Document b5X8zdOx1kdwdoNwO2wBXyZ56

AR226-3160 TRADE SECRET Study Title H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID: DuPont-2921 AUTHOR: Carol Finlay, B.A. STUDY COMPLETED ON: August 29, 2000 PERFORMING LABORATORY: E.I. du Font de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 WORK REQUEST NUMBER: SERVICE CODE NUMBER: M Page 1 of 91 Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ____ ________________DuPont-2921 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. ^ Review,dby: ^J ^ ' Zg^feZO^ Gary W. ~^ Jepson, ___ Ph.D. ______ Date Senior Research Scientist Reviewed by: QujdA^L0. \tcUDUj^^ (IJudith C Stadler,.Ph.D.. D.A.B.T. v Director 2^'^U<S -2005) Dale ^^^A, Issued by Study Director:_______(L^hf Carol FFiniln^yl,^B..B/ .A.J Staff Scientist ^ fi^ ^ -^6 Dm &- -2- Company Sannized. Does not contain TSCA CB H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLE OF CONTENTS Page CERTIFICATION........................................................................................................................^ LIST OF TABLES .........................................................................................................................4 LIST OF FIGURES.......................................................................................................................4 LIST OF APPENDICES...............................................................................................................4 STUDY INFORMATION .............................................................................................................5 STUDY PERSONNEL...................................................................................................................? SUMMARY ....................................................................................................................................8 INTRODUCTION..........................................................................................................................9 MATERIALS AND METHODS..................................................................................................9 A. Test Substance and Positive Control....................................................................................9 B. Test Species .........................................................................................................................9 C. Animal Husbandry............................................................................................................. 10 1. Housing Environment.....;....................................................................................................................... 10 2. Feed and Water.......................................................................................................................................10 3. Identification .......................................................................................................................................... 10 4. Animal Health Monitoring Program.............................................................................;......................... 10 D. Quarantine and Pretest.......................................................................................................11 E. Study Design..........................................................................................................;.. .........11 F. Assignment to Groups and Study Start..............................................................................11 G. Dosing Material Preparation and Administration ..............................................................11 1. Test Substance........................................................................................................................................11 2. Positive Controls.................................................................................................................................... 12 3. Negative Controls................................................................................................................................... 12 H. Body Weights.....................................................................................................................12 I. Mortality and Clinical Observations..................................................................................12 J. Collection and Analysis of Blood and Livers ....................................................................12 K. Treatment of Fluorine Data................................................................................................ 13 RESULTS AND DISCUSSION ..................................................................................................14 A. In-Life Toxicology............................................................................................................. 14 B. Fluorine Data .............................................;.......................,,...,,,,.,,.............--....................14 1. Factors Influencing Interpretation of Analysis .......................................................................................14 2. Positive Controls.................................................................................................................................... 14 3. Test Substance........................................................................................................................................ 15 CONCLUSIONS ..........................................................................................................................15 RECORDS AND SAMPLE STORAGE ....................................................................................16 TABLES........................................................................................................................................17 -3- IBompanySanitized. Does not contain TSCA r H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats____________________________________DuPont-2921 nGURES......................................................................................................................................23 APPENDICES ..............................................................................................................................28 LIST OF TABLES Page 1. MEAN BODY WEIGHTS...............................................................................................................................18 2. MEAN BODY WEIGHT GAINS ........................................................................................................................20 3. MEAN BLOOD FLUORINE LEVELS................................................................................................................21 4. MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE................................................22 LIST OF FIGURES Page 1. MEAN BODY WEIGHTS ...................................................................................................................................24 2. MICROMOLAR EQUIVALENTS IN RAT BLOOD..........................................................................................25 3. MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS.............................27 LIST OF APPENDICES Page A. INDIVIDUAL BODY WEIGHTS ....................................;..................................................................................29 B. INDIVIDUAL CLINICAL OBSERVATIONS....................................................................................................70 C. INDIVIDUAL FLUORINE LEVELS..................................................................................................................83 4Company Sannized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION TEST SUBSTANCE; Substance Tested: Synonyms/Codes: H-23925 Haskell Number: 23925 CompositionJjSBrt Known Impurities: None POSITIVE CONTROL; DuPont-2921 Known Impurities: Unknown - a- company Sanitized. Does not contain TSCA Cfla H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION (Continued) POSITIVE CONTROL: Substance Tested:j|^|^JU Synonyms/Codes: H-24020 DuPont-2921 Haskell Number: 24020 Composition Known Impurities: Unknown Sponsor: E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: April 30, 1999 / (see report cover page) In-Life Initiated/Completed: May 16, 1999 / August 17, 1999 -6- ompany Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY PERSONNEL Study Director: Management: Primary Technician: Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. Richard P. Mathena Blood Fluorine Data Analysis: Gary W. Jepson, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T. Toxicology Report Preparation: Wanda F. Dinbokowitz Laboratory Veterinarian: Wanda L. West, D.V.M., A.C.L.A.M. DuPont-2921 -7- Company Sanitized. Does not contain TSCA CBI SUMMARY The objective of this study was to evaluate the potential for H-23925, when administered by gavage, to be absorbed and to accumulate in a mammalian system. Six groups of 5 male Cri:CD(SD)IGS BR rats each were given 200 mg/kg/day of H-23925. The test substance was administered to one group of 5 rats for 5 consecutive days and to 5 groups for 10 days. Approximately 2 hours after the first dose, blood was collected from the orbital sinus of each rat from the 5-dose group. On selected days (5,10,13, 24, 52, 94) 5 rats per group were euthanized and the blood and livers were collected. Body weights and clinical signs were recorded on each day of dosing and then weekly, every other week, or every 3 weeks during the recovery period. Additionally, 3 negative controls, deionized water, corn oil, and corn oiVacetone (80:20), and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-23925. No deaths occurred during the study. One rat dosed with the test substance, H-23925, exhibited diarrhea on test day 4. This clinical sign is considered not to be due to administration of the test substance because it was only observed on one day in a single rat. Rats dosed with H-24019 exhibited diarrhea, salivation, alopecia, black ocular discharge, and staining of various parts of the body during the dosing period. Rats dosed with H-24020 exhibited wet perineum and diarrhea during the dosing period. Alopecia was observed during the recovery period in rats dosed with H-24019, H-24020, and deionized water. The corn oil and corn oiVacetone negative control rats exhibited no clinical signs during the study. Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive or negative controls. Accounting for the age difference at study start and the expected rate of body weight gain, it appears that the mean body weights and mean body weight gains of the rats dosed with the test substance, H-23925, are comparable to the negative controls and equal to or greater than the positive controls. A steady-state for fluorine levels in whole blood was not achieved during 10 consecutive days of dosing with 200 mg/kg H-23925. An area under the curve (estimated to infinity) was calculated and normalized for fluorine content for the test substance and each positive control. The AUCINF/D for the fluorine component of the test substance, H-23925, was 14,886.1, compared to AUCINF/D values of 566,479.1 and 70,789.6 for the positive controls, H-24019 and H-24020, respectively. Under the conditions of this study, administration of H-23925 to male rats for 10 consecutive days resulted in low absorption and retention of fluorine in the blood. Dose adjusted areas under the curve (AUCINF/D) for positive controls, H-24019 and H-24020, were approximately 38x and 5x the AUCINF/D for the test substance. -8- Cofflpany Sanitized. Does not contain TSCA C5t ^^1^|^| INTRODUCTION The objective of this study was to define the potential of H-23925 to be absorbed and to bioaccumulate in a mammalian system, as indicated by analytical determination of total fluorine in blood and liver. The test substance was compared to 2 positive controls that were materials previously shown to bioaccumulate in mammals. The daily dosage for the test substance, 200 mg/kg, was selected based on available toxicity data and the results of rangefinding studies. In the initial rangefinding study, a group of 5 male rats was dosed by oral gavage with H-23925 at a dosage of 2000 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water and served as controls. The rats dosed with H-23925 experienced an overall mean body weight loss of 29 grams. In the next rangefinding study, 2 groups of 5 male rats were dosed by oral gavage with H-23925 at a dosage of 500 or 1000 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water and served as controls. Overall mean body weight losses of 7 and 25 grams occurred in the rats dosed with the test substance. A group of 5 male rats was then dosed by oral gavage at a dosage of 125 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water and served as controls. The rats dosed at 125 mg/kg experienced an overall mean body weight gain of 13 grams. The control group had an overall mean body weight gain of 14 grams. The dosage of 200 mg/kg was chosen for the main study and was expected to produce less than a 10% difference in mean body weight when compared to controls. All blood samples were analyzed for total fluorine content. This report contains the results of those analyses. Results from analyses of selected liver samples will be presented in a supplemental report. MATERIALS AND METHODS A. Test Substance and Positive Control The test substance, H-23925, was supplied by the sponsor as an amber liquid with some flock sediment. The positive controls, H-24019 and H-24020, were supplied by the sponsor as white solids. The test substance and positive controls appeared to be stable under the conditions of the study. No evidence of instability, such as a change in color or physical state, was observed. B. Test Species Male Crl:CD(SD)IGS BR rats were received from Charles River Laboratories, me., Raleigh, North Carolina. The Crl:CD(SD)IGS BR rat was selected on the basis of extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, and low incidence of spontaneous diseases. -9- Company Sanitized. Does not contain TSCA CBI ^^\ H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 C. Animal Husbandry 1. Housing Environment Rats were housed singly in stainless steel, wire-mesh cages suspended above cage boards. Animal rooms were maintained on an approximate 12-hour light/dark cycle (fluorescent light) and at a temperature of 23 1C and a relative humidity of 50 10%. Occasional excursions outside the accepted ranges were minor and did not affect the study. 