Document OJ8b47vkvXnQMj9rBVXxoVVz1

TRADE SECRET Study Title H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID: DuPont-2922 AR226-3152 DuPont-2922 Author: Carol Finlay, B.A. Study Completed on: May 19,2000 Performing Laboratory: 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 Work Request Number - 1 Service Code Number: Page 1 o f 82 Company Sanitized. Does no! contain TSC 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 o f data obtained from this study. R eview ed b y : C -J Z ? G ary W . Jepson. P hJ 3. S enior R esearch S cientist R eview ed b y : ^ V c' d .\ B u L I j S ^ Ju d ith C . S tad i -, P h D ., D .A .B .T . D irect -, G eneral T oxicology IH M a h 2 ~ d > o t* D ate P t-M A M -Z C Q O D ate Issu ed by S tu d y D irecto r; f A A k L % C V \,L d -h C aro fF in lay , B .A .^ A ssociate S cien tist i1 - ' Zo D a te N* -2 not contain TSCA CB! Company Sanitized. Does H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLE OF CONTENTS Page CERTIFICATION..... LIST OF TABLES a ..............*................................... 4 LIST OF FIGURES................................................................. LIST OF APPENDICES.......................................................... STUDY INFORMATION......................................................... STUDY PERSONNEL.............................................................. SUMMARY........................................ ............................... ......... ....... 4 ----- 5 ....... 7 ...8 INTRODUCTION...................................................................... ----- 9 MATERIALS AND METHODS............................................. .... 9 A. Test Substance and Positive Controls......................................................... ' '..................... X B. Test Species..................................................................... ...................^ C. Animal Husbandry...................................................... ............ 1. Housing Environment...................................................................... 2. Feed and Water............................................................... 3. Identification................................................................................. .............................. 10 10 4. Animal Health Monitoring Program.................................................. ......jq D. Quarantine and Pretest................. ................................................... ..............................jq E. Study Design.............................................................................. .................j ^ F. Assignment to Groups and Study Start.............................................................................. 1 1 G. Dosing Material Preparation and Administration......................................... " ' ".............11 1. Test Substance................................................................................ 2. Positive Controls............................................................................ 3. Negative Controls................................................................................. H. Body Weights......................................................................... ............................ 11 12 ........................... 12 I. Mortality and Clinical Observations........................................................... .......................12 J. Collection and Analysis o f Blood and Livers......................................... K. Treatment o f Fluorine Data............................................................. .................. 12 RESULTS AND DISCUSSION........ ................................................................... 14 A. In-Life Toxicology............................................................ ......... . B. Fluorine Data................................................................................ ................... ^ 1. Factors Influencing Interpretation o f A nalysis............................................................................................... 14 2. Positive Controls.......................................................................... 3. Test Substance................................... ............................................... .15 .15 CONCLUSIONS....................................... .15 RECORDS AND SAMPLE STORAGE, TABLES....... ................ ....................... .15 ,17 -3 Com pany Sanitized. Does not contain TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats TABLE OF CONTENTS (Continued) F I G U R E S ___ APPENDICES LIST OF TABLES 1. MEAN BODY WEIGHTS.................................................................. 2. MEAN BODY WEIGHT G A IN S........................................................................... 3. MEAN BLOOD FLUORINE LEVELS...................................................................... 4. MEAN BLOOD FLUORINE CONCENTRATIONS NORMALIZED TO DOSE DuPont-2922 Page ...22 ...27 Page ..... 18 .....19 .... 20 .... 21 LIST OF FIGURES 1. MEAN BODY WEIGHTS....................................................................................... Page 23 2. MICROMOLAR EQUIVALENTS IN RAT BLOOD.............................................................................. 