Document Dd0eaZVn5yy8kqjeNKM9ozBEO

\ o \ 00 " sn y itt TRADE SECRET Study Title H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Laboratory Project ID : DuPont-1155 8 DuPont-11558 Author: Carol Finlay, B.A. Study Completed on: April 7,2003 Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 Company Sanitized. Does noi contain TSCA CBI Page 1 o f 82 H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study. Reviewed by: Paul M. Hinderliter, Fh.D . Research T oxicologist o 7 - t " a3 Elate Issued by Study Director: Carol pinlay, B.A . Staff T oxicologist / " " "N Company Sanitized. Does not contain T SC A CBI H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE OF CONTENTS Page CERTIFICATION................................................................................................................................. 2 LIST OF TABLES...................................................................................?............................................ 4 LIST OF FIG URES............................................................................................................................... 4 LIST OF APPENDICES...................................................................................................................... 5 STUDY INFORM ATION.................................................................... 6 STUDY PERSONNEL.......................................................................... 8 SUM M ARY............................................................................................................................................. 9 INTRODUCTION................................................................................................................................ 11 MATERIALS AND M ETHODS............................................................................... A. Test Substance and Positive Controls............................................................. B. Test Species....................................................................................................... C. Animal Husbandry............................................................................................. D. Quarantine and Pretest.............................................................................................................. 12 E. Study Design.............................................................................................................................. 13 F. Assignment to Groups and Study Start................................................................................... 13 G. Dosing Material Preparation and Administration................................................................. 13 H. Body W eights............................................................................................................................. 14 I. Mortality and Clinical Observations....................................................................................... 14 J. Collection and Analysis o f Blood, Livers, and F a t............................................................... 14 K. Treatment o f Fluorine Data.......................................................................................................15 L. Statistical A nalyses....................................................................................................................16. RESULTS AND DISCUSSION.........................................................................................................17 A. In-Life Toxicology.....................................................................................................................17 B. Liver W eights............................................................................................................................. 17 C. Fluorine D a ta ............................................................................................................................. 18 C O N C L U S IO N S ................................................................................................................................... 19 RECORDS AND SAMPLE STORAGE......................................................................................... 20 T A B L E S ................................................................................................ F I G U R E S ......................................................................................................................... APPENDICES....................................................................................................................................... 39 11 11 11 11 28 Company Sanitized. Does not contain T SCA CBI H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 LIST OF TABLES Page 1. MEAN BODY WEIGHTS................................................................................................................................................. 22 2. MEAN BODY WEIGHT GAINS.....................................................................................................................................23 3. MEAN BODY AND LIVER WEIGHTS.........................................................................................................................24 4. MEAN BLOOD FLUORINE LEVELS...........................................................................................................................25 5. MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO D O SE ................................................... 25 6. MEAN LIVER FLUORINE LEVELS.............................................................................................................................26 7. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO D O SE ......................................................26 8. MEAN FAT FLUORINE LEVELS................................................................................................................................. 27 9. MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE........................................................... 27 LIST OF FIGURES Page 1. MEAN BODY WEIGHTS............................................................................................................................................... 29 2. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE CONTROL................................................................................................................. 