Document 4a2aJvjJyjpNMyN8VXv7RERDG

TRADE SECRET Study Title H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID: DuPont-2927 AR226-3157 DuPont-2927 AUTHOR: Carol Finlay, B.A. STUDY COMPLETED ON: August 3, 2000 PERFORMING LABORATORY: E.I. du Font de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine EUcton Road, P.O. Box 50 Newark, Delaware 19714-0050 WORK REQUEST NUMBER:'p^--ss--ss--^^ SERVICE CODE NUMBER: ^^1 ^------ Page 1 of 82 Company Sanitized. Does not contain TSCA CBS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Reviewed by: ^ rGary W. Jepson, Ph.D. Senior Research Scientist Reviewed by; CJ^g^C^f. U: ^{LcL^, Judith C. Stadler, Ph.D., D.A.B.T. J Director, General Toxicology Issued by Study Director: /^fr-t J^L^ Carol Fin&y, B.A. Q Staff Scientist <7,A--& 2-000 Date S-AU&-200Q Date ^-^UL\' U6b Diitt -2. Company Sanitized. Does not contain TSCA CB? H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 TABLE OF CONTENTS Page CERTIFICATION ........................................................................................................................2 LIST OF TABLES........................................................................................................................4 LIST OF FIGURES........ LIST OF APPENDICES. STUDYINFORMATION.............................................................................................................5 STUDYPERSONNEL..................................................................................................................7 SUMMARY.................................................................................................................................... 8 INTRODUCTION.........................................................................................................................9 MATERIALS AND METHODS ................................................................................................. 9 A. Test Substance and Positive Controls................................................................................. 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................................................................................... 15 TABLES....................................................................................................................................... 16 -3- sany "SanifescJ Doss fwf wfisw TSCA CB? H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 FIGURES ............................................................................................................................ 21 APPENDICES.............................................................................................................................26 LIST OF TABLES Page 1. MEAN BODY WEIGHTS................................................................................................................................. 17 2. MEAN BODY WEIGHT GAINS........................................................................................................................ 18 3. MEAN BLOOD FLUORINE LEVELS................................................................................................................ 19 4. MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE................................................20 LIST OF FIGURES Page 1. MEAN BODY WEIGHTS................................................................................................................................... 22 2. MICROMOLAR EQUIVALENTS W RAT BLOOD..........................................................................................23 3. MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS .............................25 LIST OF APPENDICES Page A. INDIVIDUAL BODY WEIGHTS.......................................................................................................................27 B. INDIVIDUAL CLINICAL OBSERVATIONS...................................................................................................^ C. ANALYSIS OF BLOOD FLUORINE DATA .....................................................................................................73 -4- Company Sanifeed. Does not contain TSCA CBt H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION TEST SUBSTANCE: Substance Tested: Synonyms/Codes: H-24018 Haskell Number: 24018 DuPont-2927 POSITIVE CONTROL: Substance Tested^ Potassium perfluoroalkyi sulfonate Synonyms/Codes. H-24019 Haskell Number: 24019 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study m Rats DuPont-2927 Known Impurities POSITIVE CONTROL: Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt Synonyms/Codes: H-24020 Perfluorooctanoate, ammonium salt PPOA Ammonium perfluorooctanoate Haskell Number: 24020 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) m-Life Initiated/Completed: June 13, 1999 / September 14,1999 -6- -Company S.nfeed. Does not eon^. H-24018: 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-2927 eompany SwrfStea^. Doe not contain TSCA CB8 7- SfcMpmySaBHfr^ "----'eontaInTSale^ tu- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 SUMMARY The objective of this study was to evaluate the potential for H-24018, 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 exposed to 1000 mg/kg/day of H-24018. The test substance was administered to one group of five rats for 5 consecutive days and to 5 groups for 10 days. Approximately two hours after me 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 approximately every other week during the recovery period. Additionally, 2 negative controls, corn oil