24 3. MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS....................... ?6 l is t o f a ppen d ic es A. INDIVIDUAL BODY WEIGHTS.................... B. INDIVIDUAL CLINICAL OBSERVATIONS C. ANALYSIS OF BLOOD FLUORINE D A T A . Page .....28 .....62 .....73 -4 - "-- ------- Company Sanitized. D oes not contain TSCA CSI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION TEST SUBSTANTIF. Substance Tester! Synonytns/Codes: 0 H-23960 Haskell Number: 23960 DuPont-2922 POSITIVE CONTROL: Substance Tested: Potassium perfluoroalkyl sulfonate 1- 5 - ^ompany Sanitized. Doej not co! in TSCA CB H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats STUDY INFORMATION (Continued) DuPont-2922 POSITIVE CONTROL: Substance Tested: Octanoic acid, pentadecafluoro-, ammonium salt Svnonvms/Code" - m -m o n C-8 Perfluorooctanoate, ammonium salt PFOA Ammonium perfluorooctanoate Haskell Number: 24020 Sponsor: E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.SA. Study Initiated/Completed: April 30, 1999 / (see report cover page) In-Life Initiated/Completed: May 30, 1999 / August 30, 1999 -6Company Saniilzed. Does not contain TSCA C3i H-23960: 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-2922 -7 - Company Sanitized. Does not contain TSCA C3I 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 he 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 of H-23960. 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 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, 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-23960. No compound-related clinical signs of toxicity 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 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-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 o f 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 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-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 500-1000x and 50-lOOx the AUCINF/D for the test substance, respectively. - 8 - Sanitlzsd. Dess not contain TSCA C31 Company H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 INTRODUCTION The objective o f this study was to define the potential o f 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 of a rangefinding study. In the rangefinding study, a group of 5 male rats was dosed by oral gavage with H-23960 at a dosage o f 2000 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-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 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 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. The positive controls, H-24019 and H-24020, were supplied by the sponsor as white solids. The test substance and positive controls appeared to be stable under the conditions of the study. No evidence of instability, such as a change in color or physical state, was observed. B. Test Species Male Crl:CD(SD)IGS BR rats were received from Charles River Laboratories, Inc., Raleigh, North Carolina. The Crl:CD(SD)IGS BR rat was selected on the basis of extensive experience with this strain and its suitability with respect to longevity, hardiness, sensitivity, and low incidence of spontaneous diseases. -9 Com pany Sanitized. Does not contain TSCA C3t 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 o f 50 10%. Occasional excursions outside the accepted ranges were minor and did not affect the study. 2. Feed and Water Tap water was provided ad libitum. All rats were fed PMI Nutrition International, Inc. Certified Rodent LabDiet 5002 chow. The feed is guaranteed by the manufacturer to meet specified nutritional requirements and to be free of specified contaminants. 3. Identification Prior to assignment to groups, each rat was temporarily identified by cage identification. After assignment to groups, an individual identification number was marked or tattooed on the tail of each rat. The information on the cage labels included the unique 6-digit Haskell animal number assigned to each rat. 4. Animal Health Monitoring Program Haskell Laboratory has an animal health monitoring program. The following procedures are performed periodically: Water samples are analyzed for total bacterial counts, the presence of coliforms, 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 anim al 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. Q uarantine 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 - 10- Compsr.y Sanitized. Dos ;nc,t 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 Com oil Com oil:acetone Positive Controls H-24019 H-24020 Test Substance H-23960 Not applicable Com oil: acetone Com oil None Dosage (mg/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 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 o f 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 o f dosing and the test substance density of 1050 mg/mL. The test substance was stirred on a magnetic stir plate throughout the dosing procedure. Company Sanitized. Does m l certain TSCA CSl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 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 of acetone to com 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 Com oil and com 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 com oibacetone. 