30 3. COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS......................................................................................................................................................................... 31 4. MICROMOLAR EQUIVALENTS IN RAT BLOOD..................................................................................................32 5. NORMALIZED H-24743 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A 10-DAY ORAL G AVAGE............................................................................................................................................... 34 6. MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO D O S E .....................................................35 7. COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24743 AND NEGATIVE CONTROL........................................................................36 8. MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE...........................................................37 9. COMPARISON OF MEAN BLOOD, MEAN LIVER, AND MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DO SE........................................................................................................ 38 /""N Company Sanitized. Does not contain TSCA CBI -4 - H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 LIST OF APPENDICES Page A. INDIVIDUAL BODY WEIGHTS.............................................................................. 40 B. INDIVIDUAL CLINICAL OBSERVATIONS..............................................................................................................50 C. TERMS AND CALCULATIONS FACTORS INFLUENCING INTERPRETATION OF KINETIC ANALYSIS.......................................................................................................................................................................... 64 D. INDIVIDUAL FLUORINE LEVELS IN BLOOD...................................................... 68 E. INDIVIDUAL FLUORINE LEVELS IN LIVER......................................................................................................... 75 F. INDIVIDUAL FLUORINE LEVELS IN F A T .................................................... 79 /"""N Company Sanitized. Does not contain TSCA CBt rsjopersistence Screening 10-Dose Oral Gavage Study in Rats Test Substance STUDY INFORMATION DuPont-11558 Haskell Number: 24743 Positive Control /****''\ Substance Tested: Synonyms/Codes: H-24019 Haskell Number: 24019 -6- Company Sanitized. Does not contain t r o a C 0i n - z ^ w : iopersistence Screening 10-Dose Oral Gavage Study in Rats Positive Control STUDY INFORMATION (Continued) DuPont-11558 Haskell Number: 24020 Sponsor: E.I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: October 10, 2002 / (see report cover page) Company Sanitized. Does not contain TSCA CB1 -7 - H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 STUDY PERSONNEL Study Director: Carol Finlay, B.A. Management: Scott E. Loveless, Ph.D. Primary Technician: James C. Mackay II Fluorine Data Analysis: Paul M. Hinderliter, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T. Toxicology Report Preparation: Sean M. Callaghan, B.A. Management: Nancy S. Selzer, M.S. Laboratory Veterinarian: Thomas W. Mayer, D.V.M., A.C.L.A.M. .Company Sanitized. Does not contain TSCA CBI H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 SUMMARY The objective of this study was to evaluate the potential for H-24743 to be absorbed and to accumulate in a mammalian system. Two groups o f 5 male rats each were dosed by gavage with 300 mg/kg/day o f H-24743 for 10 consecutive days. One group was sacrificed on Day 10, and the other group was maintained without dosing for an additional 84 days. Blood was collected on test days 1,5, 10, 13, 24, 52, and 94. Livers and fat were collected at sacrifice. Body weights and clinical signs were recorded throughout the dosing and recovery periods. Additionally, a negative control o f deionized water, and 2 positive controls, H-24019 (10 mg/kg/day) and H-24020 (20 mg/kg/day), were tested as described for H-24743. Blood, liver, and fat samples were analyzed for total fluorine content to determine test substance absorption and retention in these tissues. No deaths occurred, and no clinical signs attributed to the test substance were observed. The mean body weight gains o f rats dosed with H-24743 were similar to the negative control rats during the dosing and recovery periods. Mean body weight gain o f rats dosed with the positive control materials was lower during the dosing period than the weight gain o f rats dosed with the test substance. During the recovery period, the mean body weight gain o f rats dosed with the positive control materials was higher than the weight gain o f rats dosed with the test substance. For the purpose o f comparing absorption and retention o f the test substance to the positive controls, all doses were normalized on a molar basis. The total fluorine in each tissue was considered as a percentage o f the fluorine delivered in the administered dose, and all units were expressed as pM equivalents. As a measure o f total dose in the blood, an area under the curve (estimated to infinity) was calculated for each test material. The AUCINF/D for the fluorine component o f H-24743 was 1.07x 103, compared to AUCINF/D values o f 5.22x10s and 8.15xl04 for H-24019 and H-24020, respectively. This parameter indicated a total dose in blood that was much less for H-24743 than for the positive controls. The mean relative liver weight (liver/body weight) of rats dosed with H-24743 was 31% higher than the negative control rats on day 10. By day 94, the mean relative liver weight o f rats dosed with H-24743 was 11% higher than the negative control. The mean relative liver weight o f rats dosed with H-24019 was 41% higher and for H-24020 was 93% higher on day 10 than the negative control group. By day 94, the mean relative liver weights o f rats dosed with H-24019 and H-24020 were similar to the negative control group. Therefore, liver weights o f rats dosed with the test substance were higher than the negative controls at both the end o f the dosing period and at the end of the recovery period. The concentration o f fluorine in the livers from rats dosed with H-24743 was 27.52 pM equivalents on day 10 and 2.26 pM equivalents on day 94. On the last day