* --k oooo ppo --k --k ooo p o poo _k _k ooo p p p Oo O _k _t --k --* --k oooooo o o p o oOo o p *_k _* _k _k __k "__k a ooooooo oppp a "__k "__k o o OO oo l_k oo pO "__k oo Op o p "__k "_k oooo p o ooo _k *_k ooo op __k oO oppp l_k L. . oooo o la o o o CO f F-3 S 2. ' <o ^ oo lo KJ IO IO lo IO IO lo IO io IO lo IO IO N3 IO lo IO IO IO IO IO lo lo IO IO lo IO IO IO IO lo IO lo IO IO IO IO NJ IO IO lo Ni IO lo IO IO ro IO ^ 3rf, d d o*0 b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b b 3 &. >a 4 J -(O %# <Q < --I -- -1 p - --I --I - i --k --l --k --k _k --k --k co b S j b c o c n - ^ c o c o c n b v i r o o c o c n c o ^ c o i o i o c o c o ^ b c n t . op cn cn vVii oopoo co IO cJok co IO co IO io vi ooO io CD io 03 ivoi O lo vi Ooop oO cn o io 03 io co io f" io 03 ivoi o co O o 4^ O O io cn o io co i0qfl o 3 *a <D *n oo e--nk e--nk eg 2 J]5I r g| o co o - o co p 03 vi Oo CO co o ai oo \tw t co o co o cn O O) o Vi o _4 o NJ 00 oo *4 co cn o co o Oo *j 4WCO IO o IO o co o*1A o *A o co -* co o 03 oo t* bai on o b vi vi o o o o o o o o o o OOo o o p o b co IO b G> bCO IO b cIOo bIO Vi b ro 00 bro b IO o> vi bro vi b cn O bIO 03 b IO co b IO bIO 03 b IO cbo o b o o IbO ai p bro 00 ke: 71 3CL M2utfi vi 03 co IcOo ccon /) X I" Cfl 03 H eo -< it o> rccooo ccon 6329159S.XLS S T U D Y # 6329-159 384 ID F54084-360 F54116-360 Serum Blk 1 Serum Blk 2 Serum Spk 1 Serum Spk 2 F54082-D22 F54111-D22 F54079-D22 F54116-D22 F54082-D28 F54111-D28 F54079-D28 F54116-D28 F54099-4 F54099-8 F54099-1 2 F54099-24 F54099-48 F54099-192 F54099-360 F54112-4 F54112-8 F54112-12 F54112-24 F54112-48 F54112-192 F54112-360 SERUM SPIKE-63-1 SERUM SPIKE-63-2 SERUM SPIKE-63-3 SERUM SPIKE-126-1 SERUM SPIKE-126-2 SERUM SPIKE-126-3 serum blank-1 serum blank-2 serum blank-3 spike 631 spike 632 spike 63-3 spike 63-4 spike 63-5 spike 63-6 spike 63 7 spike 63-8 Serum blnk 1 Serum blnk 2 Serum spike 631 Serum spike 6 3 2 mL of FC95 solutto spiked 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 Cone Actual Quantity Dl: FC95 Meter sample TJSAB % solution reading (mL or final revry (ppm) JE0m L (grams) vol (mL) (ug/ug) 0.069 0.10 2.0 0.069 0.10 2.0 0.025 0.10 2.0 0.032 0.10 2.0 63 0.069 0.10 2.0 91% 63 0.076 0.10 2.0 101% 0.046 0.10 2.0 0.041 0.10 2.0 0.040 0.10 2.0 0.045 0.10 2.0 0.048 0.10 2.0 0.036 0.10 2.0 0.033 0.10 2.0 0.036 0.10 2.0 0.028 0.10 2.0 0.026 0.10 2.0 0.074 0.10 2.0 0.087 0.10 2.0 0.047 0.10 2.0 0.033 0.10 2.0 0.029 0.10 2.0 0.038 0.10 2.0 0.059 0.10 2.0 0.066 0.10 2.0 0.051 0.10 2.0 0.066 0.10 2.0 0.059 0.10 2.0 0.041 0.10 2.0 63 0.059 0.10 2.0 78% 63 0.079 0.10 2.0 105% 63 0.074 0.10 2.0 97% 126 0.083 0.10 2.0 55% 126 0.12 0.10 2.0 77% 126 0.12 0.10 2.0 79% 0.10 0.10 2.0 0.040 0.10 2.0 0.019 0.10 2.0 63 0.058 0.10 2.0 77% 63 0.058 0.10 2.0 77% 63 0.092 0.10 2.0 121% 63 0.053 0.10 2.0 70% 63 0.055 0.10 2.0 73% 63 0.058 0.10 2.0 77% 63 0.072 0.10 2.0 95% 63 0.060 0.10 2.0 79% 0.045 0.10 2.0 0.017 0.10 2.0 63 0.12 0.10 2.0 153% 63 0.21 0.10 2.0 273% 000208 Actual ppm Fin sample 1.4 1.4 0.50 0.65 1.4 1.5 0.91 0.82 0.81 0.89 0.96 0.71 0.66 0.73 0.55 0.51 1.5 1.7 0.95 0.67 0.57 0.76 1.2 1.3 1.0 1.3 1.2 0.82 1.2 1.6 1.5 1.7 2.3 2.4 2.0 0.79 0.39 1.2 1.2 1.8 1.1 1.1 1.2 1.4 1.2 0.90 0.33 2.3 4.1 Mass spiked (ug F-) 0.15 0.15 0.15 0.15 0.15 0.30 0.30 0.30 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 0.15 Mass revrd (ug F-) 0.14 ' 0.14 0.050 0.065 0.14 0.15 0.091 0.082 0.081 0.089 0.10 0.071 0.066 0.073 0.055 0.051 0.15 0.17 0.095 0.067 0.057 0.076 0.12 0.13 0.10 0.13 0.12 0.082 0.12 0.16 0.15 0.17 0.23 0.24 0.20 0.079 0.039 0.12 0.12 0.18 0.11 0.11 0.12 0.14 0.12 0.090 0.033 0.23 0.41 00012S 6329159S.XLS S T U D Y # 6329-159 384 ID Serum blank 1 Serum blank 2 Serum spike 1 Serum spike 2 Serum spike 3 Serum spike 4 Serum spike 5 Serum spike 6 Serum spike 7 Serum blank 3 Serum blank 4 F54088-48 F54099-48 F54089-48 F54087-48 F54080-48 F54092-48 F54109-48 F54114-48 F54088 dy8 F54099 dy8 serum spike 8 serum spike 9 serum spike 10 SERUM B LK 1 SERUM BLK2 SERUM SPK 1 SERUM SPK 2 SERUM SPK 3 F54089-DY8 F54087-DY8 F54080-DY8 F54092-DY8 F54109-DY8 F54114-DY8 F54088-15 F54099-15 F54089-15 F54087-15 F54080-15 F54092-15 F54109-15 F54114-15 F54089-D22 F54087-D22 F54109-D22 F54114-D22 F54089-d28 F54087-d28 mL of Cone. Actual Quantity Dl: FC95 FC95 Meter sample T1SAB % Actual Mass Mass sorutio solution reading (mL or final revry ppm F- spiked revrd spiked (ppm) (ppm F ) (grams) voi (mL) (up/ug) in sample (ug F-) ...(v.i.ErJ,, 0.031 0.10 2.0 0.62 0.062 0.021 0.10 2.0 0.42 0.042 0.004 63 0.074 0.10 2.0 98% 1.5 0.15 0.15 0.004 63 0.066 0.10 2.0 87% 1.3 0.15 0.13 0.004 63 0.065 0.10 2.0 86% 1.3 0.15 0.13 0.004 63 0.060 0.10 2.0 80% 1.2 0.15 0.12 0.004 63 0.070 0.10 2.0 93% 1.4 0.15 0.14 0.004 63 0.069 0.10 2.0 91% 1.4 0.15 0.14 Q.004 63 0.068 0.10 2.0 90% 1.4 0.15 0.14 0.023 0.10 2.0 0.46 0.046 0.020 0.10 2.0 0.41 0.041 0.019 0.10 2.0 0.37 0.037 0.021 0.10 2.0 0.43 0.043 0.017 0.10 2.0 0.34 0.034 0.020 0.10 2.0 0.40 0.040 0.017 0.10 2.0 0.34 0.034 0.016 0.10 2.0 0.32 0.032 0.022 0.10 2.0 0.45 0.045 0.022 0.10 2.0 0.44 0.044 0.018 0.10 2.0 0.36 0.036 0.014 0.10 2.0 0.28 0.028 0.004 63 0.059 0.10 2.0 78% 1.2 0.15 0.12 0.004 63 0.060 0.10 2.0 79% 1.2 0.15 0.12 0.004 63 0.060 0.10 2.0 79% 1.2 0.15 0.12 0.031 0.10 2.0 0.62 0.062 0.016 0.10 2.0 0.32 0.032 0.004 63 0.056 0.10 2.0 74% 1.1 0.15 0.11 0.004 63 0.069 0.10 2.0 91% 1.4 0.15 0.14 0.004 63 0.072 0.10 2.0 95% 1.4 0.15 0.14 0.026 0.10 2.0 0.52 0.052 0.029 0.10 2.0 0.59 0.059 0.029 0.10 2.0 0.59 0.059 0.036 0.10 2.0 0.73 0.073 0.028 0.10 2.0 0.56 0.056 0.035 0.10 2.0 0.69 0.069 0.010 0.10 2.0 0.21 0.021 0.026 0.10 2.0 0.52 0.052 0.022 0.10 2.0 0.43 0.043 0.021 0.10 2.0 0.43 0.043 0.019 0.10 2.0 0.39 0.039 0.022 0.10 2.0 0.44 0.044 0.025 0.10 2.0 0.50 0.050 0.020 0.10 2.0 0.40 0.040 0.019 0.10 2.0 0.39 0.039 0.018 0.10 2.0 0.36 0.036 0.019 0.10 2.0 0.37 0.037 0.019 0.10 2.0 0.39 0.039 0.018 0.10 2.0 0.36 0.036 0.018 0.10 2.0 000209 0.35 0.035 O O O l^i 6329159S.XLS S T U D Y # 6329-159 384 ID F54109-<J28 F54114-d28 SERUM SPIKE 63-4 SERUM SPIKE 63-5 SERUM SPIKE 63-6 SERUM SPIKE 63-7 SERUM B LK 1 SERUM BLK 2 SERUM SPK 63-1 SERUM SPK 63-2 SERUM SPK 63-3 F54105-48 F54075-48 m L o f Cone. Actual Quantity Dl: FC95 FC95 Meter sample TiSAB solutto solution reading (mL or final spiked (ppm) (ppm F-) (grams) E o>] 0.004 0.004 0.004 0.004 63 63 63 63 0.020 0.017 0.031 0.049 0.061 0.059 0.031 0.10 0.10 0.10 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.004 0.004 0.004 63 63 63 0.017 0.061 0.078 0.062 0.021 0.026 0.10 0.10 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 2.0 2.0 % Actual Mass Mass revry ppm F- spiked revrd (ug/ug) in sample fug F-) (ug F-) 0.41 0.041' 41% 65% 81% 0.33 0.62 0.98 1.2 0.15 0.15 0.15 0.033 0.062 0.10 0.12 78% 1.2 0.15 0.12 0.62 0.34 0.062 0.034 81% 1.2 0.15 0.12 103% 1.6 0.15 0.16 82% 1.2 0.15 0.12 0.42 0.042 0.52 0.052 000210 000100 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ID serum blank-1 serum blank-2 serum spike 63-1 serum spike 63-2 serum spike 94.5-1 serum spike 94.5-2 serum spike 126-1 serum spike 126-2 serum spike 126-3 serum spike 126-4 F54081-4 F54075-4 F54107-4 F54105-4 F54074-4 Blank 1 Blank 2 Blank 3 Blank 4 Blank 5 Blank 6 Serum Spk-1 Serum Spk-2 Serum Spk-3 Serum Spk-4 Serum Spk-5 Serum Spk-6 Serum Spk-7 Serum Spk-8 Serum Spk-9 Serum Spk-10 F54085-4 F54086-4 F54 1 0 4 - 4 F54077-4 F54083-4 F54094-4 F54100-4 F54091-4 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54097-4 F54102-4 F54108-4 F54071-4 mL of Cone. Actual Quantity Dl: FC95 FC95 Meter sample T1SAB % Actual Mass Mass solutio solution reading (mL or final revry ppm F- spiked revrd spiked fppm) (ppm F-) (grams) vpl (mL) J.Y.9/Y9.L in sample J m f . M B . 0.036 0.10 2.0 ND 0.020 0.10 2.0 ND 0.004 63 0.070 0.10 2.0 93% 1.4 0.15 0.14 0.004 63 0.070 0.10 2.0 92% 1.4 0.15 0.14 0.004 94.5 0.063 0.10 2.0 56% 1.3 0.23 0.13 0.004 94.5 0.046 0.10 2.0 41% 0.92 0.23 0.092 0.004 126 0.15 0.10 2.0 101% 3.1 0.30 0.31 0.004 126 0.099 0.10 2.0 65% 2.0 0.30 0.20 0.004 126 0.15 0.10 2.0 98% 3.0 0.30 0.30 0.004 126 0.13 0.10 2.0 85% 2.6 0.30 0.26 0.040 0.10 2.0 ND ND 0.032 0.10 2.0 ND ND 0.024 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.034 0.10 2.0 ND ND 0.71 0.10 2.0 14 1.4 0.34 0.10 2.0 6.9 0.69 0.36 0.10 2.0 7.2 0.72 0.084 0.10 2.0 1.7 0.17 0.038 0.10 2.0 ND ND 0.039 0.10 2.0 ND ND 0.004 63 0.055 0.10 2.0 73% 1.1 0.15 0.11 0.004 63 0.068 0.10 2.0 90% 1.4 0.15 0.14 0.004 63 0.073 0.10 2.0 96% 1.5 0.15 0.15 0.004 126 0.10 0.10 2.0 67% 2.0 0.30 0.20 0.004 126 0.11 0.10 2.0 74% 2.2 0.30 0.22 0.004 126 0.11 0.10 2.0 73% 2.2 0.30 0.22 0.004 126 0.11 0.10 2.0 72% 2.2 0.30 0.22 0.004 126 0.098 0.10 2.0 65% 2.0 0.30 0.20 0.004 126 0.12 0.10 2.0 77% 2.3 0.30 0.23 0.004 126 0.10 0.10 2.0 68% 2.1 0.30 0.21 0.045 0.10 2.0 ND ND 0.027 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.029 0.10 2.0 ND ND 0.024 0.10 2.0 ND ND 0.004 63 0.071 0.10 2.0 94% 1.4 0.15 0.14 0.004 63 0.072 0.10 2.0 95% 1.4 0.15 0.14 0.022 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 000211 000131 // 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ED F54093-4 F54095-4 F54101-4 F54078-4 F54090-4 F54096-4 F54110-4 F54087-4 F54088-4 F54089-4 F54106-4 F54080-4 F54092-4 F54109-4 F54114-4 F54076-4 F54082-4 F54111-4 F54113-4 F54079-4 F54084-4 F54098-4 F54116-4 F54075-8 F54081-8 F54105-8 F54107-8 F54074-8 F54085-8 F54086-8 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54104-8 F54077-8 F54083-8 F54094-8 F54100-8 F54091-8 F54097-8 F54102- 8 F54108- 8 F54071-8 F54093 - 8 F54095 - 8 F54101- 8 F54078- 8 mL of Cone. Actual Quantity Dl: FC95 FC95 Meter sample T1SAB % Actual Mass Mass solutso solution reading (mL or final revry ppm F- spiked revrd spiked (ppm) {ppm F-) (grams) vol (mL) (u^/ug) in sample J m E l . (ug F-: 0.020 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.015 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.028 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.032 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.039 0.10 2.0 ND ND 0.041 0.10 2.0 ND ND 0.057 0.10 2.0 1.1 0.11 0.096 0.10 2.0 1.9 0.19 0.082 0.10 2.0 1.6 0.16 0.12 0.10 2.0 2.3 0.23 0.067 0.10 2.0 1.3 0.13 0.046 0.10 2.0 ND ND 0.10 0.10 2.0 2.0 0.20 0.025 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.015 0.10 2.0 ND ND 0.015 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.031 0.10 2.0 ND ND 0.029 0.10 2.0 ND ND 0.028 0.10 2.0 ND ND 0.004 63 0.080 0.10 2.0 106% 1.6 0.15 0.16 0.004 63 0.069 0.10 2.0 91% 1.4 0.15 0.14 0.031 0.10 2.0 ND ND 0.023 0.10 2.0 ND ND 0.018 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.014 0.10 2.0 ND ND 0.014 0.10 2.0 ND ND 0.013 0.10 2.0 ND ND 0.013 0.10 2.0 ND ND 0.015 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.026 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 000212 00013 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ID F54090-8 F54096-8 F54110-8 F54087-8 F54088-8 F54089-8 F54080-8 F54106- 8 F54092-8 F54109-8 F54114- 8 F54076-8 F54082-8 F54111-8 F54113-8 F54079-8 F54084-8 F54098-8 F54116- 8 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54075-12 F54081-12 F54105-12 F54107-12 F54074-12 F54085-1 2 F54086-12 F54086-12 F54077-12 F54083-12 F54094-12 F54100-12 F54091 -12 F54097-12 F54102-12 F54108-12 F54077-12 F54071 -12 F54093-12 F54095-1 2 F54101-12 F54110-12 F54096-12 F54090-12 F54078-12 m L o f Cone Actual Quantity Dl: FC95 FC95 Meter sample TISAB % Actual Mass Mass sofutio solution reading (m l or final revry ppnrtF- spiked revrd spiked (ppm); (ppm F-) (grams) vol (mL) (ug/ug) in sample (ug F-! 0.021 0.017 0.075 0.022 0.033 0.10 0.10 0.10 0.10 0.10 2.0 2.0 2.0 2.0 2.0 ND ND ND ND 1.5 0.15 ND ND ND ND 0.031 0.10 2.0 ND ND 0.032 0.10 2.0 ND ND 0.043 0.10 2.0 ND ND 0.037 0.10 2.0 ND ND 0.024 0.10 2.0 ND ND 0.034 0.10 2.0 ND ND 0.045 0.10 2.0 ND ND 0.063 0.10 2.0 1.3 0.13 0.044 0.10 2.0 ND ND 0.048 0.10 2.0 ND ND 0.042 0.10 2.0 ND ND 0.045 0.10 2.0 ND ND 0.050 0.10 2.0 1.0 0.10 0.17 0.10 2.0 3.4 0.34 0.012 0.10 2.0 ND ND 0.018 0.10 2.0 ND ND 0.004 0.004 63 63 0.066 0.070 0.10 0.10 2.0 87% 2.0 93% 1.3 0.15 0.13 1.4 0.15 0.14 0.028 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.026 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.018 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.038 0.10 2.0 ND ND 0.032 0.10 2.0 ND ND 0.035 0.10 2.0 ND ND 0.022 0.10 2.0 ND ND 0.029 0.10 2.0 ND ND 0.025 0.10 2.0 ND ND 0.022 0.10 2.0 ND ND 0.014 0.10 2.0 ND ND 0.046 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.012 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.014 0.10 2.0 ND ND 000213 000133 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ID F54088-12 F54106-12 F54089-1 2 F54087-12 F54080-12 F54092-1 2 F54109-12 F54114-12 F54076-1 2 F54113-12 F54082-12 F54111-12 F54098-12 F54084-12 F54079-12 F54116-12 Serum Blk-1 Serum Blk-2 Serum Spk-1 Serum Spk-2 Serum Spk-3 Serum Spk-4 Serum Spk-5 Serum Spk-6 Serum Spk-7 Serum Spk-8 F54105-24 F54075-24 F54081-24 F54107-24 F54104-24 F54086-24 F54074-24 Blank 1 Blank 2 spike 1 spike 2 spike 3 spike 4 spike 5 spike 6 Blank 3 Blank 4 F54085-24 F54083-24 F54100-24 F54077-24 F54094-24 F54091-24 mL of Cone. Actual Quantity Dl: FC95 FC95 Meter sample TISAB % Actual Mass Mass solutio solution reading (mL or final revry ppm F- spiked revrd spiked (ppm) JEEELtL (grams) yol (mL) (ug/ug) in sample (ug F-) (ug F-) 0.025 o .io 2.0 ND ND "" 0.027 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.018 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.026 0.10 2.0 ND ND 0.027 0.10 2.0 ND ND 0.022 0.10 2.0 ND ND 0.034 0.10 2.0 ND ND 0.046 0.10 2.0 ND ND 0.042 0.10 2.0 ND ND 0.044 0.10 2.0 ND ND 0.043 0.10 2.0 ND ND 0.047 0.10 2.0 ND ND 0.033 0.10 2.0 ND ND 0.030 0.10 2.0 ND ND 0.004 63 0.034 0.10 2.0 44% 0.67 0.15 0.067 0.004 63 0.040 0.10 2.0 52% 0.79 0.15 0.079 0.004 63 0.050 0.10 2.0 66% 0.99 0.15 0.10 0.004 63 0.054 0.10 2.0 72% 1.1 0.15 0.11 0.004 63 0.083 0.10 2.0 110% 1.7 0.15 0.17 0.004 63 0.053 0.10 2.0 71% 1.1 0.15 0.11 0.004 63 0.049 0.10 2.0 65% 0.99 0.15 0.10 0.004 63 0.055 0.10 2.0 73% 1.1 0.15 0.11 0.041 0.10 2.0 ND ND 0.022 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.015 0.10 2.0 ND ND 0.018 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.013 0.10 2.0 ND ND 0.031 0.10 2.0 ND ND 0.022 0.10 2.0 ND ND 0.004 63 0.033 0.10 2.0 43% 0.65 0.15 0.065 0.004 63 0.044 0.10 2.0 58% 0.88 0.15 0.088 0.004 63 0.047 0.10 2.0 63% 0.95 0.15 0.095 0.004 63 0.047 0.10 2.0 62% 0.94 0.15 0.094 0.004 63 0.052 0.10 2.0 68% 1.0 0.15 0.10 0.004 63 0.051 0.10 2.0 67% 1.0 0.15 0.10 0.036 0.10 2.0 ND ND 0.020 0.10 2.0 ND ND 0.021 0.10 2.0 ND ND 