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 o f 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 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: - 12- Compar. Sanitised. Doe: , not contain IS C A H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Group Dosing Days Tissue Collected Sampling Time DuPont-2922 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 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 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 (T1/2). 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 o f the terminal half-life were selected manually and given in units of time. Since the dosages and fluorine content for each positive control and the test material varied, all doses were normalized to 0.1 mmole/kg for comparative purposes. The accumulation index (AI, l/(l-e**)) and bioaccumulation index (BI, Cmax x AI) were calculated and reported but not further used. H-24019 and H-24020 were used as positive controls. Dosing diluents were used as negative controls, but because o f 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 o f total blood fluorine were used as received (ppm F) from Jackson Laboratory. Fluoride ion was converted to Com pany S am fcad . Does not contain TSC CBl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 micromolar (jtiM) equivalents o f 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 o f toxicity 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 o f 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 -2 3 9 6 0 , were older and heavier in weight than the com oil negative control and H - 2 4 0 2 0 positive control rats, but younger and lighter in weight than the com oil/acetone negative control and H - 2 4 0 1 9 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 g a in s o f the rats dosed with H - 2 3 9 6 0 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 o f the terminal half-life. A more complete list of considerations is shown m Appendix C. -14Company Sanifesd. Docs 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 (iU 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 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 and 70,789.6 for H-24019 and H-24020, respectively. 3. Test Substance The H-23960 normalized jM 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 o f 142.5 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-23960 accumulation index (AI) 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 of toxicity 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 o f this study, administration o f 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 500-1000x and 50-lOOx the AUCINF/D for the test substance, respectively.RECORDS AND SAMPLE STORAGE -15Compony Sanitized. Docs not contain 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 Sanitized. D oes not contalo TSCA CB H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLES -17Compsny Sar.ilized. Doss not conta'n T3CA Ci H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in 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 Negative Controls ComOil 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 - 407.8 434.5 454.9 464.7 487.0 505.1 _ 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 - 411.4 - - - 527.6 - - - 515.4 - _- -- - 539.0 -- _ _- - 556.2 . 566.9 - 562.2 - 567.8 Positive Controls H-24019 291.5 291.5 298.3 304.1 308.6 312.7 317.5 317.3 318.4 318.9 329.1 359.7 387.9 393.9 416.9 - - 438.1 - - 523.4 - - - 563.4 570.7 _ H-24020 227.5 232.4 242.0 248.9 252.4 259.8 263.3 269.8 278.5 283.9 289.1 311.3 340.9 - 402.9 425.0 438.2 - 465.6 452.3 465.8 470.9 - - - - 499.2 - - 585.3 - 597.9 - - 527.8 - 538.7 a The rats were sacrificed a day early because o f 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 - 557.2a - -18- Company Sanitized. Does not contain T5CA CBS 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 Com Oil 29.5 66.1 25.2 82.4 82.4 271.0 Com 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 -19Company Sanitized. Dcas noi contain TSCA CBS H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 TABLE 3 m ea n blood flu o rin e levels Negative Controls Positive Controls Test Substance Days Com Oil Com Oil:Acetone H-24019 H-24020 H-23960 ___1_____<_0_.p2gm_--_-_--_--_-_-________<__0._r2r_"_._-_--_-_--_-_--______2.E1gm(0_._9_)_____6_2p.6pm(_3_.2_)_______0.3PP(m0.1_) 5 <0.2 <0.2 48.8 (17.6) 71.7 (6.2) 1.0 (0.3) 10 <0.2 0.3a (0.1)b 61.7 (2.8) 54.2 (7.8) 1.5 (0.4) 13 0.2a (0.1) 0.6C(0.6) 64.5 (7.6) 26.6 (11.4) 1.0 (0.3) 24 <0.2 0.2d 42.2 (2.8) 10.5 (3.0) 0.4 (0.2) 52 0.3a (0.1) <0.2 26.9 (2.6) 0.9 (0.2) 0.4 (0.1) 93/94 0.3a (0.1) <0.2 13.2 (2.1) 0.2C (0.1) 0.3 (0.1) 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. . -20Company Sanitized. Does not contain TSG CBl 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 H-24020 juM F Equivalents liM F Equivalents Test Substance H-23960 IxM. 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) 0.17a (0.2)b 1.38 (0.5) 2.22 (0.7) 1.28 (0.5) 0.42c (0.1) 0.27 (0.2) 0.20 (0.1) 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. -21 Company Sanitized. Does not contain TSCA CBS H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 FIGURES - 22- C om psny Sanitized. Does net contain TSCA CBf H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats FIGURE 1 MEAN BODY WEIGHTS (g) DuPont-2922 Vs33as*,, N(Q!), o Owm oo 3. 