o f dosing, the mean pM equivalent concentrations of fluorine in the livers of rats dosed with the positive control materials were approximately 174-fold (H-24019) and 31-fold (H-24020) greater than values in rats treated with the test substance. By day 94, the fluorine concentrations were approximately 572x and 8x the fluorine concentration in rats treated with H-24743. Therefore, rats treated with -9 - Oompany Sanitized. Does not contain TSCA CBl H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 H-24019 or H-24020 absorbed and retained considerably more fluorine in the liver than rats treated with H-24743. The fluorine concentration in the fat from rats dosed with the test substance was 49.10 pM equivalents on day 10 and 10.87 jiM equivalents on day 94. The fluorine concentration o f one of the positive control materials, H-24019, was approximately 4x higher than H-24743 on day 10. The fluorine concentration o f the other positive control material, H-24020, was similar on day 10 to the fluorine concentration o f the test substance, H-24743. By day 94, the fluorine concentration of H-24019 was similar to the fluorine concentration in rats treated with H-24743. There was no detectable fluorine by day 94 in the fat from rats dosed with the positive control H-24020. Under the conditions o f this study, fluorine was evident in the blood during the period o f dosing with H-24743, but was quickly eliminated. The amounts o f fluorine in the livers and fat from rats dosed with the test substance were higher than levels in the blood, and the results indicate that there was evidence o f some retention o f fluorine in the liver and fat. Fluorine levels in the fat were generally slightly higher or similar to the positive control materials. Fluorine levels in the blood and liver were considerably lower than levels in rats dosed with the positive controls, H-24019 and H-24020. Company Sanitized. Does not contain TSCACBS - 10- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 INTRODUCTION The objective o f this study was to define the potential o f H-24743 to be absorbed and to bioaccumulate in a mammalian system, as indicated by analytical determination of total fluorine in blood, liver, and fat. The test substance was compared to 2 positive controls that were materials previously shown to bioaccumulate in mammals. The daily dosage o f 300 mg/kg for the test substance was selected based on existing toxicity information and the results o f a rangefinding study. Groups o f five rats were dosed at 0 (control) or 300 mg/kg for 6 consecutive days. The rats dosed with the test substance had an overall body weight gain o f 24 grams compared to 28 grams for the control rats. The dosage o f 300 mg/kg was chosen for the study and was expected to produce less than a 10% difference in mean body weight over 10 days when compared to the negative control. MATERIALS AND METHODS A. Test Substance and Positive Controls The test substance, H-24743, was supplied by the sponsor as a clear, colorless 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 o f extensive experience with this strain and its suitability with respect to longevity, hardiness, and low incidence of spontaneous diseases. C. Animal Husbandry 1. Housing Environment Rats were housed singly in stainless steel, wire-mesh cages suspended above cage boards. Animal rooms were maintained on an approximate 12-hour light/dark cycle (fluorescent light) and at a temperature o f 23 1C and a relative humidity o f 50 10%. Company Sanitized. Does not contain TSCA CBi H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 2. Feed and Water Tap water was provided ad libitum. All rats were fed PMI Nutrition International, LLC Certified Rodent LabDiet 5002 chow. The feed is guaranteed by the manufacturer to meet specified nutritional requirements and to be free o f specified contaminants. 3. Identification Each rat was assigned a unique identification number which was recorded on a card affixed to the cage. The last 3 digits o f the number were tattooed on the tail o f each rat. 4. Health Monitoring Program As specified in the Haskell Laboratory animal health and environmental monitoring program, the following procedures are performed periodically to ensure that contaminant levels are below those that would be expected to impact the scientific integrity o f the study: Water samples are analyzed for total bacterial counts, and the presence o f coliforms, lead, and other contaminants. Feed samples are analyzed for total bacterial, spore, and fungal counts. Samples from freshly washed cages and cage racks are analyzed to ensure adequate sanitation by the cagewashers. Certified animal feed is used, guaranteed by the manufacturer to meet specified nutritional requirements and not to exceed stated maximum concentrations o f key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence o f these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity o f the study. The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. Evaluation o f these data did not indicate any conditions that affected the validity o f 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 clinically apparent signs o f disease or injury. The rats were observed daily for mortality and signs o f illness, injury, or abnormal behavior. On the bases o f acceptable body weight gains and freedom from clinically apparent signs o f disease or injury, the rats were released from quarantine by the designee o f the laboratory animal veterinarian. Company Sanitized. Does not contain TSCA CB1 H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats E. Study Design Dosage Number of Substance Vehicle (mg/kg) Animals Negative Control Deionized water Not applicable 0 10 Positive Controls H-24019 Acetone/Com Oil 10 10 H-24020 Acetone/Com Oil 20 10 Test Substance H-24743 Deionized water 300 10 DuPont-11558 F. Assignment to Groups and Study Start After the quarantine period, the rats were selected on the bases o f adequate body weight gain, freedom from any clinical signs o f disease or injury. The selected rats were arbitrarily assigned to each group. After assignment to groups, each rat was housed individually. The rats were 7 weeks o f age at the time o f dosing. Dosing began on test day 1. Rats that were not assigned to the study were released for other laboratory purposes, or were sacrificed by carbon dioxide asphyxiation and discarded without pathology evaluation. G. Dosing Material Preparation and Administration 1. Test Substance H-24743 was mixed with deionized water so an accurate volume could be administered. The amount o f test substance each rat received was based on the body weight collected on each day of dosing and the mixture concentration. The rats were dosed at a volume of 1 mL/100 g of body weight. The mixture was stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. 