0.019 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 0.017 0.10 2.0 ND ND 0.015 0.10 2.0 ND ND 0.016 0.10 2.0 ND ND 000134 000214 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ID F54108-24 F54102-24 F54097-24 F54071-24 F54093-24 F54095-24 F54101-24 F54110-24 F54096-24 F54090-24 F54078-24 F54088-24 F54106-24 F54089-24 F54087-24 F54080-24 F54092-24 F54109-24 F54114-24 serum blk 1 serum blk 2 serum spk 1 serum spk 2 F54113-24 F54082-24 F54111-24 F54098-24 F54084-24 F54079-24 F54116-24 F54113-48 F54082-48 F54111-48 F54098-48 F54084-48 F54079-48 F54116-48 F54113-192 F54082-1 92 F54111-192 F54098-192 F54079-1 92 F54084-192 F54116-192 F54113-360 F54082-360 F54111-360 F54098-360 F54079-360 mL of FC95 solutto spiked 0.004 0.004 Cone. Actual Quantity Dl: FC95 Meter sample TfSAB % solution reading (mL or final revry (ppm) (ppm F-) (grams) vot (mL) (ug/ug) 0.014 0.10 2.0 0.013 0.10 2.0 0.020 0.10 2.0 0.015 0.10 2.0 0.014 0.10 2.0 0.013 0.10 2.0 0.012 0.10 2.0 0.011 0.10 2.0 0.013 0.10 2.0 0.025 0.10 2.0 0.013 0.10 2.0 0.014 0.10 2.0 0.013 0.10 2.0 0.014 0.10 2.0 0.014 0.10 2.0 0.016 0.10 2.0 0.016 0.10 2.0 0.015 0.10 2.0 0.016 0.10 2.0 0.039 0.10 2.0 0.028 0.10 2.0 63 0.071 0.10 2.0 94% 63 0.082 0.10 2.0 109% 0.064 0.10 2.0 0.055 0.10 2.0 0.063 0.10 2.0 0.064 0.10 2.0 0.061 0.10 2.0 0.059 0.10 2.0 0.064 0.10 2.0 0.068 0.10 2.0 0.066 0.10 2.0 0.073 0.10 2.0 0.067 0.10 2.0 0.091 0.10 2.0 0.059 0.10 2.0 0.085 0.10 2.0 0.080 0.10 2.0 0.073 0.10 2.0 0.063 0.10 2.0 0.067 0.10 2.0 0.068 0.10 2.0 0.077 0.10 2.0 0.065 0.10 2.0 0.065 0.10 2.0 0.043 0.10 2.0 0.063 0.10 2.0 0.054 0.10 2.0 0.063 0.10 2.0 Actual ppm Fin sample ' ' ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 1.4 1.6 1.3 1.1 1.3 1.3 1.2 1.2 1.3 1.4 1.3 1.5 1.3 1.8 1.2 1.7 1.6 1.5 1.3 1.3 1.4 1.5 1.3 1.3 ND 1.3 1.1 1.3 Mass Mass spiked revrd fug F-) (ug F-) ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 0.15 0.15 ND 0.14 0.16 0.13 0.11 0.13 0.13 0.12 0.12 0.13 0.14 0.13 0.15 0.13 0.18 0.12 0.17 0.16 0.15 0.13 0.13 0.14 0.15 0.13 0.13 0.087 0.13 0.11 0.13 000215 r' l Jt .j 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 mL of Cone. Actual Quantity Dl: FC95 FC95 Meter sample TJSAB % Actual Mass Mass sdutto solution reading (mL or final revry ppm F- spiked revrd D spiked (ppm) [ppm F-) (grams) yo l [mL) (ug/ug) in sample ug F-) (ug F-) F54084-360 0.069 0.10 2.0 1.4 o . u " F54116-360 0.069 0.10 2.0 1.4 0.14 Serum Blk 1 Serum Blk 2 Serum Spk 1 Serum Spk 2 0.025 0.10 2.0 ND ND 0.032 0.10 2.0 ND ND 0.004 63 0.069 0.10 2.0 91% 1.4 0.15 0.14 0.004 63 0.076 0.10 2.0 101% 1.5 0.15 0.15 F54082-D22 0.046 0.10 2.0 ND ND F54111-D22 0.041 0.10 2.0 ND ND F54079-D22 0.040 0.10 2.0 ND ND F54116-D22 0.045 0.10 2.0 ND ND F54082-D28 0.048 0.10 2.0 ND ND F54111-D28 0.036 0.10 2.0 ND ND F54079-D28 0.033 0.10 2.0 ND ND F54116-D28 0.036 0.10 2.0 ND ND F54099-4 F54099-8 0.028 0.026 0.10 0.10 2.0 2.0 ND ND ND ND F54099-12 0.074 0.10 2.0 1.5 0.15 F54099-24 0.087 0.10 2.0 1.7 0.17 F54099-48 0.047 0.10 2.0 ND ND F54099-192 0.033 0.10 2.0 ND ND F54099-360 0.029 0.10 2.0 ND ND F54112-4 0.038 0.10 2.0 ND ND F54112-8 0.059 0.10 2.0 1.2 0.12 F54112-12 0.066 0.10 2.0 1.3 0.13 F54112-24 0.051 0.10 2.0 1.0 0.10 F54112-48 0.066 0.10 2.0 1.3 0.13 F54112-192 0.059 0.10 2.0 1.2 0.12 F54112-360 0.041 0.10 2.0 ND ND SERUM SPIKE-63-1 0.004 63 0.059 0.10 2.0 78% 1.2 0.15 0.12 SERUM SPIKE-63-2 0.004 63 0.079 0.10 2.0 105% 1.6 0.15 0.16 SERUM SPIKE-63-3 0.004 63 0.074 0.10 2.0 97% 1.5 0.15 0.15 SERUM SPIKE-126-1 0.004 126 0.083 0.10 2.0 55% 1.7 0.30 0.17 SERUM SPIKE-126-2 0.004 126 0.12 0.10 2.0 77% 2.3 0.30 0.23 SERUM SPIKE-126-3 0.004 126 0.12 0.10 2.0 79% 2.4 0.30 0.24 serum blank-1 0.10 0.10 2.0 2.0 0.20 serum blank-2 0.040 0.10 2.0 ND ND serum blank-3 0.019 0.10 2.0 ND ND spike 63-1 0.004 63 0.058 0.10 2.0 77% 1.2 0.15 0.12 spike 63-2 0.004 63 0.058 0.10 2.0 77% 1.2 0.15 0.12 spike 63-3 0.004 63 0.092 0.10 2.0 121% 1.8 0.15 0.18 spike 63-4 0.004 63 0.053 0.10 2.0 70% 1.1 0.15 0.11 spike 63-5 0.004 63 0.055 0.10 2.0 73% 1.1 0.15 0.11 spike 63-6 0.004 63 0.058 0.10 2.0 77% 1.2 0.15 0.12 spike 63-7 spike 63-8 0.004 63 0.072 0.10 2.0 95% 1.4 0.15 0.14 0.004 63 0.060 0.10 2.0 79% 1.2 0.15 0.12 Serum blnk 1 0.045 0.10 2.0 ND ND Serum blnk 2 0.017 0.10 2.0 ND ND Serum spike 63-1 0.004 63 0.12 0.10 2.0 153% 2.3 0.15 0.23 Serum spike 63-2 0.004 63 0.21 0.10 2.0 273% 4.1 0.15 0.41 000216 O O C l-i* 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ID Serum blank 1 Serum blank 2 Serum spike 1 Serum spike 2 Serum spike 3 Serum spike 4 Serum spike 5 Serum spike 6 Serum spike 7 Serum blank 3 Serum blank 4 F54088-48 F54099-48 F54089-48 F54087-48 F54080-48 F54092-48 F54109-48 F54114-48 F54088 dy8 F54099 dy8 serum spike 8 serum spike 9 serum spike 10 SERUM BLK 1 SERUM BLK 2 SERUM SPK 1 SERUM SPK 2 SERUM SPK 3 F54089-DY8 F54087-DY8 F54080-DY8 F54092-DY8 F54109-DY8 F54114-DY8 F54088-15 F54099-15 F54089-15 F54087-15 F54080-15 F54092-15 F54109-15 F54114-15 F54089-D22 F54087-D22 F54109-D22 F54114-D22 F54089-d28 F54087-d28 mL of FC95 solutio spiked 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 0.004 Cone Actual Quantity I: FC95 Meter sample TISAB % Actual solution reading (mL or final revry ppm F- [ppm) (ppm F-) (gramsl vol [mL) M f m L m sample 0.031 0.10 2.0 ND 0.021 0.10 2.0 ND 63 0.074 0.10 2.0 98% 1.5 63 0.066 0.10 2.0 87% 1.3 63 0.065 0.10 2.0 86% 1.3 63 0.060 0.10 2.0 80% 1.2 63 0.070 0.10 2.0 93% 1.4 63 0.069 0.10 2.0 91% 1.4 63 0.068 0.10 2.0 90% 1.4 0.023 0.10 2.0 ND 0.020 0.10 2.0 ND 0.019 0.10 2.0 ND 0.021 0.10 2.0 ND 0.017 0.10 2.0 ND 0.020 0.10 2.0 ND 0.017 0.10 2.0 ND 0.016 0.10 2.0 ND 0.022 0.10 2.0 ND 0.022 0.10 2.0 ND 0.018 0.10 2.0 ND 0.014 0.10 2.0 ND 63 0.059 0.10 2.0 78% 1.2 63 0.060 0.10 2.0 79% 1.2 63 0.060 0.10 2.0 79% 1.2 0.031 0.10 2.0 ND 0.016 0.10 2.0 ND 63 0.056 0.10 2.0 74% 1.1 63 0.069 0.10 2.0 91% 1.4 63 0.072 0.10 2.0 95% 1.4 0.026 0.10 2.0 ND 0.029 0.10 2.0 ND 0.029 0.10 2.0 ND 0.036 0.10 2.0 ND 0.028 0.10 2.0 ND 0.035 0.10 2.0 ND 0.010 0.10 2.0 ND 0.026 0.10 2.0 ND 0.022 0.10 2.0 ND 0.021 0.10 2.0 ND 0.019 0.10 2.0 ND 0.022 0.10 2.0 ND 0.025 0.10 2.0 ND 0.020 0.10 2.0 ND 0.019 0.10 2.0 ND 0.018 0.10 2.0 ND 0.019 0.10 2.0 ND 0.019 0.10 2.0 ND 0.018 0.10 2.0 ND 0.018 0.10 2.0 ND Mass Mass spiked revrd (ug F-) (ug F-] 0.15 0.15 0.15 0.15 0.15 0.15 0.15 ND ND 0.15 0.13 0.13 0.12 0.14 0.14 0.14 ND ND ND ND ND ND ND ND ND ND ND ND 0.15 0.12 0.15 0.15 0.12 0.12 ND ND 0.15 0.11 0.15 0.14 0.15 0.14 ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND ND 000217 ^0013' 6329159S.XLS S T U D Y # 6329-159 S E R U M 384 ID F54109-d28 F54114-d28 SERUM SPIKE 63-4 SERUM SPIKE 63-5 SERUM SPIKE 63-6 SERUM SPIKE 63-7 SERUM BLK 1 SERUM BLK 2 SERUM SPK 63-1 SERUM SPK 63-2 SERUM SPK 63-3 F54105-48 F54075-48 mL of Cone. Actual Quantity Dl: FC95 FC95 Meter sample TISAB % Actual solutto solution reading (mL or final revry ppm F- spiked (ppm) (ppm F-) (grams) vol (mL) (ug/ug) in sample 0.020 0.10 2.0 ND 0.004 0.004 0.004 0.004 63 63 63 63 0.017 0.031 0.049 0.061 0.059 0.10 0.10 0.10 0.10 0.10 2.0 2.0 41% 2.0 65% 2.0 81% 2.0 78% ND 0.62 0.98 1.2 1.2 0.031 0.10 2.0 ND 0.004 0.004 0.004 63 63 63 0.017 0.061 0.078 0.062 0.021 0.10 0.10 0.10 0.10 0.10 2.0 2.0 81% 2.0 103% 2.0 82% 2.0 ND 1.2 1.6 1.2 ND 0.026 0.10 2.0 ND Mass Mass spiked revrd fug F-) (ug F-) 0.15 0.15 0.15 0.15 0.15 0.15 0.15 ND ND 0.062 0.10 0.12 0.12 ND ND 0.12 0.16 0.12 ND ND 000218 n f'V l'JS C ontrol F 54074 F 54075 F 54081 F 54085 F 54086 F 54104 F 54105 F 54107 5 m g/kg F 54077 F 54083 F 54091 F 54094 F 54097 F 54100 F 54102 F 54108 10 m g/kg F 54071 F 54093 F 54095 F 54101 F 54078 F 54090 F 54096 F 54110 100 m g/kg F 54080 F 54087 F 54088 F 54089 F 54092 F 54099 F 54106 F 54109 F 54114 5 0 0 m g/kg F 54076 F 54079 F 54082 F 54084 F 54098 F 54111 F 54112 F 54113 F 54116 HWI 6329-159, AMDT 042095.1 ppm F- in serum 4 hour 0 .6 7 0 0 .6 4 5 0 .8 0 0 0 .8 9 8 0 .5 4 2 0 .4 9 2 0 .5 0 9 0 .4 8 4 8 hour 0 .3 0 4 0 .4 9 0 0 .3 9 8 0 .4 2 8 0 .6 1 6 0 .6 1 6 0 .4 2 6 0 .3 0 4 12 hour 0 .3 4 6 0 .5 5 8 0 .4 1 6 0 .3 6 6 0 .4 0 8 0 .5 0 2 0 .5 2 8 0 .3 8 8 24 hour 0 .2 6 8 0 .4 4 0 0 .3 4 6 0 .4 1 2 0 .3 2 8 0 .3 6 6 0 .8 1 2 0 .3 0 4 S u m m ary o f O rion seru m an alysis w ith ou t en terin g " N D ". T h is su m m a ry is u sed to d eterm in e the relative h a lf life, th erefo re it is a p p r o p r ia te to u se v a lu e s w h ich a re less than th e d etection lim it 4 hour 0 .4 1 6 0 .4 0 0 0 .3 8 4 0 .4 1 8 0 .4 4 6 0 .4 0 8 0 .4 0 6 0 .3 1 8 8 hour 0 .4 5 8 0 .3 5 2 0 .2 8 2 0 .5 0 6 0 .2 7 8 0 .3 4 6 0 .2 5 8 0 .2 5 4 12 hour 0 .8 3 9 0 .6 3 6 0 .5 7 6 0 .7 0 8 0 .4 9 2 0 .4 4 2 0 .4 3 6 0 .2 8 8 24 hour 0 .3 3 0 0 .3 8 0 0 .3 1 4 0 .3 0 8 0 .4 0 0 0 .3 2 4 0 .2 5 0 0 .2 7 8 4 hour 0 .3 3 6 0 .4 0 8 0 .3 1 6 0 .2 9 0 0 .4 1 2 0 .3 9 0 0 .3 3 6 0 .3 9 2 8 hour 0 .3 0 2 0 .3 7 4 0 .5 1 4 0 .3 2 2 0 .4 2 4 0 .4 2 4 0 .3 4 2 1 .5 0 0 12 hour 0 .4 1 2 0 .3 2 8 0 .3 2 0 0 .4 0 4 0 .2 8 4 0 .3 4 2 0 .2 4 6 0 .3 1 2 24 hour 0 .3 0 2 0 .2 7 4 0 .2 5 6 0 .2 4 0 0 .2 6 6 0 .5 0 0 0 .2 6 4 0 .2 2 6 4 hour 0 .5 0 8 0 .4 9 6 0 .5 5 2 0 .4 0 6 0 .3 7 6 0 .5 5 2 0 .6 3 0 0 .4 1 8 0 .7 7 6 8 hour 0 .6 3 8 0 .4 3 8 0 .6 6 2 0 .6 2 4 0 .7 4 0 0 .5 1 4 0 .8 6 0 0 .4 8 8 0 .6 7 6 12 hour 0 .3 5 8 0 .3 7 2 0 .5 0 8 0 .3 8 0 0 .3 3 2 1.478 0 .5 3 6 0 .4 1 0 0 .5 2 4 24 hour 0 .3 2 0 0 .2 7 0 0 .2 7 4 0 .2 7 8 0 .3 2 2 1.748 0 .2 5 6 0 .3 0 6 0 .3 2 2 48 hour 192 hour 360 hour 528 hour 672 hour 0 .3 9 8 0 .3 3 6 0 .5 8 8 0 .5 2 4 0 .4 2 6 0 .4 3 4 0 .3 6 2 0 .3 9 0 0 .3 5 0 0 .3 6 0 0 .9 4 8 0 .4 4 6 0 .4 3 8 0 .6 6 6 0 .5 6 4 0 .6 9 4 0 .5 7 2 0 .5 0 0 0 .4 0 0 0 .3 7 4 0 .3 9 0 0 .4 0 6 0 .3 3 0 4 hour 0 .8 1 0 2 .3 2 0 1 .1 3 0 1 .3 4 4 0 .9 2 6 1 .9 1 4 0 .7 5 8 1 .6 3 8 2 .0 4 0 8 hour 0 .8 9 0 0 .8 3 8 1 .2 5 4 0 .8 9 0 1 .0 0 0 0 .8 7 8 1 .1 8 4 0 .9 5 4 3 .4 4 0 12 hour 0 .5 4 6 0 .8 5 2 0 .6 8 6 0 .8 7 0 0 .8 3 6 0 .9 1 2 1 .3 1 0 0 .4 3 6 0 .9 4 0 24 hour 1.170 1 .1 0 4 1 .2 2 2 1 .2 7 6 1.252 1 .0 2 2 1 .2 7 6 1 .2 8 6 48 hour 192 hour 360 hour 528 hour 672 hour 1 .1 8 4 1 .3 1 8 1 .8 2 4 1 .3 3 2 1 .4 5 2 1 .3 1 4 1 .3 6 4 1 .7 0 6 1 .3 6 4 1 .4 5 8 1 .5 4 6 1 .3 4 4 1 .2 5 6 1 .1 8 4 1.596 1 .2 9 2 000219 1 .2 6 6 0 .8 6 6 1.370 1.078 1.252 0 .8 2 2 1.292 1 .3 8 0 0 .8 0 8 0 .9 1 2 0 .8 2 2 0 .8 9 0 0 .6 6 0 0 .9 6 4 OOOIO0 . 7 1 0 0 .7 2 8 Figure 2 ppm F- in Serum (Group 4) Or " cOWr C oncentration (ppm F-) 000220 Ity Figure 1 ppm F- in serum (Group 5) rr O wc- C oncentration (ppm F-) 000221 0 100 200 300 400 500 600 700 T im e (H ou rs P ost D ose) 9.11.4 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. 000222 OOOIA? 'OQJ RE: 6329-159 SERUM SRMPLES RMDT 42095.1 Date o f flnalysis: May 4, 8, 9, 11,24 and May 25, 1995 Rnalyst: DDLU The samples are burned in the Dohrman at 950 C using 0.10 mL o f the serum. The gas is collected in 2.0 mL o f 1:1 TISRB/Milli-Q m ate r. The samples are then analyzed on a Skalar Segmented Flom Rnalyzer using the Ion Specific Electrode (ISE) Method. TISRB b u ffe r is added to each sample as it proceeds through the system . The sample then goes through a heated mixing coil before the potential betmeen the ion selectiue electrode and the reference electrode is measured. The signal is am plified and related to the fluoride concentration. The in s tru m e n t mas calibrated in the ranges o f 0.015 - 0.15 ppm and 0.15 - 1.50 ppm fluoride. The standard curue fo r the high range mas p lo tte d using the inuerse lo ga rith m option. The standard curue fo r the lorn range is linear. Rll standards and samples mere then calculated by the Skalar so ftm are using these curues. Rll results belom 0.0001 ppm appear on the ram data as # . # # # # . fl q u a lity control standard mas analyzed euery 10 samples to check fo r accuracy and d rift. Ram data is taken fro m the appropriate calibrated range o f the Skalar p rin to u t and summarized on an Excel spreadsheet. The final results are adjusted fo r the collection uolume and any subsequent dilutions. 000223 000143 \ <4 . SUMMARY of 6329-159 SERUM SAMPLES AMDT 42095.1 GROUP 1 Dose Level : 0 W m Mmm i F54074 F 54075 F54080 F 54085 F 54085 F 54086 F 54104 F 54105 F 54107 0.51 0.61 0.84 0.98 0.58 0.34 0.46 mmmm iWl Wl iMi ziwlmi mi . WMiSii 0.35 0.64 0.40 0.62 0.73 0.55 0.31 0.42 0.63 0.43 0.29 0.70 0.81 0.60 0.40 W m m m 0.70 0.47 0.40 0.00 0.31 0.35 0.47 1.08 0.31 0.52 0.36 ^ 'tLa__>^'_ illlg M 1 in||lp ii VMprMi. GROUP 2 Dose Level : 5 mg/kg F54077 F 54077 F 54083 F 54083 F54091 F 54094 F 54097 F54100 F 54100 F 54102 F 54108 0.53 0.33 0.36 0.36 0.40 0.39 0.38 0.20 0.58 0.43 0.12 0.45 0.05 0.41 0.29 0.01 1.22 0.39 0.98 0.90 1.03 0.67 0.62 0.46 0.20 0 .3 4 0 .0 0 0 .4 2 0.28 0 .2 7 0.48 0.00 0.31 0.18 0.27 GROUP 3 Dose Level : 10 mg/kg F54071 F 54078 F 54090 F 54093 F 54095 F 54096 F54101 F 54110 0.24 0.39 0.28 0.43 0.24 0.13 0.15 0.25 0.16 0.22 0.20 0.29 0.41 0.00 0.13 1.85 0.00 0.32 0.49 0.00 0.00 0.04 0.00 0.00 0.24 0.52 1.02 0.21 0.42 0.23 0.34 0.30 GROUP 4 D ose L evel : 100 mg/kg F54080 F 54080 F54087 F54088 F54089 F54092 F54099 F54099 F 54106 F 54109 F54114 0.56 0.57 0.50 0.40 0.36 0.28 0.63 0.68 0.32 0.86 0.73 0.41 0.64 0.75 0.87 0.60 1.05 0.67 0.80 0.53 0.64 0.79 0.65 0.44 1.79 0.96 0.64 0.72 1.21 0.81 0.48 0.99 1.02 2.15 0.88 0.78 0.64 0.28 0.38 0.31 0.22 0.19 1.14 0.47 0.43 0.46 0.65 0.66 0.32 0.49 0.73 0.74 0.34 0.56 0.73 0.21 0.35 0 .0 0 0.33 0.43 0.61 0.53 0 .5 0 0 .2 9 0.27 0.32 0.29 0.34 0.34 0.23 0.34 0.36 Page 1 000224 000144 w m m m m m m m m am 'w m M ^ .WMMMMM m SUMMARY of 6329-159 SERUM SAMPLES AMDT 42095.1 IB E ' wm&WMi llpllppjg W mM M m GROUP 5 Dose L evel : 5 0 0 mg/kg F54076 F54079 F54082 F 54084 F54098 F54111 F54112 F 54113 F 5 4 1 1-6 0.92 2.94 1.40 1.78 1.25 2.42 1.05 2.23 2.61 1.17 1.36 1.75 1.44 1.58 1.19 1.47 1.40 4.56 0.68 1.25 0.95 1.30 1.39 1.40 1.65 0.66 1.32 1.00 0.86 1.03 1.10 1.18 1.25 1.17 1.20 1.05 1.37 1.73 1.37 1.52 1.63 1.44 1.69 1.28 1.28 1.46 1.25 1.02 1.45 1.55 1.15 1.36 0.71 1.40 1.04 1.22 0 .9 0 1.17 1.92 W m wm 1.07 1.17 1.14 . 