3,,5 Vi a 5-' Mean Body Weights (g) 400 -| 350 - 300 - 250 200 150 - 100 - --O -- Negative Control (Corn Oil) -- --N e g a tiv e C o n tro l (C orn O il:A c e to n e ) --& --P o sitive C o n tro l H -24091 --X--P o sitiv e C o n tro l H -2 4 0 2 0 --X--T e s t S u b sta n c e H -2 3 9 6 0 50 - 00 2 4 6 8 10 12 Test Days 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 p M Equivalents Resulting from a 10-Day O ral Gavage Micromolar QiM) 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. Norm alized Rat Blood H -24020 pM Equivalents Resulting from a 10-Day O ralG avage Micromolar QjlM) equivalents of H-24020 in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. - 24- Company Sanitized. Cos: not contain T3C Cui H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 C. N orm alized R at Blood H-23960 jtM E quivalents Resulting from a 10-Day Oral G avage Micromolar QM) equivalents of H-23960 in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. j) -25- Cornvjarty Sanitised. D ecs not ! retain TSC CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats IP DuPont-2922 FIGURE 3 MICROMOLAR EQUIVALENTS IN TEST COMPOUND AND POSITIVE CONTROLS H-23960 arid Positive Control Normalized Blood AUCINF/D in Rats Resulting from 10-day Oral Gavage : Company Sanitized. C aos not contain TSCA CB1 -26 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDICES -27- ,,c! contain TSCACBl Sanized. DeS C om p anV H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDIX A Individual Body Weights - 28- C om pany Sanitized. Doss not contain TSCA CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats INDIVIDUAL BODY WEIGHTS EXPLANATORY NOTES ABBREVIATIONS: SD - sacrificed by design DuPont-2922 - 29- Company Sanitized. Does no? contain TSCA CBS _ H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_________________________________________ CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I ANIMAL NUMBER DAY 1 DAY 2 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-2922 Company Sanitized. D oes not contain TOGA rOa - 3 0 - 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 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 Oo n3 sa v3(aaf3l 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-2922 ;ed. D oes not contain TSCA OBI -31 - noi contain TSCA CB? 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 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 o 1 a a ao u 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 -32- DuPont-2922 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_______________________________________ ___________________________ CORN OIL (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV ANIMAL NUMBER DAY 1 DAY 2 TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 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 627791 627794 627804 627812 627817 DAY 8 274.0 286.1 267.2 278.2 277.8 DAY 9 279.1 289.5 270.6 282.7 280.4 TEST DAYS DAY 10 DAY 13 285.0 295.4 278.1 294.3 294.6 a a a a a DAY 20 334.4 377.4 345.8 356.5 368.7 DAY 24 365.6 416.7 378.1 388.4 407.3 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 DuPont-2922 Company Sanitized. Does not contain a Rat was not weighed. O> O --- -------------------- OB - 33- Company Sanitized. Does noi contain TSCA CBI 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 DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627792 627798 627813 627814 627815 235.8 241.1 214.8 217.5 230.1 240.8 253.4 219.2 220.5 242.2 244.3 257.6 220.5 223.7 243.8 247.9 266.1 233.2 225.2 255.7 252.5 272.9 239.8 233.8 265.0 264.4 281.9 243.2 236.7 270.0 268.1 286.2 246.8 240.6 274.1 ANIMAL NUMBER 627792 627798 627813 627814 627815 DAY 8 282.4 294.3 258.7 248.0 289.2 DAY 9 286.4 296.3 259.2 250.7 293.4 TEST DAYS DAY 10 DAY 13 295.2 315.7 267.9 257.3 303.3 -a DAY 20 360.0 380.2 323.6 299.3 368.3 DAY 24 389.2 403.5 342.3 319.4 396.5 DAY 34 426.1 443.1 367.9 343.7 434.9 ANIMAL NUMBER DAY 41 DAY 47 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. -34- DuPont-2922 Company Sanitized. Does not contain TSCA 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 DAYS DAY 3 DAY 4 DAY 5 DAY 6 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 a 411.1 a 455.4 a 551.7 a 529.7 a 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. DuPont-2922 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 GROUP VI TEST DAYS 627795 627811 627819 627820 627821 474.6 503.6 638.1 608.6 509.0 485.0 511.6 653.1 618.0 521.5 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 DuPont-2922 Company Sanitized. Dess not contain TSCA Ci -36- 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 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-2922 Company Sanitized. Does not contain TSCA C3i - 37- 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 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-2922 Company Sanitized. D o ss net contain TSCA CB? -38- 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 HALE 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-2922 Company Sanitized. Doss r.ot contain TCCA CEI -39- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats________________________________________ __________________________ CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP IV ANIMAL NUMBER DAY 1 DAY 2 TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 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 625354 625365 625371 625376 625381 DAY 8 336.2 370.5 340.3 331.5 321.6 DAY 9 337.0 379.3 345.9 339.0 329.1 TEST DAYS DAY 10 DAY 13 348.8 381.2 351.2 345.2 331.6 355.8 407.1 372.6 363.2 341.2 DAY 20 377.7 438.2 413.0 399.7 358.2 DAY 24 396.1 459.6 447.3 432.9 381.8 