2. Positive Controls It was necessary to dissolve H-24019 and H-24020 in acetone before suspending them in com oil. The ratio o f acetone to com oil was 20:80. The amount each rat received was based on the body weight collected on each day o f dosing and the suspension concentration. The rats were dosed at a volume 1 mL/100 g of body weight. The dosing preparations were stirred on a magnetic stir plate throughout the dosing procedure to maintain homogeneity. Company Sanitized. Does not contain TSCA C8I - 13 - /s'-'X H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 3. Negative Control Deionized water was chosen as the negative control. The rats were dosed at a volume of 1 mL/100 g o f body weight. 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 and weekly during the recovery period. I. Mortality and Clinical Observations Cage-site examinations to detect moribund or dead rats and abnormal behavior and appearance among rats were conducted at least once daily throughout the study. At every weighing, each rat was individually handled and examined for abnormal behavior and appearance. J. Collection and Analysis of Blood, Livers, and Fat At time points selected for blood sampling other than sacrifice days, approximately 1 mL of blood was collected into EDTA tubes from the orbital sinus of each rat. Rats designated for sacrifice were euthanized by carbon dioxide anesthesia and exsanguination and blood, livers, and fat were collected according to the following schedules: Group Dosing Days Tissue Collected Sampling Time I 1-10 Blood Test day 1 (2 hours post dosing) I 1-10 Blood Test day 5 (2 hours post dosing) I 1-10 Blood, Liver, and Fat Test day 10 at sacrifice (2 hours post dosing) III 1-10 Blood Test day 13 III 1-10 Blood Test day 24 III 1-10 Blood Test day 52 III 1-10 Blood, Liver, and Fat Test day 94 As much blood as possible was collected into EDTA tubes at sacrifice. The livers and fat were weighed. The fat weights were used for the calculation of fluorine levels. The liver weights were used for the calculation o f fluorine levels and for the calculation o f liver weight relative to total body weight. The blood from all rats was refrigerated and the livers were frozen. The livers and fat were appropriately packaged and shipped refrigerated to Jackson Laboratory, Deepwater, New Jersey where they were analyzed for total fluorine. The liver and fat samples remained frozen during shipping. The total fluorine content of the blood and tissue samples was determined using a Wickbold torch combustion method, followed by analysis with a fluoride ion selective electrode. The samples were decomposed or volatilized in the presence of wet oxygen and swept through an oxy-hydrogen flame in a closed quartz apparatus. The combustion products were collected in an -14jpompany Sanitized. Does not contain T SCA c a t H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 aqueous absorbing solution and analyzed with a fluoride ion selective electrode. Kinetic analysis of the data received (ppm F in each sample) was performed by Haskell Laboratory personnel for evaluation o f fluorine biopersistence. K. Treatment of Fluorine Data Since the test substance and positive controls had different toxicity profiles, it was not possible to administer a uniform mg/kg dose for all test materials. Properly conducted kinetic comparisons in situations with varied doses required the use of a normalized dose. The dosenormalization was conducted in pmolar units to accommodate test material molecular weight differences and was adjusted to an arbitrary normalized dose o f 0.1 mmoles/kg. H-24019 and H-24020 were used as positive controls and a dosing diluent was used as the negative control. The percent of fluorine and molecular weight o f the test substance and positive controls were used as provided by the sponsor. The measured total fluorine values were used as received (ppm F) from Jackson Laboratory. For the Wickbold torch method, the background fluorine level is 0.2 ppm. This is the limit o f detection (LOD) o f this method and was subtracted from each sample. The limit o f quantification (LOQ) for this method is 0.5 ppm, and any values listed as less than 0.5 ppm were excluded from further treatment. Since the dosages and molecular weight for positive controls and test material differed, the data were converted from a mass to a molar basis. In addition, the data were standardized to a single reference dose to allow comparison between compounds o f differing fluorine content. The doses were first converted from a mg o f test material basis to millimoles o f fluorine. Raw fluoride ion data (ppm F) were then normalized to a 0.1 millimole dose o f active component. Finally the molar dosage and normalized concentration were combined to yield the pmolar (pM) equivalents of active component in the tissue. Detailed calculations can be found in Appendix C. The pM equivalents can be compared across compounds provided the considerations listed in Appendix C are observed. Noncompartmental analysis was conducted on fluorine data derived from rats dosed with H-24743 and the positive controls using WinNonlin Version 4.0 software (Pharsight Corp, Mountain View, CA). WinNonlin software provided a means o f computing derived pharmacokinetic parameters from experimental data. All analysis was calculated using the pM equivalent in blood data. The maximum observed concentration in blood was Cmax (pM equivalent). Biopersistence was assessed by quantifying terminal elimination blood half-life (T./,, days). The points included in determination o f the Ty2were selected manually and included only points after apparent loglinear elimination was achieved. Internal exposure was determined by calculating the blood area-under-the-curve (AUC). AUC, which is simply the integral o f blood concentration over time, is the most common means for expressing internal dose. With the calculated