1.08 0 .8 6 1.21 0.89 1.00 Page 2 000225 000145 HWI-159A.XLS 1995-06-21 10:11 O utPut of : 9 5 0 50 4 A 1 O perator : DDW D ate of the A nalysis : 1995-05-04 0 8 :1 8 A n alysis F ile N am e : C :\S K A L A R \D A T A \IIW ID A T A \S E R U M \950504A 1 Sjtater Startar % /X8T$AI* Qtyj&jupi ^ x x ' x? ' x n' 000226 1 Tracer 1.50 2 D rift 1.50 3 W ash 4 Standard 1 0 .0 1 5 5 Standard 2 0 .0 3 6 Standard 3 0 .0 6 7 Standard 4 0 .0 9 8 Standard 5 0 .1 2 9 Standard 6 0 .1 5 10 Standard 7 0 .3 0 11 Standard 8 0 .6 0 12 Standard 9 1.20 13 Standard 10 1.50 14 D rift 1.50 15 W ash 16 B lk 1 17 B lk 2 18 Spk 63-1 19 Spk 63-2 20 Spk 94.5-1 21 Spk 94.5-2 22 Spk 126-1 23 Spk 126-2 24 Spk 126-3 25 Spk 126-4 2 6 D rift 1.50 27 W ash 28 F54081-4 29 F54075-4 30 F 54107-4 31 F54105-4 1.46 1.47 0 .0 0 0 .0 1 5 0 .0 3 0 .0 6 0 .0 9 0 .1 2 0 .1 5 0 .2 9 0 .6 1 1.23 1.47 1.47 0 .0 0 0 .0 4 0 .0 2 0 .0 7 0 .0 7 0 .0 7 0 .0 4 0 .1 7 0 .0 9 0 .1 3 0 .1 4 1.43 0 .0 0 0 .0 4 0.03 0 .0 2 0 .0 2 97% 98% 100% 101% 99% 99% 102% 99% 96% 102% 102% 98% 98% 95% 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 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 000146 Page 1 c 0 .7 7 0 .4 8 1.37 1.47 1.46 0 .8 7 3.42 1.87 2 .6 9 2 .8 6 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 9 4 .5 0 9 4 .5 0 126.00 126.00 126.00 126.00 0 .1 5 0 .1 5 0.23 0.23 0 .3 0 0 .3 0 0 .3 0 0 .3 0 0 .8 4 0.61 0 .4 6 0.34 0.14 0.15 0.15 0 .0 9 0 .3 4 0 .1 9 0 .2 7 0 .2 9 91% 97% 65% 38% 113% 62% 89% 95% S' W "? 5 rg J> f i i yj' c \i lA . iA o ^ p } HWI-159A.XLS Sampler i Skaiar v SkafcyV / % BOTSABC ; SflHXtorw S L m M m W M m m M m m . WmiiBfi Wioei Mass ' % Repayerai >Recovery, 32 F54074-4 33 Blk-1 34 Blk-2 35 Spk63-1 36 Spk63-2 37 Spk63-3 38 Drift 1.50 39 Wash 40 Spk 126-1 41 Spk 126-2 42 Spk 126-3 43 Spk 126-4 44 Spk 126-5 45 Spk 126-6 46 Spk 126-7 47 F54085-4 48 F54080-4 49 F5 4 104-4 50 Drift 1.50 51 Wash 52 F54077-4 53 F54083-4 54 F54094-4 55 F5 4 100-4 56 F54091-4 57 Blk1 58 Blk2 59 Spk 1 60 Spk2 61 F54097-4 62 Drift 1.50 63 Wash 64 F5 4 102-4 65 F54092-4 66 F54080-4 67 F54106-4 0.03 2.0 0.83 2.0 0.40 2.0 0.08 2.0 0.09 2.0 0.10 2.0 1.46 98% 0.00 0.14 2.0 0.14 2.0 0.14 2.0 0.15 2.0 0.13 2.0 0.15 2.0 0.12 2.0 0.05 2.0 0.03 2.0 0.03 2.0 1.47 98% 0.00 0.03 2.0 0.02 2.0 0.02 2.0 0.02 2.0 0.02 2.0 0.02 2.0 0.01 2.0 0.08 2.0 0.08 2.0 0.02 2.0 1.44 96% 0.00 0.02 2.0 0.01 2.0 0.03 2.0 0.03 2.0 0.10 0.51 0.10 16.56 0.10 8.04 0.10 1.53 0.004 63.00 0.15 0.10 1.86 0.004 63.00 0.15 0.10 1.93 0.004 63.00 0.15 0.10 2.79 0.004 126.00 0.30 0.10 2.83 0.004 126.00 0.30 0.10 2.74 0.004 126.00 0.30 0.10 2.93 0.004 126.00 0.30 0.10 2.58 0.004 126.00 0.30 0.10 3.07 0.004 126.00 0.30 0.10 2.36 0.10 0.98 0.10 0.56 0.10 0.58 0.10 0.53 0.10 0.33 0.10 0.36 0.10 0.39 0.10 0.36 0.10 0.44 0.10 0.26 0.10 1.60 0.10 1.67 0.10 0.40 0.10 0.38 0.10 0.28 0.10 0.57 0.10 0.68 0.15 101% 0.19 123% 0.19 127% 0.28 92% 0.28 94% 0.27 90% 0.29 97% 0.26 85% 0.31 101% 000227 000147 Page 2 ft 000145 000228 !tS t' jgj 11 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 93 94 95 * F54 lt89-4 F54/J0 8 8 - 4 F5 4 J'0 8 7 - 4 F54H 10-4 F5 4 0 9 6 - 4 F5 4 0 9 0 - 4 Drift Wash F5 4 0 7 8 - 4 F5 4 1 0 1 - 4 F5 4 0 9 5 - 4 F5 4 0 9 3 - 4 F5 4 0 7 1 - 4 F5 4 1 0 8 - 4 F5 4 1 1 6 - 4 F5 4 0 9 8 - 4 F5 4 0 8 4 - 4 F5 4 0 7 9 - 4 Drift Wash F5 4 1 1 3 - 4 F5 4 1 1 1 - 4 F5 4 0 8 2 - 4 F5 4 0 7 6 - 4 F5 4 114-4 F5 4 1 0 9 - 4 Drift Wash ,,, - 1.50 1.50 1.50 / A, " ' WM 0.02 0.02 0.02 0.01 0.01 0.01 1.46 97% 0.00 0.02 0.01 0.01 0.02 0.01 0.01 0.13 0.06 0.09 0.15 1.48 99% 0.00 0.11 0.12 0.07 0.05 0.04 0.02 1.47 98% 0.00 HWI-159A.XLS 2.0 0.10 0.39 2.0 0.10 0.15 2.0 0.10 0.24 2.0 0.10 0.43 2.0 0.10 0.24 2.0 0.10 0.20 2.0 0.10 2.61 2.0 0.10 1.25 2.0 0.10 1.78 2.0 0.10 2.94 2.0 0.10 2.23 2.0 0.10 2.42 2.0 0.10 1.40 2.0 0.10 0.92 2.0 0.10 0.86 2.0 0.10 0.32 Page 3 HWI-159B.XLS 1995-06-21 10:15 OutPutof: 950504B1 Operator :DDW DateoftheAnalysis: 1995-05-04 13:07 AnalysisFileName : C:\SKALAR\DATA\HWIDATA\SERUM\950504B1 000229 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 F54075-8 17 F54081-8 18 F54105-8 19 F54107-8 20 F54074-8 21 F54085-8 22 F54086-8 23 BLK-1 24 BLK-2 25 SPK 1 26 Drift 1.50 27 Wash 28 SPK 2 29 F54104-8 30 F54077-8 31 F54083-8 1.47 1.47 0.00 0.014 0.03 0.06 0.09 0.12 0.15 0.29 0.61 1.23 1.48 1.45 0.00 0.03 0.02 0.03 0.02 0.02 0.03 0.04 0.02 0.07 0.10 1.44 0.00 0.08 0.04 0.03 0.02 98% 98% 93% 104% 103% 97% 101% 100% 97% 101% 102% 98% 97% 96% 2.0 2.0 2.0 2.0 00..1100 00..1100 2.0 2.0 2.0 00..1100 0.10 2.0 2.0 2.0 00..1100 0.10 2.0 2.0 0.10 0.10 2.0 2.0 00..1100 Page 1 000119 0.64 0.40 0.55 0.31 0.35 0.62 0.71 0.42 1.33 2.00 0.004 63.00 0.15 1.55 0.73 0.58 0.43 0.20 132% L/TO OCZOOO ' AMjpp) 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 67 Skalar F54094-8 F54100-8 F54091-8 F54097-8 F54102-8 F54108-8 Drift Wash F54071-8 F54093-8 F54095-8 F54101-8 F54078-8 F54090-8 F54096-8 F54110-8 F54087-8 F54088-8 Drift Wash F54089-8 F54106-8 F54080-8 F54092-8 F54109-8 F54114-8 F54076-8 F54082-8 F54111-8 F54113-8 Drift Wash F54079-8 F54084-8 F54098-8 F54116-8 1.50 1.50 1.50 Skalar 0.02 0.02 0.01 0.00 0.01 0.00 1.40 0.00 0.01 0.01 0.02 0.01 0.01 0.01 0.00 0.09 0.02 0.03 1.48 0.00 0.04 0.05 0.04 0.04 0.03 0.04 0.06 0.09 0.06 0.07 1.51 0.00 0.07 0.07 0.08 0.23 HWI-159B.XLS % Di.TlSAS Qty & w p} Actual- Wii W mk 2.0 0.10 0.45 2.0 0.10 0.41 2.0 0.10 0.12 2.0 0.10 0.05 2.0 0.10 0.29 2.0 0.10 0.01 93% 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 99% 0.10 0.16 0.10 0.29 0.10 0.41 0.10 0.13 0.10 0.22 0.10 0.20 0.10 0.00 0.10 1.85 0.10 0.41 0.10 0.64 2.0 0.10 0.75 2.0 0.10 1.05 2.0 0.10 0.73 2.0 0.10 0.87 2.0 0.10 0.67 2.0 0.10 0.80 2.0 0.10 1.17 2.0 0.10 1.75 2.0 0.10 1.19 2.0 0.10 1.40 101% 2.0 0.10 1.36 2.0 0.10 1.44 2.0 0.10 1.58 2.0 0.10 4.56 Page 2 HWI-159B.XLS 68 Drift 1.50 1 .5 1 101% 6 9 W ash 0.0 0 000231 0001 Cl H1 Page 3 HWI-159C.XLS 1995-06-13 13:25 OutPut of : 9S0S08A1 Operator : DDW Date of the Analysis : 1995-05-08 07:30 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950508A1 000232 Wmmf. 1 Tracer 1.5 2 Drift 1.5 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 0.3 11 Standard 8 0.6 12 Standard 9 1.2 13 Standard 10 1.5 14 Drift 1.50 15 Wash 16 Blk 1 17 Blk 2 18 Spk 1 19 Spk2 20 F54075-12 21 F54081-12 22 F54105-12 23 F54107-12 24 F54074-12 25 F54085-12 26 Drift 1.50 27 Wash 28 F54086-12 O 29 F54104-12 30 F54077-12 TO 31 F54083-12 Cd S 1111TM I f f ll l l jP, 3 / 1.45 1.47 0.00 0.014 0.03 0.06 0.09 0.12 0.15 0.29 0.61 1.24 1.46 1.47 0.00 0.03 0.01 0.08 0.09 0.03 0.02 0.03 0.02 0.02 0.01 1.45 0.00 0.04 0.04 0.06 0.05 96% 98% 93% 104% 100% 104% 96% 103% 96% 101% 103% 98% 98% 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 2.0 0.10 Page 1 0.60 0.26 1.66 1.77 0.63 0.43 0.60 0.40 0.42 0.29 0.70 0.81 1.22 0.98 0.004 63.00 0.004 63.00 0.15 0.17 0.15 0.18 110% 117% V S Vcn>, 00015 0 0233 32 F54094-12 33 F54100-12 34 F54091-12 35 F54097-12 36 F54102-12 37 F54108-12 38 Drift 1.50 39 Wash 40 F54077-12 41 F54071-12 42 F54093-12 43 F54095-12 44 F54101-12 45 F54110-12 46 F54096-12 47 F54090-12 48 F54078-12 49 F54088-12 50 Drift 1.50 51 Wash 52 F54106-12 53 F54089-12 54 F54087-12 55 F54080-12 56 F54092-12 57 F54109-12 58 F54114-12 59 F54076-12 60 F54113-12 61 F54082-12 62 Drift 1.50 63 Wash 64 F54111-12 65 F54098-12 66 F54084-12 67 F54079-12 0.05 0.03 0.04 0.03 0.02 0.01 1.47 98% 0.00 0.02 0.00 0.00 0.00 0.00 0.00 0.00 0.02 0.02 0.04 1.46 97% 0.00 0.05 0.03 0.03 0.03 0.02 0.03 0.04 0.03 0.03 0.05 1.44 96% 0.00 0.07 0.07 0.07 0.06 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 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 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 2 1.03 0 .6 2 0 .9 0 0 .6 7 0 .4 6 0.20 0 .3 9 0.00 0.00 0.00 0.00 0.00 0 .0 4 0 .4 9 0 .3 2 0 .7 9 0 .9 6 0.65 0 .6 4 0.53 0 .4 4 0 .6 4 0.72 0.68 0.66 0 .9 5 1.40 1.39 1.30 1.25 HWI-159C.XLS 68 F54116-12 0.07 2 .0 0 .1 0 1.32 69 Drift 1.50 1.52 101% 70 Wash 0.00 000234 o 3 U Page 3 HWI-159D.XLS 1995-06-13 10:27 O utPut of : 950509A 1 O perator : DDW D ate of the A nalysis : 1995-05-09 09:4 8 A n alysis F ile N am e : C :\S K A L A R \D A T A \H W ID A T A \S E R U M \950509A 1 000235 1 Tracer 2 D rift 1.5 1.5 3 W ash 4 Standard 1 0 .0 1 5 5 Standard 2 0 .0 3 6 Standard 3 0 .0 6 7 Standard 4 0 .0 9 8 Standard 5 0 .1 2 9 Standard 6 0 .1 5 10 Standard 7 0.3 11 Standard 8 0.6 12 Standard 9 1.2 13 Standard 10 1.5 14 D rift 1.5 15 W ash 16 Serum blk 1 17 Serum blk 2 18 Serum spk 1 19 Serum spk 2 20 Serum spk 3 21 Serum spk 4 22 Serum spk 5 23 Serum spk 6 24 Serum spk 7 25 Serum spk 8 2 6 D rift 1.50 27 W ash 28 F 54105-24 29 F54075-24 30 F54081-24 31 F54107-24 1.47 1.50 0 .0 0 0 .0 1 4 0.03 0 .0 6 0 .0 9 0 .1 2 0 .1 5 0 .2 9 0.61 1.23 1.47 1.55 0 .0 0 0 .0 5 0 .0 3 0 .0 4 0 .0 5 0 .0 6 0 .0 7 0 .1 0 0 .0 7 0 .0 6 0 .0 7 1.51 0 .0 0 0 .0 5 0 .0 2 0 .0 2 0 .0 2 98% 100% 94% 106% 98% 101% 99% 102% 97% 101% 103% 98% 103% 101% 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 2.0 0.10 Page 1 000155 0 .9 0 0 .6 2 0 .8 7 0.91 1.17 1.30 2.03 1.43 1.27 1.46 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 0.15 0 .1 5 0 .1 5 0 .1 5 0.1 5 0 .1 5 0.15 0 .15 1.08 0 .4 7 0 .4 0 0.31 0 .0 9 0 .0 9 0.12 0.13 0 .2 0 0 .1 4 0.13 0 .1 5 58% 60% 77% 86% 134% 94% 84% 96% i >i s b oxy> cvm i o o o ia 000236 mm 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 67 <J) ,SkaJar Skater .% HWI-159D.XLS ' DI;TlSA8,Qiy Sathp! - '" L /C # * ' ' Cbpc a s s jy&pvi&c&gg g l l Mass F 54104-24 F 54086-24 F 54074-24 F 54085-24 F 54083-24 F 54100-24 D rift W ash B lk 1 B lk 2 Spk 1 Spk 2 Spk 3 Spk 4 Spk 5 Spk 6 B lk 3 F 54085-24 D rift W ash F 54083-24 F 54100-24 F 54077-24 F 54094-24 F 54091-24 F 54108-24 F 54102-24 F 54097-24 F 54071-24 F 54093-24 D rift W ash F 54095-24 F 54101-24 F 54110-24 F54096-24 1.50 1.50 1.50 0 .0 2 0 .0 2 0 .0 4 0 .0 0 0 .0 0 0 .0 0 1.47 0 .0 0 0.03 0.01 0 .0 4 0 .0 6 0 .0 6 0 .0 6 0 .0 7 0 .0 7 0 .0 4 0 .0 2 1.51 0 .0 0 0 .0 2 0 .0 2 0 .0 2 0.01 0.01 0.01 0.01 0 .0 2 0.01 0.01 1.49 0 .0 0 0 .0 2 0 .0 2 0 .0 2 0.01 98% 100% 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 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 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 2 0 .4 7 0 .3 5 0 .7 0 0 .0 0 0 .0 0 0 .0 0 0 .6 2 0 .2 7 0 .8 0 1.11 1.16 1.18 1.35 1.32 0 .8 2 0.31 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 0.15 0.15 0.15 0.15 0.15 0.15 0 .4 2 0 .3 1 0 .3 4 0 .2 7 0 .2 8 0 .2 7 0 .1 8 0 .4 8 0 .2 4 0.21 0.42 0 .3 4 0 .3 0 0.23 0 .0 8 0.11 0 .1 2 0 .1 2 0.13 0.13 % 53% 73% 76% 78% 89% 87% HWI-159D.XLS 68 69 70 jf l \ \ll \ 73 - ULjtT- JO Co o h* Cl Al 1VJ7 45 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94 1 2 3 4 5 6 7 8 F 54090-24 F 54078-24 F 54088-24 F 54106-24 F 54089-24 F 54087-24 Drift W ash F 54080-24 F 54092-24 F 54109-24 F 54114-24 B lk 1 Blk 2 Spk 1 Spk 2 F 54113-24 F 54082-24 Drift W ash F 54111-24 F 54098-24 F 54084-24 F 54079-24 F 54116-24 Drift W ash Tracer Drift W ash F 54106-24 F 54089-24 F 54087-24 Drift W ash 1.50 1.50 1.50 1.50 1.50 1.50 0 .0 5 0.03 0 .0 2 0.81 0 .4 4 0 .5 7 0 .2 2 0 .0 0 0 .0 6 0 .0 5 0 .0 4 0.03 0 .0 2 0.01 0 .0 9 0.11 0 .0 6 0 .0 4 1.42 0 .0 0 0 .0 6 0 .0 5 0 .0 5 0 .0 5 0 .0 6 1.46 0 .0 0 1.51 1.52 0 .0 0 0 .0 4 0 .0 5 0 .0 4 1.62 0 .0 0 15% 95% 97% 101% 101% 108% 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 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 3 1.02 0 .5 2 0 .4 8 16.10 8 .7 0 11.48 1.21 1.02 0 .7 8 0 .6 4 0.41 0 .2 9 1.75 2.13 1.17 0 .8 6 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 1.18 1.10 1.03 1.00 1.20 0 .8 8 0 .9 9 0.81 0.15 0.1 5 0 .1 7 0 .2 1 115% 141% HWI-159E.XLS 1995-06-13 10:27 OutPut of : 950511A1 ; Operator : DDW Date of the Analysis : 1995-05-11 08:16 Analysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950511A1 00023S 1 Tracer 1.50 1.47 98% 2 Drift 1.50 1.47 98% 3 Wash 0.00 4 Standard 1 0.015 0.016 109% 5 Standard 2 0.03 0.03 96% 6 Standard 3 0.06 0.06 98% 7 Standard 4 0.09 0.09 102% 8 Standard 5 0.12 0.12 100% 9 Standard 6 0.15 0.15 102% 10 Standard 7 0.30 0.29 96% 11 Standard 8 0.60 0.61 101% 12 Standard 9 1.20 1.23 103% 13 Standard 10 1.50 1.47 98% 14 Drift 1.50 1.51 101% 15 Wash 0.00 16 F54113-48 0.07 2.0 0.10 17 F54082-48 0.07 2.0 0.10 18 F54111-48 0.08 2.0 0.10 19 F54098-48 0.07 2.0 0.10 20 F54084-48 0.09 2.0 0.10 21 F54079-48 0.05 2.0 0.10 22 F54116-48 0.08 2.0 0.10 23 F54113-192 0.08 2.0 0.10 24 F54082-192 0.06 2.0 0.10 25 F54111-192 0.05 2.0 0.10 26 Drift 1.50 1.44 96% O 27 Wash o 28 F54098-192 29 F54079-192 30 F54084-192 C/l 31 F54116-192 X 0.00 0.06 0.06 0.07 0.06 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 1 , 1.44 1.37 1.52 1.37 1.73 1.05 1.69 1.55 1.28 1.02 1.25 1.28 1.46 1.15 HWI-159E.XLS 6CZ000 32 F54113-360 33 F54082-360 34 F54111-360 35 F54098-360 36 F54079-360 37 F54084-360 38 Drift 1.50 39 Wash 40 F54116-360 41 BLK 1 42 SPK 1 43 SPK 2 44 F54082-D22 45 F54111-D22 46 F54079-D22 47 F54116-D22 48 F54082-D28 49 F54111-D28 50 Drift 1.50 51 Wash 52 F54079-D28 53 F54116-D28 54 F54099-4 55 F54099-8 56 F54099-12 57 F54099-24 58 F54099-48 59 F54099-192 60 F54099-360 61 1&4UJJ_Ju l 62 Drift 1.50 63 Wash 64 F54112-4 65 F54112-8 66 F54112-12 67 F54112-24 0.06 0.04 0.06 0.05 0.07 0.07 1.48 0.00 0.10 0.02 0.07 0.10 0.06 0.06 0.05 0.05 0.06 0.04 1.52 0.00 0.04 0.05 0.03 0.03 0.09 0.11 0.06 0.04 0.03 nu.ur\ur\ 1.50 0.00 0.05 0.07 0.08 0.06 2.0 2.0 2.0 2.0 2.0 2.0 99% 0.10 0.10 0.10 0.10 0.10 0.10 101% 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 0.10 100% 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 - 2'i.UA 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 0.10 -----ttrrW-- 2.0 0.10 2.0 0.10 2.0 0.10 2.0 0.10 Page 2 1.17 01..7212 1.04 1.36 1.40 1.92 0 .3 8 1.45 2 .0 0 1.17 1.14 1.07 11 ..2018 0 .8 9 0 .0 0 4 0 .0 0 4 0.86 1.00 0.63 0 .6 0 1.79 2.15 1.14 0 .7 4 0fr6O1G---- 1.05 1.47 1.65 1.25 6 3 .0 0 6 3 .0 0 0 .1 5 0 .1 5 0 .1 5 0 .2 0 96% 132% I cuw `'jz .ih s _.T0()0 HWI-159E.XLS 68 F54112-48 69 F54112-192 70 F54112-360 71 SPK 63-1 72 SPK 63-2 73 SPK 63-3 74 Drift 1 .5 0 75 Wash 76 SPK 126-1 77 SPK 126-2 78 SPK 126-3 79 Drift 1 .5 0 80 Wash 0.08 0.07 0.05 0.08 0.10 0.07 1.47 0.00 0.11 0.15 0.15 1.50 0.00 98% 100% 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 1.63 1.45 0 .9 0 1.50 1.95 1.38 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 0 .1 0 0 .1 0 0 .1 0 2 .1 2 2 .9 0 3 .0 7 0 .0 0 4 0 .0 0 4 0 .0 0 4 1 2 6 .0 0 1 2 6 .0 0 1 2 6 .0 0 0 .1 5 0 .1 5 0 .1 5 0 .1 5 0 .1 9 0 .1 4 99% 129% 91% 0 .3 0 0 .3 0 0 .3 0 0.21 0 .2 9 0.31 70% 96% 102% 000240 0001(70 Page 3 HWI-159F.XLS 1995-06-13 12:41 O utPut of : 950524A 1 O perator : DDW D ate o f the A nalysis : 1995-05-24 11:39 A n alysis File N am e : C :\S K A L A R \D A T A \H W ID A T A \S E R U M \950524A 1 000241 1 Tracer 2 D rift 3 W ash 4 Standard 1 5 Standard 2 6 Standard 3 7 Standard 4 8 Standard 5 9 Standard 6 10 Standard 7 11 Standard 8 12 Standard 9 13 Standard 10 14 D rift 15 W ash 16 BLK 1 17 BLK 2 18 SP K 63-1 19 SPK 63-2 20 SPK 63-3 21 SPK 63-4 22 SPK 63-5 23 SPK 63-6 24 SPK 63-7 25 BLK 3 2 6 D rift 27 W ash 28 BLK 4 1.5 1.5 0 .0 1 5 0.03 0 .0 6 0 .0 9 0 .1 2 0 .1 5 0.3 0.6 1.2 1.5 1.50 1.50 1.45 1.47 0 .0 0 0 .0 1 8 0.03 0 .0 6 0 .0 9 0 .1 2 0 .1 6 0 .2 8 0.61 1.24 1.46 1.44 0 .0 0 0.03 0 .0 2 0 .0 8 0 .0 8 0 .0 8 0 .0 