and corn oil/acetone (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-24018. One rat dosed with H-24018 was found dead on test day 9. The death of this rat is considered not to be due to the test substance, since no clinical signs oftoxicity were observed and no body weight loss occurred in any of the rats during the study. The death of this rat was probably caused by a dosing injury. No compound-related clinical signs oftoxicity were observed in the rats dosed with H-24018. Rats dosed with a positive control material, 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, the other positive control substance, exhibited wet perineum and diarrhea during the dosing period. Alopecia was observed during the recovery period for rats dosed with H-24018, H-24019, and H-24020. The negative control rats exhibited no clinical signs during the study. Comparison of body weights was complicated by 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, the mean body weights and mean body weight gains of the rats dosed with H-24018 were comparable to the negative controls and equal to or greater than the positive controls. A steady-state for fluorine levels in whole blood may not have been achieved during 10 consecutive days of dosing with 1000 mg/kg H-24018. 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 H-24018 was 1402.6, compared to AUCINF/D values of 566,479.1 and 70,789.6 for H-24019 and H-24020, respectively. Under the conditions of this study, administration of H-24018 to male rats for 10 consecutive days resulted in limited 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 400x and 50x the AUCINF/D for the test material. -8. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 INTRODUCTION The objective of this study was to define the potential of H-24018 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 two positive controls that were materials previously shown to bioaccumulate in mammals. The daily dosage for the test substance was selected based on available toxicity data and the results of a rangefinding study, m the rangefinding study, a group of 5 male rats was dosed by oral gavage with H-24018 at a dosage of 1000 mg/kg for 5 consecutive days. A group of 5 male rats was dosed with deionized water for 5 consecutive days and served as controls. The rats dosed with H-24018 experienced an overall mean body weight gain of 40 grams. The control group had an overall mean body weight gain of 27 grams. A dosage of 1000 mg/kg was selected for the limit dosage for this project. The limit dosage of 1000 mg/kg was chosen for the main study and was expected to produce less than a 10% difference in mean body weight over 10 days 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 Controls The test substance, H-24018, was supplied by the sponsor as an amber brown liquid. 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. -9- ^asupeisy SaaSSse& D not wtan T^a ^ H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 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, me. 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 ofcoliforms, 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- SostpBsy SartShecS, l H-24018: Biopersistence Screening 10-Dose Oral Gavage Study m Rats DuPont-2927 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, male and female rats were released from quarantine by the laboratory animal veterinarian or designee. E. Study Design Substance Vehicle Negative Controls Corn oil Corn oil: acetone Positive Controls H-24019 H-24020 Test Substance H-24018 Not applicable Corn oil: acetone Corn oil None Dosage (mg/kg/day) 0 0 10 20 1000 Number of Animals 30 30 30 30 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 initiation 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-24018 was dosed as received. The amount of test substance each rat received was based on the body weight collected on each day of dosing and the test substance density of 1100 mg/mL. The test substance was stirred on a magnetic stir plate throughout the dosing procedure. 11- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 2. Positive Controls The solid positive control materials were suspended as emulsions in their respective vehicles. Corn oil was used as the vehicle for H-24020. It was necessary to dissolve H-24019 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 Corn oil and corn oil:acetone (80:20) were chosen as the 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 The rats were weighed on each day of dosing and then weekly or every other week 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 the blood and livers were collected according to the following schedules: Group Dosing Days Tissue Collected I 1-5 I 1-5 n 1-10 m 1-10 IV 1-10 V 1-10 VI 1-10 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 -12- ^lUpHus SssMssH. Doe not onSaSn TSGA Bi H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 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 by 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 for evaluation of fluorine biopersistence. K. Treatment of Fluorine Data Noncompartmental analysis was conducted on blood fluorine data derived from rats dosed with H-24018 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/O-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 <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 micromolar (/xM) equivalents of active component for further comparisons. The data and descriptions of data manipulation and presentation are provided in Appendix C. 13Compan^ SwSSseS. I H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats RESULTS AND DISCUSSION DuPont-2927 A. In-Life Toxicology (Tables 1 and 2; Figure 1, Appendices A and B) One rat dosed with H-24018 was found dead on test day 9. The death of this rat is considered not to be due to the test substance, since no clinical signs oftoxicity were observed and no body weight loss occurred in any of the rats during the study. The death of this rat was probably caused by a dosing injury. No compound-related clinical signs oftoxicity were observed in the rats dosed with H-24018. 