Document GQDowrGoqvOaYZ9DBGQydxY

TRADE SECRET Study Title H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID: DuPont-2927 AR226-3157 DuPont-2927 A u t h o r : Carol Finlay, B.A. S t u d y C o m p l e t e d o n : August 3,2000 P e r f o r m in g L a b o r a t o r y : E.I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 W ork R equest N um ber: Ser v ic e C o de N u m be r : Page 1 o f 82 Company Sanitized. Does not contain TSC CBF 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 o f data obtained from this study. Reviewed by: Gary W. Jepson, Ph.D. Senior Research Scientist -u1*&2.-ooo Date Reviewed by: c i c L fi. Jka u o L L k* Judith C. Stadler, Ph.D., D.A.B.T. Director, General Toxicology 3-A G -2QO Date Issued by Study Director: f d / l t r t t i n U u .............. Carol Finlay, B.A. Q Staff Scientist Utx> Dt -2Company Sanitized. Does not contain TSCA CBS 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....... 4 LIST OF APPENDICES 4 STUDY INFORMATION............................................. 5 STUDY PERSONNEL......,........................................... SUMMARY....... ............................................................. ...8 INTRODUCTION.................................. ....................... ...9 MATERIALS AND METHODS................................. A. Test Substance and Positive Controls................ B. Test Species.......................................................... C. Animal Husbandry'.............................................. 1. Housing Environment....................................................... 2. Feed and Water.................................................................. 3. Identification...................................................................... 4. Animal Health Monitoring Program............................... D. Quarantine and P retest......................................... E. Study D esign........................................................ F. Assignment to Groups and Study Start............... G. Dosing M aterial Preparation and Administration 1. Test Substance..................................................................... 2. Positive Controls................................................................ 3. Negative Controls............................................................... H. Body W eights........................................................ I. M ortality and Clinical Observations.................... J. Collection and Analysis o f Blood and Livers...... K. Treatment o f Fluorine D ata.................................. ...9 ...9 ...9 .10 ..10 ..10 ..10 ..10 11 11 11 11 ..11 .12 .12 12 12 12 13 RESULTS AND DISCUSSION........................ A. In-Life Toxicology.................................... B. Fluorine D ata............................................. 1. Factors Influencing Interpretation o f Analysis 2. Positive Controls................................................. 3. Test Substance...................................................... 14 14 14 .14 .14 .15 CONCLUSIONS 15 RECORDS AND SAMPLE STORAGE.................................................................................... 15 TABLES..................................................... ....................... ............................................................ 16 -3 o m p a iiy S a n itized . D o s s m l ecn sin TSCM. CBf H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 FIG U RES.......................................................................................................................................21 APPENDICES...... .................................................................................... ....................................26 LIST OF TABLES Page 1. MEAN BODY WEIGHTS................................................................................................................................................ 17 2. MEAN BODY WEIGHT G AINS.................................................................................................................................... 18 3. MEAN BLOOD FLUORINE LEVELS.............................................................. ...............................................................19 4. MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE......................................................20 LIST O F FIGURES Page 1. MEAN BODY W EIGHTS.................................................................................................................................................22 2. MICROMOLAR EQUIVALENTS IN RAT BLOOD.................................................. .................................................. 23 3. MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS.............................. 25 LIST OF APPENDICES Page A. INDIVIDUAL BO DY W EIGHTS.................................................................................................................................... 27 B. INDIVIDUAL CLINICAL OBSERVATIONS............................................................................................................... 62 C. ANALYSIS OF BLOOD FLUORINE D A TA ................................................................................................................ 73 Company Sanitized. Does not contain TSCA CBI >3 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORM ATION TEST SUBSTANCE: Substance Tested: DuPont-2927 Known Impurities.] 