7 0 .0 9 0 .0 8 0 .0 8 0.03 1.46 0 .0 0 0.03 97% 98% 120% 92% 95% 105% 99% 103% 95% 102% 104% 97% 96% 97% 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 2.0 0.10 00.,1100 00..1100 0.10 00..1100 0.10 0.10 0.10 OOOIGI Page 1 S> 0.66 0 .3 4 1.69 1.52 1.54 1.37 1.72 1.66 1.64 0.53 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 0.55 0.15 0.15 0.15 0.15 0.15 0.15 0 .1 5 0.17 0.15 0.15 0.14 0.17 0 .1 7 0 .1 6 111% 101% 102% 90% 114% 110% 109% HWI-159F.XLS W CV 00016 000242 tentate w JkJk SLfckt Skater % DI TJSAB Q ty Sani y '' ,, S u tu r ^, W sdp/j S& j!Jf& fy / / //// * 29 F54088-48 30 F54099-48 31 F 54089-48 32 F54087-48 33 F54080-48 34 F54092-48 35 F 54109-48 36 F54114-48 37 F54088-D Y 8 3 8 D rift 1.50 39 W ash 40 F54099-D Y 8 41 SPK 63-8 42 SPK 63-9 43 SPK 63-10 44 BLK 1 45 BLK 2 46 SPK 63-1 47 SPK 63-2 48 SPK 63-3 49 F54089-D Y 8 5 0 Drift 1.50 51 W ash 52 F54087-D Y 8 53 F54080-D Y 8 54 F54092-D Y 8 55 F54109-D Y 8 56 F54114-D Y 8 57 F54088-D 15 58 F54099-D 15 59 F54089-D 15 60 F54087-D15 61 F54080-D 15 0 .0 2 0 .0 2 0 .0 1 0 .0 2 0.01 0.01 0 .0 2 0 .0 2 0 .0 2 1.49 0 .0 0 0 .0 2 0 .0 8 0 .0 8 0 .0 8 0.03 0.01 0 .0 6 0 .0 8 0 .0 8 0 .0 2 1.47 0 .0 0 0 .0 3 0 .0 3 0 .0 4 0.03 0 .0 4 0 .0 0 0 .0 3 0 .0 2 0 .0 2 0.01 99% 98% 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 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 2.0 0.10 2.0 0.10 2.0 0.10 Page 2 0.31 0 .4 7 0.22 0 .3 8 0 .2 8 0 .1 9 0.43 0 .4 6 0 .3 2 0 .3 4 1.62 1.55 1.58 0 .5 5 0 .1 7 1.19 1.53 1.68 0 .4 9 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 0.66 0 .6 5 0.73 0 .5 6 0.73 0.00 0.53 0.33 0.35 0.21 0 .1 5 0 .1 5 0 .1 5 0 .1 6 0 .1 5 0 .1 6 107% 102% 104% 0 .1 5 0.15 0 .1 5 0 .1 2 0.15 0 .1 7 79% 101% 111% 6 2 D rift 1.50 63 W ash 64 F54092-D 15 65 F54109-D 15 66 F54114-D 15 67 F54089-D 22 68 F54087-D 22 69 F54109-D 22 70 F54114-D 22 71 F54089-D 28 72 F54087-D 28 73 F54109-D 28 7 4 D rift 1.50 7 5 W ash 76 F54114-D 28 77 SPK 63-4 78 SPK 63-5 79 SPK 63-6 80 SPK 63-7 81 F54105-48H R 82 F54075-48H R 83 D rift 1.50 84 W ash 1.47 0 .0 0 0 .0 2 0 .0 2 0 .0 1 0 .0 2 0.01 0.01 0 .0 2 0.01 0 .0 2 0 .0 2 1.46 0 .0 0 0 .0 2 0 .0 4 0 .0 6 0 .0 8 0 .0 8 0 .0 2 0.03 1.41 0 .0 0 98% 97% 94% 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 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0.43 0 .5 0 0 .2 9 0 .3 2 0 .2 7 0 ,2 9 0 .3 4 0.23 0 .3 4 0 .3 4 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .1 0 0 .3 6 0 .7 6 1.22 1.53 1.50 0 .3 6 0 .5 2 0 .0 0 4 0 .0 0 4 0 .0 0 4 0 .0 0 4 6 3 .0 0 6 3 .0 0 6 3 .0 0 6 3 .0 0 0 .1 5 0 .1 5 0 .1 5 0.15 0 .0 8 0 .1 2 0 .1 5 0 .1 5 50% 81% 101% 99% 000243 000103 Page 3 995-05-04 15:44 Output of : 950504A1 JUUoO -VMA'V* oftware : version 5 1 c l 9 9 0 ,93 perator DDW BEST COPy AVAILABLE ate of the Analysis 1 9 9 5 - 0 5 - 0 4 0 8 :1 8 nalysis File Name C:\ SKALAR\ DATA\ HWI DATA\ S E R U M \ 9 5 0 5 0 4 A 1 luoride 1.5 'a l i b r a t i o n o r d e r = Inverse L o g arithm Hope : s = #.##### r x - cl -i I ------- :esult = 1 0 L s -* x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode 2 = - 0.00000 1 = 0.00063 0 = -1.24458 luoride L alibration order = 2 C orrelation : r = 0.9 9 9 2 8 esult = a2 * X a + al * X + aO 2 = 0.00000 1 = 0.00018 0 = -0.00141 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec 1 2 0 sec, 1 sec, : Single : None : 70 mm. iluter needle Height dilution Factor dilution Volume Resample Dilution runs : 80 : 10 : 2.5 :1 :1 mm ml User file : Reproces : No . TXT 000244 000164 995-05-04 16:44 OutPut of : 950504A1 luoride 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 fffsrcopK4 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 M e a s u r e w i n d o w 75 % Filter No Regeneration No formula : output : ##. # # # luoride 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 000245 000165 13 .995-05-04 16:44 OutPut of : 950504A1 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 M e a s u r e w i n d o w 75 % Filter No Regeneration No f o r m u l a : c4:==c3 output : #.#### 000246 000166 .995-05-04 16:44 OutPut of : 950504A1 Page 1 of 2 F l u o r i d e 1. 5 Fluoride L PPM PPM 'OS Typ Ident Ch Result F Time Ch Result F Time 't iw Initial W a s h 3 0.057 t Tracer 3 1.462 d Drift 3 1.467 w Wash 3 0.057 si Standard 1 3 0.065 ) s2 Standard 2 3 0.073 > s3 Standard 3 3 0.090 s4 Standard 4 3 0.109 s5 Standard 5 3 0.132 s6 Standard 6 3 0.153 .0 s7 Standard 7 3 0.288 .. s8 Standard 8 3 0.609 .2 s9 Standard 9 3 1.226 .3 slO S t a n d a r d 10 3 1.474 .4 d Drift 3 1.472 .5 w Wash 3 0.057 .6 u Blk 1 3 0.077 .7 u Blk 2 3 0.070 .8 u Spk 63-1 3 0.095 9u Spk 63-2 3 0.098 .0 u Spk 94.5-1 3 0.098 :i u Spk 94.5-2 3 0.080 .2 u Spk 126-1 3 0.171 3u Spk 126-2 3 0 .1 1 2 .4 u Spk 126-3 3 0.142 .5 u Spk 126-4 3 0.149 .6 d Drift 3 1.425 7w Wash 3 0.057 3u F54081-4 3 0.079 9u F54075-4 3 0.073 0u F54107-4 3 0.069 1u F54105-4 3 0.066 2u F54074-4 3 0.070 3u Blk-1 3 0.828 4u Blk-2 3 0.402 3u Spk 63-1 3 0 .1 0 0 6u Spk 63-2 3 0 .1 1 1 7u Spk 63-3 3 0.113 8d Drift 3 1.464 9w Wash 3 0.057 0u Spk 126-1 3 0.146 1u Spk 126-2 3 0.147 2u Spk 126-3 3 0.144 3u 4u Spk 126-4 3 0.151 Spk 126-5 3 0.138 5u Spk 126-6 3 0.157 6u Spk 126-7 3 0.129 7u F54085-4 3 0.083 8u F54080-4 3 0.072 9u F54104-4 3 0.072 0d 1w Drift 3 1.467 Wash 3 0.057 2u F54077-4 3 0.071 3u F 54083-4 3 0.066 65 212 387 629 740 915 1087 1263 1436 1611 1787 1961 2137 2313 2487 2729 2838 3013 3188 3364 3538 3714 3888 4062 4238 4414 4588 4756 4938 5113 5291 5465 5641 5815 5989 6165 6339 6515 6689 6911 7040 7215 7390 7566 7740 7916 8092 8266 8442 8623 8790 9032 9138 9314 4 #.#### 4 1.0733 4 1.0775 4 #.#### 4 0.0150 4 0.0304 4 0.0594 4 0.0891 4 0.1221 4 0.1490 4 0.2865 4 0.5232 4 0.9080 4 1.0831 4 1.0810 4 #.#### 4 0.0385 4 0.0242 4 0.0686 4 0.0736 4 0.0732 4 0.0433 4 0.1698 4 0.0937 4 0.1343 4 0.1431 4 1.0452 4 #.#### 4 0.0420 4 0.0306 4 0.0229 4 0.0172 4 0.0255 4 0.6608 4 0.3788 4 0.0767 4 0.0929 4 0.0963 4 1.0750 4 #.#### 4 0.1397 4 0.1417 4 0.1368 4 0.1467 4 0.1292 4 0.1533 4 0.1179 4 0.0492 4 0.0278 4 0.0291 4 1.0775 4 #.#### 4 0.0265 4 0.0163 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 000247 & g Ss g oCf 00016 995-05-04 16:44 OutPut o f : 950504A1 Fluoride 1.5 Fluoride L PPM PPM OS Typ Ident Ch Result F Time Ch Result F Time 4u F54094-4 3 0.067 5u F54100-4 3 0.067 6u F54091-4 3 0.067 7u Blk 1 3 0.069 8u Blk 2 3 0.064 9u Spk 1 3 0.103 0u Spk 2 3 0.105 1u F54097-4 3 0.067 2d Drift 3 1.436 3w Wash 3 0.057 4u F54102-4 3 0.067 ju F54092-4 3 0.064 6u F54080-4 3 0.072 7u F54106-4 3 0.075 8 u 'tf- 3 0.067 9 u^ F54/088-4 3 0.067 0U F54/087-4 3 0.070 1 U ^5" 2u F54/110-4 F54096-4 3on 0.064 0.061 3u F54090-4 3 0.064 4d Drift 3 1.455 5w Wash 3 0.057 6u F54078-4 3 0.067 7u F54101-4 3 0.061 8u F54095-4 3 0.063 9u F54093-4 3 0.068 0u F54071-4 3 0.063 1u F54108-4 3 0.062 2u F54116-4 3 0.139 3u F54098-4 3 0.091 4u F 54084-4 3 0.108 5u F54079-4 3 0.152 6d Drift 3 1.478 7w Wash 3 0.057 8u F54113-4 3 0.124 9u F54111-4 3 0.131 0u 1u F54082-4 3 0.096 F54076-4 3 0.082 2u 3u F54114-4 3 0.080 F54109-4 3 0.065 4d Drift 3 1.468 5w Wash 3 0.057 t rw RunOut Wash 3 0.057 9488 9666 9842 10016 10191 10367 10543 10717 10891 11066 11241 11419 11591 11769 11942 12116 12291 12462 12642 12818 12992 13159 13342 13516 13692 13868 14042 14213 14393 14568 14744 14919 15093 15332 15444 15619 15795 15967 16145 16319 16493 16650 16968 4 0.0181 4 0.0196 4 0.0181 4 0.0220 4 0.0130 4 0.0800 4 0.0835 4 0.0200 4 1.0537 4 #.#### 4 0.0192 4 0.0141 4 0.0287 4 0.0340 4 0.0181 4 0.0200 4 0.0248 4 0.0123 4 0.0064 4 0.0138 4 1.0681 4 #.#### 4 0.0194 4 0.0076 4 0.0120 4 0.0213 4 0.0118 4 0.0102 4 0.1307 4 0.0624 4 0.0889 4 0.1471 4 1.0857 4 #.#### 4 0.1114 4 0.1210 4 0.0702 4 0.0460 4 0.0431 4 0.0160 4 1.0780 4 #.#### 4 #.#### 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 2 of 2 000248 000168 BEST COPY AVAILABLE Hay data of 9SC604A1 : hluorida i.b sc-hKii ! Fl=Hslp \ Urtl-F=fcdit peaks ! 000249 000172 Ray data of Sb0504fti - M u o r i d s 1.5 BEST COPY AVAILABLE 000250 000173 0002S1 000174 995-05-04 16:48 Output of : 950504B1 oftware : v e r s i o n 6 . 1 c l 9 9 0 ,93 perator : ddw ate of the Analysis : 1995-05-04 13:07 BEST COPY AVAILABLE nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950504B1 luoride 1.5 alibration order = Inverse Logarithm 'lo p e : s = #.##### esult r X - Cl 10 L s x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode 2 = - 0.00000 1 = 0.00062 0 = -1.24651 luoride L alibration order = 2 'o rre la tio n : r = 0.99962 esult = a2 * x 2 + al * x + aO 2 = 0.00000 1 = 0.00020 0 = -0.00460 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 m m . iluter needle Height dilution Factor dilution Volume Resample Dilution runs 80 mm 10 2.5 ml 1 1 User file : Reproces : No TXT Sc 0002S2 000175 995-05-04 16:48 OutPut of : 950504B1 luoride 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 BEST COPY AVAILABLE 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 trigger Limit 1800 Sec Peak shape Pointed stArt ignore 60 Sec eNd ignore 120 Sec M e a s u r e w i n d o w 75 % Filter No Regeneration No formula : output : ##.### luoride 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 000253 000176 995-05-04 16:48 Output Of : 950504B1 sl 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 M e a s u r e w i n d o w 75 % Filter No Regeneration No formula c 4 :=c3 output #.#### 0002S4 000177 995-05-04 16:48 OutPut of : 950504B1 F l u o r i d e 1. 5 Fluoride L PPM PPM 'OS Typ Ident Ch Result F Time Ch Result F ;t iw Initial W a s h 3 0.057 t Tracer 3 1.467 d Drift 3 1.473 w Wash 3 0.057 si S t a n d a r d 1 3 0.065 . s2 Standard 2 3 0.073 ; s3 Standard 3 3 0.089 s4 S tandard 4 3 0.105 j s5 Standard 5 3 0.128 ) s6 Standard 6 3 0.153 .0 s 7 Standard 7 3 0.291 s8 2 s9 Standard 8 3 0.605 Standard 9 3 1.228 .3 slO S t a n d a r d 10 3 1.475 .4 d Drift 3 1.448 .5 w Wash 3 0.057 .6 u F54075-8 3 0.073 7u F54081-8 3 .067 8u F54105-8 3 0.071 9u F54107-8 3 0.065 0u F54074-8 3 0.066 1u F54085-8 3 0.073 .2 u F54086-8 3 0.075 .'3 u BLK-1 3 0.068 :4 u BLK-2 3 0.092 :5 u SPK 1 3 0.113 :6 d Drift 3 1.439 :7 w Wash 3 0.057 :8 u SPK 2 3 0.098 .9 u F54104-8 3 0.075 0u F54077-8 3 0.072 1u F54083-8 3 0.063 2 u F54094-8 3 0.069 3 u F54100-8 3 0.067 >4 u F54091-8 3 0.061 >u F54097-8 3 0.060 6 u F54102-8 3 0.065 7u F54108-8 3 0.059 8 d Drift 3 1.397 ;9 w Wash 3 0.057 0 u F54071-8 3 0.062 :1 u F54093-8 3 0.065 2 u F54095-8 3 0.067 :3 u F5 4101-8 3 0.061 4 u F54078-8 3 0.063 :5 u F54090-8 3 0.063 6 u F54096-8 3 0.058 7 u F54110-8 3 0.108 3 u F54087-8 3 0.068 9 u F54088-8 3 0.073 0 d Drift 3 1.481 1w 2 -> u u Wash 3 0.057 F54089-8 3 0.076 F54106-8 3 0.084 65 212 388 629 740 910 1089 1264 1440 1614 1790 1964 2138 2314 2488 2709 2837 3013 3186 3361 3541 3712 3891 4067 4241 4415 4589 4828 4941 5112 5301 5461 5642 5813 5987 6169 6339 6513 6690 6854 7040 7214 7392 7562 7740 7916 8088 8266 8441 8615 8790 8938 9141 9315 4 #.#### 4 0.8037 4 0.8066 4 #.#### 4 0.0140 4 0.0311 4 0.0615 4 0.0877 4 0.1206 4 0.1501 4 0.2729 4 0.4486 4 0.7030 4 0.8073 4 0.7951 4 #.#### 4 0.0319 4 0.0199 4 0.0274 4 0.0154 4 0.0176 4 0.0311 4 0.0357 4 0.0211 4 0.0663 4 0.1002 4 0.7912 4 #.#### 4 0.0774 4 0.0366 4 0.0292 4 0.0217 4 0.0227 4 0.0203 4 0.0061 4 0.0027 4 0.0146 4 0.0004 4 0.7727 4 #.#### 4 0.0081 4 0.0144 4 0.0203 4 0.0063 4 0.0108 4 0.0101 4 #.#### 4 0.0923 4 0.0207 4 0.0321 4 0.8103 4 #.#### 4 0.0376 4 0.0525 0 0 00 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 oQ 0 Page 1 of 2 . wjv wT 000255 nU ow nUiI * o 995-05-04 16:48 Output of : 950504B1 Fluoride 1.5 PPM Fluoride L PPM os Typ Ident Ch Result F Time Ch Result F Time 4u 5u 6u 7u 3u 9u 0u 1u 2d 3w 4u Ju 6u 7u 3d 9w t rw F54080-8 3 0.075 F54092-8 3 0.079 F54109-8 3 0.074 F54114-8 3 0.077 F54076-8 3 0.087 F54082-8 3 0.105 F54111-8 3 0.088 F54113-8 3 0.094 Drift 3 1.512 Wash 3 0.057 F54079-8 3 0.093 F54084-8 3 0.095 F54098-8 3 0.099 F54116-8 3 0.228 Drift 3 1.512 Wash 3 0.057 RunOut Wash 3 0.057 9491 9667 9841 10015 10193 10367 10543 10717 10891 11093 11240 11416 11591 11766 11941 12097 12416 4 0.0364 4 0.0433 4 0.0333 4 0.0402 4 0.0585 4 0.0877 4 0.0593 4 0.0700 4 0.8247 4 #.#### 4 0.0681 4 0.0718 4 0.0792 4 0.2236 4 0.8245 4 #.#### 4 #.#### 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 000256 000175 1EST COPY AVAILABLE 2.181S0S8 Calibration curve of 95050481 : t Ivorida L S -Q.003333 64 M C 2 yr* 0 g 0.004603 Order Heasuraci 0.39362 000257 000180 4<2 Ca 1i b r a t i on curue o f 3bti5u41 : F lu o r id e i .5 BEST COPY AVAILABLE 000258 000181 Raw data of 3h0504il - Fluoride i.S BEST COPY AVAILABLE :CS5 ! i^e se=Exit S Fi=Help 5 CrtI-F=dit peaks ! FOOO 000259 0001 1035 'x y BEST COPY AVAILABLE Raw data of 9SG504B1 : rluorida 1.5 % r3* 4 2 HH IS i \ J 't **y ^ ^--i_;u vii ViA\ ?Jns iA\ J A\ J' T\ ,-'i ! * / \J - VJ IV, 40 f T a-, ia\ /A\ dA ia\ Ma rA I* * \ AA / \/\ J\ 0 4560 I irie sc=fcxit J Fi=Help ! Crti-P=hdit peaks \ y :i S** \ \w i'.! "a' 9560 000260 00013 BESTCOPy AVAILABLE I i.n 0.1 OI..L. 0 ;.i !"I T "! % T' U"l CoD 5 tSTOO T92000 *H 995-05-08 12:46 OutPut of : 950508A1 i O ^ 1-2.7-ltS o f t w a r e : v e r s i o n 6 . 1 c l 9 9 0 ,93 iperator : DDW i/y is s - t S e w BEST COPY AVAILABLE ate of the Analysis : 1995-05-08 07:30 .nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950508A1 luoride 1.5 :alibration order = Inverse Logarithm Uope : s = ft.##### result r x - cl 10 L s x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode a = -o.ooooo il = 0 . 0 0 0 6 6 l0 = - 1 . 2 6 7 0 4 luoride L alibration order = 2 C orrelation : r = 0.99873 esult = a2 * x 2 + al * x + aO 2 = -0.00000 1 = 0.00021 0 = -0.00833 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 120 sec. 1 sec. Single None 70 m m . 'iluter needle Height dilution Factor dilution Volume Resample Dilution runs 80 m m 10 2.5 ml. 1 1 User file : Reproces : No . TXT 000262 oooisr. 995-05-08 12:46 Output of : 950508A1 luoride 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 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 M e a s u r e win d o w 75 % Filter No Regeneration No formula : output : ##. # # # luoride 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 000263 0001< 995-05-08 12:46 OutPut of : 950508A1 sl 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 M e a s u r e w i n d o w 75 % Filter No Regeneration No formula : c4:=c3 output : #.