2. Feed and Water Tap water was provided ad libitum. All rats were fed PMI Nutrition International, Inc. Certified Rodent LabDiet 5002 chow. The feed is guaranteed by the manufacturer to meet specified nutritional requirements and to be free of specified contaminants. 3. Identification Prior to assignment to groups, each rat was temporarily identified by cage identification. After assignment to groups, an individual identification number was marked or tattooed on the tail of each rat. The information on the cage labels included the unique 6-digit Haskell animal number assigned to each rat. 4. Animal Health Monitoring Program As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to assure that contaminant levels are below those that would be expected to impact the scientific integrity of the study: Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. Feed samples are analyzed for total bacterial, spore and fungal counts. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations of key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study. The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. -10- jCompany Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 D. Quarantine and Pretest Upon arrival at Haskell Laboratory, the rats were removed from shipping cartons and quarantined for 6 days. The rats were weighed 3 times during the pretest period and examined daily for any clinically apparent signs of disease or injury. The rats were observed daily for mortality and signs of illness, injury, or abnormal behavior. On the bases of acceptable body weight gains and freedom from clinically apparent signs of disease or injury, the rats were released from quarantine by the laboratory animal veterinarian designee. E. Study Design Substance Negative Controls Deionized water Corn oil Corn oil: acetone Positive Controls H-24019 H-24020 Test Substance H-23925 Vehicle Dosage (mg/kg) Number of Animals 0 30 Not applicable 0 30 0 30 Corn oil: acetone 10 30 Corn oil 20 30 Deionized water 200 30 F. Assignment to Groups and Study Start After the quarantine period, the rats were selected on the bases of adequate body weight gain, freedom from any clinical signs of disease or injury, and a body weight within 20% of the mean. The selected rats were divided by computerized, stratified randomization into 6 groups of 5 rats, so that there were no statistically significant differences among group body weight means. After assignment to groups, each rat was housed individually. The last 3 digits of the animal number was marked or tattooed on the tail of each rat. The rats were between 7 and 9 weeks of age at the time of dosing. Dosing began on test day 1. Rats that were not assigned to the study were released for other laboratory purposes, or were sacrificed by carbon dioxide asphyxiation and discarded without pathological evaluation. G. Dosing Material Preparation and Administration 1. Test Substance H-23925 was mixed with deionized water so an accurate volume could be delivered. The amount of test substance each rat received was based on the body weight collected on each day of dosing and the dosing mixture concentration. The rats were dosed at a volume of 10 mL/kg of -11 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 body weight. The test substance was stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. 2. Positive Controls The solid positive control compounds were suspended as emulsions in their respective vehicles. Corn oil was used as the vehicle for H-24019. It was necessary to dissolve H-24020 in acetone before suspending it in corn oil. The ratio of acetone to corn oil was 20:80. The amount each rat received was based on the body weight collected on each day of dosing and the suspension concentration. The dose volumes did not exceed 1 mL/100 g of body weight. The dosing suspensions were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. 3. Negative Controls Deionized water was chosen as a negative control because it was the vehicle for the test substance. Corn oil and corn oil:acetone (80:20) were chosen as negative controls because they were the vehicles for the positive controls. Each negative control rat received 1 mL of corn oil or corn oil:acetone. These rats were dosed in a separate room from the rats dosed with the test substance or positive controls. H. Body Weights All rats were weighed on each day of dosing. The positive and negative control rats were weighed weekly or every other week during the recovery period. The rats dosed with H-23925 were weighed weekly, every other week, or every 3 weeks during the recovery period. I. Mortality and Clinical Observations Cage-site examinations to detect moribund or dead rats and abnormal behavior and appearance among rats were conducted at least once daily throughout the study. At every weighing, each rat was individually handled and examined for abnormal behavior and appearance. J. Collection and Analysis of Blood and Livers Approximately 2 hours after the first dose, 1-2 mL of blood was collected into EDTA tubes from the orbital sinus of each rat from Group I. At all other selected time points, 5 rats/group were euthanized by carbon dioxide anesthesia and exsanguination and blood and livers were collected according to the following schedules: -12- Company Sanitized. Oos not contain TSCA CBl H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Group I I n m IV v VI Dosing Days 1-5 1-5 1-10 1-10 1-10 1-10 1-10 Tissue Collected Blood Blood and Liver Blood and Liver Blood and Liver Blood and Liver Blood and Liver Blood and Liver Sampling Time Test day 1 (2 hours post dosing) Test day 5 at sacrifice (2 hours post dosing) Test day 10 (2 hours post dosing) Test day 13 Test day 24 Test day 52 Test day 94 Five to 10 mL of blood was collected into EDTA tubes at sacrifice. The livers were weighed. The blood from all rats was refrigerated and the livers were frozen. The blood was appropriately packaged and shipped refrigerated to Jackson Laboratory, Deepwater, New Jersey where it was analyzed for total fluorine. The total fluorine content of the blood samples was determined using a Wickbold torch combustion method, followed by analysis with a fluoride ion selective electrode. The liquid blood was decomposed or volatilized in the presence of wet oxygen and swept through an oxyhydrogen flame in a closed quartz apparatus. The combustion products were collected in an aqueous absorbing solution and analyzed using a fluoride ion selective electrode. The analytical data (ppm F in each sample) was supplied to Haskell Laboratory personnel for evaluation of fluorine biopersistence. K. Treatment of Fluorine Data Noncompartmental analysis was conducted on blood fluorine data derived from rats dosed with H-23925 by using WinNonlin Version 3.0 software (Pharsight Corp, Mountain View, CA). WinNonlin software provided a means of computing derived pharmacokinetic parameters from data files including area under the curve (AUCINF), Cmax and terminal half-life (Ti/z). The AUCINF (concentration x time) represents the area under the blood concentration curve from the time of dosing extrapolated to infinity. The maximum observed concentration was Cmax (concentration). The points included in determination of the terminal half-life were selected manually and given in units of time. Since the dosages and fluorine content for each positive control and the test material varied, all doses were normalized to 0.1 mmole/kg for comparative purposes. The accumulation index (AI, l^l-e^1)) and bioaccumulation index (BI, Cmax x AI) were calculated and reported but not further used. H-24019 and H-24020 were used as positive controls. Dosing diluents were used as negative controls, but because of variability and limited sensitivity of the analytical method, the background was set at 0.2 ppm fluorine. Since 0.2 ppm was the fluoride concentration limit of detection, any values listed as less than 0.2 ppm were excluded from further treatment. The percent of fluorine and molecular weight of the test substance and positive controls were used as provided by the sponsor. The measurements resulting from analysis of total blood fluorine were used as received (ppm F) from Jackson Laboratory. Fluoride ion was converted to 13Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 micromolar (^iM) equivalents of active component for further comparisons. The data and descriptions of data manipulation and presentation are provided in Appendix C. RESULTS AND DISCUSSION A. In-Life Toxicology (Tables 1-2, Figure 1, Appendices A-B) No deaths occurred during the study. One rat dosed with H-23925 exhibited diarrhea on test day 4. This clinical sign is considered not to be due to administration of the test substance because it was only observed on one day in a single rat. Rats dosed with H-24019 exhibited diarrhea, salivation, alopecia, black ocular discharge, and staining of various parts of the body during the dosing period. Rats dosed with H-24020 exhibited wet perineum and diarrhea during the dosing period. Alopecia was observed during the recovery period in rats dosed with H-24019, H-24020, and deionized water. The corn oil and corn oil/acetone negative control rats exhibited no clinical signs during the study. Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive or negative controls. This difference resulted in differences in mean body weights on test day 1. The rats dosed with the test substance, H-23925, were older and heavier in weight than the deionized water and corn oil negative control and H-24020 positive control rats, but younger and lighter in weight than the corn oil/acetone negative control and H-24019 positive control rats. Accounting for the age difference at study start and the expected rate of body weight gain, it appears that the mean body weights and mean body weight gains of the rats dosed with H-23925 are comparable to the negative controls and equal to or greater than the positive controls. B. Fluorine Data (Tables 3-4, Figures 2-3, Appendix C) 1. Factors Influencing Interpretation of Analysis The data used in the kinetic analysis were derived from a limited screen, and therefore several caveats and considerations are important. A couple of considerations of particular importance are (1) a single dose was used and kinetics may or may not be linear, (2) the kinetics apply only to blood, (3) steady-state may not have been achieved, and (4) the sample size is low and may impact calculation of the terminal half-life. A more complete list of considerations is shown in Appendix C. 2. Positive Controls The positive controls were H-24019 and H-24020. The H-24019 normalized fJM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state -14- Company Sanitized. Does not eorrfawi TSCA CBt H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 (Figure 2A). The Cmax for H-24019 was 989.85 116.90 ppm (Mean SD) with a terminal half-life of 40.5 days. The H-24019 AI was 59.0 and the BI was 58382.2. The H-24020 normalized {JIMequivalents in rat blood peaked after 5 days of dosing and then decreased throughout the dosing period (Figure 2B). The Cmax for H-24020 was 518.12 44.89 ppm (Mean SD) with a terminal half-life of 8.3 days. The H-24020 AI was 12.5 and the BI was 6497.5. For each of the positive controls, blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between positive controls and the test material. The AUCINF/D for the fluorine component was 566,479.1 for H-24019 and 70,789.6 for H-24020. 