'J POSITIVE CONTROL: Substance Tested: Potassium perfluoroalkyl sulfonate Svnonvms/Codes: H-24019 Haskell Number: 24019 Comnositio -5 - H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Known Impurities:' 2 POSITIVE CONTROL: Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt Svnonvms/Codes: H-24020 * C-8 Perfluorooctanoate, ammonium salt PFOA Ammonium perfluorooctanoate Haskell Number: 24020 Composition: Known Impurities:M | 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: June 13, 1999 / September 14,1999 Company Sanitized. Coes nofconiato TSCA cm H -24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY PERSONNEL Study Director: Carol Finlay, B.A. ' Management: Judith C. Stadler, Ph.D., D.A.B.T. Primary Technician: 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 Hompany Sarflfead. Doe* not contain TSCA CB8 -7 - pBnpoqrSanlRr''** ----contain TSCACEfej H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 SUMMARY The objective o f 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 o f 5 male Crl:CD(SD)IGS BR rats each were exposed to 1000 mg/kg/day o f H-24018. The test substance was administered to one group o f five rats for 5 consecutive days and to 5 groups for 10 days. Approximately two hours after the first dose, blood was collected from the orbital sinus o f each rat from the 5-dose group. On selected days (5,1 0 ,1 3 ,2 4 , 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 o f dosing and then approximately every other week during the recovery period. Additionally, 2 negative controls, com oil and com 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 o f this rat is considered not to be due to the test substance, since no clinical signs o f toxicity were observed and no body weight loss occurred in any o f the rats during the study. The death o f this rat was probably caused by a dosing injury. No compound-related clinical signs o f toxicity 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 o f various parts o f 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 o f 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 o f body weight gain, the mean body weights and mean body weight gains o f 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 o f 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 o f H-24018 was 1402.6, compared to AUCINF/D values o f 566,479.1 and 70,789.6 for H-24019 and H-24020, respectively. Under the conditions o f this study, administration o f H-24018 to male rats for 10 consecutive days resulted in limited absorption and retention o f 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 .Compatif Sanitized. Doss not contain TSCA e i | H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 INTRODUCTION The objective o f this study was to define the potential o f H-24018 to be absorbed and to bioaccumulate in a mammalian system, as indicated by analytical determination o f total fluorine in blood and fiver. 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 o f a rangefinding study. In the rangefinding study, a group o f 5 male rats was dosed by oral gavage with H-24018 at a dosage o f 1000 mg/kg for 5 consecutive days. A group o f 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 o f 40 grams. The control group had an overall mean body weight gain o f 27 grams. A dosage o f 1000 mg/kg was selected for the limit dosage for this project. The limit dosage o f 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 o f those analyses. Results from analyses o f selected fiver 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 o f the study. No evidence o f 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 o f extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, and low incidence o f spontaneous diseases. -9 - H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 C. Animal H usbandry 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 o f 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 o f 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 o f 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 o f the study: Water samples are analyzed for total bacterial counts, and the presence o f 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 o f key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence o f these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity o f the study. The anim al health and environmental monitoring program is administered by the attending laboratory animal veterinarian. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in 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 o f disease or injury. The rats were observed daily for mortality and signs of illness, injury, or abnormal behavior. On the bases o f acceptable body weight gains and freedom from clinically apparent signs o f 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 Com oil Com oil:acetone Positive Controls H-24019 H-24020 Test Substance H-24018 Not applicable Com oil: acetone Com oil None Dosage fmg/kg/day) 0 0 10 20 1000 Number o f Animals 30 30 30 30 30 F. Assignment to Groups and Study Start After the quarantine period, the rats were selected on the bases o f adequate body weight gain, freedom from any clinical signs o f disease or injury, and a body weight within 20% of the mean. The selected rats were divided by computerized, stratified randomization into 6 groups o f 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 o f each rat. The rats were between 7 and 9 weeks of age at the initiation o f 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 M aterial Preparation and Administration 1. Test Substance H-24018 was dosed as received. The amount o f test substance each rat received was based on the body weight collected on each day o f dosing and the test substance density o f 1100 mg/mL. The test substance was stirred on a magnetic stir plate throughout the dosing procedure. -11 - iw p a f f S an itM l Peas iwt 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. Com oil was used as the vehicle for H-24020. It was necessary to dissolve H-24019 in acetone before suspending it in com oil. The ratio o f acetone to com oil was 20:80. The amount each rat received was based on the body weight collected on each day o f dosing and the suspension concentration. The dose volumes did not exceed 1 mL/100 g o f body weight. The dosing suspensions were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. 