#### 000264 000187 995-05-08 12:46 OutPut of : 950508A1 Fluoride 1.5 Fluoride L PPM PPM 'OS T y p Ident Ch Result F Time 't iw Initial Wash 3 0.054 65 t Tracer 3 1.447 210 d Drift 3 1.466 384 ,w Wash 3 0.054 625 si Standard 1 3 0.063 734 s2 Standard 2 3 0.071 910 s3 Standard 3 3 0.086 1086 s4 Standard 4 3 0.107 1260 , s5 Standard 5 3 0.124 1436 ; s6 Standard 6 3 0.154 1611 -0 s7 Standard 7 3 0.287 1785 s8 Standard 8 3 0.607 1961 2 s9 Standard 9 3 1.239 2134 .3 slO S t a n d a r d 10 3 1.464 2309 .4 d Drift 3 1.474 2485 .5 w Wash 3 0.054 2718 5u Blk 1 3 0.071 2835 7u Blk 2 3 0.063 3010 3u Spk 1 3 0.100 3187 9u Spk 2 3 0.104 3362 .0 u F5407 5-12 3 0.071 3538 .1 u F54081-12 3 0.067 3712 2u F54105-12 3 0.071 3886 .3 u F541Q7-12 3 0.066 4057 .4 u F54074-12 3 0.066 4235 :5 u F5408 5-12 3 0.063 4402 :6 d Drift 3 1.454 4588 .1 w Wash 3 0.054 4778 :3 u F54086-12 3 0.073 4937 :9 u F54104-12 3 0.076 5110 :0 u F54077-12 3 0.087 5286 :1 u F54083-12 3 0.081 5467 ;2 u F54094-12 3 0.082 5637 3u F54100-12 3 0.071 5813 !4 u F54091-12 3 0.078 5991 3u F54097-12 3 0.072 6165 '6 u F54102-12 3 0.067 6337 ;7 u F54108-12 3 0.062 6515 .8 d Drift 3 1.467 6687 9 w Wash 3 0.054 6853 :0 u F54077-12 3 0.066 7037 :1 u _2 u F54071-12 3 0.054 A 7212 F54093-12 3 0.054 A 7387 :3 u ;4 u F54095-12 3 0.054 A 7562 F54101-12 3 0.054 A 7737 :5 u F54110-12 3 0.054 A 7912 n6f u u F54096-12 3 0.058 F54090-12 3 0.068 8088 8264 8 u F54078-12 3 0.064 8438 9 u F54088-12 3 0.075 8611 0J1. d w 12 u >u Drift Wash F54106-12 F540S9-12 3 1.457 3 0.054 3 0.080 3 0.072 8789 8995 9141 9315 Ch Result F Time 4 #.#### 4 0.7320 4 0.7393 4 #.#### 4 0.0139 4 0.0313 4 0.0597 4 0.0932 4 0.1156 4 0.1514 4 0.2640 4 0.4282 4 0.6557 4 0.7386 4 0.7425 4 #.#### 4 0.0298 4 0.0128 4 0.0829 4 0.0883 4 0.0315 4 0.0217 4 0.0298 4 0.0198 4 0.0209 4 0.0143 4 0.7348 4 #.#### 4 0.0352 4 0.0403 4 0.0610 4 0.0492 4 0.0514 4 0.0309 4 0.0448 4 0.0335 4 0.0232 4 0.0100 4 0.7399 4 #.#### 4 0.0194 4 #.#### 4 #.#### 4 #.#### 4 #.#### 4 #.#### 4 0.0021 4 0.0243 4 0.0160 4 0.0397 4 0.7356 4 #.#### 4 0.0478 4 0.0326 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 000188 000265 995-05-08 12:46 OutPut of : 950508A1 Fluoride 1.5 Fluoride L PPM ppm OS Typ Ident Ch Result F Time 4u 5u 6u 7u 8u 9u 0u 1u 2d 3w 4u ju 6u 7u 3u 9d 0w t rw F54087-12 3 0.072 F54080-12 3 0.069 F54092-12 3 0.067 F54109-12 3 0.072 F54114-12 3 0.074 F5407 6-12 3 0.073 F54113-12 3 0.072 F54082-12 3 0.080 Drift 3 1.442 Wash 3 0.054 F54111-12 3 0.092 F54098-12 3 0.092 F54084-12 3 0.089 F54079-12 3 0.088 F54116-12 3 0.090 Drift 3 1.515 Wash 3 0.054 RunOut Wash 3 0.054 9491 9664 9839 10017 10193 10367 10540 10716 10890 11058 11242 11418 11590 11764 11943 12116 12348 12591 Ch Result F Time 4 0.0320 4 0.0266 4 0.0222 4 0.0322 4 0.0362 4 0.0341 4 0.0330 4 0.0475 4 0.7301 4 #.#### 4 0.0701 4 0.0697 4 0.0650 4 0.0623 4 0.0659 4 0.7597 4 #.#### 4 #.#### 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Page 2 of 2 & V ?* 000266 0001S!> BEST COPY AVAILABLE O.iSibSfi - Calibration curve of 9bUbU8fti : M u e r ide L --- - g l B a g l ^ ^ - 0 . 0 0 7 g ? 6 ^ H I ^ f S 4 . c 0 +/ ui I c 3 2- 0.00832b ' Order : 2 Heasured r : 0.99873 900 000267 000130 1.7154138 1 Calibration curve of S50Eu8fil F iuorida i . b ---------------- F 45703m m m 4 H I .-i ! m nm km m Is `=5( _ isi c I J 1 y y /r V j.0540764 0 _ ---T ' Heasurad Orda?' i Inverse Logarithm /+ 4095 s : esos 0002G 8 000131 BESTCOPY AVAILABLE 1095 1 Haw data of SEObOofii ; rluoride 1.5 U E n m . . . . j i i ___ m r n m s A ' &* if# a 3 j3 -i \ i n . v'N li `i s n i in!ii! 1{ ! I yy^: T n \sS rt' 9j u T iriS :sc=bxit ! j-l=Help i 0rti-r'=dit peaks 1 A A V y x> l \ *** x lj\ V 5000 000269 00019- BEST COPY AVAILABLE nMjpt If If p H>|!i,|i|pi if 1 ill ^ ir\ U l T<?u f|tr-S,* [i;*" ; j U, CirT;n">i 'Coto-nXQ UCOl P 1= fe fllPl'i: O C-Ti tn XI Ul W r3ne;i o\A - > i'J,,ni sari j n-?,r& 000133 BEST COPY AVAILABLE 'I0S5 i ----------- Haw data of 95CSUSA1 ' M u o r i d a 1.5 a a ir zm sm M cA U 'l' 9 =3 JU - ^ `r . v t h A A j y/v yA UA 'Mi M- UH ^ & AA A A A ;l M 'c i y y u u y ^ 9460 T ire s-tKit I hi=Heip i Urtl-F=dii peaks ! 000271 14450 oooiy- .995-05-09 15:09 Software : version 6 . 1 OutPut of : 950509A1 cl990,93 f ^ t > T HXO^S.J - ^ St*v< )perator : DDW )ate of the A n a l y s i s : 1995-0 5 - 0 9 09:48 K M COPYAVAILABLE analysis File Name C : \ S K A L A R \ D A T A \ H W I D A T A \ S E R U M \ 9 50 50 9A1 'luoride 1.5 Calibration order = Inverse Logarithm Slope : s = #.##### Result r x - cl J10 L s x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode 12 = il = 10 = - 0.00000 0.00061 -1.24329 ^luoride L Calibration order = 2 C o rrelatio n : r = 0.99974 Result = a2 * x 2 + al * x + aO i2 = il = iO = 0.00000 0.00020 -0.00554 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. )iluter needle Height dilution Factor dilution Volume Resample Dilution runs : 80 : 10 : 2.5 :1 :1 mm ml User file : Reproces : No TXT 000272 000193 995-05-09 15:09 luoride 1 . Path number Signal type Decolor system Number diLute Resa mple dil Threshold diG output Window event OutPut of : 950509A1 3 Debubbled Yes 0 No No 4095 0 Off si sTandard Ignore s2 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 DUU DU trigger Limit 1800 Sec Peak shape Pointed stArt ignore 60 Sec eNd ignore 120 Sec M e a s u r e w i n d o w 75 SL Filter No Regeneration No formula : output : ##.### luoride 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 00273 00013* 995-05-09 15:09 OutPut of : 950509A1 s i 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 M e a s u r e wi n d o w 75 % Filter No Regeneration No formula c 4 :=c3 output #.#### 000274 00013v 9 5 - 0 5 - 0 9 1 5 : 0 9 O u t p u t o f : 9 5 0 5 0 9 A 1 Fluoride 1.5 Fluoride L PPM PPM os Typ Ident Ch Result F Time Ch Result F Time t iw Initial Wash 3 0.057 65 t Tracer 3 1.474 209 d Drift 3 1.502 384 w Wash 3 0.057 626 si S t a n d a r d 1 3 0.065 734 s2 Standard 2 3 0.074 910 s3 S t a n d a r d 3 3 0.088 1082 s4 Standard 4 3 0.107 1260 s5 Standard 5 3 0.128 1435 s 6 Standard 6 3 0.153 1610 0 s7 Standard 7 3 0.290 1787 M s8 Standard 8 3 0.606 1960 2 s9 Standard 9 3 1.233 2135 3 slO Standard 10 3 1.468 2310 4d Drift 3 1.548 2484 5w Wash 3 0.057 2726 6u Serum blk 1 3 0.080 2839 7u Serum blk 2 3 0.073 3011 8u Serum spk 1 3 0.080 3189 9u Serum spk 2 3 0.081 3361 0u Serum spk 3 3 0.088 3537 1u Serum spk 4 3 0.091 3712 2u Serum spk 5 3 0.115 3887 3u Serum spk 6 3 0.095 4062 4u Serum spk 7 3 0.090 4236 5u Serum spk 8 3 0.096 4413 6d Drift 3 1.509 4587 7w Wash 3 0.057 4813 8u F54105-24 3 0.085 4939 9u F54075-24 3 0.070 5111 0u F54081-24 3 0.063 5232 1u F54107-24 3 0.066 5464 2u F54104-24 3 0.070 5635 3u F54086-24 3 0.067 5814 4u F54074-24 3 0.075 5994 ju F54085-24 3 0.055 A 6162 6u F54083-24 3 0.055 6336 7u F54100-24 3 0.055 6508 3d Drift 3 1.472 6688 9w Wash 3 0.057 6927 0u Blk 1 3 0.073 7038 1u Blk 2 3 0.065 7210 2u Spk 1 3 0.078 7391 3u Spk 2 3 0.086 7565 4u Spk 3 3 0.087 7739 5u Spk 4 3 0.088 7912 6u Spk 5 3 0.093 8091 7u Spk 6 3 0.092 8263 8u Blk 3 3 0.078 8439 9u F 54085-24 3 0.066 8615 0d Drift 3 1.506 8789 1w Wash 3 0.057 9028 2u F54083-24 3 0.068 9142 3u F54100-24 3 0.066 9313 4 #.#### 4 0.7917 4 0.8031 4 #.#### 4 0.0141 4 0.0318 4 0.0588 4 0.0908 4 0.1193 4 0.1503 4 0.2726 4 0.4494 4 0.6983 4 0.7892 4 0.8224 4 #.#### 4 0.0451 4 0.0312 4 0.0436 4 0.0457 4 0.0584 4 0.0650 4 0.1016 4 0.0713 4 0.0633 4 0.0728 4 0.8060 4 #.#### 4 0.0541 4 0.0236 4 0.0202 4 0.0153 4 0.0236 4 0.0175 4 0.0352 4 #.#### 4 #.#### 4 #.#### 4 0.7909 4 #.#### 4 0.0310 4 0.0137 4 0.0402 4 0.0553 4 0.0578 4 0.0592 4 0.0674 4 0.0658 4 0.0412 4 0.0155 4 0.8050 4 #.#### 4 0.0208 4 0.0153 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 00013e 000275 395-05-09 15:09 O u t p u t o f ': 9 5 0 5 0 9 A 1 Fluoride 1.5 PPM Fluoride L PPM os Typ Ident Ch Result F Time Ch Result F Time 4u 5u 6u 7u 8u 9u 0u 1u 2d .3 w 4 u ju 6u 7u 3u 9u 0u 1u 2u 3u 4d 5w '5 u 1u 8u 9u 0u 1 u 2u 3u 4 u 5u 6d 7 w .8 u 9u 0 u 1 u '2 u 3 d 4 w /t rw F54077-24 3 0.067 F54094-24 3 0.065 F54091-24 3 0.065 F 54108-24 3 0.065 F54102-24 3 0.063 F54097-24 3 0.070 F54071-24 3 0.064 F54093-24 3 0.064 Drift 3 1.493 Wash 3 0.057 F54095-24 3 0.068 F54101-24 3 0.067 F54110-24 3 0.066 F54096-24 3 0.064 F54090-24 3 0.084 F54078-24 3 0.071 F54088-24 3 0.070 F54106-24 3 0.805 F54089-24 3 0.435 F54087-24 3 0.574 Drift 3 0.223 Wash 3 0.057 F54080-24 3 0.089 F54092-24 3 0.084 F54109-24 3 0.077 F54114-24 3 0.074 Blk 1 3 0.068 Blk 2 3 0.066 Spk 1 3 0.105 Spk 2 3 0.118 F54113-24 3 0.088 F54082-24 3 0.079 Drift 3 1.423 Wash 3 0.057 F54111-24 3 0.088 F54098-24 3 0.086 F54084-24 3 0.084 F54079-24 3 0.083 F54116-24 3 0.088 Drift 3 1.457 Wash 3 0.057 RunOut Wash 3 0.057 9489 9663 9842 10012 10188 10364 10540 10716 10890 11098 11236 11414 11590 11772 11943 12115 12282 12480 12655 12830 13006 13117 13335 13517 13693 13865 14042 14218 14392 14568 14743 14918 15092 15331 15442 15618 15794 15968 16145 16318 16558 16793 4 0.0169 4 0.0133 4 0.0139 4 0.0135 4 0.0090 4 0.0242 4 0.0121 4 0.0106 4 0.7997 4 #.#### 4 0.0208 4 0.0171 4 0.0151 4 0.0117 4 0.0510 4 0.0261 4 0.0242 4 0.5348 4 0.3636 4 0.4347 4 0.2195 4 #.#### 4 0.0606 4 0.0510 4 0.0389 4 0.0322 4 0.0204 4 0.0145 4 0.0873 4 0.1063 4 0.0584 4 0.0430 4 0.7713 4 #.#### 4 0.0592 4 0.0549 4 0.0514 4 0.0500 4 0.0598 4 0.7849 4 #.#### 4 #.#### 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ---- o 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 000276 oooiy?) M30787 - BEST COPY AVAILABLECalibration curve of bCbOuftl : f-luoride L 6 ~ is a g g ig m ^ p ^ 3 n g M ?M s T t* Y 5 h i '* = c a y c a y y -L--" -0.005541 0 -----i---------- r Order : 2 naasurad r : 0.S3S74 300 000277 000200 BEST COPY AVAILABLE Calibration curve of 950b09fii - Fluoride l.b i i *J if! i "T neasureti 000278 000201 BEST COPY AVAILABLE SjmlIiilii |! i j li m 'I'X'I'' w!IT rt "aDi pnJ *' cTao":* TB'G Oin iI.;InI ifc II:: /* t ....... -- HI x: terii owz PH 'S S oOOU1 . c 3 a: ;e : !>> Q' - n .mli dQi I, M! .. U r 3 vA O I' ^ ...... -- P Icf:!t ~r............ - ..... r a n r.*s?n :xt'It:l uim,, ro '?! 000279 00020 02 4GSS - BEST COPY AVAILABLE Hay da:a of SS9991 ' luoride l.b 20 j5 - ; - ' -, ? nAV-i!juA-v-.A+ A ->> A A : X I \ A J VAA A j ^ tjO *VV^HA' f A tT nn a L A2 $*$** j v / \ vA''-MJI \\'!}i ^\J \lWf \\vjl \-./ i\ i\\ H/\ ; - -i.i -.j ' ASG Ti aa sc=hMit ; Fi-Haip j CriI-P=Edit peaks I 9SP0 000280 oooao BEST COPY AVAILABLE -:DS5 I Ray data of bbubOOhi : -luoriria i.b 22 A T a '"D z ^I s30- - t 5 i! fr ?/> * i, ' ^V ^ T /. <* A fcHCM ; < i t- s. 8933 Ti tie s e -c x it : Fi-naiy 5 O r::I-P -Ed iV oaaka ; 1(r 3 1? ! 398 000281 000201 f BEST COPY AVAILABLE jprri ?li!L;!g I I m UI h'..rU iI:rTn 1t! i;CTMM ;s'il!' C'i (-li & %Ut';i N m i'M 5l11;yvi1!1 I'R"fit IM)>*'.i'i iiiii IW Nj:Mf| 1,0 3 ... r~...... t :*"!' it:' .! 00020 O I. L? <Tr N 00 ooCN o u5 995-05-09 15:59 OutPut of : 950509B1 oftware : version 6. 1 01990,93 perator DDW ty p y % ate of the Analysis 1995-05-09 15:08 .nalysis File Name C:\SKALAR\DATA\HWIDATA\SERUM\950509B1 'luor i d e 1.5 :alibration original time-date stamp : 1995-05-09 09:48 'alibration order = Inverse Logarithm >2 o p e : s = #.##### 'e s u l t r x - cl 10 s x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode .2 = - 0 . 0 0 0 0 0 il = 0 . 0 0 0 6 1 iO = - 1 . 2 4 3 2 9 luoride L alibration original time-date stamp : 1995-05-09 09:48 alibration order = 2 ".correlation : r = 0.99974 esult = a2 * x 2 + al * x + aO \2 = 0 . 0 0 0 0 0 :1 = 0 . 0 0 0 2 0 lO = - 0 . 0 0 5 5 4 ampler 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. 'i l u t e r needle Height dilution Factor dilution Volume Resample Dilution runs 80 mm 10 2.5 m l . 1 1 User file : Reproces : No . TXT 000283 00020 G 995-05-09 15:59 Output of : 950509B1 'luoride 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 so sTandard 0.150 s7 sTandard 0, 300 s3 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 M e a s u r e w i n d o w 75 Filter No Regeneration No formula : output : ##. # # # 'luoride 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 000284 00020 995-05-09 15:59 OutPut of : 950509B1 sl 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 M easure w i n d o w : 75 % Filter : No Regeneration : No formula : c4:=c3 output : #.#### 00285 00020 (?(? 995-05-09 15:59 OutPut Of : 950509B1 Fluoride 1.5 Fluoride L PPM PPM Typ Ident Ch Result F Time Ch Result F Time iw Initial Wash 3 0.057 t Tracer 3 1.509 d Drift 3 1.520 w Wash 3 0.057 u F54106-24 3 0.080 u F54089-24 3 0.083 u F54087-24 3 0.078 d Drift 3 1.619 w Wash 3 0.057 rw Run out wash 3 0.057 65 212 386 651 745 944 1093 1262 1503 1737 4 #.#### 4 0.8062 4 0.8106 4 #.#### 4 0.0438 4 0.0494 4 0.0406 4 0.8540 4 #.#### 4 #.#### 0 0 0 0 0 0 0 0 0 0 Page 1 of l 000286 r' r>rpf > r.. 995-05-09 15:44 OutPut o f . : 950509B1 Fluoride 1.5 Fluoride L PPM PPM Typ Ident Ch Result F Time Ch Result F Time iw Initial Wash 3 0.057 t Tracer 3 1.490 d Drift 3 1.482 w Wash 3 0.057 u F54106-24 3 0.113 u F54089-24 3 0.159 u F54087-24 3 0.108 d Drift 3 1.609 w Wash 3 0.057 rw Run out wash 3 0.057 65 212 386 551 751 926 1101 1262 1503 1737 4 #.#### 4 0.7985 4 0.7951 4 #. #### 4 0.0989 4 0.1577 4 0.0916 4 0.8493 4 #.#### 4 #.#### 0 0 0 0 0 0 0 0 0 0 H ' C Page 1 of l V- /La_d-^v> er-Uiy X4W.K>V>\A< ^ 't<.C.o Q `O a-v_> 000287 000210 /i Ray data of 9505U9BI : fluorida i.5 BEST COPY AVAILABLE Man 1 -.s 000288 000211 r.e 995-05-11 13:19 oftware : version 6.1 O u t P u t o f ': 9 5 0 5 1 1 A 1 cl990,93 a '3 '2 .^- I S'n perator : DDW ate of the Analysis : 1995-05-11 08:16 BEST COPY AVAILABLE nalysis File Name : C:\SKALAR\DATA\HWIDATA\3ERUM\950511A1 luoride 1.5 alibration order = Inverse Logarithm lope : s = #.##### r x - cl - esult = 10 s x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode 2 = -0.00000 1 = 0.00060 0 = -1.24775 luoride L alibration order = 2 orrelation : r = 0.99962 esult = a2 * x 2 + al * x + aO 2 = 0.00000 1 = 0.00019 0 = -0.00028 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height SA1000 1 50 sec. 1 2 0 sec. 1 sec. Single : None : 70 mm. iluter needle Height dilution Factor dilution Volume Resample Dilution runs : 80 : 10 : 2 .5 :1 :1 mm ml User file : Reproces : No . TXT 000289 000212 995-05-11 13:19 OutPut o f : 950 511A1 luoride 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 trigger Limit 1800 Sec Peak shape Pointed stArt ignore 60 Sec eNd ignore 120 Sec Measure window 75 % ' Filter No Regeneration No formula : output : ##.### luoride 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 000290 000213 995-05-11 13:19 OutPut of 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 M e a s u r e w i n d o w 75 % Filter No Regeneration No formula c 4 :=c3 output #.