Document G5od06Ea4vaRv2Lb5Em8vpJx4

AR226-3152 TRADE SECRET Study Title H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID: DuPont-2922 AUTHOR: Carol Finlay, B.A. STUDY COMPLETED ON: May 19,2000 PERFORMING LABORATORY: E.I. du Font de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 WORK REQUEST NUMBER^jl^B^ SERVICE CODE NUMBER*!------1 Page 1 of 82 Company Sanded. Does not contn TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Reviewed by: Gary W. Jqpson. Ph-D. Senior Research Scientist C^fe^_ L.^Sl^ Reviewed by: ^ //Judith C. Stadler. PbJ)., DA^.T. (/ Director, General Toxicology Issued by Study Director: ^AAlsL ^CV\.i^ Carol FFiinlliiapy,, BB..AA.J.f- Associate Scientist H MA4 2^ou Date ^MM-ZOao Date -li--M^fL-L^^O Date '<* -2- cownys.""^.-"""0"0""'"' H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLE OF CONTENTS Page CERTIFICATION........................................................................................................................! LIST OFTABLES........................................................................................................................4 LIST OF FIGURES ......................................................................................................................4 LISTOFAPPENDICES...............................................................................................................4 STUDY INFORMATION.............................................................................................................5 STUDY PERSONNEL.....................................................................,............................................? 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.......................................................................................................................................l0 3. 10 Identification.......................................................................................................................................... 4. Animal Health Monitoring Program........................................................................................................ 10 D. Quarantine and Pretest...................................................................................................... 10 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. m-Life Toxicology............................................................................................................ 14 B. Fluorine Data..................................................................................................................... 14 1. Factors Influencing Interpretation of Analysis....................................................................................... 14 2. Positive Controls.................................................................................................................................... 15 3. TestSubstance......................................................................................;.................................................15 CONCLUSIONS......................................................................................................................... 15 RECORDS AND SAMPLE STORAGE................................................................................... 15 TABLES....................................................................................................................................... 17 -3- Compa81? Sanitized. Does not contain TSCACBl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLE OF CONTENTS (Continued) Page FIGURES..................................................................................................................................... 22 APPENDICES............................................................................................................................. 27 LIST OF TABLES Page 1. MEAN BODY WEIGHTS................................................................................................................................... 18 2. MEAN BODY WEIGHT GAINS........................................................................................................................ 19 3. MEANBLOOD FLUORINE LEVELS................................................................................................................20 4. MEAN BLOOD FLUORINE CONCENTRATIONS NORMALIZED TO DOSE..............................................21 LIST OF FIGURES Page 1. MEANBODY WEIGHTS ...................................................................................................................................23 2. MICROMOLAR EQUIVALENTS IN RAT BLOOD..........................................................................................24 3. MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS.............................26 LIST OF APPENDICES Page A. INDIVIDUAL BODY WEIGHTS.......................................................................................................................28 B. INDIVIDUAL CLINICAL OBSERVATIONS....................................................................................................62 C. ANALYSIS OF BLOOD FLUORINE DATA .....................................................................................................73 -4- Comp^y Sanifeed. Does not contain TSCA CBl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION TEST SUBSTANCE Substance Tested: ||g^^^^M ^jS^^^^WwBi^ Synonyms/Codes: H-23960 Haskell Number: 23960 Composition: DuPont-2922 POSITIVE CONTROL; Substance Tested: Potassium perfluoroalkyi sulfonate Synonyms/Codes. H-2401 -5- Sardtized. Doe-s ..o1;' STUDY INFORMATION (Continued) Compositionj Known ItnpuritiesU|^ggBB|'j| POSITIVE CONTROL: Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt Synonyms/Codes: H-24020 Perfluorooctanoate, ammonium salt PFOA Ammonium perfluorooctanoate Haskell Number: 24020 Composition Known Impurities 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: May 30, 1999 / August 30, 1999 -6- Company S.