3. Test Substance The H-23925 normalized ^iM equivalents in rat blood continued to rise throughout the dosing period and did not reach steady-state (Figure 2C). The Cmax for H-23925 was 49.278 4.098 ppm (Mean SD) with a terminal half-life of 38 days. Blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The H-23925 AI was 55.4 and the BI was 2732.3. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between H-23925 and positive controls. The AUCENF/D for the fluorine component of H-23925 was 14,886.1 as compared to AUCINF/D values of 566,479.1 and 70,789.6 for H-24019 and H-24020, respectively. The data, calculations and equations are shown in Appendix C. CONCLUSIONS Rats dosed for 10 consecutive days with 200 mg/kg H-23925 exhibited no mortality or test substance-related clinical signs of toxicity and had mean body weights and mean body weight gains that were comparable to the negative control rats. A steady-state for fluorine levels in whole blood was not achieved during the 10-day dosing period. Under the conditions of this study, administration of H-23925 to male rats for 10 consecutive days resulted in low absorption and retention of fluorine in the blood. Dose adjusted areas under the curve (AUCINF/D) for positive controls, H-24019 and H-24020, were approximately 38x and 5x the AUCINF/D for the test substance. -15- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 RECORDS AND SAMPLE STORAGE Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. -16- Company Sanitized. Does not confaFn TSCA CW H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLES -17- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLE 1 MEAN BODY WEIGHTS (g) Test Days 1 2 3 4 5 6 7 8 9 10 13 18 20 24 25 27 32 34 39 41 46 47 48 52 53 55 60 61 62 66 67 68 74 76 80 81 Neigative Contnols Deionized Water Corn Oil Corn Oil: Acetone 226.8 229.8 297.6 232.8 234.1 299.6 240.3 241.4 309.4 249.0 250.4 314.3 259.1 259.3 321.5 268.3 266.8 326.0 277.1 271.2 334.9 283.6 281.9 339.2 290.2 285.2 345.2 298.3 295.9 351.7 313.6 321.1 350.7 352.6 - - - 350.5 369.8 419.4 378.3 401.0 378.7 - - - - - 400.7 - 449.5 - 463.5 - - 407.8 - 434.5 - 454.9 - 411.4 - - - - 469.7 511.0 - 528.0 464.7 - 487.0 - 505.1 527.6 - - - - - 547.7 - 558.0 - - 573.0 - 515.4 - - 539.0 - Positive Controls H-24019 291.5 291.5 298.3 304.1 308.6 312.7 317.5 317.3 318.4 318.9 329.1 - 359.7 387.9 - 393.9 - 416.9 - ' - - 438.1 - - - - 523.4 - - - - 563.4 570.7 H-24020 227.5 232.4 242.0 248.9 252.4 259.8 263.3 269.8 278.5 283.9 289.1 - 311.3 340.9 - - - 402.9 - 425.0 - 438.2 - 465.6 - 452.3 - 465.8 - - - 470.9 - - - 499.2 Test Substance H-23925 278.1 279.7 , 285.9 291.6 296.8 300.9 308.2 312.6 316.7 320.6 340.1 - 373.0 408.0 - - - - - - - - 457.2 480.3 - - - - 496.7 508.5 - - - 518.8 - -18- Company SanFHzed. Does not contain TSCA CBI TABLE 1 (Continued) MEAN BODY WEIGHTS (g) Negative Controls Positive Controls Test Deionized Corn Oil: Days Water Corn Oil Acetone H-24019 H-24020 83 88 89 90 94 In d i c a t e s t 58 58 he --9.5 - 4.6 animal was n --- 556.2 566.9 ot weighed. --562.2 567.8 --585. 597. 3 9 --52 53 - 7. 8. 8 7 Test Substance H-23925 530.1 536.5 538.9 -19- Company Sanitized. Does not contain TSCA CBI TABLE 2 MEAN BODY WEIGHT GAINS (g) Test Days 1-5 1-10 10-13 10-24 10-52 10-94 Negative Controls Deionized Water Corn Oil Corn OihAcetone 32.3 29.5 23.9 71.5 66.1 54.1 15.3 121.1 25.2 82.4 -1.0 49.3 171.4 168.8 175.9 286.3 271.0 216.1 Positive Controls H-24019 17.1 27.4 10.2 69.0 119.2 279.0 H-24020 24.9 56.4 5.2 57.0 181.7 254.8 Test Substance H-23925 18.7 42.5 19.5 87.4 159.7 218.3 -20- pompanySanitized. Does not contain TSCA CBt H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLES MEAN BLOOD FLUORINE LEVELS Test Days 1 5 10 13 24 52 94 Nesgative Contirols Deionized Corn Oil: Water Corn Oil Acetone (ppm) (ppm) (ppm) <0.2 <0.2 <0.2 <0.2 0.2' <0.2 0.90 <0.2 <0.2 <0.2 0.2^0.1) <0.2 OJ^O.l) OJ3 (0,1) <0.2 <0.2 0.38 (O.lf O^ (0.6) 0.2'' <0.2 <0.2 Positive C ontrols Test Substance H-24019 (ppm) 2.1 (0.9) 48.8 (17.6) 61.7 (2.8) 64.5 (7.6) 42.2 (2.8) 26.9 (2.6) 13.2 (2.1) H-24020 (ppm) 62.6 (3.2) 71.7 (6.2) 54.2 (7.8) 26.6 (11.4) 10.5 (3.0) 0.9 (0.2) 0.2 (0.1) H-23925 (ppm) 9.2 (3.4) 13,4 (2.2) 15.9 (1.3) 6.4 (1.1) 5.0 (0.8) 2.6 (0.4) 1.6 (0.5) a Mean of 4 of the 5 values. One of the values was below the level of detection (LOD). b Standard deviation is in parentheses. c Mean of 3 of the 5 values. Two of the values were below the LOD. d One value. Four of the values were below the LOD. e Mean of 2 of the 5 values. Three of the values were below the LOD. -21- Company Sanitized. Does not contain TSCA CBI TABLE 4 MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE Test Days 1 5 10 13 24 52 94 Positive Controls Test Substance H-24019 H-24020 H-23925 fJiM F Equivalents 28.92 (13.3)' f^M F Equivalents___pM F Equivalents 89.28 (23.4) 28.21 (10.6) 747.69 (271.5) 518.12 (44.9) 41.44 (6.9) 945.85 (43.5) 391.01 (56.8) 49.28 (4.1) 989.85 (116.9) 191.45 (82.7) 19.56 (3.6) 645.54 (42.9) 74.35 (22.1) 15.17 (2.6) 411.38 (40.6) 4.64 (1.1) 7.40 (1.2) 195.38 (32.5)_____0.24 (0.4) 4.52 (1.4) a Standard deviation is in parentheses. -22- ICompany Sanitized. Doea not contain TSCA CB8 FIGURES -23- Company Sanitized. Does not contain TSCA CBt H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY PERSONNEL Study Director: Management: Primary Technician: Carol Finlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. Richard P. Mathena Blood Fluorine Data Analysis: Gary W. Jepson, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T. Toxicology Report Preparation: Wanda F. Dinbokowitz Laboratory Veterinarian: Wanda L. West, D.V.M., A.C.L.A.M. DuPont-2921 -7- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 SUMMARY The objective of this study was to evaluate the potential for H-23925, when administered by gavage, to be absorbed and to accumulate in a mammalian system. Six groups of 5 male Crl:CD(SD)IGS BR rats each were given 200 mg/kg/day of H-23925. The test substance was administered to one group of 5 rats for 5 consecutive days and to 5 groups for 10 days. Approximately 2 hours after the first dose, blood was collected from the orbital sinus of each rat from the 5-dose group. On selected days (5, 10,13, 24, 52, 94) 5 rats per group were euthanized and the blood and livers were collected. Body weights and clinical signs were recorded on each day of dosing and then weekly, every other week, or every 3 weeks during the recovery period. Additionally, 3 negative controls, deionized water, corn oil, and corn oiVacetone (80:20), and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-23925. No deaths occurred during the study. One rat dosed with the test substance, H-23925, exhibited diarrhea on test day 4. This clinical sign is considered not to be due to administration of the test substance because it was only observed on one day in a single rat. Rats dosed with H-24019 exhibited diarrhea, salivation, alopecia, black ocular discharge, and staining of various parts of the body during the dosing period. Rats dosed with H-24020 exhibited wet perineum and diarrhea during the dosing period. Alopecia was observed during the recovery period in rats dosed with H-24019, H-24020, and deionized water. The corn oil and corn oiVacetone negative control rats exhibited no clinical signs during the study. Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive or negative controls. Accounting for the age difference at study start and the expected rate of body weight gain, it appears that the mean body weights and mean body weight gains of the rats dosed with the test substance, H-23925, are comparable to the negative controls and equal to or greater than the positive controls. A steady-state for fluorine levels in whole blood was not achieved during 10 consecutive days of dosing with 200 mg/kg H-23925. An area under the curve (estimated to infinity) was calculated and normalized for fluorine content for the test substance and each positive control. The AUCINF/D for the fluorine component of the test substance, H-23925, was 14,886.1, compared to AUCINF/D values of 566,479.1 and 70,789.6 for the positive controls, H-24019 and H-24020, respectively. Under the conditions of this study, administration of H-23925 to male rats for 10 consecutive days resulted in low absorption and retention of fluorine in the blood. Dose adjusted areas under the curve (AUCINF/D) for positive controls, H-24019 and H-24020, were approximately 38x and 5x the AUCINF/D for the test substance. 8RompanySanitized. Does not contain TSCA CB H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 INTRODUCTION The objective of this study was to define the potential of H-23925 to be absorbed and to bioaccumulate in a mammalian system, as indicated by analytical determination of total fluorine in blood and liver. The test substance was compared to 2 positive controls that were materials previously shown to bioaccumulate in mammals. The daily dosage for the test substance, 200 mg/kg, was selected based on available toxicity data and the results of rangefinding studies. In the initial rangefinding study, a group of 5 male rats was dosed by oral gavage with H-23925 at a dosage of 2000 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water and served as controls. The rats dosed with H-23925 experienced an overall mean body weight loss of 29 grams. In the next rangefinding study, 2 groups of 5 male rats were dosed by oral gavage with H-23925 at a dosage of 500 or 1000 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water and served as controls. Overall mean body weight losses of 7 and 25 grams occurred in the rats dosed with the test substance. A group of 5 male rats was then dosed by oral gavage at a dosage of 125 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water and served as controls. The rats dosed at 125 mg/kg experienced an overall mean body weight gain of 13 grams. The control group had an overall mean body weight gain of 14 grams. The dosage of 200 mg/kg was chosen for the main study and was expected to produce less than a 10% difference in mean body weight when compared to controls. All blood samples were analyzed for total fluorine content. This report contains the results of those analyses. Results from analyses of selected liver samples will be presented in a supplemental report. MATERIALS AND METHODS A. Test Substance and Positive Control The test substance, H-23925, was supplied by the sponsor as an amber liquid with some flock sediment. The positive controls, H-24019 and H-24020, were supplied by the sponsor as white solids. The test substance and positive controls appeared to be stable under the conditions of the study. No evidence of instability, such as a change in color or physical state, was observed. B. Test Species Male Crl:CD (SD)IGS BR rats were received from Charles River Laboratories, Inc., Raleigh, North Carolina. The Crl:CD(SD)IGS BR rat was selected on the basis of extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, and low incidence of spontaneous diseases. -9Company Sanitized. Does nol contain TSCA CB H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 C. Animal Husbandry 1. Housing Environment Rats were housed singly in stainless steel, wire-mesh cages suspended above cage boards. Animal rooms were maintained on an approximate 12-hour light/dark cycle (fluorescent light) and at a temperature of 23 1C and a relative humidity of 50 10%. Occasional excursions outside the accepted ranges were minor and did not affect the study. 2. Feed and Water Tap water was provided ad libitum. All rats were fed PMI Nutrition International, Inc. Certified Rodent LabDiet 5002 chow. The feed is guaranteed by the manufacturer to meet specified nutritional requirements and to be free of specified contaminants. 3. Identification Prior to assignment to groups, each rat was temporarily identified by cage identification. After assignment to groups, an individual identification number was marked or tattooed on the tail of each rat. The information on the cage labels included the unique 6-digit Haskell animal number assigned to each rat. 4. Animal Health Monitoring Program As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to assure that contaminant levels are below those that would be expected to impact the scientific integrity of the study: Water samples are analyzed for total bacterial counts, and the presence of coliforms, lead, and other contaminants. Feed samples are analyzed for total bacterial, spore and fungal counts. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations of key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study. The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. -10- Company Sanltteed. Does not contain TSCA CB? 6 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 D. Quarantine and Pretest Upon arrival at Haskell Laboratory, the rats were removed from shipping cartons and quarantined for 6 days. The rats were weighed 3 times during the pretest period and examined daily for any clinically apparent signs of disease or injury. The rats were observed daily for mortality and signs of illness, injury, or abnormal behavior. On the bases of acceptable body weight gains and freedom from clinically apparent signs of disease or injury, the rats were released from quarantine by the laboratory animal veterinarian designee. E. Study Design Substance Negative Controls Deionized water Corn oil Corn oil: acetone Positive Controls H-24019 H-24020 Test Substance H-23925 Vehicle Not applicable Dosage (mg/kg) Number of Animals 0 30 0 30 0 30 Corn oil: acetone 10 30 Corn oil 20 30 Deionized water 200 30 F. Assignment to Groups and Study Start After the quarantine period, the rats were selected on the bases of adequate body weight gain, freedom from any clinical signs of disease or injury, and a body weight within 20% of the mean. The selected rats were divided by computerized, stratified randomization into 6 groups of 5 rats, so that there were no statistically significant differences among group body weight means. After assignment to groups, each rat was housed individually. The last 3 digits of the animal number was marked or tattooed on the tail of each rat. The rats were between 7 and 9 weeks of age at the time of dosing. Dosing began on test day 1. Rats that were not assigned to the study were released for other laboratory purposes, or were sacrificed by carbon dioxide asphyxiation and discarded without pathological evaluation. G. Dosing Material Preparation and Administration 1. Test Substance H-23925 was mixed with deionized water so an accurate volume could be delivered. The amount of test substance each rat received was based on the body weight collected on each day of dosing and the dosing mixture concentration. The rats were dosed at a volume of 10 mL/kg of -11- Company Sanitized. Does not contain TSCA CBf H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 body weight. The test substance was stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. 2. Positive Controls The solid positive control compounds were suspended as emulsions in their respective vehicles. Corn oil was used as the vehicle for H-24019. It was necessary to dissolve H-24020 in acetone before suspending it in corn oil. The ratio of acetone to corn oil was 20:80. The amount each rat received was based on the body weight collected on each day of dosing and the suspension concentration. The dose volumes did not exceed 1 mL/100 g of body weight. The dosing suspensions were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. 3. Negative Controls Deionized water was chosen as a negative control because it was the vehicle for the test substance. Corn oil and corn oiLacetone (80:20) were chosen as negative controls because they were the vehicles for the positive controls. Each negative control rat received 1 mL of corn oil or corn oil:acetone. These rats were dosed in a separate room from the rats dosed with the test substance or positive controls. H. Body Weights All rats were weighed on each day of dosing. The positive and negative control rats were weighed weekly or every other week during the recovery period. The rats dosed with H-23925 were weighed weekly, every other week, or every 3 weeks during the recovery period. I. Mortality and Clinical Observations Cage-site examinations to detect moribund or dead rats and abnormal behavior and appearance among rats were conducted at least once daily throughout the study. At every weighing, each rat was individually handled and examined for abnormal behavior and appearance. J. Collection and Analysis of Blood and Livers Approximately 2 hours after the first dose, 1-2 mL of blood was collected into EDTA tubes from the orbital sinus of each rat from Group I. At all other selected time points, 5 rats/group were euthanized by carbon dioxide anesthesia and exsanguination and blood and livers were collected according to the following schedules: -12- Company Sanitized. Does not con*s;n j^f- \ H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Group I I n m IV v VI Dosing Days 1-5 1-5 1-10 1-10 1-10 1-10 1-10 Tissue Collected Blood Blood and Liver Blood and Liver Blood and Liver Blood and Liver Blood and Liver Blood and Liver Sampling Time Test day 1 (2 hours post dosing) Test day 5 at sacrifice (2 hours post dosing) Test day 10 (2 hours post dosing) Test day 13 Test day 24 Test day 52 Test day 94 Five to 10 mL of blood was collected into EDTA tubes at sacrifice. The livers were weighed. The blood from all rats was refrigerated and the livers were frozen. The blood was appropriately packaged and shipped refrigerated to Jackson Laboratory, Deepwater, New Jersey where it was analyzed for total fluorine. The total fluorine content of the blood samples was determined using a Wickbold torch combustion method, followed by analysis with a fluoride ion selective electrode. The liquid blood was decomposed or volatilized in the presence of wet oxygen and swept through an oxyhydrogen flame in a closed quartz apparatus. The combustion products were collected in an aqueous absorbing solution and analyzed using a fluoride ion selective electrode. The analytical data (ppm F in each sample) was supplied to Haskell Laboratory personnel for evaluation of fluorine biopersistence. K. Treatment of Fluorine Data Noncompartmental analysis was conducted on blood fluorine data derived from rats dosed with H-23925 by using WinNonlin Version 3.0 software (Pharsight Corp, Mountain View, CA). WinNonlin software provided a means of computing derived pharmacokinetic parameters from data files including area under the curve (AUCINF), Cmax and terminal half-life (Ti/z). The AUCINF (concentration x time) represents the area under the blood concentration curve from the time of dosing extrapolated to infinity. The maximum observed concentration was Cmax (concentration). The points included in determination of the terminal half-life were selected manually and given in units of time. Since the dosages and fluorine content for each positive control and the test material varied, all doses were normalized to 0.1 mmole/kg for comparative purposes. The accumulation index (AI, l^l-e^')) and bioaccumulation index (BI, Cmax x AI) were calculated and reported but not further used. H-24019 and H-24020 were used as positive controls. Dosing diluents were used as negative controls, but because of variability and limited sensitivity of the analytical method, the background was set at 0.2 ppm fluorine. Since 0.2 ppm was the fluoride concentration limit of detection, any values listed as less than 0.2 ppm were excluded from further treatment. The percent of fluorine and molecular weight of the test substance and positive controls were used as provided by the sponsor. The measurements resulting from analysis of total blood fluorine were used as received (ppm F) from Jackson Laboratory. Fluoride ion was converted to -13- Company Sanitized. Does not contain TSCA CB H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 micromolar (/-iM) equivalents of active component for further comparisons. The data and descriptions of data manipulation and presentation are provided in Appendix C. ^H|) ^y RESULTS AND DISCUSSION A. In-Life Toxicology (Tables 1-2, Figure 1, Appendices A-B) No deaths occurred during the study. One rat dosed with H-23925 exhibited diarrhea on test day 4. This clinical sign is considered not to be due to administration of the test substance because it was only observed on one day in a single rat. Rats dosed with H-24019 exhibited diarrhea, salivation, alopecia, black ocular discharge, and staining of various parts of the body during the dosing period. Rats dosed with H-24020 exhibited wet perineum and diarrhea during the dosing period. Alopecia was observed during the recovery period in rats dosed with H-24019, H-24020, and deionized water. The corn oil and corn oiVacetone negative control rats exhibited no clinical signs during the study. Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive or negative controls. This difference resulted in differences in mean body weights on test day 1. The rats dosed with the test substance, H-23925, were older and heavier in weight than the deionized water and corn oil negative control and H-24020 positive control rats, but younger and lighter in weight than the corn oil/acetone negative control and H-24019 positive control rats. Accounting for the age difference at study start and the expected rate of body weight gain, it appears that the mean body weights and mean body weight gains of the rats dosed with H-23925 are comparable to the negative controls and equal to or greater than the positive controls. B. Fluorine Data (Tables 3-4, Figures 2-3, Appendix C) 1. Factors Influencing Interpretation of Analysis The data used in the kinetic analysis were derived from a limited screen, and therefore several caveats and considerations are important. A couple of considerations of particular importance are (1) a single dose was used and kinetics may or may not be linear, (2) the kinetics apply only to blood, (3) steady-state may not have been achieved, and (4) the sample size is low and may impact calculation of the terminal half-life. A more complete list of considerations is shown in Appendix C. 2. Positive Controls The positive controls were H-24019 and H-24020. The H-24019 normalized pM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state 14Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 (Figure 2A). The Cmax for H-24019 was 989.85 116.90 ppm (Mean SD) with a terminal half-life of 40.5 days. The H-24019 AI was 59.0 and the BI was 58382.2. The H-24020 normalized ^M equivalents in rat blood peaked after 5 days of dosing and then decreased throughout the dosing period (Figure 2B). The Cmax for H-24020 was 518.12 44.89 ppm (Mean SD) with a terminal half-life of 8.3 days. The H-24020 AI was 12.5 and the BI was 6497.5. For each of the positive controls, blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between positive controls and the test material. The AUCINF/D for the fluorine component was 566,479.1 for H-24019 and 70,789.6 for H-24020. 3. Test Substance The H-23925 normalized /xM equivalents in rat blood continued to rise throughout the dosing period and did not reach steady-state (Figure 2C). The Cmax for H-23925 was 49.278 4.098 ppm (Mean SD) with a terminal half-life of 38 days. Blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The H-23925 AI was 55.4 and the BI was 2732.3. The total internal exposure resulting from a normalized dose was described by AUCENF/D and was the basis for comparison between H-23925 and positive controls. The AUCINF/D for the fluorine component of H-23925 was 14,886.1 as compared to AUCDMF/D values of 566,479.1 and 70,789.6 for H-24019 and H-24020, respectively. The data, calculations and equations are shown in Appendix C. CONCLUSIONS Rats dosed for 10 consecutive days with 200 mg/kg H-23925 exhibited no mortality or test substance-related clinical signs of toxicity and had mean body weights and mean body weight gains that were comparable to the negative control rats. A steady-state for fluorine levels in whole blood was not achieved during the 10-day dosing period. Under the conditions of this study, administration of H-23925 to male rats for 10 consecutive days resulted in low absorption and retention of fluorine in the blood. Dose adjusted areas under the curve (AUCINF/D) for positive controls, H-24019 and H-24020, were approximately 38x and 5x the AUCINF/D for the test substance. -15- iCompanySanitized. Does not contain TSCA CBF H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats__________________________________DuPont-2921 RECORDS AND SAMPLE STORAGE Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. -16- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study inRats DuPont-2921 TABLES -17- Company Sanitized. Does not contain TSCA CB1 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLE 1 MEAN BODY WEIGHTS (g) Test Days 1 2 3 4 5 6 7 8 9 10 13 18 20 24 25 27 32 34 39 41 46 47 48 52 53 55 60 61 62 66 67 68 74 76 80 81 Neigative Contnols Deionized Corn Oil: Water Corn Oil Acetone 226.8 229.8 297.6 232.8 234.1 299.6 240.3 241.4 309.4 249.0 250.4 314.3 259.1 259.3 321.5 268.3 266.8 326.0 277.1 271.2 334.9 283.6 281.9 339.2 290.2 285.2 345.2 298.3 295.9 351.7 313.6 321.1 350.7 352.6 - - - 350.5 369.8 419.4 378.3 401.0 378.7 - - - - - 400.7 - 449.5 - 463.5 - - 407.8 - 434.5 - 454.9 - 411.4 - - - - 469.7 511.0 - 528.0 ' 464.7 - 487.0 - 505.1 527.6 - - - - - 547.7 - 558.0 - - 573.0 - 515.4 - - 539.0 - Positive <Controls Fest Substance H-24019 291.5 291.5 298.3 304.1 308.6 312.7 317.5 317.3 318.4 318.9 329.1 - 359.7 387.9 - 393.9 - 416.9 - - - - - 438.1 - - - 523.4 - - - - 563.4 570.7 H-24020 227.5 232.4 242.0 248.9 252.4 259.8 263.3 269.8 278.5 283.9 289.1 - 311.3 340.9 - - - 402.9 - 425.0 - 438.2 - 465.6 - 452.3 - 465.8 - - - 470.9 - - - 499.2 H-23925 278.1 279.7 285.9 291.6 296.8 300.9 308.2 312.6 316.7 320.6 340.1 - 373.0 408.0 - - - - - - - - 457.2 480.3 - - - - 496.7 508.5 - - - 518.8 - - 18- Company Sanitized. Does not contain TSCA CBf TABLE 1 (Continued) MEAN BODY WEIGHTS (g) Negative Controls Positive Controls Test Deionized Corn Oil: Days Water Corn Oil Acetone H-24019 H-2402Q 83 88 89 90 94 Indicates t --589.5 584.6 he animal was n --- 556.2 566.9 ot weighed. --562.2 567.8 --585. 