#### 950511A1 000291 000214 995-05-11 13:19 OutPut of : 950511A1 Fluoride 1.5 Fluoride L PPM PPM OS Typ Ident Ch Result F Time Ch Result F Time t iw Initial Wash 3 0.057 t Tracer 3 1.472 d Drift 3 1.472 w Wash 3 0.057 si S t a n d ar d 1 3 0.064 s2 Standard 2 3 0.070 s3 S t a n d ar d 3 3 0.085 s4 Standard 4 3 0.107 s5 S t a n d ar d 5 3 0.127 s6 Standard 6 3 0.153 0 s7 Standard 7 3 0.289 s8 Standard 8 3 0.607 2 s9 Standard 9 3 1.233 3 slO Standard 10 3 1.468 4d Drift 3 1.509 5w Wash 3 0.057 6u F54113-48 3 0.094 7 wo u u F54082-48 3 0.091 F54111-48 3 0.096 9u F54098-48 3 0.091 0u F54084-48 3 0.103 1u 2 3-i u u F54079-48 3 0.082 F54116-48 3 0.101 F54113-192 3 0.097 4u F54082-192 3 0.089 5u F54111-192 3 0.081 5d Drift 3 1.435 7w Wash 3 0.057 8u 9u F54098-192 3 0.088 F 5 4 0 7 9 - 1 9 2 -kJ5 0.089 0u iu F54084-192 O 0.094 F54116-192 3 0.085 2u F54113-360 3 0.085 3u F54082-360 3 0.073 4u F54111-360 3 0.087 ju F54098-360 3 0.082 H61 u u F54079-360 3 0.091 F54084-360 3 0.092 3d Drift 3 1.481 9w Wash 3 0.057 0u F54116-360 3 0.109 1u BLK 1 3 0.065 2u SPK 1 3 0.094 3u SPK 2 3 0.112 4u F54082-D22 3 0.085 5u 6u F54111-D22 3 0.084 F54079-D22 3 0.083 7u F54116-D22 3 0.083 3u F 54032-D28 3 0.087 9u F54111-D28 3 0.078 0d Drift J 1.519 1w Wash 3 0.057 2u F54079-D2S 3 0.077 1 J u F54116-D23 3 0.081 65 208 382 624 729 913 1085 1260 1435 1610 1784 1959 2133 2309 2484 2725 2836 3011 3185 3359 3537 3711 3887 4061 4237 4413 4585 4732 4936 5111 5286 5458 5638 5810 5984 6164 6336 6513 6686 6926 7039 7213 7387 7565 7739 7912 8085 8263 8438 8614 8787 9027 9138 9314 4 #.#### 4 0.7342 4 0.7342 4 #.#### 4 0.0164 4 0.0287 4 0.0586 4 0.0921 4 0.1194 4 0.1498 4 0.2642 4 0.4285 4 0.6527 4 0.7329 4 0.7476 4 #.#### 4 0.0722 4 0.0685 4 0.0758 4 0.0685 4 0.0863 4 0.0523 4 0.0844 4 0.0775 4 0.0639 4 0.0512 4 0.7213 4 #.#### 4 0.0626 4 0.0639 4 0.0729 4 0.0573 4 0.0586 4 0.0353 4 0.0611 4 0.0519 4 0.0681 4 0.0699 4 0.7374 4 #.#### 4 0.0959 4 0.0188 4 0.0727 4 0.1002 4 0.0586 4 0.0569 4 0.0536 4 0.0540 4 0.0607 4 0.0447 4 0.7510 4 #.#### 4 0.0431 4 0.0502 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 2 000 15 000292 995-05-11 13:19 OutPut of : 950511A1 Page of Fluoride 1.5 Fluoride L OS Typ Ident PPM Ch Result F Time PPM ftS T C O p Ch Result F Time 4u 5u 5U 7U 3U 9U 0U iU 2d 3w 4U JU 6U 7U 3U 9U 0U 1U 2U 3U 4d 5w 0U 7u 3U 9d 0w t- rV/ F54099-4 F54099-8 F54099-12 F54099-24 F54099-48 F54099-192 F54099-360 r >4 {J'J'J "J 0 1 Drift Wash F54112-4 F54112-8 F54112-12 F54112-24 F54112-48 F54112-192 F54112-360 SPK 63-1 SPK 63-2 SPK 63-3 Drift Wash SPK 126-1 SPK 126-2 SPK 126-3 Drift Wash RunOut Wash 3 0.071 9490 3 0.070 9662 3 0.105 9840 3 0.118 10015 3 0.084 10190 3 0.074 10362 3 0.070 10538 n J -- U.ITT" A J /i J 3 1.504 10888 3 0.057 11129 3 0.032 11237 3 0.094 11415 3 0 . 1 0 0 11591 3 0.088 11765 3 0 . 1 0 0 11941 3 0.094 12115 3 0.078 12289 3 0.095 12465 3 0 . 1 1 0 12641 3 0.092 12817 3 1.469 12989 3 0.057 13166 3 0.117 13342 3 0 .149 13518 3 0.157 13692 3 1.496 13865 3 0.057 14107 3 0.057 14340 4 0.0317 4 0.0298 4 0.0894 4 0.1075 4 0.0569 4 0.0371 4 0.0306 4 ft itiftttt 4 0.7457 4 #.#### 4 0.0527 4 0.0735 4 0.0827 4 0.0624 4 0.0815 4 0.0725 4 0.0452 4 0.0750 4 0.0973 4 0.0689 4 0.7334 4 #.#### 4 0.1061 4 0.1452 4 0.1537 4 0.7427 4 #.#### 4 #.#### 0 0 0 0 0 0 0 0 0 - O. - __j< ts-rs O v w : 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 000293 0 0 0 2 1 ' BEST COPY AVAILABLE J.1 72b Calibration curve of SCubiifil : fluoride L m j.n n n a^g ?m m s s im i! S |; - 1- V Q: -i: u .yyuvH4 Urdar Measured 0.3333: 30u 000294 00021'/ EST COPY AVAILABLE 'loo/bo " C a l i b ration curve or 3bl;t>iiA1 H u o r i d e 1 ' g Qj1e~nQ jQ liia S f P g 5-4 0 1 1J y 3 u --T j .0585334 -------- ,-----------:--------- 1---------- i---------- '------U Measured Order : Invarsa Locarithi-, 563-1 403t 000295 00021'S ;iavi eist 3 cf SFSiiFi ` F l u o r ida i .5 F iis l Fi-He in i Crtl-F=Edit peaks l BEST COPY AVILABLE sono i 000296 000219 ~:Vo5 1 'y I Haw data cf ShOhiiAi : rluGride i.5 igjj 3*30 ir r m & & s- " i /' /i M h i ,, \ * f *,M *J : - .y \ J -a -.j 4 M3 & * /! r w :h n Hi \U \ ! \ \ ! \ \ \ '-. ! `-' "-! '> \ 1 \ ' V '. ! w/yiA 4560 -----,------------- r Ti?ia 9560 :-Exit ! rI-Help ! Crii-P=dit peaks 1 BEST COPY AVAILABLE 000297 OOOO -'095 -j fksw tlats of 950b i i Ai : t-Iuoride i .5 -qn-i A* <A yj 2 5 ., : aA a r? A -/ $2. *? iM,n.','-j/rt;1\ ^'i i9\ s i /!!I n i ! AA m \ SI n9\ w A /m1 U' j 3 ^ (f\ l x <? ^ .i h h m m y uy u uyy y yv y j y 3935 T ira e-txi ; i-l=Heip t Orti-F=Erfit peaks ! 3955 EST COPY AVAILABLE 000298 000221 4095 Haw d a ta o f 3 b 0b ii.fil - f l u o r i d e i . b 1 m m m m m W ^ W i2 ^ S S W M ^ S ~ ~ B 2 1 J?0 0 1_______ _________________________________ ! !i!i 1293b Tina se=xii i ft-HaIp I Crtl-P=dit peaks ? :743b BEST COPY AVAILABLE 000299 0002 995- UP 24 :lb :C:i OutPat of : 950524A1 OOO oitware : version b.l clS90,93 perator : DDW BEST COPY AVAILABLE u I 7.1. i< \S m-LOSS-.j Sow- ate of the Analysis : 1995-05-24 11:39 nalysis File Name : C:\SKALAR\DATA\HWIDATA\SERUM\950524A1 luoride 1.5 alibration order = Inverse Logarithm >l o p e : s = #.#### esult = 10 L s J x = corrected value of the sample cl = corrected value of the concentration 1 s = Slope of the electrode 2 = - 0.00000 1 = 0.00064 0 = -1.22588 luoride L alibration order = 2 'orrelation : r = 0.99786 esult = a2 * x 2 + al * x + aO 2 = -0.00000 1 = 0.00025 0 = -0.00699 ampler Type Number Sample Time Wash Time Air Time Take up sPecial needle Height 3A1000 1 50 s e c . 120 sec. 1 sec. Single None 70 m m . iluter needle Height dilution Factor dilution Volume Resample Dilution runs 30 mm 10 2 .5 mi . 1 1 User file : Reproces : No . TXT 000300 0003 995-05-24 16:01 OutPut of : 950524A1 luoride 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 s i 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 M e a s u r e w i n d o w 75 Filter No Rgnrt ion No formula : outDut : ## ### luoride 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 ******* 000301 0002 995-05-24 16:01 OutPut of : 950524A1 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 1300 Sec Peak shape Pointed stArt ignore 60 Sec eNd ignore 120 Sec Measure window 75 % Filter No Regeneration No formula : c4:=c3 output : #.#### 000302 0002iif 995-05-24 16:01 OutPut of : 950524A1 Fluoride 1.5 Fluoride L PPM PPM os Typ Ident Ch Result F Time Ch Result F Time t iw Initial Wash 3 0.059 t Tracer 3 1.452 d Drift 3 1.469 w Wash 3 0.059 sl S t a n d a r d 1 3 0.069 s2 Standard 2 3 0.073 s3 s4 Standard 3 Standard 4 n3J 0.087 0.109 s5 Standard 5 3 0.127 s6 Standard 6 3 0.155 0 s7 Standard 7 3 0.284 s8 Standard 8 3 0.612 2 3 s9 slO Standard 9 Standard 10 3Ti 1.242 1.458 4d Drift 3 1.442 5w Wash 3 0.059 5u BLK 1 3 0.075 7u 3u BLK 2 3 0.068 SPK 63-1 3 0.103 9u SPK 63-2 3 0.098 0u SPK 63-3 o 0.098 1u SPK 63-4 3 0.093 2u SPK 63-5 3 0.104 3u SPK 63-6 3 0 . 1 0 2 4u SPK 63-7 3 0 . 1 0 1 5u 6d BLK 3 3 0.072 Drift 3 1.455 7w Wash 3 0.059 3u BLK 4 3 0.073 9u F54088-48 3 0.068 0u F54099-48 3 0.071 1u F54089-48 3 0.066 LO IO u F54087-48 3 0.069 u F54080-48 3 0.067 u F54092-48 3 0.066 u F 5 4 1 0 9 -4 8 3 0.070 5u 7u F54114-48 3 0.071 F54088-DY8 3 0.068 3d Drift 3 1.485 9w Wash 3 0.059 0u F54099-DY8 3 0.069 1u 2u SPK 63-8 3 0 . 1 0 1 SPK 63-9 3 0.099 3u SPK 63-10 3 0 . 1 0 0 4u 5u BLK 1 3 0.073 BLK 2 3 0.065 6u SPK 63-1 3 0 .osa 7u 3u SPK 63-2 0.098 SPK63-3 3 0.102 9u F 54089-DY8 3 0.072 O d Drift 3 1.471 1w Wash 3 0.059 2u u F54087-DY8 3 0.075 F54080-DYS 3 0.075 65 215 389 621 740 916 1090 1269 1442 1618 1790 1965 2140 2314 2490 2728 2841 3015 3191 3365 3539 3715 3891 4067 4243 4415 4591 4831 4939 5118 5290 5466 5644 5812 5988 6168 6344 6517 6692 6930 7041 7218 7393 7568 7741 7915 8093 8267 8443 8619 87 93 9028 9143 9319 4 #.#### 4 0.4057 4 0.4054 4 #.#### 4 0.0180 4 0.0276 4 0.0569 4 0.0945 4 0.1182 4 0.1498 4 0.2417 4 0.3431 4 0.4037 4 0.4056 4 0.4058 4 #.#### 4 0.0330 4 0.0168 4 0.0343 4 0.0761 4 0.0768 4 0.0683 4 0.0860 4 0.0329 4 0.0821 4 0.0264 4 0.4056 4 #.#### 4 0.0273 4 0.0156 4 0.0233 4 0.0110 4 0.0192 4 0.0141 4 0.0095 4 0.0216 4 0.0228 4 0.0158 4 0.4052 4 #.#### 4 0.0170 4 0.0812 4 0.0774 4 0.0790 4 0.0276 4 0.0083 4 0.0597 4 0.0765 4 0.0838 4 0.0247 4 0.4054 4 #.#### 4 0.0328 4 0.0323 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 0002 000303 9 9 5 - 0 5 - 2 4 .16:01 OutPut of': 950524A1 Fluoride 1.5 Fluoride L PPM PPM ->S Typ Ident Ch Result F Time i. u 5u iu 7u 9u 9j\ u u !u 2d 3w iu 5u 7u 3u 3u 3u Lu 2u 3u 4d 3w }u 7u 3u 3u 3t u u Ju 3d 4w rw F54092-DY8 F5410 9-DY3 F54114-DY3 F54088-D15 F54099-D15 F54089-D15 F54087-D15 F54080-D15 Drift Wash F54092-D15 F54109-D15 F54114-D15 F54089-D22 F54087-D2 2 F54109-D22 F54114-D22 F54089-D28 F54087-D28 F 54109-D28 Drift Wash F54114-D28 SPK 63-4 SPK 63-5 SPK 63-6 SPK 63-7 F54105-48HR F5407 5-48HR Drift Wash RunOut Wash 3 0.077 3 0.073 3 0.077 3 0.061 3 0.073 3 0.068 3 0.069 3 0.066 3 1.469 3 0.059 3 0.070 3 0.072 3 0.068 3 0.068 3 0.067 3 0.068 3 0.069 3 0.066 3 0.068 3 0.069 3 1.455 3 0.059 3 0.069 3 0.078 3 0.089 3 0.098 3 0.097 3 0.069 3 0.072 o 1.411 o3-> 0.059 0.059 9493 9667 9843 10022 10194 10370 10542 10718 10893 11133 11244 11418 11594 11767 11944 12118 12296 12466 12642 12822 12994 13222 13345 13515 13695 13872 14045 14224 14393 14569 14810 15044 Ch Result F Time 4 0.0365 4 0.0280 4 0.0365 4 # .#### 4 0.0266 4 0.0165 4 0.0175 4 0.0107 4 0.4054 4 #.#### 4 0.0216 4 0.0249 4 0.0146 4 0.0158 4 0.0134 4 0.0146 4 0.0170 4 0.0114 4 0.0168 4 0.0170 4 0.4056 4 #.#### 4 0.0182 4 0.0379 4 0.0610 4 0.0763 4 0.0750 4 0.0180 4 0.0261 4 0.4060 4 #.#### 4 #.#### 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 000304 0 0 0 2 2 *7 socooo 3W 7IWIVAd031S38 0=j9 ________ O>DO - pS.ifr^eBy 'j. f: , i _________ i __________ j__________ !__________ i__________ i __________ ___________i________ ! __________ 7 sseoc' 0 fj 0 - l =r?- r * 4 ! ; i ~j B p k J O t i \ ; )-H [r7 ;'5 rj5 5 * 0 B J H D U O i B J u '.: ; = ;> -------Cal ibra :cn curve or :S C S24f)i : F l vo ri de l.v ------------------- 7^6F30v ^ f S ~~. ; li-i:>-Ji:n.":i-sa u " rii 1 o :n _..ubHev/S ir^ ;rder Featured Irvet'S LcuaHtr;- ___ j aOSb K r Copy 4KAI14BIE 000306 000225J BEST COPY AVAILABLE 000307 000230 nati data of 9^0524^1 : rluoria i .5 i i ungi -*^V \ J \ A. /V a A. a 4 A i\ a '"-J \j ">/ \; \ 4HS0 T?M:- c->at S Fl--Haip I Crti-r-E'c peaks BEST COPY AVAILABLE -K::-" 1 000308 O OO^'Jl P o* : 60C000 W lIV A U d O O iS jg if;3C- J.: --|i ..j ; i)\--- ;- :V>' y--: I- i __i__ ,& J) bL * ! ii i.i w : i rr I ;: vili__ 5 ' -'* !-"i ; 1 -: j...r i ~ .:~- ; .: n ^ M --------------- - :.`tv 7. A -a: IV n " _i \; xN ^N V A* AA ^ S S A< 9> - i v.x~ r e q g *o 1'Vu A a s BEST COPY AVAILABLE 000310 oooi:''- HAZLSTOM WISCONSIN OX 7545 707-7545 en ro Sponsor: 3M St. Paul, Minnesota a CORNING Company FINAL REPORT Study Title: Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Author: Steven M. Glaza Study Completion Date: September 14, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Pro.iect Identification: HWI 6329-159 P h o n e 6 0 3 - : 4 i - 4 4 7 i____ EXPRESS -MAIl DELIVERY Page 1 of 38 3301 KI NSMAN BLVD 000311 Fax 6 0 8 - 2 4 1 - 7 2 2 7 MADISON, W I 53704 Page 2 of 38 HWI 6329-159 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 Phase Date Reported to Date to Study Director Management 03/22/95 04/10/95 04/27/95 07/05/95 09/12/95 03/22/95 04/10/95 04/27/95 07/10/95 09/12/95 Protocol Review Protocol Amendment Animal Observation Data/Report Review Report Rereview 03/22/95 04/10/95 04/27/95 07/10/95 09/12/95 03/10/95 05/10/95 05/10/95 08/10/95 10/10/95 9 /y> Date 000312 Page 3 of 38 STUDY IDENTIFICATION Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits HWI 6329-159 Test Material T-6246 Sponsor 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. 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.O. Box 33220 St. Paul, MN 55133-3220 (612) 733-1962 Study Director Steven M. Glaza Hazleton Wisconsin, Inc. P.O. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Study Location Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, WI 53704 Study Timetable Study Initiation Date Experimental (In-life) Start Date In-life End Date Experimental Termination Date Study Completion Date March 30, 1995 April 4, 1995 May 1, 1995 September 14, 1995 September 14, 1995 000313 Page 4 of 38 HWI 6329-159 KEY PERSONNEL Acute Toxicoloqv Oualitv Assurance Steven M. Glaza Study Director Manager Sherry R. W. Petsel Manager Francis (Bud) W. McDonald Study Coordinator Patricia Padgham In-life Supervisor Laboratory Animal Medicine Cindy J. Cary, DVM Dipl ornate, ACLAM Supervisor Rose M. Bridge Report Supervisor Anatomical Patholoav Toxicoloqv SuDDort Thomas E. Palmer, PhD Anatomical Pathologist Kathy Myers Manager Calvin L. Horton Supervisor Jack Serfort/ Deborah L. Pirkel Supervisors Necropsy Anne Mosher Supervisor Pathology Data 000314 Page 5 of 38 CONTENTS Quality Assurance Statement Study Identification Key Personnel Summary Objective Regulatory Compliance Test and Control Materials Test System Procedures Results Discussion Signature Pathology Report Table 1 Individual Body Weights (g) 2 Individual Clinical Signs 3 Individual Pathology Comments 4 Individual Animal Tissue Weights and Bile Volumes Appendix A Protocol Deviations Protocol TP8084.PK Protocol Amendment No. 1 HWI 6329-159 Page 2 3 4 6 8 8 8 9 10 13 13 14 15 16 18 21 22 25 26 27 37 000315 Page 6 of 38 SUMMARY HWI 6329-159 This study was done to assess the level of systemic exposure of T-6246 when administered by a single intravenous injection to rabbits. The study was conducted using four male and four female acclimated rabbits of the Hra:(NZW)SPF strain for each treatment group as follows: Group 1 (Control ) 2 (Low) 3 (Medium) 4 (High) 5 (High) Test Material Sterile water T-6246 T-6246 T-6246 T-6246 Dose Level Number iof Animals (mq/kq) Mai es Females 04 54 10 4 100 4* 500 4* 4 4 4 4 4 * One animal sacrificed after treatment due to a possible broken back and replaced with another male animal. The animals received a single intravenous injection of the test material at the indicated dose level into the marginal ear vein of the right ear. The dose volume was 0.5 mL/kg of body weight for Groups 1 through 4 and was 0.75 mL/kg for Group 5. Two animals/sex/dose level were sacrificed on Day 15 and the remaining animals (two animals/sex/dose level) were sacrificed on Day 28. Clinical observations were conducted predose and at approximately 0.5, 2, and 4 hours after intravenous injection. Additional clinical observations and twice a day mortality checks were conducted daily thereafter until the scheduled sacrifice interval (Day 15 or Day 28). Body weights were determined on Day -7 for randomization purposes, before test or control material administration (Day 1), and at the scheduled sacrifice interval (Day 15 or Day 28). A blood sample (approximately 4 mL) was collected from a marginal ear vein (left ear) of the animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. An approximate 4-mL blood sample was also collected on Days 15 and 22 for the animals scheduled for sacrifice on Day 28. In addition, at the time of the scheduled sacrifice (Day 15 or Day 28), approximately 20 mL of blood was obtained from each animal. All samples were centrifuged, separated into serum and cellular fractions, and sent to the Sponsor. On Day 15 or 28, the animals were anesthetized with sodium pentobarbital, bled via the posterior vena cava, and exsanguinated. An abbreviated gross necropsy examination was not done, however, tissues were collected. The whole liver, bile, and both kidneys from each animal were collected, weighed (volume only determined for bile), and sent frozen to the Sponsor. 