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 DuPont-2922 Company Sanitized. Coes not contain TSCA CBS -40- Company Sanitized. Does not contain ISC CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625356 625359 625360 625372 625374 DAY 1 291.7 297.9 296.8 294.8 295.9 DAY 2 291.2 303.3 300.7 298.8 302.2 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 7 304.5 314.0 308.5 310.6 312.1 307.0 315.0 312.7 317.4 321.7 316.9 323.5 319.1 324.0 323.4 319.8 323.5 332.2 329.0 328.3 323.9 333.9 339.5 335.6 337.2 ANIMAL NUMBER 625356 625359 625360 625372 625374 DAY 8 328.8 339.1 347.9 344.7 338.0 DAY 9 334.7 346.2 354.3 348.8 344.6 TEST DAYS DAY 10 DAY 13 341.3 353.6 363.4 355.1 350.8 361.9 372.9 381.7 373.1 371.5 DAY 20 396.7 410.0 419.8 395.1 401.3 DAY 24 __a --a -- -- DAY 27 431.3 440.9 465.9 432.1 447.2 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. - 41 - DuPont-2922 Company Sanitized. D oss 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 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 625353 625369 625373 625375 625377 DAY 8 344..7 307..3 340..6 324..7 325 .0 DAY 9 352.7 311.9 348.2 336.0 333.7 TEST DAYS DAY 10 DAY 13 358.1 317.2 352.6 336.9 340.7 371.7 331.1 371.5 350.1 353.5 DAY 20 399.1 347.6 402.3 387.9 376.3 DAY 24 __a _a _a __a __a DAY 27 427.7 374.9 444.6 432.3 405.2 ANIMAL NUMBER 625353 625369 625373 625375 625377 DAY 34 453.3 388.2 477.1 455.6 419.1 DAY 52 _a _a __a _a _a TEST DAYS DAY 76 DAY 90 DAY 94 544.9 483.8 628.8 567.3 470.3 562.8 505.3 661.3 589.8 492.0 565.3 514.8 673.7 588.8 496.6 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 o o o 3' a Rat was not weighed. DuPont-2922 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 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-2922 Company Sanitized. Dess not contain TSCA CBi -43- 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 II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625320 625330 625332 625341 625350 279.3 286.7 300.0 304.9 257.1 283.0 286.1 295.9 309.0 268.5 287.7 291.1 304.1 317.5 267.3 288.4 296.5 308.8 320.0 272.2 295.9 304.0 315.0 325.1 272.6 299.4 302.3 322.4 327.5 276.3 300.6 306.7 320.7 327.4 283.6 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 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-2922 Company Sanitized. Does not contain TSCA C3I -44- 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 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-2922 Sanitized. Does not contain Company Cn'J 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 IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 625325 625328 625338 625340 625352 282.8 315.6 264.6 304.0 285.5 285.9 315.2 261.9 309.2 283.9 291.7 323.9 268.5 316.9 289.1 295.5 327.7 272.6 326.0 299.6 303.2 335.0 275.7 329.8 307.9 309.2 340.4 281.9 334.1 308.2 308.3 340.1 284.4 338.4 312.4 ANIMAL NUMBER 625325 625328 625338 625340 625352 DAY 8 318.6 352.0 287.4 347.0 316.9 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-2922 Company Sanitized. Does net contain T3CA - 46- O 53 Compan.y.." -nitirod. DOSS HG 00. 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 303.0 318.9 292.4 274.6 262.9 298.5 316.3 291.2 277.0 262.2 305.8 326.5 297.8 286.4 269.0 312.1 335.4 304.1 290.6 273.5 320.1 338.3 307.7 292.0 274.5 322.8 346.7 312.0 300.1 284.1 326.8 351.6 313.5 304.2 - ANIMAL NUMBER 625324 625329 625333 625334 625344 DAY 8 332.5 352.5 315.8 292.6 280.7 DAY 9 337.9 360.1 316.7 302.2 285.2 TEST DAYS DAY 10 DAY 13 336.1 357.0 319.3 305.8 289.3 299.6 364.3 325.8 312.0 295.8 DAY 20 336.0 396.4 349.2 338.6 309.7 DAY 24 __a DAY 27 380.3 423.3 375.2 368.8 330.4 ANIMAL NUMBER DAY 34 DAY 52 TEST DAYS 625324 625329 625333 625334 625344 397.3 442.7 384.6 384.2 348.3 447.4 498.1 427.4 426.5 391.0 SD test day 52 SD test day 52 SD test day 52 SD test day 52 SD test day 52 , a Rat was not weighed. 5" H _ _ O ' fi tu -47- DuPont-2922 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 625326 625335 625346 625347 625351 DAY 1 313.2 301.0 290.2 266.0 310.6 DAY 2 317.1 303.6 293.4 275.6 310.9 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP VI TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 325.5 302.3 300.2 287.7 321.1 330.9 310.4 309.5 300.0 329.6 336.2 315.1 313.5 308.5 338.2 336.0 299.6 318.6 316.6 341.6 _ 293.6 326.0 326.9 340.7 DuPont-2922 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 DAY 34 DAY 52 TEST DAYS DAY 62 DAY 76 DAY 80 DAY 90 DAY 94 625326 625335 625346 625347 625351 427.5 427.0 407.4 466.4 483.3 _a _a _a _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 day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 Company Sanitized. Dosa not contain TSCA cBi a Rat was not weighed. -48- 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) 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-2922 Company Sanitized. Coes net contain TSCA -49OCl not contain TSCA CB 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) OF MALE RATS GROUP II TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627768 627774 627776 627782 627785 236.9 258.6 212.0 221.6 220.3 244.1 261.6 216.3 228.3 221.7 245.2 275.8 223.7 237.3 234.7 256.5 283.4 226.6 239.0 238.4 256.2 289.3 229.6 238.8 241.8 265.5 301.5 233.7 244.7 250.5 271.4 303.2 236.9 246.7 255.6 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 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 TM3 <3< ms3a VS N 0e, oos -50- DuPont-2922 