597. 3 9 --52 53 7 8 . . 8 7 Test Substance H-23925 530.1 536.5 538.9 -19- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLE 2 MEAN BODY WEIGHT GAINS (g) Test Days 1-5 1-10 10-13 10-24 10-52 10-94 Negative Controls Deionized Water Corn Oil Corn OiLAcetone 32.3 29.5 23.9 71.5 66.1 54.1 15.3 121.1 25.2 82.4 -1.0 49.3 171.4 168.8 175.9 286.3 271.0 216.1 Positive Controls H-24019 17.1 27.4 10.2 69.0 119.2 279.0 H-24020 24.9 56.4 5.2 57.0 181.7 254.8 Test Substance H-23925 18.7 42.5 19.5 87.4 159.7 218.3 -20- Company Sanitized. Does nol contain TSCA CBl H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLES MEAN BLOOD FLUORINE LEVELS Test Days 1 5 10 13 24 52 94 Ne;eative Contirols Deionized Corn Oil: Water Corn Oil Acetone (ppm) (ppm) (ppm) <0.2 <0.2 <0.2 <0.2 0.2s <0.2 0.9" <0.2 <0.2 <0.2 0.2" (0.1) <0.2 0.3' (0.1) 0.3'(0.1) <0.2 <0.2 O^O.l)" 0.6C (0.6) 0.2'' <0.2 <0.2 Positive 0 ontrols Test Substance H-24019 (ppm) 2.1 (0.9) 48.8 (17.6) 61.7 (2.8) 64.5 (7.6) 42.2 (2.8) 26.9 (2.6) 13.2 (2.1) H-24020 (ppm) 62.6 (3.2) 71.7 (6.2) 54.2 (7.8) 26.6 (11.4) 10.5 (3.0) 0.9 (0.2) 0.2 (0.1) H-23925 (ppm) 9.2 (3.4) 13.4 (2.2) 15.9 (1.3) 6.4 (1.1) 5.0 (0.8) 2.6 (0.4) 1.6 (0.5) a Mean of 4 of the 5 values. One of the values was below the level of detection (LOD). b Standard deviation is in parentheses. c Mean of 3 of the 5 values. Two of the values were below the LOD. d One value. Four of the values were below the LOD. e Mean of 2 of the 5 values. Three of the values were below the LOD. -21- Company Sanitized. Does not contain TSCA CW H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TABLE 4 MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE Test Days 1 5 10 13 24 52 94 Positive Controls Test Substance H-24019 H-24020 H-23925 /xM F Equivalents 28.92 (13.3)3 {fM F Equivalents____ptM F Equivalents 89.28 (23.4) 28.21 (10.6) 747.69 (271.5) 518.12 (44.9) 41.44 (6.9) 945.85 (43.5) 391.01 (56.8) 49.28 (4.1) 989.85 (116.9) 191.45 (82.7) 19.56 (3.6) 645.54 (42.9) 74.35 (22.1) 15.17 (2.6) 411.38 (40.6) 4.64 (1.1) 7.40 (1.2) 195.38 (32.5)_____0.24 (0.4) 4.52 (1.4) a Standard deviation is in parentheses. -22- Company Sanitized. Does not contain TSCA CBi H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 FIGURES -23- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats 400 -i 350 } 300 -\ " 250 H a Ia >> 200 -\ a o ca g 150 -| 100 -\ FIGURE 1 MEAN BODY WEIGHTS (g) 0 Negative Control (De D Negative Control (Co A Negative Control (Co --<--Positive Control H-2 -9K-Positive Control H-2 -e-Test Substance H-23 Test Days 10 12 -24- FIGURE 2 MICROMOLAR EQUIVALENTS IN RAT BLOOD A. Normalized Rat Blood H-24019 (iM Equivalents Resulting from a 10-Day 0 ralG avage Test Days Micromolar (fiM) equivalents ofH-24019 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. B. Normalized Rat Blood H-24020 fi M Equivalents Resulting from a 10-Day 0 ralG avage Micromolar (fiM) equivalents of H-24020 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. -25- Company Santtfeed. Does not confafn TSCA CW H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 C. Normalized Rat Blood H-23925 fi M Equivalents Resulting from a 10-Day 0 ralG avage S 30 } Micromolar (/xM) equivalents of H-23925 (test substance) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. -26- Company SanRfzed. Does nol conlafn TSCA CBl H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURES MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS -27- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 APPENDICES -28- Company Sanitized. Does not contain TSCA CBi H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 APPENDIX A Individual Body Weights -29- Company Sanitized. Does not contain TSCA CBI INDIVIDUAL BODY WEIGHTS ABBREVIATIONS: SD - sacrificed by design EXPLANATORY NOTES -30- Company Sanitized. Does not contain TSCA CW H-23925; Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632649 632650 632651 632652 632653 DAY 1 208.8 223.3 238.6 243.4 210.5 DAY 2 208.2 231.1 243.1 240.0 210.5 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I TEST DAYS DAY 3 DAY 4 DAY 5 219.8 239.3 253.6 253.0 217.6 225.1 243.7 260.8 257.3 223.0 240.4 261.7 276.7 272.4 236.0 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -31 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632654 632655 632656 632657 632658 ANIMAL NUMBER 632654 632655 632656 632657 632658 DAY 1 210 .1 234 .4 236 .4 251 .5 220 .8 DAY 8 282 .8 295 .4 286 .5 317 .0 274 .3 DAY 2 217 .0 238 .1 243 .5 255 .0 227 .5 DAY 9 286 .6 307 .1 286 .1 324 .3 278 .2 DEIONI2ED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS TEST DAYS DAY 3 DAY 4 GROUP II DAY 5 DAY 6 DAY 7 227. 9 248. 1 250. 2 265. 0 235. 1 236. 0 258. 7 258. 0 277. 0 240. 8 248. 5 265. 7 261. 9 292. 0 249. 3 259. 9 278. 0 272. 2 297. 3 256. 7 270.6 289.3 280.3 308.0 264.1 TEST DAYS DAY 10 303. 1 315. 8 294. 5 329. 6 286. 0 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -32- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632659 632660 632661 632662 632663 Day 1 199. 9 232. 6 238. 1 248. 1 219. 4 Day 2 208. 0 238. 9 243. 1 255. 5 227. 9 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS TEST DAYS . Day 3 Day 4 GROUP III Day 5 Day 6 Day 7 211. 0 240. 7 247. 4 261. 5 229. 9 220.9 251.8 258.1 269.7 241.7 231.0 260.8 265.7 279.8 249.9 238.9 268.0 273.5 286.2 257.7 247. 8 275. 7 285. 0 292. 8 265. 2 ANIMAL NUMBER 632659 632660 632661 632662 632663 Day 8 254. 1 279. 4 291. 8 297. 0 272. 0 Day 9 261. 5 286. 1 302. 4 302. 4 276. 4 TEST DAYS Day 10 Day 13 270. 0 296. 3 304. 5 309. 2 285. 1 293.1 309.1 334.9 331.1 299.9 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 -33- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632664 632665 632666 632667 632668 Day 1 208. 2 228. 3 245. 8 254. 3 218. 7 Day 2 213. 9 236. 6 251. 0 262. 3 226. 8 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TEST DAYS Day 3 Day 4 Day 5 Day 6 Day 7 219 .9 244 .9 263 .4 272 .0 238 .5 226.1 252.4 273.8 285.6 249.3 236.6 264.0 282.8 293.6 260.3 242. 9 274. 3 297. 4 300. 6 270. 1 252.6 282.1 310.0 315.2 282.0 ANIMAL NUMBER 632664 632665 632666 632667 632668 Day 8 256. 0 288. 0 314. 6 319. 9 291. 6 Day 9 262. 8 297. 9 326. 9 330. 0 301: 7 TEST DAYS Day 10 Day 13 273 .2 a 306 .5 a 334 .0 a 342 .1 a 314 .7 a Day 18 318.2 367.1 398.1 414.4 385.7 Day 24 349. 0 408. 5 441. 8 460. 4 437. 5 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 i Rats not weighed. -34. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632669 632670 632671 632672 632673 Day 1 194 .4 224 .5 226 .8 256 .3 217 .3 Day 2 197. 8 228. 6 230. 9 267. 1 224. 8 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS Day 3 Day 4 Day 5 Day 6 Day 7 202 .5 237 .4 237 .2 272 .4 230 .8 209.5 246.7 243.5 281.3 241.3 215.1 251.9 249.0 295.1 248.2 221.4 259.9 256.9 304.6 258.6 228.1 268.8 262.0 315.4 265.7 ANIMAL NUMBER 632669 632670 632671 632672 632673 Day 8 234 .3 271 .0 268 .0 319 .4 268 .9 Day 9 232. 6 271. 0 273. 6 328. 4 276. 4 TEST DAYS Day 10 Day 13 239 .6 a 280 .9 a 276 .0 a 334 .8 a 285 .2 a Day 18 278.1 328.3 308.4 389.8 335.6 Day 24 a -a -a Day 25 301.2 361.5 346.2 441.1 370.8 ANIMAL NUMBER 632669 632670 632671 632672 632673 Day 32 322 .8 389 .9 378 .5 377 .5 400 .9 Day 39 355. 5 415. 7 421. 9 517. 0 433. 8 TEST DAYS Day 46 Day 52 366 .6 434 .9 434 .6 542 .7 457 .3 392.4 449.1 456.4 566.6 484.2 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rats not weighed. -35. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632674 632675 632676 632677 632678 Day 1 197. 0 228. 8 238. 3 238. 1 211. 3 Day 2 205 .1 239 .3 243 .2 240 .7 227 .6 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS Day 3 Day 4 Day 5 Day 6 Day 7 214 .1 243 .1 252 .0 248 .2 233 .9 221.7 255.3 259.8 255.9 244.4 232 .7 264 .9 270 .3 261 .7 253 .6 242.0 271.8 280.9 276.0 262.6 250.9 278.0 289.4 282.0 266.0 ANIMAL NUMBER 632674 632675 632676 632677 632678 Day 8 261. 7. 285. 7 294. 1 289. 0 278. 3 Day 9 266 .0 291 .6 300 .3 298 .0 286 .3 TEST DAYS Day 10 Day 13 276 .7 a 296 .7 a 309 .8 a 302 .7 a 291 .4 a Day 18 336. 2 352. 9 368. 3 356. 9 351. 4 Day 24 a a a a a Day 25 380.4 389.6 411.3 397.5 387.6 ANIMAL NUMBER 632674 632675 632676 632677 632678 Day 32 419. 5 433. 4 441. 0 423. 9 419. 3 Day 39 465 .7 482 .3 484 .3 458 .8 460 .1 TEST DAYS Day 46 Day 52 471 .8 485 .0 502 .4 472 .8 467 .2 508.2 536.0 530.2 492.8 487.8 Day 53 529 .6 540 .7 552 .8 514 .7 502 .0 Day 60 545.8 584.8 565.0 525.1 518.0 Day 67 566.2 592.2 571.8 536.4 523.5 a Rats not weighed. -36- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 632674 632675 632676 632677 632678 Day 74 581.9 612.8 587.0 544.2 539.3 Day 81 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS Day 88 Day 94 583.5 645.4 605.1 562.0 551.6 548.9 652.9 603.2 569.4 548.5 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 a Rats not weighed. -37. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627789 627801 627805 627810 627818 DAY 1 222.6 215.9 249.4 230.5 229.7 DAY 2 220.7 209.1 250.1 231.6 228.6 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I TEST DAYS DAY 3 DAY 4 DAY 5 223.5 223.5 263.3 241.9 239.1 236.9 233.5 273.6 254.4 252.4 247.7 240.1 283.7 265.0 260.2 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -38. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627793 627797 627799 627800 627808 ANIMAL NUMBER 627793 627797 627799 627800 627808 DAY 1 214. 8 239. 9 228. 9 246. 9 235. 6 DAY 8 270. 6 3.03. 9 280. 2 312. 0 303. 6 DAY 2 224 .4 245 .9 231 .4 252 .8 241 .6 DAY 9 274 .9 305 .1 284 .3 315 .4 305 .7 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TE ST I )AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 232 .1 257 .3 242 .3 265 .2 250 .1 243. 1 267. 5 246. 2 279. 3 262. 1 TEST DAYS DAY 10 249 .9 280 .7 258 .9 283 .6 270 .4 257. 0 288. 0 261. 5 295. 5 284. 1 262.0 290.9 266.4 300.1 287.4 283 .5 324 .2 294 .7 331 .3 321 .8 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -39 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627796 627806 627807 627809 627816 ANIMAL NUMBER 627796 627806 627807 627809 627816 DAY 1 239. 8 227. 3 221. 5 234. 2 235. 0 DAY 2 249. 7 234. 2 225. 9 240. 4 238. 6 DAY 8 285. 9 282. 8 270. 7 297. 9 291. 8 DAY 9 290. 1 285. 8 274. 7 300. 2 296. 4 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III TE ST I )AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 253 .6 239 .4 237 .1 252 .5 250 .0 252 .5 242 .9 244 .1 260 .3 258 .9 267. 7 258. 5 254. 9 272. 8 265. 1 277. 2 270. 4 257. 7 277. 9 274. 5 289 .8 275 .2 260 .3 282 .6 278 .6 TEST DAYS DAY 10 DAY 13 300 .0 302 .8 280 .7 311 .9 305 .7 322 .7 320 .3 305 .2 336 .2 330 .2 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 -40- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627791 627794 627804 627812 627817 ANIMAL NUMBER 627791 627794 627804 627812 627817 DAY 1 231 .1 224 .4 224 .7 221 .9 233 .5 DAY 8 274 .0 286 .1 267 .2 278 .2 277 .8 DAY 2 235 .4 233 .0 228 .8 231 .5 231 .3 DAY 9 279 .1 289 .5 270 .6 282 .7 280 .4 -^ CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TE ST I )AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 241 .2 238 .1 230 .6 236 .3 229 .0 246 .8 247 .8 243 .0 245 .7 244 .8 TEST DAYS DAY 10 DAY 13 255 .9 260 .0 251 .7 255 .5 255 .4 DAY 20 262 .6 267 .4 252 .9 264 .9 266 .4 DAY 24 268. 4 270. 1 259. 5 269. 5 270. 