000316 Page 7 of 38 HWI 6329-159 Intravenous injection of T-6246 did not result in any test material-related changes in body weight gain. All animals appeared clinically normal throughout the study with the exception of one Group 4 and one Group 5 male animals that were sacrificed on Days 2 and 1, respectively, due to injury (possible broken backs). These animals were replaced and the replacement animals appeared clinically normal throughout the study. 000317 Page 8 of 38 HWI 6329-159 OBJECTIVE The objective of this study was to assess the level of systemic exposure to the test material, T-6246, when administered as a single intravenous injection to 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, with the exception that analysis of the test mixtures for concentration, homogeneity/solubility, and stability was not conducted. All procedures used in this study were 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 material was identified as T-6246 and described as a clear, colorless liquid. The control material was Sterile Water for Injection, USP (Abbott Laboratories, Lot No. 86-748-DM-02; Exp. April 1, 1996), 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). Analysis of the test material mixtures for concentration, homogeneity/solubility, and stability was not conducted or requested by the Sponsor. The purity and stability of the control material were considered to be adequate for the purposes of this study. Storage and Retention The test material was stored at room temperature. The control material was stored refrigerated. Any unused test material was returned to the Sponsor after completion of all in-life testing according to Hazleton Wisconsin (HWI) Standard Operating Procedure (SOP). Any remaining vehicle may be used for other testing and will not be discarded after issuance of the final report. 000318 Page 9 of 38 Safety Precautions HWI 6329-159 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 received from HRP, Inc., Kalamazoo, Michigan on March 8, 1995 and maintained at the Hazleton Wisconsin facility at 3301 Kinsman Boulevard, 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 -7. 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. 000319 Page 10 of 38 Study Design HWI 6329-159 Animals weighing from 2,523 to 3,276 g at initiation of treatment were placed into the following study groups: GrouD Dose Test/Control Dose Level Volume Number of Animals Materi al (mq/kg) (mL/kq) Mal es Females 1 (Control) 2 (Low) 3 (Medium) 4 (High) 5 (High) Sterile water T-6246 T-6246 T-6246 T-6246 0 5 10 100 500 0.5 0.5 0.5 0.5 0.75 4 4 4 4* 4* 4 4 4 4 4 * One animal sacrificed after treatment due to a possible broken back and replaced with another male animal, a Two animals/sex/dose level were sacrificed on Day 15. The remaining animals (two animals/sex/dose level) were sacrificed on Day 28. 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 Dose Preparation and Administration The test material was diluted with sterile water to achieve a specific concentration for each dose level in Groups 2 to 5. An individual dose of each respective test solution or control was calculated for each animal based on its body weight on the day of treatment. The respective test solution was administered by intravenous injection into the marginal ear vein of the right ear over approximately 22 to 60 seconds. The prepared test solutions were stored at room temperature until administered. After administration, any remaining test solutions were discarded. Reason for Route of Administration Intravenous injection is an acceptable route to assess systemic exposure. 000320 Page 11 of 38 Observations of Animals HWI 6329-159 Clinical observations were conducted predose and at approximately 0.5, 2, and 4 hours after intravenous injection. Additional clinical observations and twice a day mortality checks were conducted daily thereafter until the scheduled sacrifice interval (Day 15 or Day 28). Body weights were determined on Day -7 for randomization purposes and before test or control material administration (Day 1). Additional body weights were determined at the scheduled sacrifice interval (Day 15 or Day 28) or at unscheduled sacrifices (when survival exceeded 1 day). Sample Collections A blood sample (approximately 4 mL) was collected from the marginal ear vein (left ear) of the animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. An approximate 4-mL blood sample was also collected on Days 15 and 22 for the animals scheduled for sacrifice on Day 28. In addition, at the time of necropsy, approximately 20 mL of blood was obtained from the posterior vena cava of each animal. All samples were stored at room temperature, and then centrifuged, 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 The animals sacrificed on Days 1 and 2 due to apparent broken backs were necropsied in the same manner as the animals surviving to the scheduled sacrifices with the exception that they received an abbreviated gross necropsy examination and any abnormalities were recorded. On Day 15, the first two animals/sex assigned to each dose level (based on the group assignment randomization) were anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the posterior vena cava, and exsanguinated. An abbreviated gross necropsy examination was not done, however, tissues were collected. The whole liver, bile, and both kidneys from each animal were collected, weighed (volume only determined for bile), and immediately placed in a freezer set to maintain a temperature of -20C 10C. After tissue/bile collection, the animals were discarded. The remaining two animals/sex/dose level were anesthetized, bled, and exsanguinated on Day 28 in the same manner as the animals sacrificed on Day 15. 000321 Page 12 of 38 Shipment of Tissues HWI 6329-159 After completion of the in-life phase, the blood samples (serum and cellular fractions), livers, bile, 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), along with their corresponding weights or volumes. The Sponsor is responsible for the retention and disposition of the samples. HWI does not accept any responsibility for the analysis of the 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. 000322 Page 13 of 38 HWI 6329-159 RESULTS Body Weights Individual body weights are in Table 1. There was no meaningful effect on body weight gain during the study. Clinical Observations Individual clinical signs are in Table 2. All animals appeared normal throughout the study with the following exceptions: One Group 4 male (No. F54106) treated with T-6246 at 100 mg/kg appeared to have injured its back on Day 2 and was sacrificed, necropsied, and replaced with No. F54099. The replacement animal appeared normal throughout the study. One Group 5 male (No. F54076) treated with T-6246 at 500 mg/kg appeared to have injured its back on the day of treatment and was sacrificed, necropsied, and replaced with No. F54112. The replacement animal appeared normal throughout the study. Pathology Individual animal pathology comments are presented in Table 3. Individual animal tissue weights and bile volumes are in Table 4. The necropsy of animal No. F54106 revealed the skeletal muscle surrounding the lumbar spinal cord was diffusely dark red. In animal No. F54076, the skeletal muscle surrounding both hind limbs and the sacral spinal cord was diffusely dark red. The remaining animals survived to their scheduled sacrifice and were not examined grossly, although tissues were saved. Page 15 contains a pathology report by the study pathologist. DISCUSSION The level of systemic exposure of T-6246 was evaluated in male and female albino rabbits when administered as a single intravenous injection at levels of 5, 10, 100, and 500 mg/kg. There were no test material-related effects in any of the animals following administration of this material. 000323 Steven M. Glaza Study Director Acute Toxicology Page 14 of 38 SIGNATURE Date HWI 6329-159 000324 Page 15 of 38 PATHOLOGY REPORT HWI 6329-159 There were two male rabbits sacrificed on Day 1 or Day 2 and necropsied. At necropsy, the skeletal muscle surrounding the lumbar spinal cord in Animal No. F54106 (100 mg/kg of body weight) was diffusely dark red. In Animal No. F54076 (500 mg/kg of body weight), the skeletal muscle surrounding both hind limbs and the sacral spinal cord was diffusely dark red. The findings in these animals are indicative of injury of undetermined etiology to the caudal region of the back. The liver, bile, and both kidneys from these animals were collected as required by protocol. After necropsy, the animals were discarded. Pathologist (6329-159.Ihm) 051295 Date 000325 Page 16 of 38 HWI 6329-159 Table 1 Individual Body Weights (g) Random- Animal ization Initial Terminal Sex Number (Dav -7) (Dav 1) Dav 15 Dav 28 Group 1 (Control) - Sterile Water for In.iecti on (0 mo/ka) Mal e F54105 F54075 F54081 F54107 2,603 2,531 2,373 2,544 2,761 2,539 2,526 2,731 2,933 2,703 - - _ 2,874 3,056 Female F54104 F54086 F54074 F54085 2,665 2,526 2,540 2,553 2,890 2,632 2,703 2,667 3,130 2,775 - - _ 2,990 3,014 Male Female F54083 F54100 F54077 F54094 F54091 F54108 F54102 F54097 Group 2 - T-6246 (5 mg/kq) 2,381 2,675 2,459 2,466 2,598 2,869 2,592 2,601 2,538 3,030 - - 2,717 2,412 2,690 2,439 2,785 2,622 2,839 2,533 2,996 2,790 - - _ 2,859 2,916 - 3,290 2,928 Male Female F54071 F54093 F54095 F54101 F54110 F54096 F54090 F54078 Group 3 - T-6246 (10 mq/kq) 2,487 2,652 2,662 2,622 2,523 2,776 2,772 2,642 2,709 2,993 - - 2,719 2,772 2,542 2,710 2,802 2,914 2,740 2,842 2,959 3,022 - - _ _ 3,126 3,162 . - 3,022 3,244 - Not required. 000326 Sex Male Female Page 17 of 38 HWI 6329-159 Table 1 (Continued) Individual Body Weights (g) Animal Number Randomization (Day -7) Ini ti al (Dav 11 Terminal Dav 15 Dav 28 GrouD 4 - T-6246 f100 ma/kal F54088 F54106a F54089 F54087 F54099 F54080 F54092 F54109 F54114 2,494 2,677 2,597 2,578 2,360 2,498 2,400 2,592 2,776 2,582 2,833 2,662 2,665 2,764 2,622 2,540 2,657 2,933 2,771 2,803(2) - - 3,004 2,835 2,667 - - - 2,911 3,060 - _ - 2,979 3,296 Male Female GrouD 5 - T-6246 f500 ma/ka) F54076b F54113 F54082 F54111 F54112 2,655 2,667 2,734 2,547 2,996 2,678 2,753 2,843 2,784 3,276 - 2,896 - - 3,530 F54098 F54084 F54079 F54116 2,565 2,510 2,468 2,442 2,679 2,600 2,526 2,611 2,856 2,821 - - - 3,216 3,228 - - 2,959 3,002 Not required. a Animal No. F54106 was originally selected by the randomization program.for use in the study and was treated. This animal was sacrificed on Day 2 due to a broken back and was replaced with No. F54099. ( ) Number in superscripted parentheses indicates the day the body weight was taken. b Animal No. F54076 was originally selected by the randomization program for use in the study and was treated. This animal was sacrificed on Day 1 due to a broken back and was replaced with No. F54112. 000327 Page 18 of 38 HWI 6329-159 Table 2 Individual Clinical Signs Animal Sex Number Observation Hour (Day 1) __________ Day 0.5 2 4 2 3 to 15 16 to 28 Mai e GrouD 1 (Control) - Sterile Water for In.iec:tion (0 mo/kal F54105 Appeared normal //// / F54075 Appeared normal //// / * * F54081 Appeared normal //// / / Female F54107 F54104 F54086 Appeared normal Appeared normal Appeared normal //// //// //// / / / * * F54074 Appeared normal //// / / F54085 Appeared normal //// / / Male F54083 F54100 GrouD 2 - T--6246 (5 mq/kql Appeared normal // Appeared normal /// / / / / * * F54077 Appeared normal //// / / Female F54094 F54091 F54108 Appeared normal Appeared normal Appeared normal //// /// //// / J / / * * F54102 Appeared normal //// / / F54097 Appeared normal /// / / * Animal sacrificed on Day 15. / Condition existed. 000328 Sex Male Female Male Female Page 19 of 38 HWI 6329-159 Table 2 (Continued) Individual Clinical Signs Animal Number Observation Hour (Dav 11 0.5 2 4 Dav 2 3 to 15 16 to 28 F54071 F54093 Group 3 - T-6246 (10 mg/kg). Appeared normal //// Appeared normal / / / / * * F54095 Appeared normal /// / / / F54101 F54110 F54096 Appeared normal Appeared normal Appeared normal // / / // / / // / / / / / * * F54090 Appeared normal // / / / F54078 Appeared normal /// / / F54088 GrouD 4 - T- 6246 f100 ma/kq) Appeared normal //// / * F54106 Appeared normal // / - Broken back /- - - Moribund sacrifice - - - / F54089 Appeared normal // / / / / F54087 Appeared normal F54099a Appeared normal F54080 Appeared normal F54092 Appeared normal //// // / / /// / /// / J / / / / * * * F54109 Appeared normal // / / / F54114 Appeared normal //// / / * Anima'1 sacrificed on Day 15. / Condition existed. - Condition not evident. a Replacement animal for Animal Nci. F54106. 000329 Sex Male Female Page 20 of 38 Table 2 (Continued) Individual Clinical Signs Animal Number Observation Hour (Day 0.5 2 4 2 GrouD 5 - T-6246 (500 ma/kql F54076 Appeared normal /// Broken back t- - Moribund sacrifice - - t F54113 Appeared normal //// F54082 Appeared normal //// F54111 Appeared normal /// F541123 Appeared normal /// F54098 Appeared normal //// F54084 Appeared normal //// F54079 Appeared normal /// F54116 Appeared normal /// HWI 6329-159 Dav 3 to 15 16 to 28 /* // // /* /* /* // // * Animal sacrificed on Day 15. / Condition existed. - Condition not evident. f Condition evident at time of 12-hour bleeding interval, a Replacement animal for Animal No. F54076. 