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 for rats dosed with H-24019 and H-24020. The negative control rats exhibited no clinical signs during the study. The mean body weights and mean body weight gains of the rats dosed with H-24018 were comparable to the corn oil controls throughout the study. The mean body weights and mean body weight gains of the rats with H-24018 were greater than the H-24020 positive control rats. Comparison of body weights between the H-24018 rats and the H-24019 positive control and corn oiVacetone negative control rats was complicated by the fact that there was a difference in age on test day 1. This difference resulted in differences in mean body weights on test day 1. The rats dosed with the test substance, H-24018, were 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-24018 was comparable to the corn oiVacetone negative controls and equal to or greater than the H-24019 positive controls. B. Fluorine Data (Tables 3 and 4; Figures 2 and 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 IJI.M equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state 14- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 (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 jjiM 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-24018 normalized /zM equivalents for fluorine in rat blood continued to rise throughout the dosing period and may not have reached steady-state (Figure 2C). The Cmax for H-24018 was 6.140 1.389 ppm (Mean SD) with a terminal half-life of 29.1 days. Blood was sampled at seven time points throughout the study, with only four of those 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-24018 accumulation index (AI) was 42.5 and the bioaccumulation index (BI) was 261.1 ppm. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between H-24018 and positive controls. The AUCINF/D for the fluorine component of H-24018 was 1402.6 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 1000 mg/kg H-24018 exhibited no test substance-related mortality or clinical signs oftoxicity and had mean body weight gains that were comparable to the negative control rats. A steady state for fluorine in the blood may not have been achieved for H-24018 during the 10-day dosing period. Under the conditions of this study, administration of H-24018 to male rats for 10 consecutive days resulted in limited 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 400x and 50x the AUCINF/D for the test material.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. -15 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 TABLES -16- Bft^dHSF^nRtod, De@B Mfi Mnlsh TSGA eat H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 TABLE 1 MEAN BODY WEIGHTS (g) Test Days 1 2 3 4 5 6 7 8 9 10 13 20 24 27 34 38 41 47 48 52 55 61 62 68 69 76 80 81 82 88 90 94 Negiitive Controls Corn Oil 229.8 234.1 Corn OihAcetone 297.6 299.6 241.4 250.4 259.3 266.8 271.2 281.9 285.2 295.9 321.1 309.4 314.3 321.5 326.0 334.9 339.2 345.2 351.7 350.7 350.5 378.3 369.8 401.0 - - 407.8 411.4 - - 434.5 - 454.9 - - - 464.7 487.0 505.1 527.6 - - - 515.4 - - - - 539.0 - - - - - - - 556.2 566.9 - 562.2 567.8 Indicates that the animal was not weighed. 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 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 - - 438.1 - - 523.4 - 465.6 452.3 465.8 - 470.9 - - 563.4 570.7 - - 499.2 - - - 585.3 597.9 - 527.8 538.7 Test Substance H-24018 234.6 240.8 250.5 258.1 267.2 275.9 283.1 285.3 298.6 306.0 333.8 362.2 406.6 - 432.5 453.9 - - 483.5 488.9 511.6 530.4 - - 545.5 - - - 570.6 583.1 - 596.5 -17. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 TABLE 2 MEAN BODY WEIGHT GAINS (g) Test Days 1-5 1-10 10-13 10-24 10-52 10-94 Negative Controls Corn Oil 29.5 66.1 Corn Oil:Acetone 23.9 54.1 25.2 82.4 168.8 271.0 -1.0 49.3 175.9 216.1 Positive Controls H-24019 H-24020 17.1 27.4 10.2 69.0 119.2 279.0 24.9 56.4 5.2 57.0 181.7 254.8 Test Substance H-24018 32.6 71.4 27.8 100.6 182.9 290.5 -18 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rate DuPont-2927 TABLES MEAN BLOOD FLUORINE LEVELS Test Days 1 5 10 13 24 52 94 Negative Controls Corn Oil ppm <0.2 <0.2 <0.2 0.2s (0.1) <0.2 0.3^0.1) 0.3a(0.l) (^om Oil:Acetone ppm <0.2 00..23s (0.1)'' 0.6s (0.6) O^ 0.2 0.2 Positive Controls 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) 0.20 (0.1) a Mean of 4 of the 5 values. One of the values was below the 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. Test Substance H-24018 ppm 2.9(1.1) 9.0 (0.6) 10.6 (2.4) 5.7 (0.9) 2.5 (0.2) 6.6 (0.3) 0.6 (0.3) -19- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 TABLE 4 MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE Test Days 1 5 10 13 24 52 94 Positive Controls Test Substance H-24019 /zM F Equivalents 28.92 (13.3) 747.69 (271.5) 945.85 (43.5) 989.85(116.9) 645.54 (42.9) 411.38 (40.6) 195.38 (32.5) H-24020 fJiM F Equivalents 89.28 (23.4) 518.12 (44.9) 391.01 (56.8) 191.45(82.7) 74.35 (22.1) 4.64 (1.1) 0.24 (0.4) H-24018 /AM F Equivalents 1.6 (0.62)" 5.2 (0.33) 6.1 (1.39) 3.2 (0.52) 1.