0 285 .0 a 334 .4 365 .6 SD test day 24 295 .4 a 377 .4 416 .7 SD test day 24 278 .1 a 345 .8 378 .1 SD test day 24 294 .3 a 356 .5 388 .4 SD test day 24 294 .6 a 368 .7 407 .3 SD test day 24 a Rats not weighed. -41 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627792 627798 627813 627814 627815 ANIMAL NUMBER 627792 627798 627813 627814 627815 ANIMAL NUMBER 627792 627798 627813 627814 627815 DAY 1 235 .8 241 .1 214 .8 217 .5 230 .1 DAY 2 240 .8 253 .4 219 .2 220 .5 242 .2 DAY 8 282 .4 294 .3 258 .7 248 .0 289 .2 DAY 9 286 .4 296 .3 259 .2 250 .7 293 .4 DAY 41 456 .5 470 .7 395 .0 358 .7 462 .6 DAY 47 482 .3 496 .9 407 .2 372 .7 487 .3 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TE ST I )AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 244 .3 257 .6 220 .5 223 .7 243 .8 247.9 266.1 233.2 225.2 255.7 TEST DAYS DAY 10 DAY 13 252 .5 272 .9 239 .8 233 .8 265 .0 DAY 20 264 .4 281 .9 243 .2 236 .7 270 .0 DAY 24 268 .1 286 .2 246 .8 240 .6 274 .1 DAY 34 295 .2 a 315 .7 a 267 .9 a 257 .3 a 303 .3 a TEST DAYS DAY 52 360 .0 380 .2 323 .6 299 .3 368 .3 389 .2 403 .5 342 .3 319 .4 396 .5 426 .1 443 .1 367 .9 343 .7 434 .9 496 .8 517 .3 416 .8 386 .9 505 .9 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rats not weighed. -42. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627795 627811 627819 627820 627821 ANIMAL NUMBER 627795 627811 627819 627820 627821 ANIMAL NUMBER 627795 627811 627819 627820 627821 DAY 1 218 .0 228 .6 248 .8 225 .5 227 .5 DAY 2 219 .5 226 .3 255 .3 228 .9 233 .2 DAY 8 261 .4 273 .0 304 .5 281 .3 272 .6 DAY 9 265 .0 276 .7 308 .0 282 .9 275 .3 DAY 41 384 .2 422 .0 505 .9 - 478 .0 411 .8 DAY 47 398 .0 439 .3 534 .9 504 .1 426 .1 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TE ST I )AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 225 .3 236 .7 264 .9 239 .2 240 .7 234 .3 246 .1 274 .4 247 .9 246 .1 TEST DAYS DAY 10 DAY 13 239 .4 251 .5 280 .4 255 .6 251 .2 DAY 20 246 .2 259 .6 288 .0 261 .3 259 .8 DAY 24 250 .1 263 .6 290 .3 265 .3 263 .9 DAY 34 270 .7 a 285 .6 a 324 .3 a 292 .4 a 281 .8 a TEST DAYS DAY 52 DAY 55' a 411 .1 a 455 .4 a 551 .7 a 529 .7 a 439 .1 308 .0 337 .8 399 .7 367 .2 330 .1 DAY 61 435 .7 463 .0 574 .4 547 .1 459 .0 329 .8 360 .6 431 .2 393 .0 352 .9 DAY 68 440 .7 473 .6 588 .6 558 .6 463 .3 362 .0 396 .4 472 .5 445 .5 385 .6 a Rats not weighed. -43 .^|j|_,..,._._^.---^^^-- m^- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 90 DAY 94 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS 627795 627811 627819 627820 627821 474.6 503.6 638.1 608.6 509.0 485.0 511.6 653.1 618.0 521.5 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 -44. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 DAY 3 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I TEST DAYS DAY 4 DAY 5 625361 625363 625364 625380 625383 322.6 273.7 291.1 289.4 304.7 325.0 271.4 290.9 286.2 299.7 337.6 278.4 299.9 291.5 302.7 340.3 281.7 306.0 300.1 310.7 349.7 286.7 312.0 305.5 313 .1 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -45 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625355 625362 625367 625368 625378 ANIMAL NUMBER 625355 625362 625367 625368 625378 DAY 1 280 .0 290 .3 301 .5 312 .3 309 .6 DAY 8 321 .7 323 .9 349 .1 364 .2 350 .1 DAY 2 282 .0 294 .1 307 .8 318 .0 306 .4 DAY 9 320 .9 332 .3 352 .4 380 .8 354 .2 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 293.0 303.5 315.6 326.0 327.3 297.9 308.9 320.1 336.4 328.4 TEST DAYS DAY 10 306. 1 313. 8 327. 7 343. 7 333. 7 311. 2 315. 8 333. 7 347. 5 336. 6 315 .7 323 .8 345 .6 360 .8 346 .9 333.6 332.9 363.8 386.7 365.4 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -46 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 CORN OIL:ACETONE (NEGATIVE Control) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS TEST DAY 3 DAYS DAY GROUP III 4 DAY 5 DAY 6 DAY 7 625357 625358 625370 625384 625385 ANIMAL NUMBER 311 .9 285 .7 307 .9 294 .2 297 .6 DAY 8 308. 8 286. 3 316. 7 300. 0 304. 6 DAY 9 320.3 295.5 329.5 312.0 313.0 328 .4 296 .4 337 .9 309 .3 320 .4 TEST DAYS DAY 10 DAY 13 340.5 306.0 346.9 316.3 331.9 343 .3 302 .7 349 .8 314 .1 334 .0 361 .1 309 .2 364 .1 321 .4 345 .4 625357 625358 625370 625384 625385 364 .2 319 .8 373 .4 325 .6 347 .0 370. 3 321. 6 374. 7 331. 0 349. 5 376.9 320.3 385.6 340.6 358.9 405 .6 340 .7 407 .0 357 .1 377 .6 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 -47- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625354 625365 625371 625376 625381 ANIMAL NUMBER 625354 625365 625371 625376 625381 DAY 1 303. 0 327. 7 294. 5 289. 7 297. 8 DAY 2 304. 1 328. 7 299. 0 289. 9 297. 5 DAY 8 336. 2 .370. 5 340. 3 331. 5 321. 6 DAY 9 337. 0 379. 3 345. 9 339. 0 329. 1 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 310 .5 337 .9 313 .4 302 .2 301 .7 312 .2 343 .6 318 .3 304 .9 305 .6 TEST DAYS DAY 10 DAY 13 319 .3 357 .7 321 .8 313 .2 314 .4 DAY 20 324 .7 357 .2 329 .6 317 .0 314 .4 DAY 24 331. 2 364. 3 338. 5 322. 8 321. 0 348 .8 381 .2 351 .2 345 .2 331 .6 355 .8 407 .1 372 .6 363 .2 341 .2 377 .7 438 .2 413 .0 399 .7 358 .2 396 .1 459 .6 447 .3 432 .9 381 .8 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 -48. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625356 625359 625360 625372 625374 ANIMAL NUMBER 291 .7 297 .9 296 .8 294 .8 295 .9 DAY 8 291 .2 303 .3 300 .7 298 .8 302 .2 DAY 9 304.5 314.0 308.5 310.6 312.1 307 .0 315 .0 312 .7 317 .4 321 .7 TEST DAYS DAY 10 DAY 13 316 .9 323 .5 319 .1 324 .0 323 .4 DAY 20 319 .8 323 .5 . 332 .2 329 .0 328 .3 323. 9 333. 9 339. 5 335. 6 337. 2 DAY 24 DAY 27 625356 625359 625360 625372 625374 ANIMAL NUMBER 328 .8 334 .7 341.3 361 .9 396 .7 a 339 .1 346 .2 353.6 372 .9 410 .0 a 347 .9 354 .3 363.4 381 .7 419 .8 a 344 .7 348 .8 355.1 373 .1 395 .1 a 338 .0 344 .6 350.8 371 .5 401 .3 a DAY 34 DAY 52 TEST DAYS 431 .3 440 .9 465 .9 432 .1 447 .2 625356 625359 625360 625372 625374 466 .2 467 .7 484 .1 - 454 .7 466 .8 533 .7 530 .3 555 .4 506 .2 512 .3 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rats not weighed. .49. H-23925: Biopersistence Screening 10-Dose Oral.Gavage Study in Rats ANIMAL NUMBER 625353 625369 625373 625375 625377 ANIMAL NUMBER 625353 625369 625373 625375 625377 ANIMAL NUMBER 625353 625369 625373 625375 625377 DAY 1 305.9 281.7 294.6 289.1 293.9 DAY 2 307.3 279.9 301.3 291.0 294.8 DAY 8 344.7 307.3 340.6 324.7 325.0 DAY 9 352.7 311.9 348.2 336.0 333.7 DAY 34 453.3 388.2 477.1 - 455.6 419.1 DAY 52 a a a a a CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 316.8 289.7 312.7 298.0 302.9 325.0 286.9 317.0 307.0 312.0 TEST DAYS DAY 10 DAY 13 331.7 292.8 325.4 309.6 318.0 DAY 20 358.1 317.2 352.6 336.9 340.7 371.7 399.1 331.1 347.6 371.5 402.3 350.1 . 387.9 353.5 376.3 TEST DAYS DAY 76 DAY 90 DAY 94 330.1 295.3 . 329.0 314.2 317.4 342.5 303.8 334.6 323.5 325.0 DAY 24 a a a a a DAY 27 427.7 374.9 444.6 432.3 405.2 544.9 483.8 628.8 567.3 470.3 562.8 505.3 661.3 589.8 492.0 565.3 514.8 673.7 588.8 496.6 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 a Rats not weighed. -50- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625322 625323 625336 625343 625349 DAY 1 290.3 285.0 316.5 288.2 303.6 DAY 2 283.7 285.5 313.5 281.9 295.9 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I TEST DAYS DAY 3 DAY 4 DAY 5 291.5 290.2 320.1 286.8 308.2 298.9 290.9 327.1 291.0 310.1 298.0 294.0 329.0 293.7 318.1 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -51 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625320 625330 625332 625341 625350 ANIMAL NUMBER 279 .3 286 .7 300 .0 304 .9 257 .1 DAY 8 283. 0 286. 1 295. 9 309. 0 268. 5 DAY 9 287 .7 291 .1 304 .1 317 .5 267 .3 288. 4 296. 5 308. 8 320. 0 272. 2 TEST DAYS DAY 10 295 .9 304 .0 315 .0 325 .1 272 .6 299 .4 302 .3 322 .4 327 .5 276 .3 300 .6 306 .7 320 .7 327 .4 283 .6 625320 625330 625332 625341 625350 308 .5 314 .0 324 .1 332 .4 279 .8 306. 3 312. 6 321. 8 320. 3 282. 5 312 .1 SD test day 10 , 310 .1 SD test day 10 318 .3 SD test day 10 325 .6 SD test day 10 284 .3 SD test day 10 -52- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625321 625327 625342 625345 625348 ANIMAL NUMBER 625321 625327 625342 625345 625348 DAY 1 307 .0 311 .4 273 .7 291 .2 265 .5 DAY 8 337 .8 3-34 .3 304 .3 317 .3 302 .7 DAY 2 307. 8 306. 6 275. 5 291. 7 258. 6 DAY 9 339. 6 337. 0 305. 9 312. 9 306. 5 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 311 .4 312 .5 282 .3 297 .3 268 .0 319 .6 317 .8 285 .8 303 .3 275 .2 TEST DAYS DAY 10 DAY 13 322. 3 318. 3 290. 8 306. 4 280. 1 328 .4 319 .3 294 .2 310 .4 285 .2 335 .2 327 .6 326 .6 316 .1 290 .6 346 .1 336 .3 307 .8 315 .0 309 .7 371 .2 355 .3 324 .4 336 .9 336 .4 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 -53. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625325 625328 625338 625340 625352 ANIMAL NUMBER 625325 625328 625338 625340 625352 DAY 1 282 .8 315 .6 264 .6 304 .0 285 .5 DAY 8 318 .6 352 .0 287 .4 347 .0 316 .9 DAY 2 285 .9 315 .2 261 .9 309 .2 283 .9 DAY 9 321 .9 351 .7 291 .3 352 .2 322 .3 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TEST DAYS . DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 291.7 323.9 268.5 316.9 289.1 295 .5 327 .7 272 .6 326 .0 299 .6 TEST DAYS DAY 10 DAY 13 303 .2 335 .0 275 .7 329 .8 307 .9 DAY 20 309 .2 340 .4 281 .9 334 .1 308 .2 DAY 24 308. 3 340. 1 284. 4 338. 4 312. 4 327.4 357.5 290.2 359.2 321.0 331 .6 367 .4 291 .1 368 .1 329 .6 355 .8 400 .2 320 .4 389 .3 355 .7 381 .3 423 .9 339 .9 419 .0 375 .3 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 -54. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625324 625329 625333 625334 625344 ANIMAL NUMBER 303 .0 318 .9 292 .4 274 .6 262 .9 DAY 8 298 .5 316 .3 291 .2 277 .0 262 .2 DAY 9 305.8 326.5 297.8 286.4 269.0 312 .1 335 .4 304 .1 290 .6 273 .5 TEST DAYS DAY 10 DAY 13 320 .1 338 .3 307 .7 292 .0 274 .5 DAY 20 322.8 346.7 312.0 300.1 284.1 326 .8 351 .6 313 .5 304 .2 - DAY 24 DAY 27 625324 625329 625333 625334 625344 ANIMAL NUMBER 332 .5 337 .9 336.1 299 .6 336 .0 a 380 .3 352 .5 360 .1 357.0 364 .3 396 .4 a 423 .3 315 .8 316 .7 319.3 325 .8 349 .2 a 375 .2 292 .6 302 .2 305.8 312 .0 338 .6 a 368 .8 280 .7 285 .2 289.3 295 .8 309 .7 a 330 .4 DAY 34 DAY 52 TEST DAYS 625324 625329 625333 625334 625344 397 .3 442 .7 384 .6 384 .2 348 .3 447 .4 498 .1 427 .4 426 .5 391 .0 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rats not weighed. -55. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625326 625335 625346 625347 625351 ANIMAL NUMBER 625326 625335 625346 625347 625351 ANIMAL NUMBER 625326 625335 625346 625347 625351 DAY 1 313 .2 301 .0 290 .2 266 .0 310 .6 DAY 2 317 .1 303 .6 293 .4 275 .6 310 .9 DAY 8 311 .5 294 .2 292 .9 . 330 .5 351 .1 DAY 9 308 .9 285 .3 290 .8 330 .1 357 .8 DAY 34 427 .5 427 .0 407 .4 466 .4 483 .3 DAY 52 a a a a a H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 325 .5 302 .3 300 .2 287 .7 321 .1 330 .9 310 .4 309 .5 300 .0 329 .6 TEST DAYS DAY 10 DAY 13 336 .2 315 .1 313 .5 308 .5 338 .2 DAY 20 336 .0 299 .6 318 .6 316 .6 341 .6 DAY 24 293 .6 326 .0 326 .9 340 .7 DAY 27 297 .5 274 .0 288 ..4 333 .0 350 .5 274 .2 266 .3 296 .9 355 .7 380 .0 TEST DAYS DAY 62 DAY 76 335 .4 340 .1 352 .5 402 .1 413 .7 DAY 80 a a a a a DAY 90 390 .7 393 .8 388 .2 436 .2 452 .4 DAY 94 497 .5 506 .0 485 .8 568 .6 559 .0 540 .0 538 .6 523 .7 617 .8 596 .8 547 .9 547 .7 532 .8 618 .1 607 .2 565 .2 562 .1 542 .4 632 .6 624 .0 577 .5 579 .5 556 .9 638 .9 636 .8 SD test day SD test day SD test day SD test day SD test day a Rats not weighed. -56. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627756 627767 627771 627773 627777 DAY 1 227.2 227.0 219.5 246.8 225.4 DAY 2 226.4 231.8 214.7 244.4 221.4 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I TEST DAYS DAY 3 DAY 4 DAY 5 233.4 237.9 226.1 252.7 232.9 238.6 245.8 233.5 263.8 239.4 237.5 248.6 239.6 262.3 236.0 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -57- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627768 627774 627776 627782 627785 ANIMAL NUMBER 627768 627774 627776 627782 627785 DAY 1 236. 9 258. 6 212. 0 221. 6 220. 3 DAY 8 272. 3 319. 8 246. 1 251. 0 260. 7 DAY 2 244 .1 261 .6 216 .3 228 .3 221 .7 DAY 9 291 .9 326 .3 252 .3 264 .0 268 .5 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 245.2 275.8 223.7 237.3 234.7 256.5 283.4 226.6 239.0 238.4 TEST DAYS DAY 10 256. 2 289. 3 229. 6 238. 8 241. 8 265 .5 301 .5 233 .7 244 .7 250 .5 271 .4 303 .2 236 .9 246 .7 255 .6 293.5 338.2 254..4 268.2 267.9 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -58- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627758 627763 627764 627787 627788 ANIMAL NUMBER 627758 627763 627764 627787 627788 DAY 1 235. 1 212. 1 230. 0 235. 1 219. 4 DAY 2 240. 3 218. 3 235. 0 241. 0 222. 3 DAY 8 280. 6 255. 2 275. 8 221. 7 274. 1 DAY 9 283. 6 261. 2 281. 9 235. 7 276. 9 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 248 .6 226 .5 246 .1 250 .1 232 .2 255 .8 234 .3 251 .3 257 .2 239 .6 TEST DAYS DAY 10 DAY 13 261. 