-.:'ii(ized. Doe's not contain TSCA G3E H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY PERSONNEL Study Director: Management: Primary Technician: Carol Pinlay, B.A. Judith C. Stadler, Ph.D., D.A.B.T. Richard P. Mafhena 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-2922 7Company Sanitized. Does not contain TSCA C31 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 SUMMARY The objective of this study was to evaluate the potential for H-23960, when administered by gavage, to te absorbed and to accumulate in a mammalian system. Six groups of 5 male CrLCD (SD)IGS BR rats each were exposed to 1000 mg/kg/day of H-23960. 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 the first dose, blood was collected from the orbital sinus of each rat from the 5-dose group. On selected days (5,10,13,24, 52,93/94) 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-23960. No compound-related clinical signs oftoxicity were observed in the rats dosed with H-23960. 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-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-23960 were comparable to the negative controls and equal to or greater than the positive controls. A steady-state for fluorine levels in whole blood was not achieved during 10 consecutive days of dosing with 1000 mg/kg H-23960. 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-23960 was 857.7 as 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-23960 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 500-lOOOx and 50-lOOx the AUCINF/D for the test substance, respectively. -8- c,^^'""0-"'0"0^'"'5'"' H-23960: Biopersistence Screening 10-DoseOral Gavage Study in Rats DuPont-2922 INTRODUCTION The objective of this study was to define the potential of H-23960 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 the results ofarangefinding study. m the rangefinding study, a group of 5 male rats was dosed by oral gavage with H-23960 at a dosage of 2000 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-23960 experienced an overall mean body weight gain of 23 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-23960, was supplied by the sponsor as an off-white milky liquid. positive controls, H-24019 and H-24020, were supplied by the sponsor as white solids. substance and positive controls appeared to be stable under the conditions of the study. evidence of instability, such as a change in color or physical state, was observed. The The test No B. Test Species Male Crl:CD(SD)IGS BR rats were received from Charles River Laboratories, me., Raleigh, North Carolina. The Crl:CD(SD)IGS BR rat was selected on the basis of extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, and low incidence of spontaneous diseases. -9- Company Sanitized. Doss not contain TSGA C31 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 C. Animal Husbandry 1. Housing Environment Rats were housed singly in suspended, 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 Haskell Laboratory has an animal health monitoring program. The following procedures are performed periodically: Water samples are analyzed for total bacterial counts, the presence ofcoliforms, lead, and other contaminants. Feed samples are analyzed for the presence of bacteria and fungi. Samples from freshly washed cages and cage racks are analyzed to assure proper sanitation by the cagewashers. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and to be free of impurities which might influence the results of the study. The animal health monitoring program is administered by the laboratory animal veterinarian; data are maintained separately from study records and are not included in the final report. Evaluation of these data did not indicate any conditions that affected the validity of the study. 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 r- -10. Company Samfaed. Does noS contain H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 clinically apparent signs of disease or injury. The rats were observed daily for mortality and signs of illness, injury, or abnormal behavior. On the bases of acceptable body weight gains and freedom from clinically apparent signs of disease or injury, the rats were released from quarantine by the laboratory animal veterinarian or designee. E. Study Design Substance Vehicle Negative Controls Corn oil Corn oil: acetone Positive Controls H-24019 H-24020 Test Substance H-23960 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-23960 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 1050 mg/mL. The test substance was stirred on a magnetic stir plate throughout the dosing procedure. 