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) OF MALE RATS GROUP III TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627758 627763 627764 627787 627788 235.1 212.1 230.0 235.1 219.4 240.3 218.3 235.0 241.0 222.3 248.6 226.5 246.1 250.1 232.2 255.8 234.3 251.3 257.2 239.6 261.2 230.8 254.8 258.6 247.6 272.0 235.5 261.7 237.6 256.7 276.1 238.0 264.7 240.3 257.9 ANIMAL NUMBER DAY 8 DAY 9 TEST DAYS DAY 10 DAY 13 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 SD test day 13 SD test day 13 SD test day 13 DuPont-2922 Company Sanitized. Does not contain TSCA CBI -51 - 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) OF MALE RATS GROUP IV TEST DAYS DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 627759 627761 627770 627779 627786 226.6 240.7 235.6 231.5 211.6 232.9 247.4 244.3 237.9 218.2 241.4 257.1 253.6 252.6 222.5 250.9 262.5 260.1 261.4 230.4 254.2 266.7 267.7 262.3 230.6 262.1 259.2 277.3 273.8 240.6 265.4 262.6 279.8 277.5 244.1 ANIMAL NUMBER 627759 627761 627770 627779 627786 DAY 8 277.8 240.1 291.4 289.0 251.3 DAY 9 284.9 257.4 301.3 293.4 255.8 TEST DAYS DAY 10 DAY 13 288.0 266.9 306.4 301.0 257.0 __a _a __a _a _a DAY 20 348.0 335.7 355.8 374.4 304.4 DAY 24 380.1 359.4 379.8 406.2 326.1 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 DuPont-2922 '"Company Sanitized. D oes net contain TC <V}j a Rat was not weighed. Company Sanitized. Does noi contain TSC CSI 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) 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 DAY 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 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 . aRat was not weighed. 53- DuPont-2922 Oo 3o & v3i o38 fap4a aoW 3a ou*s3a** 3' 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) 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 627762 627769 627775 627778 627783 ANIMAL NUMBER 627762 627769 627775 627778 627783 DAY 8 285.9 302.0 286.6 244.8 256.6 DAY 41 438.7 479.7 433.2 356.8 409.6 DAY 9 299.1 309.6 296.0 250.8 259.2 DAY 47 439.9 474.6 451.1 373.5 426.9 TEST DAYS DAY 10 DAY 13 301.5 312.5 304.5 255.1 264.3 --a __a _a _a _a TEST DAYS DAY 52 DAY 55 __a 453.5 _a _a _a 512.2 460.8 386.0 449.0 DAY 20 356.5 386.6 353.3 294.0 311.3 DAY 61 473.0 511.4 477.4 393.3 473.8 DAY 24 381.5 414.4 377.4 320.0 340.9 DAY 68 471.6 531.4 478.6 398.3 474.8 DAY 34 419.6 456.5 413.9 347.1 380.0 a Rat was not weighed. DuPont-2922 H-23960: 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 GROUP VI TEST DAYS DAY 94 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-2922 Company Sanitized. Does not contain TSC CBI - 55- 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 I DAY 3 DAY 4 DAY 5 626831 626837 626846 626858 626860 258.3 250.3 224.7 264.8 239.8 265.4 248.5 227.5 271.5 243.2 273.1 257.1 237.1 276.7 257.6 282.6 259.2 243.6 287.4 265.0 287.8 267.7 254.9 294.8 268.0 SD test day 5 SD test day 5 SD test day 5 SD test day 5 SD test day 5 DuPont-2922 Company Sanitized. D o ss not contain i S C k Ci S3 5 6 - 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 II DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 626832 626841 626842 626851 626859 ANIMAL NUMBER 249.8 232.4 274.3 257.9 251.4 DAY 8 253.9 241.2 280.2 265.8 257.8 264.9 246.5 289.0 271.3 269.8 DAY 9 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 626832 626841 626842 626851 626859 297.4 277.0 316.0 298.4 307.3 301.5 282.0 323.5 304.0 310.5 312.1 283.9 331.2 307.8 318.5 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 DuPont-2922 P IIP H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats________________________________________ H-23960 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP III ANIMAL NUMBER DAY 1 DAY 2 DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 626836 626839 626845 626849 626856 ANIMAL NUMBER 257.7 251.8 265.9 241.3 248.3 DAY 8 261.7 254.1 272.3 252.1 261.0 273.4 266.0 276.6 258.3 271.3 279.8 269.7 278.8 262.9 275.3 DAY 9 DAY 10 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 626836 626839 626845 626849 626856 315.5 289.5 313.3 283.6 310.3 319.9 296.3 324.3 283.0 321.6 329.3 304.3 324.6 285.7 328.2 335.8 314.4 348.2 297.6 343.0 SD test day- 13 SD test day 13 SD test day 13 SD test day 13 SD test day 13 DuPont-2922 Company Sanitized. Does not contain TSCA CBS -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 626843 626844 626850 626853 626857 244.0 222.3 264.0 248.3 253.2 DAY 8 306.6 269.6 319.1 309.0 295.3 250.2 232.3 270.6 252.6 264.2 263.2 243.1 278.1 261.3 267.5 270.1 248.3 284.8 271.2 271.8 279.2 255.5 300.4 281.3 277.6 286.4 264.9 309.7 288.7 287.3 291.9 268.0 309.0 300.0 288.3 DAY 9 312.1 282.4 326.9 326.3 302.5 DAY 10 318.1 290.9 332.2 328.0 308.0 DAY 13 _a _a _a _a _a DAY 24 395.7 358.9 423.0 445.4 386.3 SD test day 24 SD test day 24 SD test day 24 SD test day 24 SD test day 24 DuPont-2922 Company Sanitized. D oes nei contain TSC CBi a Rat was not weighed. -59- Company Sanizad. Does H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER DAY 1 DAY 2 H-23'960 (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 626833 626835 626852 626861 626862 ANIMAL NUMBER 282.2 252.8 239.4 249.7 257.9 DAY 8 348.9 312.9 299.1 284.5 328.8 DAY 52 291.3 261.5 249.9 252.9 265.6 299.1 272.0 257.0 254.3 278.7 309.9 280.5 266.4 262.3 289.6 320.9 291.7 274.6 270.3 302.3 326.1 299.3 286.1 276.6 311.5 333.9 304.6 289.5 280.8 320.0 DAY 9 356.9 319.7 310.6 