3. Negative Controls Com oil and com oiliacetone (80:20) were chosen as the negative controls because they were the vehicles for the positive controls. Each negative control rat received 1 mL o f com oil or com oil:acetone. These rats were dosed in a separate room from the rats dosed with the test substance or positive controls. H. Body W eights The rats were weighed on each day o f dosing and then weekly or every other week during the recovery period. I. M ortality and C linical 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 o f 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 Sampling Time 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 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- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 Five to 10 mL o f 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 o f 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 o f 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 o f 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 o f computing derived pharmacokinetic parameters from data files including area under the curve (AUCINF), Cmax and terminal half-life (Tl/2). The AUCINF (concentration x time) represents the area under the blood concentration curve from the time o f dosing extrapolated to infinity. The maximum observed concentration was Cmax (concentration). The points included in determination o f the terminal half-life were selected manually and given in units o f 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 _kt)) 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 o f variability and limited sensitivity o f the analytical method, the background was set at 0.2 ppm fluorine. Since 0.2 ppm was the fluoride concentration limit o f detection, any values listed as <0.2 ppm were excluded from further treatment. The percent o f fluorine and molecular weight o f the test substance and positive controls were used as provided by the sponsor. The measurements resulting from analysis o f total blood fluorine were used as received (ppm F) from Jackson Laboratory. Fluoride ion was converted to micromolar (piM) equivalents o f active component for further comparisons. The data and descriptions o f data manipulation and presentation are provided in Appendix C. -13- HoiHpanySanitised. 0am not contain T86AGSS 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 o f this rat is considered not to be due to the test substance, since no clinical signs o f toxicity were observed and no body weight loss occurred in any o f the rats during the study. The death o f this rat was probably caused by a dosing injury. No compound-related clinical signs o f toxicity were observed in the rats dosed with H-24018. Rats dosed with H-24019 exhibited diarrhea, salivation, alopecia, black ocular discharge, and staining o f various parts o f 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 com oil controls throughout the study. The mean body weights and mean body weight gains o f the rats with H-24018 were greater than the H-24020 positive control rats. Comparison o f body weights between the H-24018 rats and the H-24019 positive control and com oil/acetone 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 com oil/acetone negative control and H-24019 positive control rats. Accounting for the age difference at study start and the expected rate o f body weight gain, it appears that the mean body weights and mean body weight gains o f the rats dosed with H-24018 was comparable to the com oil/acetone 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 o f Analysis The data used in the kinetic analysis were derived from a limited screen, and therefore several caveats and considerations are important. A couple o f considerations o f 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 o f the terminal half-life. A more complete list o f considerations is shown in Appendix C. 2. Positive Controls The positive controls were H-24019 and H-24020. The H-24019 normalized fiM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state -14- ISompwpSawfflsed B&mnotrafatm7&m BL 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 o f 40.5 days. The H-24019 AI was 59.0 and the BI was 58382.2. The H-24020 normalized jlM equivalents in rat blood peaked after 5 days o f 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 o f 8.3 days. The H-24020 AI was 12.5 and the BI was 6497.5. For each o f the positive controls, blood was sampled at seven time points throughout the study, with only four o f 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 fiM 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 o f 29.1 days. Blood was sampled at seven time points throughout the study, with only four o f 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 o f H-24018 was 1402.6 as compared to AUCINF/D values o f 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 o f toxicity 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 o f this study, administration o f H-24018 to male rats for 10 consecutive days resulted in limited absorption and retention o f 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- OEtxpmg Santtfeed. Does no ***# H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________ DuPont-2927 TABLES -16- !IW rnm m esnlfifft TSCAC il 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 Negative Controls Com Oil 229.8 234.1 241.4 250.4 259.3 266.8 271.2 281.9 285.2 295.9 321.1 350.5 378.3 Com Oil:Acetone 297.6 299.6 309.4 314.3 321.5 326.0 334.9 339.2 345.2 351.7 350.7 369.8 401.0 - 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- jjSpmpanf to iilto d O&mnot ewrtaln YS e f t 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 Com Oil 29.5 66.1 25.2 82.4 168.8 271.0 Com Oil:Acetone 23.9 54.1 - 1.0 49.3 175.9 216.1 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-24018 32.6 71.4 27.8 100.6 182.9 290.5 - 18- Pompanjr SanHtasd. Bom nocontain TSCA CB H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rate DuPont-2927 TABLE 3 MEAN BLOOD FLUORINE LEVELS Test Days 1 5 10 13 24 52 94 Negative Controls Cora Oil ppm <0.2 <0.2 <0.2 0.2a (0.1) <0.2 0.3a (0.1) 0.3a (0.1) Com Oil:Acetone ppm <0.2 <0.2 0.3a (0.1)b 0.6C(0.6) 0.2d <0.2 <0.2 Positive Controls 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-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) a Mean o f 4 o f the 5 values. One o f the values was below the LOD. b Standard deviation is in parentheses. c Mean o f 3 o f the 5 values. Two o f the values were below the LOD. d One value. Four o f the values were below the LOD. - 19- 0@not eoniaSn TSCA(TOI 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 fiM 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 fiM 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 fiM F Equivalents 1.6 (0.62)a 5.2 (0.33) 6.1 (1.39) 3.2 (0.52) 1.4b (0.12) 0.7 (0.15) 0.3 (0.15) a Standard deviation is in parentheses. b Mean o f 4 o f the 5 values. One o f the rats was found dead on test day 9. -20- @tpanf SmHteed. Bom swSeomtaSss TSC c m H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 FIGURES -21 - Igempwf H w ftted Benot