11Company Sanitized. Doss not ewtasn TSCA ""31 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study m Rats DuPont-2922 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 All rats were weighed on each day of dosing. The positive and negative control rats were weighed weekly or every other week during the recovery period. The rats dosed with H-23960 were weighed 3 weeks after the beginning of the recovery period and then weekly or every other week thereafter. 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: -12- Compar.y cS- -j,wii;-"ss"d- D""oe-s not cordain TSCA C.3 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 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 93/94 Five to 10 mL of blood was collected into EDTA tubes at sacrifice. The livers were weighed. The blood from all rats was refrigerated, and the livers were frozen. The blood was appropriately packaged and shipped refrigerated to Jackson Laboratory, Deepwater, New Jersey where it was analyzed for total fluorine. The total fluorine content of the blood samples was determined 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-23960 by using WinNonlin Version 3.0 software (Pharsight Corp. Mountain View, CA). WinNonlin software provided a means of computing, derived pharmacokinetic parameters from data files including area under the curve (AUCINF), Cmax and terminal half-life (Ti/z). The AUCINF (concentration x time) represents the area under the blood concentration curve from the time of dosing extrapolated to infinity. The maximum observed concentration was Cmax (concentration). The points included in determination of the terminal half-life were selected manually and given in units of time. Since the dosages and fluorine content for each positive control and the test material varied, all doses were normalized to 0.1 mmole/kg for comparative purposes. The accumulation index (AI, l^l-e'^)) and bioaccumulation index (BI, Cmax x AT) were calculated and reported but not farther 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 form analysis of total blood fluorine were used as received (ppm F) from Jackson Laboratory. Fluoride ion was converted to -13- Company Sanifcad. Does not contain TSCA CBt H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 micromolar QtM) equivalents of active component for further comparisons. The data and descriptions of data manipulation and presentation are provided in Appendix C. RESULTS AND DISCUSSION A. In-Life Toxicology (Tables 1 and 2; Figure 1, Appendices A and B) No deaths occurred during the study. No compound-related clinical signs oftoxicity were observed in the rats dosed with H-23960. One rat exhibited an umbilical hernia on test days 4-10. The hernia was not present on test day 13. This clinical sign is not considered to be test substance-related since it was only seen in one animal. 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. Comparison of body weights was complicated by the fact that there was a difference in age on test day 1 between the rats dosed with the test substance and those dosed with the positive or negative controls. This difference resulted in differences in mean body weights on test day 1. The rats dosed with the test substance, H-23960, were older and heavier in weight than the corn oil negative control and H-24020 positive control rats, but younger and lighter in weight than the corn oil/acetone negative control and H-24019 positive control rats. Accounting for the age difference at study start and the expected rate of body weight gain, the mean body weights and mean body weight gains of the rats dosed with H-23960 was comparable to the negative controls and equal to or greater than the 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. -14- Compsny Sanifesd. Doss not contain TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 2. Positive Controls The positive controls were H-24019 and H-24020. The H-24019 normalized fJiM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state (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 j^M equivalents in rat blood peaked after 5 days of dosing and then decreased throughout the dosing period (Figure 2B). The Cmax for H-24020 was 518.12 44.89 ppm (Mean SD) with a terminal half-life of 8.3 days. The H-24020 AI was 12.5 and the BI was 6497.5. For each of the positive controls, blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCENF/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 and 70,789.6 for H-24019 and H-24020, respectively. 3. Test Substance The H-23960 normalized fJiM equivalents for fluorine in rat blood continued to rise throughout the dosing period and did not reach steady-state (Figures 2 and 3). The Cmax for H-23960 was 2.224 0.668 (Mean SD) with a terminal half-life of 142.5 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 me derived terminal halflife for comparative purposes. The H-23960 accumulation index (AT) was 204.6 and the bioaccumulation index (BI) was 455 ppm. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between H-23960 and positive controls. The AUCINF/D for the fluorine component of H-23960 was 857.7 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-23960 exhibited no 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 was not achieved for H-23960 during the 10-day dosing period. Under the conditions of this study, administration of H-23960 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 500-lOOOx and 50-lOOx me AUCINF/D for the test substance, respectively.RECORDS AND SAMPLE STORAGE -15- Compsny Sanitized. Docs not conSa!n TSCA CBS H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. -16- Company Sanifcsd. Does not contai.