2 230. 8 254. 8 258. 6 247. 6 272 .0 235 .5 261 .7 237 .6 256 .7 276 .1 238 .0 264 .7 240 .3 257 .9 295 .8 272 .5 288 .0 234 .7 284 .8 308 .7 285 .8 306 .7 234 .9 309 .4 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 -59- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627759 627761 627770 627779 627786 ANIMAL NUMBER 627759 627761 627770 627779 627786 DAY 1 226 .6 240 .7 235 .6 231 .5 211 .6 DAY 8 277 .8 240 .1 291 .4 289 .0 251 .3 DAY 2 232 .9 247 .4 244 .3 237 .9 218 .2 DAY 9 284 .9 257 .4 301 .3 293 .4 255 .8 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 241.4 257.1 253.6 252.6 222.5 250 .9 262 .5 260 .1 261 .4 230 .4 TEST DAYS DAY 10 DAY 13 254 .2 266 .7 267 .7 262 .3 230 .6 DAY 20 262 .1 259 .2 277 .3 273 .8 240 .6 DAY 24 265. 4 262. 6 279. 8 277 . 5 244. 1 288.0 266.9 306.4 301.0 257.0 a 348 .0 380 .1 SD test day 24 a 335 .7 359 .4 SD test day 24 -a 355 .8 379 .8 SD test day 24 a . 374 .4 406 .2 SD test day 24 a 304 .4 326 .1 SD test day 24 a Rats not weighed. -60- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627757 627760 627765 627772 627781 ANIMAL NUMBER 627757 627760 627765 627772 627781 ANIMAL NUMBER 627757 627760 627765 627772 627781 DAY 1 221 .2 226 .2 228 .3 245 .2 219 .0 DAY 2 225 .4 232 .0 235 .3 256 .4 226 .3 DAY 8 269 .1 262 .6 271 .5 300 .9 258 .1 DAY 9 276 .7 280 .3 282 .4 307 .5 266 .8 DAY 41 402 .8 437 .7 422 .9 - 478 .4 389 .9 DAY 47 419 .7 448 .9 444 .8 494 .3 408 .2 -^ H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 236 .3 244 .4 247 .1 266 .2 236 .5 241.1 251.7 251.6 277.0 242.9 TEST DAYS DAY 10 DAY 13 247 .9 260 .2 256 .4 276 .9 247 .2 DAY 20 252 .2 254 .5 261 .1 285 .0 251 .6 254 .4 258 .3 264 .7 291 .0 256 .1 DAY 24 DAY 34 279 .7 a 287 .3 a 288 .1 a ' 317 .0 a 271 .2 a TEST DAYS DAY 52 324 .3 348 .6 349 .8 375 .5 315 .1 343 .0 379 .9 382 .7 404 .0 338 .8 372 .3 419 .4 400 .9 446 .9 372 .0 436 .7 456 .5 455 .6 513 .5 421 .0 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rats not weighed. -61 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627762 627769 627775 627778 627783 ANIMAL NUMBER 627762 627769 627775 627778 627783 ANIMAL NUMBER 627762 627769 627775 627778 627783 DAY 1 227 .2 231 .1 224 .7 213 .1 216 .4 DAY 2 235 .0 241 .1 235 .7 217 .4 218 .2 DAY 8 285 .9 302 .0 286 .6 244 .8 256 .6 DAY 41 438 .7 479 .7 433 .2 356 .8 - 409 .6 DAY 9 299 .1 309 .6 296 .0 250 .8 259 .2 DAY 47 439 .9 474 .6 451 .1 373 .5 426 .9 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 248 .7 255 .9 246 .8 221 .3 226 .6 257 .3 264 .2 254 .7 225 .5 232 .0 TEST DAYS DAY 10 DAY 13 263 .0 272 .3 265 .7 230 .1 238 .0 DAY 20 275 .1 286 .3 276 .7 236 .9 243 .9 DAY 24 279.7 289.9 280.2 240.4 247.6 DAY 34 301 .5 a 312 .5 a 304 .5 a 255 .1 a 264 .3 a TEST DAYS DAY 52 DAY 55 356 .5 386 .6 353 .3 294 .0 311 .3 DAY 61 381 .5 414 .4 377 .4 320 .0 340 .9 DAY 68 419.6 456.5 413 .9 347.1 380.0 a 453 .5 473 .0 471 .6 a 512 .2 511 .4 531 .4 a 460 .8 477 .4 478 .6 a 386 .0 393 .3 398 .3 a 449 .0 473 .8 474 .8 a Rats not weighed. -62 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627762 627769 627775 627778 627783 DAY 81 500.3 566.2 510.4 410.1 508.8 DAY 90 523.3 603.2 537.5 436.5 538.5 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS DAY 94 528.6 609.6 552.8 445.3 557.2 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 -63 H-23925; Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL. NUMBER 626399 626407 626426 626427 626428 DAY 1 290.2 248.9 280.4 305.4 272.8 DAY 2 286.0 245.1 278.1 304.6 267.3 H-23925 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I TEST DAYS . DAY 3 DAY 4 DAY 5 296.0 247.4 290.2 309,5 276.4 304.2 254.6 295.6 315.6 275.4 307.9 256.1 298.6 321.7 282.9 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -64. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626396 626397 626403 626406 626421 ANIMAL NUMBER 626396 626397 626403 626406 626421 DAY 1 299. 7 234. 7 272. 1 280. 1 297. 8 DAY 8 341. 6 265. 8 311. 3 291. 0 350. 6 DAY 2 299 .5 236 .0 270 .5 283 .2 299 .0 DAY 9 346 .4 271 .1 317 .9 292 .7 354 .7 H-23925 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 307 .7 242 .4 283 .2 278 .9 312 .3 320. 2 246. 5 285. 2 284. 3 315. 8 TEST DAYS DAY 10 319. 0 251. 2 292. 0 283. 1 328. 3 325. 8 253. 6 297. 6 285. 3 337. 1 331 .6 261 .1 305 .0 290 .2 341 .0 345 .6 277 .7 320 .3 300 .4 364 .9 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -65 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626402 626404 626409 626417 626418 ANIMAL NUMBER 626402 626404 626409 626417 626418 DAY 1 253. 4 280. 2 291. 5 271. 9 283. 0 DAY 8 307. 8 315. 7 311. 5 301. 0 319. 2 DAY 2 261 .1 281 .7 292 .1 276 .5 287 .8 DAY 9 318 .5 314 .4 316 .8 307 .7 320 .8 -^ H-23925 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 273 .4 287 .3 291 .5 277 .8 294 .4 280 .6 289 .3 300 .1 280 .7 296 .9 TEST DAYS DAY 10 DAY 13 289. 5 298. 5 303. 0 290. 8 302. 7 292 .2 299 .6 303 .3 292 .6 311 .6 296.5 308.4 311.4 300.2 318.9 319 .2 317 .2 319 .0 316 .9 326 .7 342 .7 335 .7 337 .0 333 .8 347 .7 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD.test day 13 -66- H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626405 626414 626419 626423 626424 ANIMAL NUMBER 626405 626414 626419 626423 626424 DAY 1 284. 4 285. 5 276. 5 299. 8 279. 0 DAY 2 293. 9 283. 6 276. 1 301. 4 284. 3 DAY 8 326. 2 318. 6 307. 3 338. 6 308. 4 DAY 9 331. 2 323. 7 315. 3 345. 2 314. 9 H-23925 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 296.7 294.3 282.3 313.8 290.0 305 .0 301 .2 288 .3 318 .5 290 .5 TEST DAYS DAY 10 DAY 13 312 .1 307 .6 294 .3 330 .8 299 .7 DAY 20 311 .3 309 .3 293 .9 328 .2 298 .9 DAY 24 323. 8 316. 7 304. 5 338. 7 305. 5 333.5 325.9 322.3 352.1 317.2 355 .5 348 .0 338 .9 370 .8 336 .0 387 .6 370 .4 369 .5 403 .5 368 .1 417 .7 402 .5 395 .6 427 .7 396 .6 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 -67 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626411 626415 626416 626422 626425 ANIMAL NUMBER 626411 626415 626416 626422 626425 ANIMAL NUMBER 626411 626415 626416 626422 626425 DAY 1 245 .0 265 .1 287 .7 283 .5 275 .3 DAY 2 249 .1 267 .3 297 .0 287 .6 277 .3 DAY 8 277 .7 292 .1 327 .6 331 .7 312 .7 DAY 9 276 .1 293 .2 330 .8 337 .2 314 .6 DAY 52 430 .1 440 .0 508 .3 - 509 .7 513 .6 H-23925 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 255.5 267.9 295.5 292.3 283.6 262 .0 274 .4 307 .8 302 .9 293 .2 TEST DAYS DAY 10 DAY 13 261 .4 275 .4 311 .0 313 .3 293 .6 DAY 20 266 .0 287 .4 313 .7 316 .0 301 .1 DAY 24 273.6 294.7 320.7 324.9 306.6 DAY 48 279.0 292 .5 325 .7 a 297.1 310 .0 347 .9 a 335.2 359 .6 399 .0 a 342.0 361 .1 395 .8 a 324.6 345 .2 374 .2 a TEST DAYS 411.0 416.6 481.4 481.8 481.4 a Rats not weighed. -68 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626398 626401 626408 626410 626413 ANIMAL NUMBER 626398 626401 626408 626410 626413 ANIMAL NUMBER 626398 626401 626408 626410 626413 DAY 1 276 .5 264 .0 275 .6 264 .8 316 .8 DAY 2 277 .4 266 .6 268 .8 271 .5 321 .4 DAY 8 312 .8 2-84 .4 302 .4 302 .8 356 .2 DAY 9 311 .8 284 .9 308 .0 311 .9 357 .8 DAY 52 a a a a a DAY 62 474 .1 421 .9 455 .3 505 .1 627 .0 H-23925 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TE ST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 287 .6 268 .3 276 .6 275 .5 328 .7 290 .2 270 .6 281 .9 283 .6 332 .2 TEST DAYS DAY 10 DAY 13 295.0 274.0 287.9 288.5 333.4 DAY 20 297.8 276.7 . 291.9 290.5 342.2 306 .9 283 .9 300 .7 293 .8 345 .1 DAY 24 DAY 48 315 .7 285 .3 308 .6 311 .3 357 .0 339 .1 304 .6 332 .4 334 .0 377 .2 TEST DAYS DAY 66 DAY 76 379.6 319.3 359.6 373.8 421.0 DAY 83 a a a a a DAY 89 454 .5 391 .1 424 .1 469 .6 560 .1 DAY 94 484 .9 432 .6 464 .4 515 .4 645 .1 491 .3 442 .0 476 .8 521 .6 662 .1 496.7 450.3 487.1 531.6 684.8 504.8 451.3 494.6 538.0 694.0 505 .5 455 .6 493 .4 542 .4 697 .5 SD test day SD test day SD test day SD test day SD test day a Rats not weighed. -69. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 APPENDIX B Individual Clinical Observations -70- Pompany Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Animal Number 632649 632650 632651 632652 632653 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Observation_______________ No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 632654 632655 632656 632657 632658 GROUP II Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-10 1-10 1-10 1-10 1-10 Animal Number 632659 632660 632661 632662 632663 GROUP III Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-13 1-13 1-13 1-13 1-13 -71Company SanRTzetf. Does not contain T8CA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Animal Number 632664 632665 632666 632667 632668 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation__________________Test Day No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected 1-24 1-24 1-24 1-24 1-24 Animal Number 632669 632670 632671 632672 632673 GROUP V Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-52 1-52 1-52 1-52 1-52 Animal Number 632674 632675 632676 632677 632678 GROUP VI Observation No Hair No Hair No No Hair No abnormalities detected Loss, Forelimb, Bilateral abnormalities detected Loss, Forepaw, Bilateral abnormalities detected abnormalities detected Loss, Forepaw, Bilateral abnormalities detected Test Day 1-88 94 1-81 88-94 1-94 1-53 60-94 1-94 -72Company Sanitized. Does nol contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 CORN OIL (NEGATIVE CONTROL) Animal Number 627789 627801 627805 627810 627818 INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Observation ___ No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 627793 627797 627799 627800 627808 GROUP II Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-10 1-10 1-10 1-10 1-10 Animal Number 627796 627806 627807 627809 627816 Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-13 1-13 1-13 1-13 1-13 -73- Company Sanitized. Does nol contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Stiidy in Rats DuPont-2921 Animal Number 627791 627794 627804 627812 627817 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 627792 627798 627813 627814 627815 GROUP V Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-52 1-52 1-52 1-52 1-52 Animal Number 627795 627811 627819 627820 627821 GROUP VI Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day ' 1-94 1-94 1-94 1-94 1-94 -74- Company Sanitized. Does not contafn TSCA ^e>t H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 CORN OIL:ACETONE (NEGATIVE CONTROL) Animal Number 625361 625363 625364 625380 625383 INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I ___ Observation___ No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected __________Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 625355 625362 625367 625368 625378 GROUP II Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-10 1-10 1-10 1-10 1-10 Animal Number 625357 625358 625370 625384 625385 Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-13 1-13 1-13 1-13 1-13 Company Sanitized. Does not contain TSCACBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 CORN OIL:ACETONE (NEGATIVE CONTROL) Animal Number 625354 625365 625371 625376 625381 INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation_________________ No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 625356 625359 625360 625372 625374 GROUP V Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-52 1-52 1-52 1-52 1-52 Animal Number 625353 625369 625373 625375 625377 Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-94 1-94 1-94 1-94 1-94 -76- Co"Pany Sanded. Does not contain TSCACBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Animal Number 625322 625323 625336 625343 625349 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Observation___________________Test Day No abnormalities detected 1-5 No abnormalities detected 1-5 No abnormalities detected 1-5 No abnormalities detected 1-5 No abnormalities detected 1-5 Animal Number 625320 625330 625332 625341 625350 GROUP II Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Diarrhea No abnormalities detected Test Day 1-10 1-10 1-10 1-4, 6-10 5 1-10 Animal Number 625321 625327 625342 625345 625348 Observation No abnormalities detected Diarrhea No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-4, 6-13 5 1-13 1-13 1-13 1-13 -77- cwv>m!SM"'^ ^,----^c H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Animal Number 625325 625328 625338 625340 625352 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation ________ No abnormalities detected No abnormalities detected No abnormalities detected Salivation No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-6, 8-24 7 1-24 1-24 Animal Number 625324 625329 625333 625334 625344 Observation No abnormalities detected No abnormalities detected Salivation No abnormalities detected Salivation No abnormalities detected No abnormalities detected Test Day 1-52 1-6, 8-52 7 1-6, 8-52 7 1-52 1-52 Animal Number 625326 625335 625346 625347 625351 Observation No abnormalities detected Salivation Alopecia both front legs Alopecia abdomen No abnormalities detected Salivation Black ocular discharge Yellow-stained perineum Red-stained chin Brown-stained chin No abnormalities detected No abnormalities detected Salivation Test Day 1-6, 9-76, 77-94 7-8 76 7, 8, 9, 10, 13, 20, 27 1-6, 11-94 7-8 8 8 8 9-10 1-94 1-6, 8-94 7 -78- Pompany Sanitized. Does not contain TSCA CB H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS DuPont-2921 Animal Number 627756 627767 627771 627773 627777 GROUP I Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 627768 627774 627776 627782 627785 GROUP II Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-10 1-10 1-10 1-10 1-10 Animal Number 627758 627763 627764 627787 627788 GROUP III Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Wet perineum No abnormalities detected Diarrhea Test Day 1-13 1-13 1-13 1-5, 7-13 6 1-8, 10-13 9 -79- Cwnpany Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS DuPont-2921 Animal Number 627759 627761 627770 627779 627786 GROUP IV Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 627757 627760 627765 627772 627781 Animal Number 627762 627769 627775 627778 627783 Observation No abnormalities detected No abnormalities detected No abnormalities detected Wet perineum No abnormalities detected Wet perineum No abnormalities detected GROUP VI Observation No abnormalities detected Diarrhea No abnormalities detected No abnormalities detected Alopecia perineum No abnormalities detected Alopecia both front paws Alopecia right front leg Test Day 1-52 1-52 1. 