half-life, the AUC can be extrapolated to infinity (AUCINF) to reflect the elimination o f the compound. AUCINF normalized to dose (AUCINF/D) can be used to compare the relative exposures of different compounds and dosages. -15- pjompany Sanitized. Does not contain T SC A CB H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats L. Statistical Analyses Descriptive statistics (e.g. mean, standard deviation) were used. DuPont-11558 Gompany Sanitized. Does not contain TSCA CB1 - 16- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 /r* \ RESULTS AND DISCUSSION A. In-Life Toxicology (Tables 1-2, Figure 1, Appendices A-B) No deaths occurred. Hair loss observed in one rat during the recovery period was not considered to be test substance-related because it was only seen in one rat. The mean body weight gains o f rats dosed with H-24743 were similar to the negative control rats during the dosing and recovery periods. Mean body weight gain o f rats dosed with the positive control materials was lower during the dosing period than the weight gain o f rats dosed with the test substance. During the recovery period, the mean body weight gain o f rats dosed with the positive control materials was higher than the weight gain o f rats dosed with the test substance. B. Liver Weights (Table 3, Figures 2, 3, and 7) 1. Test Substance The mean relative liver weight (liver/body weight) o f rats dosed with H-24743 was 31 % higher than the negative control rats on day 10. By day 94, the mean relative liver weight o f rats dosed with H-24743 was 11% higher than the negative control. 2. Positive Controls The mean relative liver weights o f rats dosed with one o f the positive controls, H-24019, were similar on days 10 and 94 to the liver weights o f rats dosed with the test substance, H-24743. The mean relative liver weight o f rats dosed with the other positive control, H-24020, was 44% higher at day 10 than the liver weight o f rats dosed with H-24743. By day 94, the weights were similar. The mean relative liver weight o f rats dosed with H-24019 was 38% higher at day 10 than the negative control group. By day 94, the mean relative liver weight o f rats dosed with H-24019 was similar to the negative control group. The mean relative liver weight of rats dosed with H24020 was 88% higher on day 10 than the negative control group. By day 94, the weights were similar. Therefore, the relative liver weights o f rats dosed with the test substance, H-24743, were higher than the negative controls at both the end o f the dosing period and at the end o f the recovery period. jPginpany Sanitized. Does not contain TSCA CBI -17- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 C. Fluorine Data (Tables 4-9, Figures 4-9, Appendices C-F) 1. Factors Influencing Interpretation o f Analysis The data used in the kinetic analysis were derived from a limited screen, and therefore several caveats and considerations are important. A couple o f considerations o f particular importance are (1) a single dose was used and kinetics may or may not be linear, (2) the kinetics apply only to blood, (3) steady-state may not have been achieved, and (4) the sample size is low and may impact calculation o f the terminal half-life. A more complete list o f considerations is shown in Appendix C. 2. Positive Controls The positive controls were H-24019 and H-24020. The H-24019 and H-24020 normalized pM equivalents in rat blood continued to rise throughout the dosing period and may not have reached steady-state (Figure 4A and 4B). The Cmax for H-24019 was 1043.08 54.57 pM equivalents (mean S.D.) with a terminal half-life of 42.2 days. The Cmax for H-24020 was 541.45 50.37 pM equivalents (mean S.D.) with a terminal half-life o f 15.1 days. For each of the positive controls, blood was sampled at 7 time points throughout the study, with only 4 of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between positive controls and the test substance. The AUCINF/D for the fluorine component was 5.22 x 105 for H-24019 and 8.15 x 104 for H-24020. The concentrations o f fluorine in the livers on day 10 from rats dosed with the positive control materials were 4802.15 and 866.67 pM equivalents for H-24019 and H-24020, respectively. By day 94, the concentrations were 1292.92 and 17.10 pM equivalents. The concentrations o f fluorine in the fat on day 10 from rats dosed with the positive control materials were 194.46 and 57.25 pM equivalents for H-24019 and H-24020, respectively. By day 94, the concentration o f fluorine in the fat from rats dosed with H-24019 was 15.38 pM equivalents. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020. 3. Test Substance The H-24743 normalized uM equivalents in rat blood rose rapidly and appears to reach steadystate (Figure 4C). The Cmax for H-24743 was 4.39 5.66 uM equivalents (Mean SD) with a terminal half-life o f 9.5 days. Blood was sampled at seven time points throughout the study, with only four of them occurring post-dose. The small sample size and analytical variability should be taken into account when using the derived terminal half-life for comparative purposes. The total internal exposure resulting from a normalized dose was described by AUCINF/D and was the basis for comparison between H-24743 and positive controls. The - 18figppanjf Sanitized. Does not contain T SC A fiat H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 AUCINF/D for the fluorine component o f H-24743 was 1.07xl03 as compared to AUCINF/D values of 5.22xl05 and 8.15xl04 for H-24019 and H-24020, respectively. Levels o f total fluorine in livers from rats dosed with the test substance, H-24743, were lower than the levels in livers from rats dosed with the positive control materials. The total fluorine concentration in the liver from rats dosed with H-24743 was 27.52 uM equivalents at day 10 and 2.26 pM equivalents at day 94. For the positive control H-24019, the liver concentrations were approximately 174x higher (day 10) and approximately 572x higher (end o f study) than H24743. For H-24020, the liver concentrations were approximately 3lx higher (end of dosing) and 8x higher (end o f study) than H-24743. Levels of total fluorine in fat from rats dosed with the test substance were slightly higher than or similar to the levels in fat from rats dosed with the positive control materials. The fluorine concentration in the fat from rats dosed with the test