sfiata W CA Bt H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 4 00 -, 350 - 300 - 3 250 - I? "&O 200 eS I 150- 100 - 50 - 00 FIGURE 1 MEAN BODY WEIGHTS (g) ...O..N eg a tiv e C ontrol (Corn O il) --O--N e g a tiv e C ontrol (Corn O ih A c e to n e ) --A--P o sitiv e C ontrol H -2 4 0 1 9 X Positive Control H -24020 --X - T est S ub stan ce H -2 4 0 1 8 -, I | j 24 6 8 Test Days |---------------j 10 12 -22- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 FIGURE 2 MICROMOLAR EQUIVALENTS IN RAT BLOOD A. N orm alized R at Blood H-24019 ftM Equivalents Resulting from a 10-Day O ral Gavage Micromolar (jM ) equivalents o f 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. N o rm a liz e d R a t Blood H-24020 ftM E q u iv a le n ts R esulting from a 10-Day O ral G avage Micromolar (/xM) equivalents o f 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- offlipwsf Swfcfdl" D@@ ondate TS6A CBS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 C . N orm alized R at Blood H -24018 ftM E quivalents R esulting from a 10-Day Oral Gavage Micromolar (/M) equivalents o f 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- Company Sanitized. Does not contain TS^a cm H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ________________ DuPont-2927 FIGURE 3 MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS '.^ t',H "24018 and P o sitiv e C o n tro l N ^ ^ iajfeed Blood AUCiNF/D in Ra ts Resulting ,,'I-: from 10-day Oral Gavage Company Sanitized. Does not contain TSC C3I - -25 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDICES -26- wpany fSanfflged. D@$not contain TSC cm H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDIX A Individual Body Weights -27- Oompany Sanitized. Does not contato TS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats INDIVIDUAL BODY WEIGHTS EXPLANATORY NOTES ABBREVIATIONS: FD - found dead SD - sacrificed by design DuPont-2927 -28- 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 I DAY 3 DAY 4 DAY 5 627789 627801 627805 627810 627818 222.6 215.9 249.4 230.5 229.7 220.7 209.1 250.1 231.6 228.6 223.5 223.5 263.3 241.9 239.1 236.9 233.5 273.6 254.4 252.4 247.7 240.1 283.7 265.0 260.2 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 DuPont-2927 8fey>W3SAMppsw |0H 8@Sfl " -29- 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 II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627793 627797 627799 627800 627808 214.8 239.9 228.9 246.9 235.6 224.4 245.9 231.4 252.8 241.6 232.1 257.3 242.3 265.2 250.1 243.1 267.5 246.2 279.3 262.1 249.9 280.7 258.9 283.6 270.4 257.0 288.0 261.5 295.5 284.1 262.0 290.9 266.4 300.1 287.4 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 627793 627797 627799 627800 627808 270.6 303.9 280.2 312.0 303.6 274.9 305.1 284.3 315.4 305.7 283.5 324.2 294.7 331.3 321.8 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 DuPont-2927 te a TOSA u|B|uo3 6usaofl -30- taovs& L 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 III TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627796 627806 627807 627809 627816 239.8 227.3 221.5 234.2 235.0 249.7 234.2 225.9 240.4 238.6 253.6 239.4 237.1 252.5 250.0 252.5 242.9 244.1 260.3 258.9 267.7 258.5 254.9 272.8 265.1 277.2 270.4 257.7 277.9 274.5 289.8 275.2 260.3 282.6 278.6 ANIMAL NUMBER 627796 627806 627807 627809 627816 DAY 8 285.9 282.8 270.7 297.9 291.8 DAY 9 TEST DAYS DAY 10 DAY 13 290.1 285.8 274.7 300.2 296.4 300.0 302.8 280.7 311.9 305.7 322.7 320.3 305.2 336.2 330.2 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 I 1 -31 - DuPont-2927 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 IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627791 627794 627804 627812 627817 231.1 224.4 224.7 221.9 233.5 235.4 233.0 228.8 231.5 231.3 241.2 238.1 230.6 236.3 229.0 246.8 247.8 243.0 245.7 244.8 255.9 260.0 251.7 255.5 255.4 262.6 267.4 252.9 264.9 266.4 268.4 270.1 259.5 269.5 270.0 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 627791 627794 627804 627812 627817 274.0 286.1 267.2 278.2 277.8 279.1 289.5 270.6 282.7 280.4 285.0 295.4 278.1 294.3 294.6 __a _a --a __a _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 DuPont-2927 a Rat was not weighed. H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627792 627798 627813 6 2 /814 627815 ANIMAL NUMBER 627792 627798 627813 627814 627815 ANIMAL NUMBER 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 DAY 47 tf CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF HALE RATS GROUP V TEST DAYS 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 .7 252.5 272.9 239.8 233.8 265.0 264.4 281.9 243.2 236.7 270.0 268.1 286.2 246.8 240.6 274.1 TEST DAYS DAY 10 DAY :13 DAY 20 DAY 24 DAY 34 295.2 315.7 267.9 257.3 303.3 __a __a _a __a __a 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 TEST DAYS DAY 52 627792 627798 627813 627814 627815 456.5 470.7 395.0 358.7 462.6 482.3 496.9 407.2 372.7 487.3 496.8 517.3 416.8 386.9 505.9 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. DuPont-2927 Ooes not conta1 TSCA C 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 DAYS 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 627795 627811 627819 627820 627821 DAY 8 261.4 273.0 304.5 281.3 272.6 DAY 9 265.0 276.7 308.0 282.9 275.3 TEST DAYS DAY 10 DAY 13 270.7 285.6 324.3 292.4 281.8 _a _a _a __a _a DAY 20 308.0 337.8 399.7 367.2 330.1 DAY 24 329.8 360.6 431.2 393.0 352.9 DAY 34 362.0 396.4 472.5 445.5 385.6 ANIMAL NUMBER 627795 627811 627819 627820 627821 DAY 41 384.2 422.0 505.9 478.0 411.8 DAY 47 398.0 439.3 534.9 504.1 426.1 TEST DAYS DAY 52 DAY 55 __a 411.1 --a _a _a _a 455.4 551.7 529.7 439.1 DAY 61 435.7 463.0 574.4 547.1 459.0 DAY 68 440.7 473.6 588.6 558.6 463.3 a Rat was not weighed. -34- DuPont-2927 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 90 DAY 94 W 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 SD test SD test SD test SD test DuPont-2927 D@not contain TSC CM -35- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 ilP r CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I DAY 3 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 DuPont-2927 w a il &iipwm Q -36- 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 II test days DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625355 625362 625367 625368 625378 280.0 290.3 301.5 312.3 309.6 282.0 294.1 307.8 