9 000330 Page 21 of 38 Table 3 Individual Pathology Comments HWI 6329-159 Animal Number ______ Test Day_____ Sex Died Sacrificed Necroosv Observation Group 4 - T-6246 (100 ma/kql F54106 M 2 The skeletal muscle surrounding the lumbar spinal cord is diffusely dark red. F54076 M Group 5 - T-6246 (500 ma/kq) - 1 The skeletal muscle surrounding both hind limbs and sacral spinal cord is diffusely dark red. 000331 Page 22 of 38 HWI 6329-159 Table 4 Individual Animal Tissue Weights and Bile Volumes Animal Sacri fice Weiaht (a) Bi 1e Sex Number ___Day___ Liver Kidnevs Volume (mL) GrouD 1 (Control) - Sterile Water for In.iection (0 mq/ka) Mal e F54105 15 86.35 14.92 0.6 F54075 15 60.25 15.97 0.1 F54081 28 80.597 14.764 1.2 F54107 28 80.649 15.620 0.4 Female F54104 15 98.40 16.03 0.7 F54086 15 77.30 15.23 0.7 F54074 28 82.895 18.793 1.6 F54085 28 80.950 14.925 1.3 Mal e Female F54083 F54100 F54077 F54094 F54091 F54108 F54102 F54097 GrouD 2 - T-6246 (5 ma/kq) 15 73.15 14.87 15 77.90 17.62 28 75.699 17.864 28 78.689 17.213 15 84.90 14.92 15 79.90 15.40 28 101.494 18.308 28 73.621 16.214 0.7 0.7 0.3 0.8 1.6 0.4 1.7 1.4 000332 Sex Male Female Page 23 of 38 HWI 6329-159 Table 4 (Continued) Individual Animal Tissue Weights and Bile Volumes Animal Number Sacrifice Day Weioht (q) Liver Kidnevs Bile Volume I GrouD 3 - T-6246 ilO ma/kq) F54071 15 77.10 15.88 0.5 F54093 15 81.40 15.50 0.8 F54095 28 85.504 15.561 1.2 F54101 28 85.668 16.120 1.0 F54110 15 83.68 15.79 1.4 F54096 15 78.10 18.70 1.1 F54090 28 76.220 15.878 2.1 F54078 28 83.460 18.130 1.0 Mai e Female GrouD 4 - T-6246 H O O ma/kal F54088 15 84.26 16.98 F54106 2 82.301 16.550 F54089 28 71.796 16.434 F54087 28 83.710 18.591 F54099a 15 82.143 16.038 F54080 15 75.42 18.32 F54092 15 78.36 16.68 F54109 28 73.958 16.915 F54114 28 80.966 19.404 0.4 2.0 1.0 0.6 0.5 1.2 * 1.6 2.0 a Replacement animal for Animal No. F54106. * This animal had no gallbladder, so no bile was collected. 000333 Page 24 of 38 Table 4 (Continued) Individual Animal Tissue Weights and Bile Volumes HWI 6329-159 Animal Sacri fice Weight (q) Bile Sex Number Day Liver Kidnevs Volume GrouD 5 - T-6246 (500 ma/kal Mai e F54076 F54113 1 15 78.058 91.52 14.695 16.57 2.0 1.0 F54082 28 88.965 19.431 1.6 F54111 28 83.823 19.405 1.5 F541123 15 97.824 21.069 1.4 Female F54098 15 78.95 14.72 1.2 F54084 15 66.42 13.18 0.8 F54079 28 83.117 14.785 1.8 F54116 28 71.962 15.793 2.0 a Replacement animal for Animal No. F54076. 000334 Page 25 of 38 APPENDIX A Protocol Deviations Protocol TP8084.PK. Protocol Amendment No. 1 HWI 5329-159 000335 Page 26 of 38 Protocol Deviations HWI 6329-159 Protocol Page 6, 7. Experimental Design, C. Dosing Procedures, (1) Dosing Route. Intravenous injection into the marginal ear vein of the right ear over approximately 30 to 60 seconds. Actual Procedure The rate of injection for one Group 5 female (No. F54098) was 22 seconds. Also, the time for dose administration for one male animal (Group 4, No. F54087) was incorrectly recorded thus the exact duration of dosing can not be determined. These deviations are not considered to have had an adverse effect on the outcome of the study. 000336 0 HAZLETON WISCONSIN P O S T O F F I C E B O X 7 5 4 ft M A D I S O N . W I S .1 I 0 : 7 `j 4 r> Page 27 of 38 a C O R N IN G Company Sponsor: 3M , St. Paul, Minnesota PROTOCOL TP8084.EXT Study Title: Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits Date: March 30, 1995 Performing Laboratory: Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, Wisconsin 53704 Laboratory Project Identification: HWI 6329-159 000337 P h o n e 6 0 8 2 4 i -i 4 ' l E X P R E S S N/ A I L D E I i V E n Y 330 1 KINSMAN BIVD F a x 6 0 a 2 4 1- 7 2 2 7 MADISON. W I 53704 Page 28 of 38 STUDY IDENTIFICATION TP8084.EXT Page 2 Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits HWI No. Test Material Sponsor Sponsor's Representative Study Director Study Location Proposed Study Timetable Experimental Start Date Experimental Termination Date Draft Report Date 5329-159 T-6246 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 John L. Butenhoff, PhD 3M Toxicology Service Medical Department 3M Center, Bldg. 220-2E-02 P.O. Box 33220 St. Paul, MN 55133-3220 (612) 733-1962 Steven M. Glaza Hazleton Wisconsin, Inc. P.O. Box 7545 Madison, WI 53707-7545 (608) 241-7292 Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, WI 53704 April 4, 1995 May 1, 1995 June 12, 1995 000338 Page 29 of 38 TP8084.EXT Page 3 1. Study Single-Dose Intravenous Pharmacokinetic Study in Rabbits 2. Purpose To assess the level of systemic exposure when the test material is administered as a single intravenous injection to rabbits 3. Regulatory Compliance This study will be conducted in accordance with the following Good Laboratory Practice Regulations/Standards/Guidelines with the exception that analysis of the test material mixtures for concentration, solubility, homogeneity, and stability will not be conducted: [ ] Conduct as a Nonregulated Study [X] 21 CFR 58 (FDA) [ ] 40 CFR 150 (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. Quality Assurance The protocol, study conduct, and the final report will be audited by the Quality Assurance Unit in accordance with Hazleton Wisconsin (HWI) Standard Operating Procedures (SOPs) and policies. 5. Test Material A. Identification T-6246 B. Physical Description (To be documented in the raw data) C . Purity and .Stability The Sponsor assumes responsibility for purity and stability determinations (including under test conditions). Samples of test material/vehicle mixture(s) for concentration, solubility, homogeneity, and stability analyses will be taken before administration if requested by the Sponsor. These samples (if taken) will be sent to the Sponsor after experimental termination. D. Storage Room temperature 000339 Page 30 of 38 TP8084.EXT Page 4 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 maintain a temperature of -20C 10C 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 material 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 Sterile water for injection B. Physical Description Clear, colorless liquid C. Purity and Stability The purity and stability of this USP grade material is considered adequate for the purposes of this study. D. Storage Refrigerated 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 7. Experimental Design Animals (1) Species Rabbit (2) Strain/Source Hra:(NZW)SPF/HRP, Inc. 000340 Page 31 of 38 TP8084.EXT Page 5 (3) Age at Initiation Adult (4) Weight at Initiation 2.5 to 3.5 kg (5) Number and Sex 20 males and 20 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 #5325 (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, specified microbiological content, 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 19C to 23C, a relative humidity of 50% +20%, and a 12-hour 1ight/12-hour dark cycle. (f) Acclimati on 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. 000341 Page 32 of 38 TP8084.EXT Page 6 (9) 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. B. Dose Administration (1) Test Groups GrouD Test/Control Material Dose Level Number of Animals (mg/kg)a Mal es Females 1 (Control) Sterile water 0 4 4 2 (Low) T-6246 5 4 4 3 (Medium) T-6246 10 4 4 4 (High) T-6246 100 4 4 5 (High) T-6246 500 4 4 a The dose volume will be 0.5 mL/kg of body weight, b Two animals/sex/dose level will be sacrificed on Day 15. The remaining animals (two animals/sex/dose level) will be sacrificed on Day 28. C . Dosing Procedures (1) Dosing Route Intravenous injection into the marginal ear vein of the right ear over approximately 30 to 60 seconds. (2) Reason for Dosing Route Intravenous injection is an acceptable route to assess systemic exposure. (3) Dosing Duration Single dose (4) Dose Preparation The day of treatment will be designated as Day 1. The Group 1 animals will be treated with sterile water at a dose volume of 0.5 mL/kg. The test material will be diluted with sterile water to achieve a specific concentration for each dose level in Groups 2-5. Individual doses will be calculated based on the animal's body weight taken just before test material administration. The prepared test mixtures will be stored at room temperature until administration. J 000342 Page 33 of 38 TP8084.EXT Page 7 D. Observation of Animals (1) Clinical Observations The animals will be observed for clinical signs of toxicity before test or control material administration, at approximately 0.5, 2.0, 4.0 hours post-injection (Day 1), and daily thereafter for clinical signs and twice daily (a.m. and p.m.) for mortality until the scheduled sacrifice interval (Day 15 or Day 28). Observations may be extended when directed by the Study Director. (2) Body Weights For randomization, before test or control material injection (Day 1), at the scheduled sacrifice interval (Day 15 or Day 28), and at unscheduled death and sacrifices (when survival exceeds 1 day) (3) Sample Collections (a) Frequency 4-, 8-, 12-, 24-, and 48-hours post-injection, on Days 8, 15, 22, and at the scheduled sacrifice interval (Day 15 or Day 28) (b) Method of Collection/Number of Animals Blood samples (approximately 4 ml) will be collected from the marginal ear vein (left ear) of all animals at 4-, 8-, 12-, 24-, and 48-hours post-injection, and on Day 8. Additional samples will be collected on Days 15 and 22 for the animals scheduled for sacrifice on Day 28. Approximately 20 mL of blood (actual volume to be documented in the raw data) will be obtained from the posterior vena cava at the time of the scheduled sacrifice (Day 15 or Day 28). Approximately 20 mL of blood will be collected from moribund animals during the study, also, if possible. 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 a temperature of -20C 10C. The separated serum and cellular fractions will be sent frozen on dry ice to the Sponsor after experimental termination. The Sponsor is responsible for the retention and disposition of the samples. 000343 TP8084.EXT Page 8 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 his alternate will be notified by telephone at (612) 778-5294 prior to the shipment of the samples. Termination (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. Tissues, as described in section 7.E. (3) Sample Collection, will be collected from any animal dying during the study or sacrificed in a moribund condition. After necropsy, the animals will be discarded. (2) Scheduled Sacrifices On Day 15, the first two animals/sex assigned to each dose level (based on the group assignment randomization) will be anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the vena cava, and exsanguinated. The remaining two animals/sex/dose level will be anesthetized with sodium pentobarbital (via injection in the marginal ear vein), bled via the vena cava, and exsanguinated on Day 28. An abbreviated gross necropsy examination will not be done, however, tissues (as described in section 7.E. (3) Sample Collection) will be collected. (3) Sample Collection The whole liver, bile, and both kidneys from each animal will be collected, weighed (volume only determined for bile), and immediately placed in a freezer set to maintain a temperature of -20C 10C. After sample collection, the animals will be discarded. The samples (liver, bile, and kidneys) will be sent frozen on dry ice to the Sponsor after experimental termination. The samples and their corresponding weights or volumes will be shipped to the person listed in Section 7.D.(3).(b). The Sponsor is responsible for the retention and disposition of the samples. 000344 Page 35 of 38 TP8084.EXT Page 9 F. 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 Description of any toxic effects Gross pathology findings (if applicable) Gross pathology report (if applicable and requested by the Study Director) Individual animal tissue weights and bile volumes 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 Sample collection records Shipping records Pathology 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 j 000345 Page 36 of 38 PROTOCOL APPROVAL TP8084.EXT Page 10 John L. Butenhoff, PhD Sponsor's Representative 3M Date Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. (j 4\4t~ {HudL_________ Representative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-159.protdsk2) Date z-zo-^s- Date ) 000346 Page 37 of 38 0 HAZLETON WISCONSIN P O S T O F F I C E B O X 7 54 5 M A D I S O N . Wl 5 3 7 0 7 75 4 5. a C ORN ING Company I PROTOCOL TP8084.EXT Single-Dose Intravenous Pharmacokinetic Study of T-6246 in Rabbits HWI 6329-159 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 Contractor Hazleton Wisconsin, Inc. 3301 Kinsman Boulevard Madison, WI 53704 Study Director Steven M. Glaza Amendment No. 1 This amendment modifies the following portions of the protocol: Effective April 4, 1995 1. Page 6, 7. Experimental Design: B. Dose Administration: (II Test Groups. The test material mixture for Group 5 could not be prepared at the concentration needed to utilize a dose volume of 0.5 mL/kg. Modify footnote "a" in this section with the following underlined addition: a The dose volume will be 0.5 mL/kg of body weight for Groups 1-4 and 0.75 mL/kg for Group 5. P h_o_n e 6 0 8 - 2 4 1 - 4 4 7 1 EXPRESS-MAIL DELIVERY 000347 3301 KINSMAN BLVD _________F a x __ .6.08. - 2 4 1 - 7 2 2.7 MADISON. Wl 53704 Page 38 of 38 Amendment No. HWI 6329-159 Page 2 Effective April 5, 1995 2. Page 6. 7. Experimental Design; B. Dose Administration; (11 Test Groups. Animal No. F54076 (Group 5 male) and Animal No. F54106 (Group 4 male) were sacrificed on Days 1 and 2, respectively, due to injuries (apparent broken backs). Add the following paragraph to this section: Due to the sacrifice on Day 1 of one Group 5 male (Animal No. F54076) and on Day 2 of one Group 4 male (Animal No. F54106) because of injuries (apparent broken backs), replacement animals will be treated at the same dose levels in the same manner as for the initial animals in the study. The observations (clinical observations, body weights and sample collections) and the termination of the animals (unscheduled sacrifices and deaths, scheduled sacrifices, and sample collection) will be conducted in the same manner as for the other animals in the study terminating on Day 15. PROTOCOL AMENDMENT APPROVAL John L. Butenhoff, PhD Sponsor's Representative 3M Toxicology Service Medical Department Date Steven M. Glaza Study Director Acute Toxicology Hazleton Wisconsin, Inc. iM Representative Quality Assurance Unit Hazleton Wisconsin, Inc. (6329-159.Ami.dsk3) Date _______ i t - Date 000348 ISSI733 i Calibration curve of 95b04A : Fluor ida L M -C b4 affi 0.001409 Or d a r 0.93928 000349 000169 i .7131803 i Calibration curve of 36Ub04Ai - fluoride i .5 0.057^42711H s i H W O'O.fvcti&.n. t: ir a t: i o n d. 0563488 u 1 t----------r Measured Order : Inverse Logarithm; 4095 6174 000350 000170 409b Hay data o f 3S0S u4h : r lu o r id e i .5 194 f ^ f g w `2J-" H 5- 1 ?/ Ut s a.il ai ^ ;\ ] 1 ! ,. \n\ ii i: 'i \ ii Ui iI i V '-S 'J if ' j - t f ,i i ' a o i II ( u AjlAiUllll : \ J 'g Z?ni, \Xf* f( i1 0 T ine :sc=xit ! Fi=Halp ! Crii-r^hdit peaks ; 5000 000351 0001V1