substance was 49.10 pM equivalents on day 10 and 10.87 pM equivalents on day 94. The fluorine concentration o f one o f the positive control materials, H-24019, was approximately 4x higher than H-24743 on day 10. The fluorine concentration of the other positive control material, H-24020, was similar on day 10 to the fluorine concentration of the test substance, H-24743. By day 94, the fluorine concentration o f H-24019 was similar to the fluorine concentration in rats treated with H-24743. There was no detectable fluorine by day 94 in the fat from rats dosed with H-24020. The uM equivalents in the liver and fat were higher than levels in the blood. CONCLUSIONS Rats dosed for 10 consecutive days with 300 mg/kg H-24743 exhibited no mortality, no test substance-related clinical signs, and no body weight effects. Liver weights were affected in rats dosed with the test substance. A steady-state for fluorine in the blood appeared to be achieved during the 10-day dosing period with the test substance. Dose-adjusted areas under the curve (AUCINF/D) for positive controls, H-24019 and H-24020, were approximately 490x and 75x the AUCINF/D for the test substance. Under the conditions o f this study, there was limited absorption and no retention of fluorine in the blood following dosing with H-24743. Administration o f the test substance, H-24743, to male rats for 10 consecutive days resulted in some absorption and retention o f fluorine in the liver. There was some absorption and retention o f fluorine in the fat. Levels in the fat in rats dosed with the test substance were slightly higher than or similar to levels in the positive control materials. However, levels in blood and liver in rats dosed with H-24743 were considerably lower than the fluorine levels in rats dosed with the positive control materials, H-24019 and H-24020. Company Sanitized. Does no! contain TSCA CBI H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 RECORDS AND SAMPLE STORAGE Specimens (if applicable), raw data, and the final report will be retained at Haskell Laboratory, Newark, Delaware, or at Iron Mountain Records Management, Wilmington, Delaware. .Company Sanitized. D oe, not contain tsca CUI - 20- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLES Company Sanitized. Does not contain TSCA CBi H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE 1 MEAN BODY WEIGHTS DAYS ON TEST ___________________MEAN BODY WEIGHTS (g)__________________ Negative Control_______ Positive Controls__________ Test Substance Deionized Water H-24019 H-24020 H-24743 1 189.6 184.4 184.1 192.9 2 195.5 189.2 187.8 196.0 3 204.6 199.5 197.8 205.5 4 211.7 206.4 204.8 216.3 5 222.1 216.2 212.5 229.4 6 225.0 222.0 216.0 233.9 7 235.2 229.4 223.6 244.2 8 242.3 233.7 226.9 250.3 9 248.3 240.4 234.2 258.8 10 257.0 246.5 243.0 264.6 17 293.3 290.1 297.4 306.2 24 325.5 313.3 338.6 338.9 31 359.3 348.4 381.9 360.7 38 380.9 370.8 404.9 383.3 45 407.4 403.2 434.4 409.8 52 420.5 422.8 460.6 429.9 59 436.4 439.6 483.1 449.9 67 452.8 455.6 502.7 - 66 - - - 463.7 73 472.6 480.1 525.6 482.5 80 489.6 496.5 542.1 500.2 87 497.7 512.2 546.2 504.7 94 511.7 524.6 570.5 515.8 Indicates the animals were not weighed Company Sanitized. Does not contain TSCA CB - 22- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE 2 MEAN BODY WEIGHT GAINS DOSING DAYS ON TEST MEAN BODY WEIGHT GAINS (g) Negative Control Positive Controls Test Substance Deionized Water H-24019 H-24020 H-24743 1-5 32.5 31.8 28.4 36.5 5-10 34.9 30.3 30.5 35.2 1-10 67.4 62.1 58.9 71.7 RECOVERY DAYS ON TEST MEAN BODY WEIGHT GAINS (g) Negative Control Positive Controls Test Substance Deionized Water H-24019 H-24020 H-24743 10-17 17-24 24-52 52-94 36.3 43.6 54.4 41.6 32.2 23.2 41.2 32.7 95.0 109.5 122.0 91.0 91.2 101.8 109.9 85.9 10-94 254.7 278.1 327.5 251.2 Company Sanitized. Does not contain TSCA C8 -23 - /****\ H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE 3 MEAN BODY AND LIVER WEIGHTS DAYS ON TEST NEGATIVE CONTROL - DEIONIZED WATER WEIGHT (g) Absolute Mean Relative Liver Weight Body Weight Liver Weight (Liver/Body Weight) 10 258.8 10.814 94 511.7 17.868 0.042 0.035 DAYS ON TEST 10 94 POSITIVE CONTROL - H-24019 WEIGHT (g) Absolute Mean Relative Liver Weight Body Weight Liver Weight (Liver/Body Weight) 243.9 524.6 14.205 19.296 0.058 0.037 DAYS ON TEST 10 94 POSITIVE CONTROL - H-24020 WEIGHT (g) Absolute Mean Relative Liver Weight Body Weight Liver Weight (Liver/Body Weight) 243.1 570.5 19.174 19.590 0.079 0.034 DAYS ON TEST 10 94 TEST SUBSTANCE - H-24743 WEIGHT (g) Absolute Mean Relative Liver Weight Body Weight Liver Weight (Liver/Body Weight) 260.7 515.8 14.333 20.212 0.055 0.039 Company Sanitized. Does not contain TfiCA reti - 24- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE 4 MEAN BLOOD FLUORINE LEVELS DAYS ON TEST MEAN FLUORINE LEVELS (ppm) Negative Control Positive Controls Test Substance Deionized Water H-24019 H-24020 H-24743 1 0.60` 2.60 (0.1)b 9.40 (2.4) 9.07 (4.23) 5 C 31.32 (1.1) 74.92 (7.0) 9.152 (11.52) 10 1.10: 68.00 (3.5) 61.76 (5.5) 9.1 (4.1) 13 c 53.98 (1.2) 29.5.2 (4.9) 6.4 (1.36) 24 0.5a 39.62 (3.4) 11.18 (2.9) 3.536 (0.54) 52 c 23.56 (2.1) 2.26 (1.1) 0.58a 94 c 12.60 (1.2) 0.85 (0.1)d C a One o f 5 values. Four o f the values were below the limit o f quantification (LOQ). b Standard deviation (S.D.) is in parentheses, c All values were below the LOQ. d Mean o f 2 o f the 5 values. Three o f the values were below the LOQ. TABLE 5 MEAN BLOOD FLUORINE CONCENTRATION NORMALIZED TO DOSE DAYS ON TEST MEAN FLUORINE CONCENTRATION (pM F Equivalents) Positive Controls Test Substance H-24019 H-24020 H-24743 1 36.92 ( l.l) a 66.67 (17.5) 4.35 (2.08) 5 478.77 (17.6) 541.45 (50.4) 4.39 (5.66) 10 1043.08 (54.6) 446.09 (39.7) 4.37 (2.02) 13 827.38 (19.0) 212.46 (35.6) 3.04 (0.67) 24 606.46 (53.1) 79.57 (20.8) 1.64 (0.27) 52 359.38 (32.7) 14.93 (7.7) 94 190.77 (19.2) 4.71 (0.5)b 0.19C _d a Standard deviation (S.D.) is in parentheses. b Mean o f 2 o f the 5 values. Three o f the values were below the limit o f quantification (LOQ). c One o f 5 values. Four o f the values were below the LOQ. d All values were below the LOQ. - 25jggmpany Sanitized. Does not contain tsca '**! H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE 6 MEAN LIVER FLUORINE LEVELS DAYS ON TEST MEAN FLUORINE LEVELS (ppm) Negative Control Positive Controls Test Substance Deionized Water H-24019 H-24020 H-24743 10 0.90 (0.2)a 312.34 (19.7) 119.80 (3.5) 56.25 (22.64) 94 0.78 (0.0) 84.24 (7.4) 2.56 (1.3) 4.81 (0.89) a Standard deviation (S.D.) is in parentheses. TABLE 7 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE DAYS ON TEST MEAN FLUORINE CONCENTRATION (pM F Equivalents) Positive Controls Test Substance H-24019 H-24020 H-24743 10 4802.15 (303.8)a 866.67 (25.5) 27.52 (11.12) 94 1292.92. (114.1) 17.10 (9.8) 2.26 (0.44) a Standard deviation (S.D.) is in parentheses. Z0*"-,, Company Sanitized. Does not contain TSCA CBI - 26- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TABLE 8 MEAN FAT FLUORINE LEVELS DAYS ON TEST MEAN FLUORINE LEVELS (ppm) Negative Control Positive Controls Test Substance Deionized Water H-24019 H-24020 H-24743 10 a 12.84 (1.7)b 8.10 (1.1) 100.21 (55.64) 94 a 1.20 (0.3) a 22.34 (10.43) a All values were below the limit o f quantification (LOQ) or non-detectable. b Standard deviation (S.D.) is in parentheses. TABLE 9 MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE DAYS ON TEST MEAN FLUORINE CONCENTRATION (gM F Equivalents) Positive Controls Test Substance H-24019 H-24020 H-24743 10 94 194.46 (26.8)a 15.38 (4.1) 57.25 b(7.8) 49.10 (27.32) 10.87 (5.12) a Standard deviation (S.D.) is in parentheses. b All values were below the limit o f quantification (LOQ) or non-detectable. .Company Sanitized. Does not contain TSCA or| - 27- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 FIGURES not contain TSCACB1 Company Sanitized.Does - 28- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats )) _____________________________________________________________ DuPont-11558 FIGURE 1 MEAN BODY WEIGHTS .Company Sanitized. Does not contain TSCA CBI - 29- Company Sanitized. Does not contain TSCA CB1 )>> H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats________________________________________________________ _____________ __________________________ DuPont-11558 FIGURE 2 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND NEGATIVE CONTROL -30- > H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) FIGURE 3 COMPARISON OF MEAN RELATIVE LIVER WEIGHTS: TEST SUBSTANCE AND POSITIVE CONTROLS ) DuPont-11558 Company Sanitized. Does not contain TSCACB1 10 94 Test Days -31 - H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 FIGURE 4 MICROMOLAR EQUIVALENTS IN RAT BLOOD A . N o r m a l i z e d R a t B l o o d H -2 4 0 1 9 jj M E q u i v a l e n ts R e s u l t i n g f r o m a 1 0 - D a y O r a l G a v a g e Mean pM Equivalents Micromolar (pM) equivalents o f H-24019 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. B . N o r m a l i z e d R a t B l o o d H >2 4 0 2 0 p M E q u i v a l e n t s R e s u l t i n g f r o m a 1 0 - D a y O r a l G a v a g e Mean pM Equivalents Micromolar (pM) equivalents o f H-24020 (positive control) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. Company Sanitized. Does not contain rqra nqi H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 C. Normalized Rat Blood H-24743 uM Equivalents Resulting from a 10-Day Oral Gavage Micromolar (pM) equivalents o f H-24743 (test substance) in rat blood resulting from a 10-day oral gavage exposure. Values are means and error bars are standard deviation. .Company Sanitized. Does not contain TSCA CBt -33 - Company Sanitized. Does not contain TSCA CBI ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) 600000 FIGURE 5 NORMALIZED H-24743 AND POSITIVE CONTROL BLOOD AUCINF/D RESULTING FROM A 10-DAY ORAL GAVAGE 500000 400000 uz. <o3OOD 300000 oC4r-Qca 200000 100000 ) DuPont-11558 AUCINF/D H-24019 5.22E+05 H-24020 8.15&-04 H-24743 1.07B-03 -34- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) FIGURE 6 MEAN LIVER FLUORINE CONCENTRATION NORMALIZED TO DOSE ) D uPont-115 5 8 H-24019 (Positive Control) H-24020 (Positive Control) H-24743 (Test Substance) IPpmpany Sanitized, Does not contain TSCA CBI Day 10 Test Day -35- Day 94 Company Sanitized. Does not contain TSCA CRt ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) ) DuPont-11558 FIGURE 7 COMPARISON OF RELATIVE LIVER WEIGHT AND MEAN LIVER FLUORINE CONCENTRATION FOR H-24743 AND NEGATIVE CONTROL 0.06 0.05 0.04 $5 =Q>) $o- CD 0.03 TO C or O) can>s O-- 0.02 S 0.01 Day 10 Test Days Day 94 60.00 c 50.00 o dco> 40.00 o o .5 a. 30.00 o3 -- U. 'c3>roa> 20.00 S 10.00 0.00 Day 10 Test Day Day 94 Deionized Water (Negative Control) H-24743 (Test Substance) - 36- Company Sanitized. Does not contain TSCA CB! ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_______ ) FIGURE 8 MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE ) DuPont-11558 H-24019 (Positive Control) H-24020 (Positive Control) H H-24743 (Test Substance) 4<c0t>t^/s)> o*3 A ll values fo r H-24020 (Positive Control) ui w ere non-detectable a. Day 10 Test Day Day 94 ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) FIGURE 9 COMPARISON OF MEAN BLOOD, MEAN LIVER, AND MEAN FAT FLUORINE CONCENTRATION NORMALIZED TO DOSE DuPont-11558 oo 3m3<o 330 a a 1& "338* i 3r 9*1 a> Blood / Liver / Fat H-24019 (Positive Control) Blood / Liver / Fat H-24020 (Positive Control) Blood / Liver / Fat H-24743 (Test Substance) - 38- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 APPENDICES Company Sanitized. Does not contain TSCA CB1 - 39- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 Z*'"***'-. APPENDIX A Individual Body Weights G&mpany Sanitized. Does not contain TSCA CBl -40- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats INDIVIDUAL BODY WEIGHTS ABBREVIATIONS: EXPLANATORY NOTES SD - sacrificed by design DuPont-11558 Company Sanitized. Does noi coniata Tr>a /of fipnipsny Sanitized. Does no! confain TSCA CBl H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 646932 646933 646934 646935 646936 Day 1 197.1 191.9 182.6 203.0 196.8 Day 2 202.1 195.4 188.9 208.9 201.4 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP I TEST DAY Day 4 Day 5 Day 6 212.9 205.0 195.4 218.0 208.6 221.5 213.2 207.0 228.1 204.0 233.6 226.5 213.9 239.7 216.7 234.1 219.3 215.9 243.0 218.6 Day 7 242.4 232.9 224.2 257.2 229.9 ANIMAL NUMBER Day 10 646932 646933 646934 646935 646936 264.7 254.4 248.6 278.0 248.1 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 TEST DAY DuPont-11558 Day 8 251.1 239.5 230.9 265.7 234.9 Day 9 256.1 246.4 238.1 270.5 240.3 jpniphy Sanitized. Does not contain TR^a cm H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 646937 646938 646939 646940 646941 Day 1 184.5 163.7 193.3 191.5 