3-52 2 1, 3-52 2 1-52 Test Day 1-8, 10-94 9 1-94 1-94 81, 90, 94 1-23, 94 24, 34, 41, 47, 55, 61, 74, 81, 90 -80Company Sanitized. Does not conteh tSCSAC81 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 H-23925 (TEST SUBSTANCE) Animal Number 626399 626407 626426 626427 626428 INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 626396 626407 626426 626427 626428 Animal Number 626402 626404 626409 626417 626418 Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected GROUP III Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Diarrhea Test Day 1-10 1-10 1-10 1-10 1-10 Test Day 1-13 1-13 . 1-13 1-13 1-3,5-13 4 -81- Company Sanitized. Does not contain TSCA CB H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Animal Number 626405 626414 626419 626423 626424 H-23925 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 626411 626415 626416 626422 626425 Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-52 1-52 1-52 1-52 1-52 Animal Number 626398 626401 626408 626410 626413 Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-94 1-94 1-94 1-94 1-94 -82Company Sanifeed. Does not contafn TSCA C8 H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 APPENDIX C Individual Fluorine Levels -83- Company Sanitized. Does not contain TSCA CBI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 Terms: Active % Active Mol Wt Active Formulation Dose % F in Active Mol Wt F TERMS AND CALCULATIONS Fluorine containing compound The % of formulation that is made up of fluorine containing components The molecular weight of the fluorine containing components (g/mole) The mg of formulation given per kg of animal body weight The % fluorine in the fluorine containing components of the formulation (weight basis) The molecular weight of fluorine g/mol Compound Calculations: Dose Active (mg/kg) The mg of fluorine containing compound administered per kg of animal body weight. = (% active/100) x Formulation Dose Dose Active (mmole/kg) The mmole of fluorine containing compound administered per kg of animal body weight = dose [mg/kg] / Mol Wt Active [mg/mmol] Dose F (mg/kg) The mg Fluorine administered per kg of animal body weight = (% F in active/100) x Dose Active [mg/kg] Dose F (mmol/kg) The mmole of fluorine administered per kg of animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol] Molar Ratio (Active/F) The moles of fluorine containing compound per mole of fluorine = Dose Active [mmol/kg] / Dose F [mmol/kg] Formulation Dose Normalization Factor The formulation dose that would be required to administer the amount of active needed to achieve the normalized dose = (Normalized dose of Active [mmol/kg] / Dose Active [mmol/kg]) x Formulation Dose -84- Company Sanitized. Does not contain TSCA CB) H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 TERMS AND CALCULATIONS Individual Animal Measurement: ppm F in blood The ppm fluoride measured in blood Individual Animal Calculations: ppm F in Blood minus Bkg 0.2 ppm ppm F in Blood normalized to 0.1 mmol/kg Dose The ppm fluoride measured in plasma minus the background fluoride measured in control animal blood. In this case the value was established at 0.2 ppm. The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead of the actual active dose. This assumes linearity between administered dose and blood fluorine levels, but is needed because different doses of active were used in the study. = (0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in blood minus background) /xmolar equivalents of active in blood The /zmolar [/imol/L] concentration of fluorine containing compound in plasma based on the ppm fluorine in plasma normalized to 0.1 mmol/kg active dose. This assumes that all plasma fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm - 1 mg/L = (Normalized ppm [mg/L] fluorine in blood /' Mol Wt F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x 1000 /xmol/mmol 6 -85- Company Sanitized. Does not contain TSCA CB( H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2921 _____FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS_____ Considerations: - The data used for kinetic analysis was from a limited screen and extrapolation should be done cautiously. - Sample size is low - Analytical data used without validation - Steady-state not reached - Terminal phase may not be reached - Some compounds are mixtures of fluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - The kinetics apply only to plasma, not to specific tissues or the whole body - Each compound may have very different potency for producing toxicity Assumptions: (May or may not be justified in all cases) - Plasma fluorine concentrations are linear with respect to dose - Analytical method is appropriate for all types of compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of individual components - Background plasma Fluorine is 0.2 ppm - % F data is the % Fluorine of the active (Fluorine containing component(s) in the formulation - Molecular weight is the molecular weight of the active component in the formulation -86- CompanySanded. Does nof contain TSCA CW H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24019 DuPont-2921 Given: Mol Wt. Active (g/mole): 497 Formulation Dose (mg/kg): 10 % Active (F Containing) in Formulation: 100 %F in Active: 65 Mol Wt. F (g/mol): 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 10 0.020 49.7 Dose F (mmol/kg): 0.342 Molar Ratio (Active/F): 0.059 Dose F (mg/kg):_____6.5 Rat Number Test Day Sample ppmF in Blood Group I 625322 1 1.0 625323 1 1.3 625336 1 2.6 625343 1 2.6 625349 1 2.9 Group I 625322 5 51.7 625323 5 38.0 625336 5 38.7 625343 5 78.5 625349 5 37.1 Group II 625320 10 625330 10 625332 10 625341 10 625350 10 65.6 61.1 60.5 63.1 58.1 ppm F in Blood Minus Bkg 0.2 ppm 0.8 1.1 2.4 2.4 2.7. 51.5 37.8 38.5 78.3 36.9 65.4 60.9 60.3 62.9 57.9 ppm F in Blood Normalized to 0.1 nunoles/kg Dose 3.98 5.47 11.93 11.93 13.42 255.96 187.87 191.35 389.15 183.39 325.04 302.67 299.69 312.61 287.76 ^imolar Equivalents of Active in Blood 12.31 16.92 36.92 36.92 41.54 792.31 581.54 592.31 1204.62 567.69 1006.15 936.92 927.69 967.69 890.77 -87- Company Sanitized. Does not contain TSCA CRI H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat Number Test Day Sample ppmF in Blood Group III 625321 13 625327 13 625342 13 625345 13 625348 13 77.2 61.5 64.6 57.0 62.4 Group IV 625325 24 625328 24 625338 24 625340 24 625352 24 40.3 46.3 43.2 39.1 41.9 Group V 625324 52 625329 52 625333 52 625334 52 625344 52 24.7 29.8 27.3 29.1 23.8 Group VI 625326 94 12.4 625335 94 16.4 625346 94 13.1 625347 94 11.0 625351 94 11.6 ppm F in Blood Minus Bkg 0.2 ppm 77.0 61.3 64.4 56.8 62.2 40.1 46.1 43.0 38.9 41.7 24.5 29.6 27.1 28.9 23.6 12.2 16.2 12.9 10.8 11.4 ppm F in BIlood Normalize!ito 0.1 mmolef,/kg Dose 382.69 304.66 320.07 282.30 309.13 199.30 229.12 213.71 193.33 207.25 121.77 147.11 134.69 143.63 117.29 60.63 80.51 64.11 53.68 56.66 DuPont-2921 fimolar Equivalents of Active in Blood 1184.62 943.08 990.77 873.85 956.92 616.92 709.23 661.54 598.46 641.54 376.92 455.38 416.92 444.62 363.08 187.69 249.23 198.46 166.15 175.38 Company SanWzed. Does not confam TSCA CBt H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H- 24020 DuPont-2921 Given: Mol Wt. Active (g/mole): 426 Formulation Dose (mg/kg): 20 % Active (F Containing) in Formulation: 100 % F in Active: 69 Mol Wt. F (g/mol): 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 20 0.047 42.6 Dose F (mmol/kg): 0.726 Molar Ratio (Active/F): 0.065 Dose F (mg/kg):_____13.8 Rat Number Test Day Sample ppm F in Blood Group I 627773 1 13.2 627771 1 8.1 627767 1 16.5 627756 1 14.1 627777 1 10.7 Group I 627773 5 627771 5 627767 5 627756 5 627777 5 75.3 66.8 64.8 80.0 71.6 Group II 627768 10 627774 10 627776 10 627782 10 627785 10 56.3 51.0 54.8 43.6 65.1 ppm F in Blood Minus Bkg 0.2 ppm 13.0 7.9 16.3 13.9 10.5 75.1 66.6 64.6 79.8 71.4 56.1 50.8 54.6 43.4 64.9 ppm F in Blood Normalized to 0.1 mmoles/kg Dose 27.69 16.83 34.72 29.61 22.37 159.96 141.86 137.60 169.97 152.08 119.49 108.20 116.30 92.44 138.24 /irnolar Equivalents of Active in Blood 94.20 57.25 118.12 100.72 76.09 544.20 482.61 468.12 578.26 517.39 406.52 368.12 395.65 314.49 470.29 -89- Company Sanitized. Does not contain TSCA cat H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat Number Test Day Sample ppmF in Blood Group III 627758 13 627764 13 627787 13 627763 13 627788 13 43.5 20.7 13.2 30.8 24.9 Group IV 627759 24 9.2 627779 24 14.9 627770 24 11.6 627761 24 9.9 627786 24 6.7 Group V 627757 52 0.9 627781 52 0.6 627760 52 1.0 627765 52 0.9 627772 52 0.8 Group VI 627778 94 627783 94 627762 94 627775 94 627769 94 0.3 <0.2 0.2 0.2 <0.2 * = Below LOD (Level of Detection) ppm F in Blood Minus Bkg 0.2 ppm 43.3 20.5 13.0 30.6 24.7 9.0 14.7 11.4 9.7 6.5 0.7 0.4 0.8 0.7 0.6 0.1 * 0.0 0.0 * ppm F in BHood Normalize dto 0.1 nunoles/kg Dose DuPont-2921 /imolar Equivalerits of Active in Blood 92.23 43.67 27.69 65.18 52.61 313.77 148.55 94.20 221.74 178.99 19.17 31.31 24.28 20.66 13.85 65.22 106.52 82.61 70.29 47.10 1.49 0.85 1.70 1.49 1.28 5.07 2.90 5.80 5.07 4.35 0.21 * 0.00 0.00 * 0.72 * 0.00 0.00 * -90- Company SanWzed. Does not contain TSCA Cffl H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-23925 DuPont-2921 Given: Mol Wt. Active (g/mole): 669.5 % P in Active: 55 Formulation Dose (mg/kg): 200 Mol Wt. F (g/mol): 19 % Active (F Containing) in Formulation: 29 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 58 0.087 230.9 Dose F (mmol/kg): 1.679 Molar Ratio (Active/F): 0.052 Dose F (mg/kg):_____31.9 Rat Number Test Day Sample ppmF in Blood Group I 626399 10.4 626407 12.6 626426 4.9 626427 11.7 626428 6.4 Group I 626399 5 626407 5 626426 5 626427 5 626428 5 14.7 12.9 10.7 16.4 12.4 Group II 626396 10 16.7 626397 10 13.9 626403 10 15.3 626406 10 16.7 626421 10 17.0 ppm F in Blood Minus Bkg 0.2 ppm 10.2 12.4 4.7 11.5 6.2 14.5 12.7 10.5 16.2 12.2 16.5 13.7 15.1 16.5 16.8 ppm F in Blood Normalized to 0.1 mmoles/kg Dose 11.77 14.31 5.43 13.27 7.16 16.74 14.66 12.12 18.70 14.08 19.05 15.81 17.43 19.05 19.39 ^molar Equivalents of Active in Blood 31.97 38.87 14.73 36.05 19.44 45.45 39.81 32.92 50.78 38.24 51.72 42.95 47.34 51.72 52.66 -91- CompanySanffaed. Does not contain TSCA W. H-23925: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat Number Test Day Sample ppmF in Blood Group III 626402 13 5.2 626404 13 5.9 626409 13 6.1 626417 13 8.2 626418 13 6.8 Group IV 626405 24 5.6 626414 24 5.5 626419 24 5.8 626423 24 4.3 626424 24 4.0 Group V 626411 52 3.0 626415 52 2.7 626416 52 2.2 626422 52 2.8 626425 52 2.1 Group VI 626398 94 1.6 626401 94 1.5 626408 94 2.4 626410 94 1.5 626413 94 1.2 ppm F in Blood Minus Bkg 0.2 ppm 5.0 5.7 5.9 8.0 6.6 5.4 5.3 5.6 4.1 3.8 2.8 2.5 2.0 2.6 1.9 1.4 1.3 2.2 1.3 1.0 ppm F in I Hood Normalize;d to 0.1 mmoles/kg Dose 5.77 6.58 6.81 9.23 7.62 6.23 6.12 6.46 4.73 4.39 3.23 2.89 2.31 3.00 2.19 1.62 1.50 2.54 1.50 1.15 DuPont-2921 ^tmolar Equivalents of Active in Blood 15.67 17.87 18.50 25.08 20.69 16.93 16.61 17.55 12.85 11.91 8.78 7.84 6.27 8.15 5.96 4.39 4.08 6.90 4.08 3.13 -92- Company Sanitized. Does nol contain TSCA CB)