^(0.12) 0.7 (0.15) 0.3 (0.15) a Standard deviation is in parentheses. b Mean of 4 of the 5 values. One of the rats was found dead on test day 9. -20- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 FIGURES -21- QyMpsssy Saartteed. Vwss BoaewtehB weA 81 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study m Rats 400 350 -i 300 @ 250 *-* to S ?. 200 S 150 100 FIGURE 1 MEAN BODY WEIGHTS (g) --^--Negative Control (Corn O --B--Negative Control (Corn O --A--Positive Control H-24019 --^--Positive Control H-24020 --9K-Test Substance H-24018 Test Days 10 12 -22. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 FIGURE 2 MICROMOLAR EQUIVALENTS IN RAT BLOOD A. Normalized Rat Blood H-24019 fiM Equivalents Resulting from a 10-Day 0 ral Gavage Micromolar (^M) equivalents of H-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 f(M Equivalents Resulting from a 10-Day 0 ralG avage Micromolar (jaM) 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. -23- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 C. Normalized Rat Blood H-24018 itM Equivalents Resulting from a 10-Day Oral Gavage Micromolar (/AM) equivalents of H-24018 (test substance) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. -24- H-24018: Biopersistence Screening 10-Dose Oral Oavage Study in Rats FIGURES MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS -25- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDICES -26. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDIX A Individual Body Weights -27- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats INDIVIDUAL BODY WEIGHTS ABBREVIATIONS: PD SD - found dead sacrificed by design EXPLANATORY NOTES DuPont-2927 -28- H-24018: 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 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 -29- H-24018: 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 2 224 .4 245 .9 231 .4 252 .8 241 .6 DAY 8 270. 6 303. 9 280. 2 312. 0 303. 6 DAY 9 274 .9 305 .1 284 .3 315 .4 305 .7 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS GROUP II TEST r )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 -30- H-24018: 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 CORK OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS GROUP III TEST 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 -31 H-24018: 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 2 235.4 233.0 228.8 231.5 231.3 DAY 8 274 .0 286 .1 267 .2 278 .2 m .8 DAY 9 279.1 289.5 270.6 282.7 280.4 CORK OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TEST r )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 295.4 ^a 278.1 a 294.3 ^a 294.6 a 334.4 377.4 345.8 356.5 368.7 365.6 416.7 378.1 388.4 407.3 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 a Rat was not weighed. -32. ''/-' H-24018: 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 TEST E>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 Rat was not weighed. -33. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST I:)AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627795 627811 627819 627820 627821 218.0 228.6 248.8 225.5 227.5 219.5 226.3 255.3 228.9 233.2 225.3 236.7 264.9 239.2 240.7 234.3 246.1 274.4 247.9 246.1 239.4 251.5 280.4 255.6 251.2 246.2 259.6 288.0 261.3 259.8 250.1 263.6 290.3 265.3 263.9 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 DAY 34 627795 627811 627819 627820 627821 261.4 265.0 270.7 a 273.0 276.7 285.6 a 304.5 308.0 324.3 a 281.3 282.9 292.4 ^a 272.6 275.3 281.8 a 308.0 337.8 399.7 367.2 330.1 329.8 360.6 431.2 393.0 352.9 362.0 396.4 472.5 445.5 385.6 ANIMAL NUMBER DAY 41 DAY 47 TEST DAYS DAY 52 DAY 55 DAY 61 DAY 68 627795 627811 627819 627820 627821 384.2 398.0 a 411.1 435.7 440.7 422.0 439.3 a 455.4 463.0 473.6 505.9 534.9 a 551.7 574.4 588.6 478.0 504.1 a 529.7 547.1 558.6 411.8 426.1 a 439.1 459.0 463.3 a Rat was not weighed. -34- ^ -i''^'.' ^.sLy'' H-24018: 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 -35. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study m Rats ANIMAL NUMBER 625361 625363 625364 625380 625383 DAY 1 322.6 273.7 291.1 289.4 304.7 DAY 2 325.0 271.4 290.9 286.2 299.7 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I DAY 3 DAY 4 DAY 5 337.6 278.4 299.9 291.5 302.7 340.3 281.7 306.0 300.1 310.7 349.7 286.7 312.0 305.5 313.1 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -36- H-24018: 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 2 282 .0 294 .1 307 .8 318 .0 306 .4 DAY 8 321 .7 323 .9 349 .1 364 .2 350 .1 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 -37- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625357 625358 625370 625384 625385 ANIMAL NUMBER 625357 625358 625370 625384 625385 DAY 1 311 .9 285 .7 307 .9 294 .2 297 .6 DAY 2 308 .8 286 .3 316 .7 300 .0 304 .6 DAY 8 364 .2 319 .8 373 .4 325 .6 347 .0 DAY 9 370 .3 321 .6 374 .7 331 .0 349 .5 CORN OIL:ACETONE (NEGATIVE Control) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 320.3 295.5 329.5 312.0 313.0 328.4 296.4 337.9 309.3 320.4 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 .4 376.9 320.3 385.6 340.6 358.9 405.6 340.7 407.0 357.1 377.6 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 -38- H-24018: 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 TEST 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 -39- H-24018: 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 DAY 24 323.9 333.9 339.5 335.6 337.2 DAY 27 625356 625359 625360 625372 625374 ANIMAL NUMBER 625356 625359 625360 625372 625374 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 34 466.2 467.7 484.1 454.7 466.8 DAY 52 533.7 530.3 555.4 506.2 512.3 TEST DAYS SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 431.3 440.9 465.9 432.1 447.2 a Rat was not weighed. -40- H-24018: 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 20 330.1 295.3 329.0 314.2 317.4 DAY 24 342. 