297.0 340.6 DAY 10 367.6 330.4 316.1 298.1 357.2 DAY 13 _a _a _a _a _a DAY 24 _a _a __a _a _a DAY 34 481.7 432.6 417.3 392.4 519.1 DAY 48 536.9 485.1 452.8 450.4 593.3 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 nu~Ji oo I B2" a Rat was not weighed. CO DuPont-2922 > 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 VI DAY 3 DAY 4 DAY 5 DAY 6 DAY 7 DuPont-2922 626847 626848 626854 626855 626863 ANIMAL NUMBER 626847 626848 626854 626855 626863 `o ANIMAL NUMBER 235.7 252.1 220.5 272.0 247.4 DAY 8 277.5 302.3 255.9 328.0 312.9 242.5 258.9 224.5 279.5 252.5 247.4 269.0 226.9 286.6 270.8 254.4 277.7 235.0 297.0 270.6 258.3 284.0 241.9 302.9 281.6 269.2 290.5 245.0 313.8 292.4 272.0 299.5 247.0 317.9 298.9 DAY 9 284.0 308.8 260.3 331.5 318.5 DAY 10 291.3 316.5 261.6 339.9 325.6 DAY 13 _a _a _a _a DAY 24 --a _a __a _a _a DAY 34 381.7 396.0 347.9 476.6 452.3 DAY 48 428.8 456.6 411.0 541.1 510.7 DAY 52 DAY 62 DAY 69 DAY 75 DAY 83 DAY 89 DAY 93 626847 626848 626854 626855 626863 437.7 471.5 412.9 552.3 535.2 448.9 485.8 430.0 577.9 561.4 460.6 505.4 441.8 599.0 584.1 475.8 516.1 454.2 614.4 588.8 489.4 535.0 459.6 625.4 606.2 498.1 544.3 460.9 637.1 604.5 501.0 553.4 473.8 641.0 616.7 SD test day 93 SD test day 93 SD test day 93 SD test day 93 SD test day 93 MMipany Sauteed. Doss r,i couiain TSCA a Rat was not weighed. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDIX B Individual Clinical Observations - 62- Does net contain TSCA CBl Company Sanitized. H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 CORN OIL (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number- 627789 627801 627805 627810 627818 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 __________________ 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 -63- Company Sanitized. Does it contain A CBS H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 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 - 64- SanXized. Does nct contain Company Zh C H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 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 -65Company SaaAized. .e s noi contain TOCk C1 H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 CORN OIL:ACETONE (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number 625354 625365 625371 625376 625381 GROUP IV Observation No' abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-24 1-24 1-24 1-24 1-24 Animal Number 625356 625359 625360 625372 625374 GROUP V Observation No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected Test Day 1-52 1-52 1-52 1-52 1-52 Animal Number 625353 625369 625373 625375 625377 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 SarriCzed. Does not contais 1CCA CBl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP I Observation________________________ Test Day 625322 625323 625336 625343 625349 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 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- C om pany Sanitized. Doss net contain CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP IV Observation________________________ Test Day 625325 625328 625338 625340 625352 No abnormalities detected No abnormalities detected No abnormalities detected Salivation No abnormalities detected No abnormalities detected 1-24 1-24 1-6, 8-24 7 1-24 1-24 Animal Number 625324 625329 625333 625334 625344 GROUP V Observation No abnormalities detected No abnormalities detected Salivation No abnormalities detected Salivation No abnormalities detected No abnormalities detected Test Day 1-52 1-6, 8-52 7 1-6, 8-52 7 1-52 1-52 Animal Number 625326 625335 625346 625347 625351 GROUP VI Observation No abnormalities detected Salivation Alopecia both front legs Alopecia abdomen No abnormalities detected Salivation Black ocular discharge Yellow-stained perineum Red-stained chin Brown-stained chin No abnormalities detected No abnormalities detected Salivation Test Day 1-6, 9-76, 77-94 7-8 76 7, 8, 9, 10, 13, 20, 27 1-6, 11-94 7-8 8 8 8 9-10 1-94 1-6, 8-94 7 -68Sanitized. Does \ yi contait TC---D C o mp-'-'tf H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 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, 7-13 6 1-8, 10-13 9 -69anitized. Does not contain TSCA CB1 Comps"y S H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP IV Observation Test Day 627759 627761 627770 627779 627786 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 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. D oes not contain TSCA CBl H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 H-23960 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP I Observation________________________ Test Day 626831 626837 626846 626858 626860 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 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 Com pany Sanitized. D oes not contain TSCA CBf H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 H-23960 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Number GROUP IV Observation __________ Test Day 626843 626844 626850 626853 626857 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 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____________;____________ Test Day No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected No abnormalities detected 1-93 1-93 1-93 1-93 1-93 Company Sanitized. Does not contain TSC CBI H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-2922 