-s TSCA CB" H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLES -17- Company Sar.^zed. Doas not conta'.i T3CA CI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study m Rats DuPont-2922 TABLE 1 MEAN BODY WEIGHTS (g) Test Days 1 2 3 4 5 6 7 8 9 10 13 20 24 27 34 41 47 48 52 55 61 62 68 69 75 76 80 81 83 89 90 93 94 N egative Controls Corn 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 Corn OiLAcetone 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 - - 407.8 434.5 454.9 411.4 - - - 464.7 487.0 505.1 527.6 - - - 515.4 - - - - - - 539.0 - - - - - - - 556.2 - 566.9 - 562.2 - 567.8 Positive iControls 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 a The rats were sacrificed a day early because of a laboratory event on test day 94. Indicates that the animal was not weighed. Test Substance H-23960 250.3 256.8 265.5 272.0 280.1 289.3 293.8 302.3 309.8 316.3 327.8 - 401.9 - 429.8 - - 486.7 502.0 - - 500.8 - 518.2 529.9 - - - 543.1 549.0 - 557.22 - -18- Comsany SanE'czed. Does not contain TSCA CBt H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLE 2 MEAN BODY WEIGHT GAINS (g) Test Days 1-5 1-10 10-13 10-24 10-52 10-93/94 Negative Controls Corn Oil 29.5 66.1 25.2 82.4 82.4 271.0 Corn OihAcetone 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-23960 29.8 66.0 11.5 85.6 185.7 240.9 Company SaniiizecS. Duas noi contain TSCA CSS H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLES MEAN BLOOD FLUORINE LEVELS Test Days 1 5 10 13 24 52 93/94 Negative Controls Corn Oil ppm 0.2 0.2 0.2 Q.T (0.1) 0.2 0.3s1 (0.1) OJ^O.l) Corn OiLAcetone ppm 0.2 0.2 0.3' (O.I)1' O^ (0.6) 0.2'' 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 26.9 13.2 (2.8) (2.6) (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) 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. Pour of the values were below the LOD. Test Substance H-23960 ppm 0.3 (0.1) 1.0(0.3) 1.5 (0.4) "1.0(0.3) 0.4 (0.2) 0.4(0.1) 0.3 (0.1) 20Company Sanitized. Does not contain TSCA CB8 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLE 4 MEAN BLOOD FLUORINE CONCENTRATIONS NORMALIZED TO DOSE Test Days Positive Controls H-24019 jtiM F Equivalents H-24020 juM F Equivalents Test Substance H-23960 IJIM. F Equivalents 1 5 10 13 24 52 93/94 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) 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) 0.17''(0.2)'' 1.38 (0.5) 2.22 (0.7) 1.28 (0.5) 0.42 (0.1) 0.27 (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. -21 Compsny Sanitized. Does not contain TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study m Rats DuPont-2922 FIGURES -22- Company SanEllzed. Doss nsl contain TSC;i CBf H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats 400 -i 350 -\ 300 " a 250 JS M *33 ?, 200 9 0 ca | 150 s 100 50 ^ FIGURE 1 MEAN BODY WEIGHTS (g) -0--Negative Control -0-- Negative Control -A--Positive Control -X--Positive Control -X--Test Substance H Test Days 10 12 -23- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 FIGURE 2 MICROMOLAR EQUIVALENTS IN RAT BLOOD A. Normalized Rat Blood H-24019 fiM Equivalents Resulting from a 10-Day 0 ral Gavage 1200 -] I Micromolar (/iM) equivalents of H-24019 in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. B. NormalizedRatBloodH-24020(iM Equivalents Resulting from a 10-Day 0 ralG avage Micromolar QxM) equivalents ofH-24020 in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. -24- Company Sanitised. Doss not contain T3CA CD! H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 C. Normalized Rat Blood H-23960 iiM Equivalents Resulting from a 10-Day Oral Gavage 30 50 70 80 90 100 Test Days Micromolar (^M) equivalents of H-23960 in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. -25- Conipany Senltisad. De-is not contain TSCA! H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURES MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS -26- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDICES -27- Does not contain T! Company ,San,,^-tiezeda. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDIX A Individual Body Weights -28- Cosnpany Sanitized, "oes not eonta'n TSCA CB8 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats INDIVIDUAL BODY WEIGHTS ABBREVIATIONS: SD - sacrificed by design EXPLANATORY NOTES DuPont-2922 -29- Company Sanitized. Uioes not contain TSCA CBS H-23960: 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 -30. H-23960: 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) OF MALE RATS GROUP II TEST E>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 -31- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627796 627806 627807 627809 627816 ANIMAL NUMBER 627796 627806 627807 627809 627816 DAY 1 239 .8 227 .3 221 .5 234 .2 235 .0 DAY 2 249 .7 234 .2 225 .9 240 .4 238 .6 DAY 8 285 .9 282 .8 270 .7 297 .9 291 .8 DAY 9 290 .1 285 .8 274 .7 300 .2 296 .4 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) 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 .32. H-23960: 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 .2 277 .8 DAY 9 279 .1 289 .5 270 .6 282 .7 280 .4 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS GROUP IV TEST I 1AYS 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. -33. H-23960: 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 V TEST r )AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627792 627798 627813 627814 627815 ANIMAL NUMBER 235 .8 241 .1 214 .8 217 .5 230 .1 DAY 8 240.8 253.4 219.2 220.5 242.2 DAY 9 244.3 257.6 220.5 223.7 243.8 247.9 266.1 233.2 225.2 255.7 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 627792 627798 627813 627814 627815 ANIMAL NUMBER 282 .4 294 .3 258 .7 248 .0 289 .2 DAY 41 286.4 296.3 259.2 250,7 293.4 DAY 47 295.2 315.7 267.9 257.3 303.3 ^a ^a a a 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 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. -34. H-23960: 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 E1AYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627795 627811 627819 627820 627821 ANIMAL NUMBER 218.0 228.6 248.8 225.5 227.5 DAY 8 219.5 226.3 255.3 228.9 233.2 DAY 9 225.3 236.7 264.9 239.2 240.7 234.3 246.1 274.4 247.9 246.1 TEST DAYS DAY 10 DAY 13 239.4 251.5 280.4 255.6 251.2 DAY 20 246.2 259.6 288.0 261.3 259.8 DAY 24 250.1 263.6 290.3 265.3 263.9 DAY 34 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. -35. H-23960: 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 TEST DAYS GROUP VI 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 .36. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in 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 -37- H-23960: 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) OP 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 -38- H-23960: 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 -39- w H-23960: 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 -40- H-23960: 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. -41 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats AMIMAL NUMBER DAY 1 DAY 2 CORM 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 ANIMAL NUMBER 305 .9 281 .7 294 .6 289 .1 293 .9 DAY 8 307 .3 279 .9 301 .3 291 .0 294 .8 DAY 9 316.8 289.7 312.7 298.0 302.9 325.0 286.9 317.0 307.0 312.0 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 625353 625369 625373 625375 625377 ANIMAL NUMBER 344 .7 352 .7 358.1 371.7 399.1 ^a 307 .3 311 .9 317.2 331.1 347.6 --a 340 .6 348 .2 352.6 371.5 402.3 a 324 .7 336 .0 336.9 350.1 387.9 ^a 325 .0 333 .7 340.7 353.5 376.3 a. DAY 34 DAY 52 TEST DAYS DAY 76 DAY 90 DAY 94 427 .7 374 .9 444 .6 432 .3 405 .2 625353 625369 625373 625375 625377 453 .3 ^a 388 .2 a 477 .1 a 455 .6 a 419 .1 ^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. -42- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625322 625323 625336 625343 625349 DAY 1 290 .3 285 .0 316 .5 288 .2 303 .6 DAY 2 283 .7 285 .5 313 .5 281 .9 295 .9 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I 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 -43. H-23960: 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) OP MALE RATS GROUP II TE,ST 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 -44- H-23960: 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 -45- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625325 625328 625338 625340 625352 ANIMAL NUMBER 625325 625328 625338 625340 625352 DAY 1 282 .8 315 .6 264 .6 304 .0 285 .5 DAY 2 285 .9 315 .2 261 .9 309 .2 283 .9 DAY 8 318 .6 352 .0 287 .4 347 .0 316 .9 DAY 9 321 .9 351 .7 291 .3 352 .2 322 .3 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 291.7 323.9 268.5 316.9 289.1 295 .5 327 .7 272 .6 326 .0 299 .6 TEST DAYS DAY 10 DAY 13 303 .2 335 .0 275 .7 329 .8 307 .9 DAY 20 309.2 340.4 281.9 334.1 308.2 DAY 24 308.3 340.1 284.4 338.4 312.4 327.4 357.5 290.2 359.2 321.0 331 .6 367 .4 291 .1 368 .1 329 .6 355 .8 400 .2 320 .4 389 .3 355 .7 381.3 423.9 339.9 419.0 375.3 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 -46. H-23960: 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 31.2.1 335.4 304.1 290.6 273.5 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. .47. H-23960: 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 & 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 d SD test d SD test d SD test d SD test d a Rat was not weighed. -48- H-23960: 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) OP 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 -49. H-23960: 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) OP 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 -50- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627758 627763 627764 627787 627788 ANIMAL NUMBER 627758 627763 627764 627787 627788 DAY 1 235 .1 212 .1 230 .0 235 .1 219 .4 DAY 2 240 .3 218 .3 235 .0 241 .0 222 .3 DAY 8 280 .6 255 .2 275 .8 221 .7 274 .1 DAY 9 283 .6 261 .2 281 .9 235 .7 276 .9 H-24020 (POSITIVE CONTROL) INDIVIDUAL.BODY WEIGHTS (g) OF MALE RATS GROUP III 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 -51- H-23960: 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. -52. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 627757 627760 627765 627772 627781 ANIMAL NUMBER 627757 627760 627765 627772 627781 ANIMAL NUMBER 627757 627760 627765 627772 627781 DAY 1 221 .2 226 .2 228 .3 245 .2 219 .0 DAY 2 225.4 232.0 235.3 256.4 226.3 DAY 8 269 .1 262 .6 271 .5 300 .9 . 