318.0 306.4 293.0 303.5 315.6 326.0 327.3 297.9 308.9 320.1 336.4 328.4 306.1 313.8 327.7 343.7 333.7 311.2 315.8 333.7 347.5 336.6 315.7 323.8 345.6 360.8 346.9 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 625355 625362 625367 625368 625378 321.7 323.9 349.1 364.2 350.1 320.9 332.3 352.4 380.8 354.2 333.6 332.9 363.8 386.7 365.4 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 DuPont-2927 Sanitized. Does not contain TSCA CW -37- J 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 III TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625357 625358 625370 625384 625385 311.9 285.7 307.9 294.2 297.6 308.8 286.3 316.7 300.0 304.6 320.3 295.5 329.5 312.0 313.0 328.4 296.4 337.9 309.3 320.4 340.5 306.0 346.9 316.3 331.9 343.3 302.7 349.8 314.1 334.0 361.1 309.2 364.1 321.4 345.4 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 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 DuPont-2927 Oeno*contata TSCCffi -38- 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 IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625354 625365 625371 625376 625381 303.0 327.7 294.5 289.7 297.8 304.1 328.7 299.0 289.9 297.5 310.5 337.9 313.4 302.2 301.7 312.2 343.6 318.3 304.9 305.6 319.3 357.7 321.8 313.2 314.4 324.7 357.2 329.6 317.0 314.4 331.2 364.3 338.5 322.8 321.0 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 625354 625365 625371 625376 625381 336.2 370.5 340.3 331.5 321.6 337.0 379.3 345.9 339.0 329.1 348.8 381.2 351.2 345.2 331.6 355.8 407.1 372.6 363.2 341.2 377.7 438.2 413.0 399.7 358.2 396.1 459.6 447.3 432.9 381.8 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 DuPont-2927 WslS M|8su0*WMm m psiMs - 39- ffl^W@SI.WJ8p 0S P 68800S l" H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 CORN OILACETONE (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 291.7 297.9 296.8 294.8 295.9 291.2 303.3 300.7 298.8 302.2 304.5 314.0 308.5 310.6 312.1 307.0 315.0 312.7 317.4 321.7 316.9 323.5 319.1 324.0 323.4 319.8 323.5 332.2 329.0 328.3 323.9 333.9 339.5 335.6 337.2 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 DAY 27 625356 625359 625360 625372 625374 328.8 339.1 347.9 344.7 .338.0 334.7 346.2 354.3 348.8 344.6 341.3 353.6 363.4 355.1 350.8 361.9 372.9 381.7 373.1 371.5 396.7 410.0 419.8 395.1 401.3 __a _a _a _a 431.3 440.9 465.9 432.1 447.2 ANIMAL NUMBER 625356 625359 625360 625372 625374 DAY 34 466.2 467.7 484.1 454.7 466.8 DAY 52 TEST DAYS 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 Rat was not weighed. 40 DuPont-2927 gffwagjLMtBPQai8898^ 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 VI TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625353 625369 625373 625375 625377 305.9 281.7 294.6 289.1 293.9 307.3 279.9 301.3 291.0 294.8 316.8 289.7 312.7 298.0 302.9 325.0 286.9 317.0 307.0 312.0 331.7 292.8 325.4 309.6 318.0 330.1 295.3 329.0 314.2 317.4 342.5 303.8 334.6 323.5 325.0 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 DAY 27 625353 625369 625373 625375 625377 344.7 307.3 340.6 324.7 325.0 352.7 311.9 348.2 336.0 333.7 358.1 317.2 352.6 336.9 340.7 371.7 331.1 371.5 350.1 353.5 399.1 347.6 402.3 387.9 376.3 --a __a _a _a 427.7 374.9 444.6 432.3 405.2 ANIMAL NUMBER DAY 34 DAY 52 TEST DAYS DAY 76 DAY 90 DAY 94 625353 625369 625373 625375 625377 453.3 388.2 477.1 455.6 419.1 _a __a --a _a 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 - DuPont-2927 H-24018: B iopersistence 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 I DAY 3 DAY 4 DAY 5 625322 625323 625336 625343 625349 290.3 285.0 316.5 288.2 303.6 283.7 285.5 313.5 281.9 295.9 291.5 290.2 320.1 286.8 308.2 298.9 290.9 327.1 291.0 310.1 298.0 294.0 329.0 293.7 318.1 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 DuPont-2927 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 II TEST 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 8 283.0 286.1 295.9 309.0 268.5 287.7 291.1 304.1 317.5 267.3 288.4 296.5 308.8 320.0 272.2 DAY 9 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 308.5 314.0 324.1 332.4 279.8 306.3 312.6 321.8 320.3 282.5 312.1 310.1 318.3 325.6 284.3 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 DuPont-2927 -43- 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 III TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625321 625327 625342 625345 625348 307.0 311.4 273.7 291.2 265.5 307.8 306.6 275.5 291.7 258.6 311.4 312.5 282.3 297.3 268.0 319.6 317.8 285.8 303.3 275.2 322.3 318.3 290.8 306.4 280.1 328.4 319.3 294.2 310.4 285.2 335.2 327.6 326.6 316.1 290.6 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 625321 625327 625342 625345 625348 337.8 334.3 304.3 317.3 302.7 339.6 337.0 305.9 312.9 306.5 346.1 336.3 307.8 315.0 309.7 371.2 355.3 324.4 336.9 336.4 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 DuPont-2927 S anK M . @ m m l Mutate TS6 -44- ! H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats w 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 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 .316.9 285.9 315.2 261.9 309.2 283.9 291.7 323.9 268.5 316.9 289.1 295.5 327.7 272.6 326.0 299.6 303.2 335.0 275.7 329.8 307.9 309.2 340.4 281.9 334.1 308.2 308.3 340.1 284.4 338.4 312.4 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 321.9 351.7 291.3 352.2 322.3 327.4 357.5 290.2 359.2 321.0 331.6 367.4 291.1 368.1 329.6 355.8 400.2 320.4 389.3 355.7 381.3 423.9 339.9 419.0 375.3 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 DuPont-2927 |ou m m 103 -45- H30V3S1. UJBUOOp u s0 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625324 625329 625333 625334 625344 ANIMAL NUMBER 625324 625329 625333 625334 625344 ANIMAL NUMBER DAY 1 303.0 318.9 292.4 274.6 262.9 DAY 2 298.5 316.3 291.2 277.0 262.2 DAY 8 332.5 352.5 315.8 292.6 280.7 DAY 9 337.9 360.1 316.7 302.2 285.2 DAY 34 DAY 52 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 305.8 326.5 297.8 286.4 269.0 312.1 335.4 304.1 290.6 273.5 320.1 338.3 307.7 292.0 274.5 322.8 346.7 312.0 300.1 284.1 326.8 351.6 313.5 304.2 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 DAY 27 336.1 357.0 319.3 305.8 289.3 299.6 336.0 364.3 396.4 325.8 349.2 312.0 ' 338.6 295.8 309.7 --a _a _a __a _a 380.3 423.3 375.2 368.8 330.4 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 Rat was not weighed. 