APPENDIX C Analysis of Blood Fluorine Data -73Cotnpany amtaecf. ua** co.aa^i i A Cbk Company Sanitized. Does* not contain T8CA E H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Terms and Calculations DuPont-2922 Terms: Active . Fluorine containing compound % Active The % of formulation that is made up o f fluorine containing components Mol Wt Active The molecular weight o f the fluorine containing components (g/mole) Formulation Dose % F in Active The mg o f formulation given per kg o f 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 o f animal body weight = (% F in active/100) * Dose Active [mg/kg] Dose F (mmol/kg) The mmole o f 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 o f Active [mmol/kg] / Dose Active [mmol/kg]) * Formulation Dose -74- fszed.Dcasr.oic unlain TwCA C Company G 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. 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 -75 - Company zed. D oes not contain TSCA CE; H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Factors Influencing Interpretation of Kinetic Analysis DuPont-2922 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 o f compounds - Elimination kinetics can be determined based on total fluorine rather than on concentrations of individual components - Background plasma Fluorine is 0.2 ppm - % F data is the % Fluorine of the active (Fluorine containing component(s) in the formulation - Molecular weight is the molecular weight of the active component in the formulation - 7 6 - Jompany Sanitized. Does not c ia '.n TSC CBS H-23960: 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-2922 65 19 0.342 0.059 6.5 R at# 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 F in blood minus Bkg 0.2 ppm ppm F in blood Normalized to 0.1 mmoles/kg Dose 0.8 1-1 . 2.4 2.4 2.7 3.98 5.47 11.93 11.93 13.42 51.5 255.96 37.8 187.87 38.5 191.35 78.3 389.15 36.9 183.39 65.4 325.04 60.9 302.67 60.3 299.69 62.9 312.61 57.9 287.76 unolar 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 Company Sanitized. net contain TSCft C" l H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat # Test ppm F Day in Sample blood Group III 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 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 molar equivalents of active in blood 1184.62 943.08 990.77 873.85 956.92 616.92 709.23 661.54 598.46 641.54 376.92 455.38 416.92 444.62 363.08 187.69 249.23 198.46 166.15 175.38 78- ;.3S not contain rscA C ji Compaia Samtized . D H-23960: Biopersistence Screening 10-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-2922 69 19 0.726 0.065 13.8 R at# 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 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 /t m o la r 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 ,, Pi?"* 79- H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats____________________________________________________________ DuPont-2922 R at# Test ppm F Day in Sample blood ppm F in blood minus Bkg 0.2 ppm ppm F in blood Normalized to 0.1 mmoles/kg Dose /molar equivalents of active in blood Group III 627758 627764 627787 627763 627788 13 13 13 13 13 43.5 20.7 13.2 30.8 24.9 43.3 20.5 13.0 30.6 24.7 92.23 43.67 27.69 65.18 52.61 313.77 148.55 94.20 221.74 178.99 Group IV 627759 627779 627770 627761 627786 24 24 24 24 24 9.2 14.9 11.6 9.9 6.7 9.0 14.7 11.4 9.7 6.5 19.17 31.31 24.28 20.66 13.85 65.22 106.52 82.61 70.29 47.10 Group V 627757 627781 627760 627765 627772 52 52 52 52 52 0.9 0.6 1.0 0.9 0.8 0.7 0.4 0.8 0.7 0.6 1.49 5.07 0.85 2.90 1.70 5.80 1.49 5.07 1.28 4.35 Group VI 627778 627783 627762 627775 627769 94 94 94 94 94 0.3 <0.2 0.2 0.2 <0.2 0.1 * 0.0 0.0 * 0.21 0.72 ** 0.00 0.00 0.00 0.00 ** * = Below LOD (Level o f Detection) -80- Company Sanitized. D oes not contain TSC CBS H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H- 23960 DuPont-2922 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 27500 1000 18 % F in Active: Mol Wt. F (g/mol): 33 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Formulation Dose Normalization Factor: 180 0.007 15277.8 Dose F (mmol/kg): Molar Ratio (Active/F): Dose F (mg/kg): 3.126 0.002 59.4 R at# Test ppm F Day in Sample 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 626837 626846 626858 626860 5 5 5 5 5 1.4 0.9 0.9 1.2 0.7 Group II 626832 626841 626842 626851 626859 10 10 10 10 10 1.7 1.0 1.4 2.1 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 blood Normalized to 0.1 mmoles/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 - not contain TSCft CBt Company Sanitized- D ss H-23960: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Rat # Test ppm F Day in Sample blood Group III 626836 626839 626845 626849 626856 13 13 13 13 13 0.7 1.1 1.0 0.6 1.4 G roup IV 626843 626844 626850 626853 626857 24 24 24 24 24 0.5 0.4 <0.2 <0.2 0.2 Group V 626833 626835 626852 626861 626862 52 52 52 52 52 0.3 0.4 0.2 0.4 0.5 Group VI 626847 626848 626854 626855 626863 93 93 93 93 93 0.3 0.2 0.4 0.4 0.3 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 * = Below LOD (Level o f Detection) ppm F in blood Normalized to 0.1 mmoles/kg Dose DuPont-2922 limolar equivalents of active in blood 7.64 13.75 12.22 6.11 18.33 0.84 1.52 1.35 0.67 2.02 4.58 0.51 3.06 0.34 ** ** 0.00 0.00 1.53 . 3.06 0.00 3.06 4.58 0.17 0.34 0.00 0.34 0.51 1.53 0.17 0.00 0.00 3.06 0.34 3.06 0.34 1.53 0.17 Company Sanitized. D oes not contain TSCA CB1