258 .1 DAY 9 276.7 280.3 282.4 307.5 266.8 DAY 41 402 .8 437 .7 422 .9 478 .4 389 .9 DAY 47 419.7 448.9 444.8 494.3 408.2 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 236.3 244.4 247.1 266.2 236.5 241.1 251.7 251.6 277.0 242.9 TEST DAYS DAY 10 DAY 13 247.9 260.2 256.4 276.9 247.2 DAY 20 252.2 254.5 261.1 285.0 251.6 DAY 24 254.4 258.3 264.7 291.0 256.1 DAY 34 279.7 a 287.3 ^a 288.1 a 317.0 a 271.2 a. TEST DAYS DAY 52 324.3 348.6 349.8 375.5 315.1 343.0 379.9 382.7 404.0 338.8 372.3 419.4 400.9 446.9 372.0 436.7 456.5 455.6 513.5 421.0 SD test day 52 SD test day 52 . SD test day 52 SD test day 52 SD test day 52 a Rat was not weighed. -53. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OP 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 453.5 473 .0 471.6 479.7 474.6 a 512.2 511 .4 531.4 433.2 451.1 a 460.8 477 .4 478.6 356.8 373.5 a 386.0 393 .3 398.3 409.6 426.9 a 449.0 473 .8 474.8 a Rat was not weighed. 54. H-23960: 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 -55. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626831 626837 626846 626858 626860 DAY 1 . 258 .3 250 .3 224 .7 264 .8 239 .8 DAY 2 265 .4 248 .5 227 .5 271 .5 243 .2 H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I DAY 3 DAY 4 DAY 5 273 .1 257 .1 237 .1 276 .7 257 .6 282 .6 259 .2 243 .6 287 .4 265 .0 287 .8 267 .7 254 .9 294 .8 268 .0 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 -56. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626832 626841 626842 626851 626859 ANIMAL NUMBER 626832 626841 626842 626851 626859 DAY 1 249. 8 232. 4 274. 3 257. 9 251. 4 DAY 8 297. 4 277. 0 316. 0 298. 4 307. 3 DAY 2 253 .9 241 .2 280 .2 265 .8 257 .8 DAY 9 301 .5 282 .0 323 .5 304 .0 310 .5 H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS GROUP II DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 264.9 246.5 289.0 271.3 269.8 DAY 10 269.3 252.5 290.0 278.6 274.8 283 .0 259 .4 297 .0 286 .9 281 .6 291 .8 263 .9 306 .9 290 .0 289 .0 292.5 269.2 307.9 292.7 294.1 312.1 283.9 331.2 307.8 318.5 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 -57- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626836 626839 626845 626849 626856 ANIMAL NUMBER 626836 626839 626845 626849 626856 DAY 1 257 .7 251 .8 265 .9 241 .3 248 .3 DAY 8 315 .5 289 .5 313 .3 283 .6 310 .3 DAY 2 261 .7 254 .1 272 .3 252 .1 261 .0 DAY 9 319 .9 296 .3 324 .3 283 .0 321 .6 H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OP MALE RATS GROUP III DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 273 .4 266 .0 276 .6 258 .3 271 .3 DAY 10 279.8 269.7 278.8 262.9 275.3 DAY 13 288. 1 277. 6 283. 0 267. 4 282. 6 298.5 282.2 295.8 274.9 292.7 303.1 283.9 304.7 275.3 299.2 329 .3 304 .3 324 .6 285 .7 328 .2 335.8 314.4 348.2 297.6 343.0 SD test day 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 .58. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 626843 626844 626850 626853 626857 ANIMAL NUMBER 244.0 222.3 264.0 248.3 253.2 DAY 8 250.2 232.3 270.6 252.6 264.2 DAY 9 263 .2 243 .1 278 .1 261 .3 267 .5 DAY 10 270.1 248.3 284.8 271.2 271.8 DAY 13 279.2 255.5 300.4 281.3 277.6 DAY 24 286.4 264.9 309.7 288.7 287.3 291.9 268.0 309.0 300.0 288.3 626843 626844 626850 626853 626857 306.6 312.1 318 .1 a 395.7 SD test day 24 269.6 282.4 290 .9 a 358.9 SD test day 24 319.1 326.9 332 .2 a 423.0 SD test day 24 309.0 326.3 328 .0 a 445.4 SD test day 24 295.3 302.5 308 .0 a 386.3 SD test day 24 a Rat was not weighed. -59- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP V DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 626833 626835 626852 626861 626862 ANIMAL NUMBER 282 .2 252 .8 239 .4 249 .7 257 .9 DAY 8 291. 3 261. 5 249. 9 252. 9 265. 6 DAY 9 299 .1 272 .0 257 .0 254 .3 278 .7 DAY 10 309.9 280.5 266.4 262.3 289.6 DAY 13 320.9 291.7 274.6 270.3 302.3 DAY 24 326 .1 299 .3 286 .1 276 .6 311 .5 DAY 34 333 .9 304 .6 289 .5 280 .8 320 .0 DAY 48 626833 626835 626852 626861 626862 ANIMAL NUMBER 348 .9 356. 9 367 .6 a 312 .9 319. 7 330 .4 a 299 .1 310. 6 316 .1 a 284 .5 297. 0 298 .1 a 328 .8 340. 6 357 .2 a DAY 52 a 481. 7 536. 9 a 432. 6 485. 1 --a 417. 3 452. 8 a 392. 4 450. 4 -a 519. 1 593. 3 626833 626835 626852 626861 626862 558 .8 505 .1 466 .7 462 .3 617 .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. -60. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 626847 626848 626854 626855 626863 ANIMAL NUMBER 626847 626848 626854 626855 626863 ANIMAL NUMBER 626847 626848 626854 626855 626863 DAY 1 235 .7 252 .1 220 .5 272 .0 247 .4 DAY 8 277 .5 302 .3 255 .9 328 .0 312 .9 DAY 2 242.5 258.9 224.5 279.5 252.5 DAY 9 284.0 308.8 260.3 331.5 318.5 DAY 52 437 .7 471 .5 412 .9 552 .3 535 .2 DAY 62 448.9 485.8 430.0 577.9 561.4 H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 247 .4 269 .0 226 .9 286 .6 270 .8 DAY 10 254.4 277.7 235.0 297.0 270.6 DAY 13 258 .3 284 .0 241 .9 302 .9 281 .6 DAY 24 269 .2 290 .5 245 .0 313 .8 292 .4 DAY 34 272.0 299.5 247.0 317.9 298.9 DAY 48 291 .3 a 316 .5 a 261 .6 a 339 .9 a 325 .6 a a 381..7 428.8 a 396 .0 456.6 a 347 .9 411.0 a 476.6 541.1 a 452.3 510.7 DAY 69 460 .6 505 .4 441 .8 599 .0 584 .1 DAY 75 475.8 516.1 454.2 614.4 588.8 DAY 83 489 .4 535 .0 459 .6 625 .4 606 .2 DAY 89 498 .1 544 .3 460 .9 637 .1 604 .5 DAY 93 501.0 553.4 473.8 641.0 616.7 SD test da SD test da SD test da SD test da SD test da a Rat was not weighed. -61 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDIX B Individual Clinical Observations -62- not c^nTSCACBl Company .S-afnzAe^dsa-D"oes H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number- 627789 627801 627805 627810 627818 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 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- Sanitized. Do88 ^-coniai" TCCA CBS Company H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number 627791 627794 627804 627812 627817 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 627792 627798 627813 627814 627815 GROUP V Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-52 1-52 1-52 1-52 1-52 Animal Number 627795 627811 627819 627820 627821 GROUP VI Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-94 1-94 1-94 1-94 1-94 -64- Rc^con^""33^81 Co^S^'26^085" H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Numbel" 625361 625363 625364 625380 625383 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-5 1-5 1-5 1-5 1-5 Animal Number 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 Sbaa^^i-ez-do". --"oes no^ontain TGC*. Cui H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 CORN 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 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 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 -66Company Sa^zed. Does not contain 7SCA CBi H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number 625322 625323 625336 625343 625349 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I __________Observation___________________Test Day No abnormalities detected 1-5 No abnormalities detected 1-5 No 'abnormalities detected 1-5 No abnormalities detected 1-5 No abnormalities detected 1-5 Animal Number 625320 625330 625332 625341 625350 GROUP II Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Diarrhea No abnormalities detected Test Day 1-10 1-10 1-10 1-4, 6-10 5 1-10 Animal Number 625321 625327 625342 625345 625348 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- CompE.'.y Sanifced. Doss nci contain T=GACBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number 625325 625328 625338 625340 625352 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV __________Observation____ _____ ______ No abnormalities detected No abnormalities detected No abnormalities detected Salivation No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-6, 8-24 7 1-24 1-24 Animal Number 625324 625329 625333 625334 625344 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, 7 8-52 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- c^;^^.-."0""-1"""'-"6"3" H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number 627756 627767 627771 627773 627777 H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I _____ No No No No No Observation__________________Test Day abnormalities detected 1-5 abnormalities detected 1-5 abnormalities detected 1-5 abnormalities detected 1-5 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- CW~.y Sanitized. Does not contain TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number 627759 627761 627770 627779 627786 H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV __ Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected __Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 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. Company Sanitized. Does noi contain TSCA CBl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number' 626831 626837 626846 626858 626860 H-23960 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I __________Observation__________________Test Day No abnormalities detected 1-5 Mo abnormalities detected 1-5 No abnormalities detected 1-5 No abnormalities detected 1-5 No abnormalities detected 1-5 Animal Number 626832 626841 626842 626851 626859 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 626836 626839 626845 626849 626856 GROUP III Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Umbilical hernia Test Day 1-13 1-13 1-13 1-13 1-3,11-13 4-10 -71- Con^pany Sanitized. Does n3t contain TSCA CBt H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 Animal Number 626843 626844 626850 626853 626857 H-23960 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP IV __________Observation___ No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 626833 626835 626852 626861 626862 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 626847 626848 626854 626855 626863 GROUP VI Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-93 1-93 1-93 1-93 1-93 -72. Company San.feed. Does not contain TSCA CB1 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDIX C Analysis of Blood Fluorine Data -73- CGinpany aanrtaao. yo^-> "ul cu.*Eci.i< s aCA C&i Company Ssnittee^- ^^e? not con^!" TSCA CE H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Terms and Calculations DuPont-2922 Terms: Active . Fluorine containing compound % Active Mol Wt Active The % of formulation that is made up of fluorine containing components 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/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 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 -74. C,n,p..,s^-D""ic--t"i"T3CAC" H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Individual Animal Measurement: ppm F in blood The ppm fluoride measured in blood DuPont-2922 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 mmol/kg The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead of the actual active dose. This assumes linearity between