46 DuPont-2927 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 f 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 DuPont-2927 625326 625335 625346 625347 625351 313.2 301.0 290.2 266.0 310.6 317.1 303.6 293.4 275.6 310.9 325.5 302.3 300.2 287.7 321.1 330.9 310.4 309.5 300.0 329.6 336.2 315.1 313.5 308.5 338.2 336.0 299.6 318.6 316.6 341.6 293.6 326.0 326.9 340.7 ANIMAL NUMBER 625326 625335 625346 625347 625351 DAY 8 311.5 294.2 292.9 330.5 351.1 DAY 9 308.9 285.3 290.8 330.1 357.8 TEST DAYS DAY 10 DAY 13 297.5 274.0 288.4 333.0 350.5 274.2 266.3 296.9 355.7 380.0 DAY 20 335.4 340.1 352.5 402.1 413.7 DAY 24 __a _a _a _a --a DAY 27 390.7 393.8 388.2 436.2 452.4 ANIMAL NUMBER 625326 625335 625346 625347 625351 DAY 34 427.5 427.0 407.4 466.4 483.3 DAY 52 _a --a _a __a _a TEST DAYS DAY 62 DAY 76 497.5 506.0 485.8 568.6 559.0 540.0 538.6 523.7 617.8 596.8 DAY 80 547.9 547.7 532.8 618.1 607.2 DAY 90 565.2 562.1 542.4 632.6 624.0 DAY 94 577.5 579.5 556.9 638.9 636.8 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 P i 883,tosa uffuoo m a Rat was not weighed. -47- 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 I DAY 3 DAY 4 DAY 5 627756 627767 627771 627773 627777 227.2 227.0 219.5 246.8 225.4 226.4 231.8 214.7 244.4 221.4 233.4 237.9 226.1 252.7 232.9 238.6 245.8 233.5 263.8 239.4 237.5 248.6 239.6 262.3 236.0 SD test day- 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 DuPont-2927 1 Does notcontain TSCA CB -48- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL DAY 1 DAY 2 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 627768 627774 627776 627782 627785 ANIMAL NUMBER 236.9 258.6 212.0 221.6 220.3 DAY 8 244.1 261.6 216.3 228.3 221.7 245.2 275.8 223.7 237.3 234.7 256.5 283.4 226.6 239.0 238.4 DAY 9 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 627768 627774 627776 627782 627785 272.3 319.8 246.1 251.0 260.7 291.9 326.3 252.3 264.0 268.5 293.5 338.2 254.4 268.2 267.9 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 DuPont-2927 anfflzed. Boss not enn TSCA cm -49- 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 HALE RATS GROUP III TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 627758 627763 627764 627787 627788 ANIMAL NUMBER 235.1 212.1 230.0 235.1 219.4 DAY 8 240.3 218.3 235.0 241.0 222.3 248.6 226.5 246.1 250.1 232.2 255.8 234.3 251.3 257.2 239.6 DAY 9 TEST 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 627758 627763 627764 627787 627788 280.6 255.2 275.8 221.7 274.1 283.6 261.2 281.9 235.7 276.9 295.8 272.5 288.0 234.7 284.8 308.7 285.8 306.7 234.9 309.4 SD test day 13 SD test day 13 SDtest day 13 SD test day .13 SDtest day 13 DuPont-2927 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 IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627759 627761 627770 627779 627786 ANIMAL NUMBER 226.6 240.7 235.6 231.5 211.6 DAY 8 232.9 247.4 244.3 237.9 218.2 241.4 257.1 253.6 252.6 222.5 250.9 262.5 260.1 261.4 230.4 254.2 266.7 267.7 262.3 230.6 262.1 259.2 277.3 273.8 240.6 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 265.4 262.6 279.8 277.5 244.1 627759 627761 627770 627779 627786 277.8 240.1 291.4 289.0 251.3 284.9 257.4 301.3 293.4 255.8 288.0 266.9 306.4 301.0 257.0 _a __a _a __a _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 DuPont-2927 a Rat was not weighed. / H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in R ats______________ 1C? ANIMAL NUMBER DAY 1 DAY 2 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627757 627760 627765 627772 627781 221.2 226.2 228.3 245.2 219.0 225.4 232.0 235.3 256.4 226.3 236.3 244.4 247.1 266.2 236.5 241.1 251.7 251.6 277.0 242.9 247.9 260.2 256.4 276.9 247.2 252.2 254.5 261.1 285.0 251.6 254.4 258.3 264.7 291.0 256.1 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY1 20 DAY 24 DAY 34 627757 627760 627765 627772 627781 269.1 262.6 271.5 300.9 258.1 276.7 280.3 282.4 307.5 266.8 279.7 287.3 288.1 317.0 271.2 _a __a __a __a __a 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 ANIMAL NUMBER DAY 41 DAY 47 TEST DAYS DAY 52 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 IBS V381 "I"0 G a Rat was not weighed. ! -52- DuPont-2927 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 _____________ __ ___________________________________________________________________ DuPont-2927 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 227.2 231.1 224.7 213.1 216.4 235.0 241.1 235.7 217.4 218.2 248.7 255.9 246.8 221.3 226.6 257.3 264.2 254.7 225.5 232.0 263.0 272.3 265.7 230.1 238.0 275.1 286.3 276.7 236.9 243.9 279.7 289.9 280.2 240.4 247.6 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 DAY 20 DAY 24 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 47 301.5 312.5 304.5 255.1 264.3 -a -a -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 438.7 479.7 433.2 356.8 409.6 439.9 474.6 451.1 373.5 426.9 _a _a _a __a _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 IP@ W a S A UtP0 P 0 0 a Rat was not weighed. -53- 1*? H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 81 DAY 90 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS TEST DAYS DAY 94 GROUP VI 627762 627769 627775 627778 500.3 566.2 510.4 410.1 523.3 603.2 537.5 436.5 528.6 609.6 552.8 445.3 SD test day 94 SD test day 94 SD test day 94 SD test day 94 DuPont-2927 Bgg> tgggpM p tt *G -54- 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 I DAY 3 DAY 4 DAY 5 627252 627253 627260 627273 627274 222,5 237.4 232.6 261.1 221.4 224.6 241.2 231.0 259.7 224.9 233.5 251.9 243.4 275.3 234.2 244.2 260.6 254.2 285.1 243.3 247.9 271.4 265.0 293.7 249.3 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 DuPont-2927 Itavira g j, m$m|0&8bQ -55- H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627246 627258 627261 627268 627269 ANIMAL NUMBER 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 DAY 9 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 627246 627258 627261 627268 627269 265,.1 260,.9 298..7 315..2 266,.8 274,,1 262,,4 305..0 327,.9 266,,4 283.0 272.7 314.3 338.6 273.7 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 DuPont-2927 IQU*Q 109 WM -56- 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 III DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627249 627262 627265 627266 627277 252.3 229.8 233.4 221.4 250.8 260.7 235.6 234.8 230.1 259.7 270.2 243.5 242.4 238.9 264.2 281.2 253.6 252.4 248.3 275.1 295.0 261.4 256.6 255.4 287.0 301.5 268.7 261.9 261.3 296.3 312.3 277.8 272.9 268.8 308.5 ANIMAL NUMBER DAY 8 DAY 9 DAY 10 DAY 13 627249 627262 627265 627266 627277 317.4 280.5 276.3 269.7 309.1 328.4 289.2 281.5 281.3 329.2 334.2 299.9 286.7 289.7 337.8 357.2 322.7 312.7 309.1 367.3 SD test day- 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 DuPont-2927 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 211.7 240.0 250.5 232.5 231.5 228.1 251.6 260.6 235.8 236.5 233.7 259.5 271.4 272.9 243.4 251.6 269.5 280.2 255.7 247.8 259.5 279.6 285.5 261.4 253.8 266.9 291.4 295.0 267.6 260.4 271.7 296.5 305.2 278.5 269.4 ANIMAL NUMBER DAY 8 DAY 9 DAY 10 DAY 13 DAY 20 DAY 24 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 DuPont-2927 a Rat was not weighed. 