5 303. 8 334. 6 323. 5 325. 0 DAY 27 358.1 371. 7 399.1 a 317.2 331. 1 347.6 a 352.6 371. 5 402.3 ^a 336.9 350. 1 387.9 a 340.7 353. 5 376.3 a TEST DAYS DAY 76 DAY 90 DAY 94 427 .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 Rat was not weighed. -41 w H-24018: B iopersistence 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 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 -42- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625320 625330 625332 625341 625350 ANIMAL NUMBER 625320 625330 625332 625341 625350 DAY 1 279 .3 286 .7 300 .0 304 .9 257 .1 DAY 2 283 .0 286 .1 295 .9 309 .0 268 .5 DAY 8 308 .5 314 .0 324 .1 332 .4 279 .8 DAY 9 306 .3 312 .6 321 .8 320 .3 282 .5 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 287.7 291.1 304.1 317.5 267.3 288.4 296.5 308.8 320.0 272.2 TEST DAYS DAY 10 295 .9 304 .0 315 .0 325 .1 272 .6 299 .4 302 .3 322 .4 327 .5 276 .3 300.6 306.7 320.7 327.4 283.6 312.1 310.1 318.3 325.6 284.3 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -43- H-24018: 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 2 307 .8 306 .6 275 .5 291 .7 258 .6 DAY 8 337 .8 334 .3 304 .3 317 .3 302 .7 DAY 9 339 .6 337 .0 305 .9 312 .9 306 .5 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE KATS GROUP III TEST 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 -44. ' WH-24018: 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 IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625325 625328 625338 625340 625352 ANIMAL NUMBER 625325 625328 625338 625340 625352 282.8 315.6 264.6 304.0 285.5 DAY 8 318.6 352.0 287.4 347.0 .316.9 285.9 315.2 261.9 309.2 283.9 DAY 9 321.9 351.7 291.3 352.2 322.3 291.7 323.9 268.5 316.9 289.1 295.5 327.7 272.6 326.0 299.6 TEST DAYS DAY 10 DAY 13 327.4 357.5 290.2 359.2 321.0 331.6 367.4 291.1 368.1 329.6 303.2 335.0 275.7 329.8 307.9 DAY 20 355.8 400.2 320.4 389.3 355.7 309 .2 340 .4 281 .9 334 .1 308 .2 308.3 340.1 284.4 338.4 312.4 DAY 24 381 .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 -45- H-24018: 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 DAY 24 326.8 351.6 313.5 304.2 DAY 27 625324 625329 625333 625334 625344 ANIMAL NUMBER 332.5 337.9 336.1 299.6 336 .0 ^a 352.5 360.1 357.0 364.3 396 .4 a 315.8 316.7 319.3 325.8 349 .2 a 292.6 302.2 305.8 312.0 338 .6 ^a 280.7 285.2 289.3 295.8 309 .7 a DAY 34 DAY 52 TEST DAYS 380.3 423.3 375.2 368.8 330.4 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 Rat was not weighed. -46. H-24018: 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 VI TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625326 625335 625346 625347 625351 ANIMAL NUMBER 313.2 301.0 290.2 266.0 310.6 DAY 8 317.1 303.6 293.4 275.6 310.9 DAY 9 325.5 302.3 300.2 287.7 321.1 330.9 310.4 309.5 300.0 329.6 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 625326 625335 625346 625347 625351 ANIMAL NUMBER 311.5 294.2 292.9 330.5 351.1 DAY 34 308.9 285.3 290.8 330.1 357.8 DAY 52 297.5 274.0 288.4 333.0 350.5 274.2 266.3 296.9 355.7 380.0 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 625326 625335 625346 625347 625351 427.5 a 427.0 a 407.4 a 466.4 a 483.3 --a 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 da SD test da SD test da SD test da SD test da a Rat was not weighed. -47- ^aar H-24018: 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 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 -48- H-24018: 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 2 244 .1 261 .6 216 .3 228 .3 221 .7 DAY 8 272 .3 319 .8 246 .1 251 .0 260 .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 -49- H-24018: 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 HATS GROUP III TEST 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 50- H-24018: 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 2 232.9 247.4 244.3 237.9 218.2 DAY 8 277.8 240.1 291.4 289.0 251.3 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 a 266.9 ^a 306.4 ^a 301.0 a 257.0 -a 348.0 335.7 355.8 374.4 304.4 380.1 359.4 379.8 406.2 326.1 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 a Rat was not weighed. -51 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE HATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627757 627760 627765 627772 627781 ANIMAL NUMBER 221.2 226.2 228.3 245.2 219.0 DAY 8 225.4 232.0 235.3 256.4 226.3 DAY 9 236.3 244.4 247.1 266.2 236.5 241.1 251.7 251.6 277.0 242.9 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 DAY 24 254.4 258.3 264.7 291.0 256.1 DAY 34 627757 627760 627765 627772 627781 ANIMAL NUMBER 269.1 262.6 271.5 300.9 258.1 DAY 41 276.7 280.3 282.4 307.5 266.8 DAY 47 279.7 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 627757 627760 627765 627772 627781 402.8 437.7 422.9 478.4 389.9 419.7 448.9 444.8 494.3 408.2 436.7 456.5 455.6 513.5 421.0 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rat was not weighed. -52. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627762 627769 627775 627778 627783 ANIMAL NUMBER 227.2 231.1 224.7 213.1 216.4 DAY 8 235.0 241.1 235.7 217.4 218.2 DAY 9 248.7 255.9 246.8 221.3 226.6 257.3 264.2 254.7 225.5 232.0 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 627762 627769 627775 627778 627783 ANIMAL NUMBER 285.9 302.0 286.6 244.8 256.6 DAY 41 299.1 309.6 296.0 250.8 259.2 DAY 47 301.5 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 627762 627769 627775 627778 627783 438.7 439.9 a 479.7 474.6 a 433.2 451.1 ^a 356.8 373.5 ^a 409.6 426-9 a 453.5 512.2 460.8 386.0 449.0 473.0 511.4 477.4 393.3 473.8 471.6 531.4 478.6 398.3 474.8 a Rat was not weighed. -53. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627762 627769 627775 627778 DAY 81 DAY 90 500.3 566.2 510.4 410.1 523.3 603.2 537.5 436.5 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS TEST DAYS DAY 94 GROUP VI 528.6 609.6 552.8 445.3 SD test day 94 SD test day 94 SD test day 94 SD test day 94 -54- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627252 627253 627260 627273 627274 DAY 1 222.5 237.4 232.6 261.1 221.4 DAY 2 224.6 241.2 231.0 259.7 224.9 H-24018 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS GROUP I DAY 3 DAY 4 DAY 5 233.5 251.9 243.4 275.3 234.2 244.2 260.6 254.2 285.1 243.3 247.9 271.4 265.0 293.7 249.3 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -55- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats AMIMAL NUMBER 627246 627258 627261 627268 627269 ANIMAL NUMBER 627246 627258 627261 627268 627269 DAY 1 223 .7 226 .4 243 .0 260 .7 216 .2 DAY 2 229 .4 231 .9 251 .8 266 .9 223 .6 DAY 8 265 .1 260 .9 298 .7 315 .2 266 .8 DAY 9 274 .1 262 .4 305 .0 327 .9 266 .4 H-24018 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 233 .9 238 .0 262 .6 277 .8 232 .1 245.6 247.4 286.5 238.9 236.5 252 .9 253 .6 275 .8 296 .9 245 .6 260.2 263.0 283.5 305.9 254.2 262.4 257.8 295.3 314.5 264.1 DAY 10 283 .0 272 .7 314 .3 338 .6 273 .7 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -56. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627249 627262 627265 627266 627277 ANIMAL NUMBER 627249 627262 627265 627266 627277 DAY 1 252 .3 229 .8 233 .4 221 .4 250 .8 DAY 2 260 .7 235 .6 234 .8 230 .1 259 .7 DAY 8 317 .4 280 .5 276 .3 269 .7 309 .1 DAY 9 328 .4 289 .2 281 .5 281 .3 329 .2 H-24018 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III DAY 3 DAY 4 DAY ;5 DAY 6 DAY 7 270 .2 243 .5 242 .4 238 .9 264 .2 281 .2 253 .6 252 .4 248 .3 275 .1 295. 0 261. 4 256. 6 255. 4 287. 0 301 .5 268 .7 261 .9 261 .3 296 .3 312 .3 277 .8 272 .9 268 .8 308 .5 DAY 10 334 .2 299 .9 286 .7 289 .7 337 .8 DAY 13 357 .2 322 .7 312 .7 309 .1 367 .3 SD test day 13 SD test day 13 SD test day '.13 SD test day 13 SD test day ,13 57- H-24018; Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24018 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627247 627250 627255 627257 627259 ANIMAL NUMBER 211.7 240.0 250.5 232.5 231-5 DAY 8 228.1 251.6 260.6 235.8 236.5 233.7 259.5 271.4 272.9 243.4 251.6 269.5 280.2 255.7 247.8 259.5 279.6 285.5 261.4 253.8 266 .9 291 .4 295 .0 267 .6 260 .4 DAY 9 DAY 10 DAY 13 DAY 20 DAY 24 271. 7 296. 5 305. 2 278. 5 269. 4 627247 627250 627255 627257 627259 275.4 296.7 298.8 a 357.3 397 .3 SD test day 24 296.7 318.9 326.3 a 386.5 419 .9 SD test day 24 306.4 317.5 327.1 a 391.5 430 .7 SD test day 24 280.5 287.7 289.2 a 344.4 378 .3 SD test day 24 273.1 FD test day 9 a Rat was not weighed. -58. va^ H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24018 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627256 627264 627271 627275 627278 ANIMAL NUMBER 222 .7 229 .4 238 .7 238 .3 245 .9 DAY 8 230 .6 239 .2 244 .0 244 .0 249 .8 236.7 247.8 254.0 254.0 261.8 241.6 259.2 259.2 263.3 275.4 252 .6 264 .2 266 .0 272 .7 281 .0 258.1 277.8 274.9 276.5 286.6 260.7 283.3 281.4 282.2 295.8 DAY 9 DAY 10 DAY 13 DAY 20 DAY 24 DAY 34 627256 627264 627271 627275 627278 263 .6 279 .4 278.8 a 318 .9 -" 401.6 285 .4 302 .2 310.5 3. 364 .8 a 430.8 283 .0 296 .7 297.8 a 354 .2 422.1 283 .9 296 .8 302.0 a 341 .1 -a 395.9 . 297 .6 312 .3 318.8 a 369 .7 --a 461.0 ANIMAL NUMBER DAY 38 DAY 48 DAY 52 627256 627264 627271 627275 627278 419 .3 450 .2 444 .0 420 .4 484 .7 455 .6 481 .7 481 .1 444 .7 521 .5 468.5 489.0 495.9 455.6 535.5 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 a Rat was not weighed. -59- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627248 627251 627254 627263 627270 ANIMAL NUMBER DAY 1 240 .7 239 .7 234 .7 217 .7 230 .3 DAY 2 250 .0 248 .2 242 .6 219 .7 237 .3 DAY 8 DAY 9 H-24018 (TiEST SU.BSTA]NCE) INDIVIDUAL BODY WE:IGHTS (g) 'OP MALE RA'rs DAY 3 GRi3UP VI DAY 4 DAY 5 DAY 6 DAY 7 257.6 257.3 249.7 229.0 243.6 266.0 268.9 262.0 238.3 252.6 276 .8 282 .4 270 .5 243 .3 259 .2 287.5 293.7 283.9 252.4 269.4 295 .3 301 .0 289 .8 255 .6 277 .6 DAY 10 DAY 13 DAY 20 DAY 24 DAY 34 627248 627251 627254 627263 627270 ANIMAL NUMBER 627248 627251 627254 627263 627270 297 .1 314 .4 326.9 a 302 .6 326 .2 342.3 a 290 .7 306 .7 321.0 a 258 .0 274 .5 280.0 ^a 279 .2 290 .5 294.3 a 377 .8 a 401 .4 a 381 .9 ^a 330 .3 a 351 .6 -a 443. 6 485. 5 470. 6 396. 1 417.9 DAY 38 463 .9 509 .1 496 .3 410 .9 439 .8 DAY 48 489 .0 537 .6 523 .5 433 .9 466 .7 DAY 52 a a a a ^a DAY 55 510.0 559.7 552.2 451.8 484.4 DAY 61 531 .7 582 .5 565 .7 470 .6 501 .4 DAY 69 543.7 599.9 581.9 478.2 524.0 a Rat was not weighed. -60- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 82 DAY 88 H-24018 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI DAY 94 627248 627251 627254 627263 627270 565 .1 627 .7 608 .3 503 .4 548 .4 574 .9 643 .4 622 .6 512 .6 562 .0 591 .0 651 .5 628 .7 531 .5 579 .6 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 -61. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDIX B Individual Clinical Observations -62 'Gwsspwy SanNteed. Qow noS ew8M WftA Cffl H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Animal Number 627789 627801 627805 627810 627818 CORN OIL (NEGATIVE 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 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 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 -63 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Animal Number 627791 627794 627804 627812 627817 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IM 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 -64- Comapan^ Sanitized. Does 1 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study mRats DuPont-2927 CORN OIL:ACETONE (NEGATIVE CONTROL) Animal Number 625361 625363 625364 625380 625383 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 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 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 -65- Company Sanitized. Does not contain TSCA CB1 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 COHN OIL:ACETONE (NEGATIVE CONTROL) Animal Number 625354 625365 625371 625376 625381 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 625356 625359 625360 625372 625374 GROUP V Observation No abnormalities detected No abnormalities detected Mo 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 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 -66- Companif Sanitized. Does not contain TSCA CBl H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Animal Number 625322 625323 625336 625343 625349 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MAIjE 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 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 GROUP III 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 -67-Coiapahy Samteed. uoes *n c^.d&n 1 SCA CBi H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Animal Number 625325 625328 625338 625340 625352 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation Test Day No abnormalities detected No abnormalities detected No abnormalities detected Salivation No abnormalities detected No abnormalities detected 1-24 1-24 1-6, 8-24 7 1-24 1-24 Animal Number 625324 625329 625333 625334 625344 GROUP V 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 GROUP VI 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 -68- Company banned. Does nui contain TSCA CW H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Animal Number 627756 627767 627771 627773 627777 H-24020 (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 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 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 -69- Company Samtaeet. uoes <un c^a.n H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rate DuPont-2927 Animal Number 627759 627761 627770 627779 627786 H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV __ ____Observation__________________Test Day No abnormalities detected 1-24 No abnormalities detected No abnormalities detected 1-24 1-24 No abnormalities detected No abnormalities detected 1-24 1-24 Animal Number 627757 627760 627765 627772 627781 GROUP V Observation No abnormalities detected No abnormalities detected No abnormalities detected Wet perineum No abnormalities detected Wet perineum No abnormalities detected Test Day 1-52 1-52 1, 3-52 2 1, 3-52 2 1-52 Animal Number 627762 627769 627775 627778 627783 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 1-8, 10-94 9 1-94 1-94 81, 90, 94 1-23, 94 24, 34, 41. 47, 55. 61, 74, 81, 90 -70- Sanifced.DoesnotcontainTSCACBa Coropany H-24018: Biopersistence Screening 10-Dose Oral Gavage Study m Rats DuPont-2927 Animal Miunbet' 627252 627253 627260 627273 627274 H-24018 (TEST SUBSTANCE) 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 627246 627258 627261 627268 627269 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 627249 627262 627265 627266 627277 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 -71 Company Sanitized. Does not contain TSCA GBS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Animal Number 627247 627250 627255 627257 627259 H-24018 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Found dead ____Test Day 1-24 1-24 1-24 1-24 9 Animal Number 627256 627264 627271 627275 627278 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 627248 627251 627254 627263 627270 GROUP VI Observation No abnormalities detected Alopecia perineum No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Alopecia both front paws Test Day 1-82 88, 94 1-94 1-94 1-94 1-38, 69-94 48, 55, 61 -72. Company Sanitized, uoes noi contain TSCA CB1 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDIX C Analysis of Blood Fluorine Data "73 ' Company Samteed. Does nw contain TSCA CB1 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study inRats Terms and Calculations DuPont-2927 Terms: Active Fluorine containing compound % Active The % of formulation that is made up of fluorine containing components Mol Wt Active The molecular weight of the fluorine containing components (g/mole) Formulation Dose % F in Active The mg of formulation given per kg of animal body weight The % fluorine in the fluorine containing components of the formulation (weight basis) Mol Wt F 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/I 00) * 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/I 00) * 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]) * Formulation Dose 74Gompany Sanitized. Does not contain TSCACB8 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study m Rats Individual Animal Measurement: ppm F in blood The ppm fluoride measured in blood DuPont-2927 Individual Animal Calculations: ppm F in Blood minus Bkg 0.2 ppm The ppm fluoride measured in plasma minus the background fluoride measured in control animal blood, m this case the value was established at 0.2 ppm. ppm F in Blood normalized to 0.1 Dose The ppm fluoride minus background that would be expected if the mmol/kg 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]) * (ppm F in blood minus background) fJimolar equivalents of active in blood The ^molar [^mol/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]) * molar ratio active/F [mmol active/mmol F] * 1000 /imol/mmol -75- Company Sanitized. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 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 offluorinated 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 components) in the formulation - Molecular weight is the molecular weight of the active component in the formulation -76- Company Sanitized. Does not certain TSCA CB8 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H- 2 4019 Given: Mol^^t. Active (;g/mole): Fonnulation Dos<i (mg/kg): % Actiive (F Containing) in Formulation: 497 % Fin Active: 10 MolWt.F(g/mol): 100 Calculate!i Values: Dose^active (mg/kg): Dose./active(mm ole/kg): Formullation Dose! Normalisation Factor: 10 0.020 49.7 Dose F (mmol/kg): Molar Ratio (Active/P): Dose F (mg/kg): DuPont-2927 65 19 0.342 0.059 6.5 Rat# 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 625323 5 625336 5 625343 5 625349 5 51.7 38.0 38.7 78.5 37.1 Group II 625320 10 625330 10 625332 10 625341 10 625350 10 65.6 61.1 60.5 63.1 58.1 ppmF 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 mmoles/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 pimolar 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 -77Company Sanifced. Does not contain TSCA CB8 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat# Test Day Sample ppmF in blood Group in 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 625335 94 625346 94 625347 94 625351 94 12.4 16.4 13.1 11.0 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 blood Normalized to 0.1 mmoles/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-2927 ^moiar 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 -78Company Sanitized. Does not contain T8CA CBI H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H- 24K)20 Given: MorWt. Active (jg/mole): Form.ulation DOS(3 (mg/kg): %Ac;tive (F Coni;aimng)in Form ulation: 426 % F in Active: 20 Mol Wt. F (g/mol): 100 Calculated Values: Dose Active (mg/leg): Dose Active (mm.ole/kg): Formulation Dose; Normalization Factor: 20 0.047 42.6 Dose F (mmol/kg): Molar Ratio (Active/F): Dose F (mg/kg): DuPont-2927 69 19 0.726 0.065 13.8 Rat# Test Day Sample ppmF in blood Group I 627773 1 627771 1 627767 1 627756 1 627777 1 13.2 8.1 16.5 14.1 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 iainus 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 /(molar equivalenits 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 -79CompawSaniteed.Doesi H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat# Test Day Sample ppmF in blood Group ffl 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) ppmF 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 b lood Normalize dto 0.1 mmoles/kg Dose 92.23 43.67 27.69 65.18 52.61 19.17 31.31 24.28 20.66 13.85 1.49 0.85 1.70 1.49 1.28 0.21 * 0.00 0.00 * DuPont-2927 jamolar equivalents of active in blood 313.77 148.55 94.20 221.74 178.99 65.22 106.52 82.61 70.29 47.10 5.07 2.90 5.80 5.07 4.35 0.72 * 0,00 0.00 * -80- Conipany Sanitized. Does not contain TSCACBI H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-2-4018 Given: MolVft. Active (,g/mole): Fonnuilation Dos<3 (mg/kg): % Active (F Coni:ammg)m Formuilation: Calculated Values: DoSGjtVctive (mg/kg): Dose./Vctive (mmole/kg): Formulation Dose! Normalizalion Factor: 845 1000 34 340 0.402 248.5 % F in Active: Mol Wt. F (g/mol): Dose F (mmol/kg) : Molar Ratio (Acthre/F): Dose F (mg/kg): DuPont-2927 50 19 8.947 0.045 170 Rat# Group I 627252 627253 627260 627273 627274 Group I 627252 627253 627260 627273 627274 Group n 627246 627258 627261 627268 627269 Test Day Sample ppmF in blood 1 4.1 1 3.5 1 3.1 1 2.7 1 1.3 5 9.9 5 9.0 5 9.0 5 8.7 5 8.4 10 8.7 10 12.3 10 7.8 10 11.0 10 13.4 ppm F in blood minus Bkg 0.2 ppm 3.9 3.3 2.9 2.5 1.1 9.7 8.8 8.8 8.5 8.2 8.5 12.1 7.6 10.8 13.2 ppm F in blood Normalized to 0.1 mmoles/kg Dose 0.97 0.82 0.72 0.62 0.27 2.41 2.19 2.19 2.11 2.04 2.11 3.01 1.89 2.68 3.28 pimolar equivalents of active in blood 2.29 1.94 1.71 1.47 0.65 5.71 5.18 5.18 5.00 4.82 5.00 7.12 4.47 6.35 7.76 -81 Company Sanitized. Does not contain TSCA CBI H-24018: Biopersistence Screening 10-Dose Oral Gavage Study mRate Rat# Test Day Sample ppmF in blood Group in 627249 13 6.8 627262 13 6.4 627265 13 5.1 627266 13 5.2 627277 13 4.8 Group IV 627247 24 2.3 627250 24 2.8 627255 24 2.5 627257 24 2.5 627259 24 NA Group V 627256 52 1.7 627264 52 1.2 627271 52 1.3 627275 52 1.4 627278 52 1.0 Group VI 627248 94 0.2 627251 94 0.8 627254 94 0.7 627263 94 0.7 627270 94 0.8 NA Animal found dead on test day 9. No samples collected ppmF in blood minus Bkg 0.2 ppm 6.6 6.2 4.9 5.0 4.6 2.1 2.6 2.3 2.3 - 1.5 1.0 1.1 1.2 0.8 0.0 0.6 0.5 0.5 0.6 ppm F in blood Normalized to 0.1 mmoles/kg Dose 1.64 1.54 1.22 1.24 1.14 0.52 0.65 0.57 0.57 - 0.37 0.25 0.27 0.30 0.20 0.00 0.15 0.12 0.12 0.15 DuPont-2927 jtmolai equivaleiits of active in blood 3.88 3.65 2.88 2.94 2.71 1.24 1.53 1.35 1.35 - 0.88 0.59 0.65 0.71 0.47 0.00 0.35 0.29 0.29 0.35 -82- Coropany Sanitized. Does not eontain TSCA CBS