administered dose and blood fluorine levels, but is needed because different doses of active were used in the study. = ( 0.1 [mmol/kg] / Active dose [mmol/kg]) * (ppm F in blood minus background) jiimolar equivalents of active in blood The /xmolar [^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- .........edOoesnolcor.lalnTSCACSl Company S--""-"-1"- -"- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 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' ;on,p.n, S,.,d. ""=-"" TSCA CB1 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: Data for H- 24019 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/1F): Dose F (mg/kg): DuPont-2922 65 19 0.342 0.059 6.5 Rat # Test Day Sample ppm F 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 ppm F in blood iminus 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 ^molar 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 -77. annized.D^nctcontainTSCACBl Company S H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat# Test Day Sample ppmF in blood Group III 625321 13 625327 13 625342 13 625345 13 625348 13 77.2 61.5 64.6 57.0 62.4 Group IV 625325 24 625328 24 625338 24 625340 24 625352 24 40.3 46.3 43.2 39.1 41.9 Group V 625324 52 625329 52 625333 52 625334 52 625344 52 24.7 29.8 27.3 29.1 23.8 Group VI 625326 94 12.4 625335 94 16.4 625346 94 13.1 625347 94 11.0 625351 94 11.6 ppm F in blood minus Bkg 0.2 ppm 77.0 61.3 64.4 56.8 62.2 40.1 46.1 43.0 38.9 41.7 24.5 29.6 27.1 28.9 23.6 12.2 16.2 12.9 10.8 11.4 ppm F in bliood Normalizeldto 0.1 mmolefi/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-2922 ^molai equivaleiits 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 -78- contain TSCAC.I Corripan? Sanmzed.D^not H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: Data for H- 24020 426 % F in Active: 20 Mol Wt. F (g/mol): 100 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose NormalizEition Factor: 20 0.047 42.6 Dose F (mmol/kg): Molar Ratio (Active/]F): Dose F (mg/kg): DuPont-2922 69 19 0.726 0.065 13.8 Rat # Test Day Sample ppm F in blood Group I 627773 1 13.2 627771 1 8.1 627767 1 16.5 627756 1 14.1 627777 1 10.7 Group I 627773 5 627771 5 627767 5 627756 5 627777 5 75.3 66.8 64.8 80.0 71.6 Group II 627768 10 627774 10 627776 10 627782 10 627785 10 56.3 51.0 54.8 43.6 65.1 ppm F in blood minus Bkg 0.2 ppm 13.0 7.9 16.3 13.9 10.5 75.1 66.6 64.6 79.8 71.4 56.1 50.8 54.6 43.4 64.9 ppm F in blood Normalized to 0.1 mmoles/kg Dose 27.69 16.83 34.72 29.61 22.37 159.96 141.86 137.60 169.97 152.08 119.49 108.20 116.30 92.44 138.24 ^motar 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 -79- 'tC3nta5nTSCAC":>S Company Sanitized. Does no' H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat# Test Day Sample ppmF in blood Group III 627758 13 627764 13 627787 13 627763 13 627788 13 43.5 20.7 13.2 30.8 24.9 Group IV 627759 24 9.2 627779 24 14.9 627770 24 11.6 627761 24 9.9 627786 24 6.7 Group V 627757 52 0.9 627781 52 0.6 627760 52 1.0 627765 52 0.9 627772 52 0.8 Group VI 627778 94 627783 94 627762 94 627775 94 627769 94 0.3 0.2 0.2 0.2 0.2 * =1 Below LOD (Level of Detection) ppm F in blood minus Bkg 0.2 ppm 43.3 20.5 13.0 30.6 24.7 9.0 14.7 11.4 9.7 6.5 0.7 0.4 0.8 0.7 0.6 0.1 * 0.0 0.0 * ppm F in 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-2922 jitmolar 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- Company Sanitized. Does not contain TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats D;ata for H- 2:3960 Given: MolW't. Active ({^/mole): Formulation Dose; (mg/kg): % Active (F Containing) in Fonnulation: 27500 1000 18 % F in Adtive: Mol Wt. F (g/mol) : Calculate!i Values: Dose/active (mg/kg): Dose/active (mmole/kg): Formul ation Dose Nonnali2';ati<an Factor: 180 0.007 15277.8 Dose F (numol/kg);: Molar Rati o (Acti\^e/F): Dose F (mg/kg): DuPont-2922 33 19 3.126 0.002 59.4 Rat# Test Day Sample ppmF in blood Group I 626831 1 0.4 626837 1 0.2 626846 1 0.4 626858 1 0.2 626860 1 0.2 Group I 626831 5 1.4 626837 5 0.9 626846 5 0.9 626858 5 1.2 626860 5 0.7 Group II 626832 10 1.7 626841 10 1.0 626842 10 1.4 626851 10 2.1 626859 10 1.4 ppm F in blood minus Bkg 0.2 ppm 0.2 0.0 0.2 0.0 * 1.2 0.7 0.7 1.0 0.5 1.5 0.8 1.2 1.9 1.2 ppm F in b lood Normalize dto 0.1 mmole;s/kg Dose 3.06 0.00 3.06 0.00 * 18.33 10.69 10.69 15.28 7.64 22.92 12.22 18.33 29.03 18.33 ^molar equivalents of active in blood 0.34 0.00 0.34 0.00 * 2.02 1.18 1.18 1.68 0.84 2.53 1.35 2.02 3.20 2.02 -81- S.nBz.-.B------1"'1181^3""' Company H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat# Test Day Sample ppmF in blood Group III 626836 13 0.7 626839 13 1.1 626845 13 1.0 626849 13 0.6 626856 13 1.4 Group IV 626843 24 626844 24 626850 24 626853 24 626857 24 0.5 0.4 0.2 0.2 0.2 Group V 626833 52 0.3 626835 52 0.4 626852 52 0.2 626861 52 0.4 626862 52 0.5 Group VI 626847 93 0.3 626848 93 0.2 626854 93 0.4 626855 93 0.4 626863 93 0.3 ! Below LOD (Level of Detection) ppm F in blood minus Bkg 0.2 ppm 0.5 0.9 0.8 0.4 1.2 0.3 0.2 * * 0.0 0.1 0.2 0.0 0.2 0.3 0.1 0.0 0.2 0.2 0.1 ppm F in b'lood Normalize dto 0.1 mmoles/kg Dose 7.64 13.75 12.22 6.11 18.33 4.58 3.06 * * 0.00 1.53 3.06 0.00 3.06 4.58 1.53 0.00 3.06 3.06 1.53 DuPont-2922 (imolsr equivalents of active in blood 0.84 1.52 1.35 0.67 2.02 0.51 0.34 * * 0.00 0.17 0.34 0.00 0.34 0.51 0.17 0.00 0.34 0.34 0.17 -82- Company Samfeed. Does not contain TSCA CB1