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 222.7 229.4 238.7 238.3 245.9 230.6 239.2 244.0 244.0 249.8 236.7 247.8 254.0 254.0 261.8 241.6 259.2 259.2 263.3 275.4 252.6 264.2 266.0 272.7 281.0 258.1 277.8 274.9 276.5 286.6 260.7 283.3 281.4 282.2 295.8 ANIMAL NUMBER DAY 8 DAY 9 DAY 10 DAY 13 DAY 20 DAY 24 DAY 34 627256 627264 627271 627275 627278 263.6 285.4 283.0 283.9 297.6 279.4 302.2 296.7 296.8 312.3 278.8 310.5 297.8 302.0 318.8 _a _a _a __a 318.9 364.8 354.2 341.1 369.7 __a _a _a _a __a 401.6 430.8 422.1 395.9 461.0 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. DuPont-2927 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 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS DAY 3 GROUP 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 297.1 302.6 290.7 258.0 279.2 314.4 326.2 306.7 274.5 290.5 326.9 342.3 321.0 280.0 294.3 _a __a __a __a _a 377.8 401.4 381.9 330.3 351.6 --a _a _a _a _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 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. DuPont-2927 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 t e s t day 94 SD t e s t d ay 94 SD t e s t d ay 94 SD t e s t d ay 94 SD t e s t d ay 94 DuPont-2927 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDIX B Individual Clinical O bservations - 62smpanf Sanitized. Deenot contain dS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP I _________ Observation ______________ ________ Test Day 627789 627801 627805 627810 627818 No abnormalities detected No abnormali ties detected No abnormalities detected No abnormalities detected No abnormalities detected 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 GROUP III _______ Observation _________________ Test Day No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected 1-13 1-13 1-13 1-13 1-13 - 63Company SanHlMd. 9oes m iecnrtain TSC H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP IV ____ Observation________________________ Test Day 627791 627794 627804 627812 627817 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 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 Company Sanitized. Does not contain IS C A CBI H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ______________________ DuPont-2927 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP I _________ Observation___________ _____________ Test Day 625361 625363 625364 625380 625383 , No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected 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 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 T S C A CBS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal NuTTiH^r-________________ GROUP IV Observation________________________ Test Day 625354 625365 625371 625376 625381 . No abnormalities No abnormalities No abnormalities No abnormalities No abnormalities detected detected detected detected 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 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 GROUP VI _________Observation________________________ Test Day No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected 1-94 1-94 1-94 1-94 1-94 Company Sanitized. D oes not contain TSC A CB1 H -24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN HALE RATS Animal Number 625322 625323 625336 625343 625349 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 - 67cenami SCACB Gompstaf Sanitized, uoes m H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number 625325 625328 625338 625340 625352 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 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 Company b a n n e d . Does not contain T S C C B i H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP I _______________ Observation______________ __________ Test Day 627756 627767 627771 627773 627777 No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected 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- 6 1-8, 10- 9 -69Sanitized. Does 01 couiam 1 S C A CB1 Compan# H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rate DuPont-2927 H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Animal Number____________________ Observation________________________ Test Day 627759 627761 627770 627779 627786 No abnormalities No abnormalities No abnormalities No abnormalities No abnormalities detected detected detected detected detected 1-24 1-24 1-24 1-24 1-24 Animal Number 627757 627760 627765 627772 627781 Animal Number 627762 627769 627775 627778 627783 GROUP V ________ Observation ___________________Test Day No abnormalities detected No abnormalities detected No abnormalities detected Wet perineum No abnormalities detected Wet perineum No abnormalities detected 1-52 1-52 1, 3-52 2 1, 3-52 2 1-52 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 \ \) not contain TSCA CBS Company Sanitized. Does H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 H-24018 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number 627252 627253 627260 627273 627274 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 627246 627258 627261 627268 627269 GROUP II 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 CBi H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 H-24018 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP IV Observation_________________ _______Test Day 627247 627250 627255 627257 627259 No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Found dead 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. Does not contain TSCA OBI H -24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2927 APPENDIX C Analysis of Blood Fluorine Data Company Sanitized. Does noi contain TSC OBI H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Terms and Calculations DuPont-2927 Terms: Active % Active Mol Wt Active Formulation Dose % F in Active Mol Wt F Fluorine containing compound The % of formulation that is made up o f fluorine containing components The molecular weight o f the fluorine containing components (g/mole) The mg of formulation given per kg of animal body weight The % fluorine in the fluorine containing components o f 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) * 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 o f animal body weight = (% F in active/100) * Dose Active [mg/kg] Dose F (mmol/kg) The mmole of fluorine administered per kg o f animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol] Molar Ratio (Active/F) The moles of fluorine containing compound per mole o f 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 o f Active [mmol/kg] / Dose Active [mmol/kg]) * Formulation Dose -74Company Sanitized. Does not contain TSCACBI H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in 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. In this case the value was established at 0.2 ppm. ppm F in Blood The ppm fluoride minus background that would be expected if the normalized to 0.1 mmol/kg active dose was 0.1 mmol/kg instead of the actual active dose. This Dose 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) /molar 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 /mol/mmol -75Company Sanitized. Does not contain TSC A CBS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Factors Influencing Interpretation of Kinetic Analysis DuPont-2927 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 bejustified 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 contain TSCA CBS H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H- 24019 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 497 % F in Active: 10 Mol Wt. F (g/mol): 100 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 10 0.020 49.7 Dose F (mmol/kg): Molar Ratio (Active/F): Dose F (mg/kg): DuPont-2927 65 19 0.342 0.059 6.5 Rat # Test ppm F Day in Sample blood Group I 625322 625323 625336 625343 625349 1 1 1 1 1 1.0 1.3 2.6 2.6 2.9 Group I 625322 625323 625336 625343 625349 5 5 5 5 5 51.7 38.0 38.7 78.5 37.1 Group II 625320 625330 625332 625341 625350 10 10 10 10 10 65.6 61.1 60.5 63.1 58.1 ppm Fin 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 /xm o la r 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 C o m p ly Sanitized. Does not contain TSC A OB! H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat # Test ppm F Day in Sample blood Group IH 625321 625327 625342 625345 625348 13 13 13 13 13 77.2 61.5 64.6 57.0 62.4 Group IV 625325 625328 625338 625340 625352 24 24 24 24 24 40.3 46.3 43.2 39.1 41.9 Group V 625324 625329 625333 625334 625344 52 52 52 52 52 24.7 29.8 27.3 29.1 23.8 Group VI 625326 625335 625346 625347 625351 94 94 94 94 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 DuPont-2927 pim o la r equivalents of active in blood 382.69 304.66 320.07 282.30 309.13 1184.62 943.08 990.77 873.85 956.92 199.30 229.12 213.71 193.33 207.25 616.92 709.23 661.54 598.46 641.54 121.77 147.11 134.69 143.63 117.29 - 376.92 455.38 416.92 444.62 363.08 60.63 80.51 64.11 53.68 56.66 187.69 249.23 198.46 166.15 175.38 -78Company Sanitized. Does no, contain TSC A CBI H-24018: Biopersistence Screening 1Q-Dose Oral Gavage Study in Rats Data for H- 24020 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 426 % F in Active: 20 Mol Wt. F (g/mol): 100 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/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 ppm F Day in Sample 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 627771 627767 627756 627777 5 5 5 5 5 75.3 66.8 64.8 80.0 71.6 Group II 627768 627774 627776 627782 627785 10 10 10 10 10 56.3 51.0 54.8 43.6 65.1 ppm F in blood minus Bkg -2 PPm____ ppm F in blood Normalized to 0.1 mmoles/kg Dose 13.0 27.69 7.9 16.83 16.3 34.72 13.9 29.61 10.5 22.37 75.1 159.96 66.6 141.86 64.6 137.60 79.8 169.97 71.4 152.08 56.1 119.49 50.8 108.20 54.6 116.30 43.4 92.44 64.9 138.24 /molar 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 ` compan. Sanitized. Does not contain TSCAGB H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat # Test ppm F Day in Sample blood Group HI 627758 627764 627787 627763 627788 13 13 13 13 13 43.5 20.7 13.2 30.8 24.9 Group IV 627759 627779 627770 627761 627786 24 24 24 24 24 9.2 14.9 11.6 9.9 6.7 Group V 627757 627781 627760 627765 627772 52 52 52 52 52 0.9 0.6 1.0 0.9 0.8 Group VI 627778 627783 627762 627775 627769 94 94 94 94 94 0.3 <0.2 0.2 0.2 <0.2 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 * * = Below LOD (Level o f Detection) ppm F in blood Normalized to 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 jumolar 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 * " Company Sanitiaad. Doan no, contain TSC A C B 1 H-24018: Biopersistence Screening 10-Dose Oral Gavage Study in Rats______________________ Data for H-24018 DuPont-2927 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 845 1000 34 340 0.402 248.5 % F in Active: Mol Wt. F (g/mol): Dose F (mmol/kg): Molar Ratio (Active/F): Dose F (mg/kg): 50 19 8.947 0.045 170 R at# Test ppm F Day in Sample blood Group I 627252 627253 627260 627273 627274 1 1 1 1 1 4.1 3.5 3.1 2.7 1.3 Group I 627252 627253 627260 627273 627274 5 5 5 5 5 9.9 9.0 9.0 8.7 8.4 Group H 627246 627258 627261 627268 627269 10 10 10 10 10 8.7 12.3 7.8 11.0 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 /molar 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 in Rats R at# Test ppm F Day In Sample blood Group in 627249 627262 627265 627266 627277 13 13 13 13 13 6.8 6.4 5.1 5.2 4.8 Group IV 627247 627250 627255 627257 627259 24 24 24 24 24 2.3 2.8 2.5 2.5 NA Group V 627256 627264 627271 627275 627278 52 52 52 52 52 1.7 1.2 1.3 1.4 1.0 Group VI 627248 627251 627254 627263 627270 94 94 94 94 94 0.2 0.8 0.7 0.7 0.8 ppm F 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 NA Animal found dead on test day 9. - N o samples collected 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 /molar equivalents 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- Company Sanitized. Does not contain TSCA CBI