191.6 ANIMAL NUMBER Day 10 646937 646938 646939 646940 646941 249.8 237.2 277.9 244.5 266.3 ANIMAL NUMBER Day 73 646937 646938 646939 646940 646941 504.3 417.4 540.1 407.3 493.7 Day 2 186.8 173.8 204.2 197.1 195.9 Day 17 282.6 270.4 329.7 271.5 312.4 Day 80 520.5 431.0 568.9 424.0 503.5 DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP III TEST DAY Day 4 Day 5 Day 6 199.0 180.9 214.6 203.8 207.9 205.9 189.3 221.1 209.8 216.8 213.6 200.6 234.4 218.5 223.1 221.0 204.1 240.7 222.1 231.2 Day 7 229.6 215.9 248.9 229.3 241.3 Day 24 322.3 296.5 372.8 291.3 344.8 TEST DAY Day 31 Day 38 366.1 325.8 415.2 317.9 371.6 395.0 342.9 443.2 332.8 390.7 Day 45 425.0 367.5 475.2 356.3 412.8 Day 52 443.9 373.7 484.1 367.0 433.6 Day 87 533.7 432.1 578.1 430.8 513.8 TEST DAY Day 94 545.7 450.6 586.9 449.4 525.8 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 DuPont-11558 Day 8 238.7 221.8 259.2 232.0 249.1 Day 9 241.4 228.3 267.3 237.5 257.0 Day 59 461.7 378.7 505.7 380.3 455.8 Day 67 478.7 401.3 516.4 392.5 475.3 - 43- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 646910 646911 646912 646913 646914 Day 1 184.3 184.4 180.5 186.4 185.5 Day 2 185.3 187.1 185.9 189.7 189.6 ) H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP I TEST DAY Day 4 Day 5 Day 6 197.3 195.7 195.9 198.1 198.8 204.8 202.0 202.5 205.7 203.8 212.7 211.7 211.9 221.8 215.4 216.3 217.2 217.7 225.0 222.7 ANIMAL NUMBER Day 10 646910 646911 646912 646913 646914 241.1 236.6 241.9 257.0 243.0 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 TEST DAY Day 7 226.4 225.2 227.2 228.6 226.7 ") DuPont-11558 Day 8 232.3 225.6 232.4 238.4 231.5 Day 9 237.5 231.9 238.5 245.8 238.8 Company Sanitized Does not contain TSCA ''BI -44- Company Sanitized Does not contain TSCA CBl ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 646915 646916 646917 646918 646919 Day 1 182.7 185.0 187.8 175.5 192.0 Day 2 188.3 192.7 194.3 181.9 196.9 H-24019 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP III TEST DAY Day 4 Day 5 Day 6 200.1 205.5 205.8 189.8 207.9 206.3 211.6 210.4 197.6 218.9 211.1 220.0 221.8 207.0 228.3 220.6 226.9 227.5 212.4 234.0 Day 7 225.8 239.5 233.9 220.2 240.8 ) DuPont-11558 Day 8 231.1 240.1 235.8 224.1 245.8 Day 9 236.8 249.3 244.3 228.2 252.9 ANIMAL NUMBER Day 10 646915 646916 646917 646918 646919 240.4 257.0 250.2 237.1 260.2 Day 17 283.4 310.0 280.3 270.6 306.1 Day 24 302.9 334.4 308.2 292.7 328.5 TEST DAY Day 31 Day 38 326.7 383.2 339.7 323.2 369.2 337.9 411.9 362.8 350.8 390.4 Day 45 371.1 450.3 387.9 379.4 427.3 Day 52 399.7 469.8 409.0 396.7 439.0 Day 59 412.5 494.8 420.0 418.8 451.7 Day 67 433.1 511.8 441.5 433.8 457.6 ANIMAL NUMBER Day 73 646915 646916 646917 646918 646919 461.2 544.2 457.5 448.1 489.5 Day 80 475.0 565.5 464.7 472.5 504.8 Day 87 498.1 578.5 479.5 480.8 524.2 TEST DAY Day 94 513.3 590.9 495.7 500.3 522.9 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 -45- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 646921 646922 646923 646924 646925 Day 1 204.5 178.3 185.9 188.7 172.2 Day 2 204.9 180.9 185.1 194.6 176.0 ) H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS GROUP I Day 3 TEST DAY Day 4 Day 5 Day 6 215.7 188.9 196.0 209.5 183.9 222.2 197.4 202.3 212.6 193.4 233.3 197.7 203.5 230.4 202.7 238.8 202.4 204.9 225.9 202.7 ANIMAL NUMBER Day 10 646921 646922 646923 646924 646925 262.6 218.6 240.6 259.1 234.8 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 TEST DAY Day 7 244.3 202.4 219.1 236.2 215.1 ) DuPont-11558 Day 8 244.9 204.0 224.9 236.5 221.5 Day 9 252.2 207.7 231.7 250.9 228.7 Sanitized. Does not contain r q c A o 3 -46- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 646926 646927 646928 646929 646930 Day 1 182.0 172.2 190.4 178.6 187.9 ANIMAL NUMBER Day 10 646926 646927 646928 646929 646930 250.8 243.3 254.1 225.5 240.3 ANIMAL NUMBER Day 73 646926 646927 646928 646929 646930 535.3 551.2 540.3 502.6 498.5 Day 2 187.8 178.8 195.3 182.5 191.6 Day 17 297.2 306.2 310.6 280.9 292.3 Day 80 551.9 574.5 557.0 509.6 517.6 ) H-24020 (POSITIVE CONTROL) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP III TEST DAY Day 4 Day 5 Day 6 197.0 191.1 204.5 190.7 201.0 203.6 195.1 212.3 198.3 210.4 206.0 206.6 222.1 203.5 219.6 213.2 213.2 230.1 204.2 224.4 Day 7 222.9 218.2 236.1 214.6 226.7 Day 24 349.3 355.1 349.4 323.2 316.2 TEST DAY Day 31 Day 38 383.8 404.0 392.8 376.0 352.7 412.1 424.8 422.4 396.8 368.4 Day 45 432.1 463.2 443.5 426.4 406.8 Day 52 467.8 484.0 464.8 450.2 436.0 Day 87 564.4 541.3 568.3 526.7 530.3 TEST DAY Day 94 580.6 602.5 581.9 544.0 543.4 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 ) DuPont-11558 Day 8 229.9 228.7 235.8 210.3 232.6 Day 9 239.7 233.1 243.4 219.2 235.3 Day 59 493.2 511.7 492.2 463.8 454.7 Day 67 509.5 527.2 518.6 484.2 474.2 Sanitized. Does not contain TSCA CBl - 47- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ANIMAL NUMBER 665511 665512 665513 665514 665515 Day 1 186.0 194.8 186.1 193.5 193.9 Day 2 189.4 200.2 192.0 202.5 195.3 H-24743 (TEST SUBSTANCE) INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP I TEST DAY Day 4 Day 5 Day 6 194.2 207.4 201.8 210.8 207.1 205.2 219.7 213.0 218.6 213.0 221.7 230.5 226.6 234.5 227.2 223.2 233.8 230.3 239.0 230.3 Day 7 234.1 242.2 242.2 249.3 238.0 DuPont-11558 Day 8 242.0 247.6 250.9 257.7 243.2 Day 9 246.0 252.5 256.0 266.3 251.5 ANIMAL NUMBER Day 10 665511 665512 665513 665514 665515 251.6 260.3 265.0 271.5 255.3 SD test day 10 SD test day 10 SD test day 10 SD test day 10 SD test day 10 TEST DAY Company Sanitized. Does not O 3 3na?t >OC0 mo )) ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats__________________________________________________________________________________________________DuPont-11558 H-24743 (TEST SUBSTANCE) ANIMAL NUMBER Day 1 Day 2 INDIVIDUAL BODY WEIGHTS (g) OF MALE RATS Day 3 GROUP III TEST DAY Day 4 Day 5 Day 6 Day 7 Day 8 Day 9 S65516 665518 665519 665520 665521 194.8 185.8 195.3 195.5 202.8 198.8 189.8 189.0 201.4 201.8 208.6 199.8 202.7 208.1 214.4 222.1 210.9 215.9 217.0 227.5 234.0 223.4 227.1 235.5 233.4 238.5 227.0 235.1 240.5 241.0 252.2 235.2 247.5 249.5 251.5 258.9 239.7 254.8 256.8 251.8 269.6 249.3 266.2 265.1 265.1 ANIMAL NUMBER Day 10 665516 665518 665519 665520 665521 274.4 256.0 270.6 274.5 266.6 Day 17 318.6 282.3 320.9 308.0 301.1 Day 24 354.7 308.7 352.7 337.6 341.0 TEST DAY Day 31 Day 38 371.6 332.1 377.5 361.2 361.2 397.0 346.2 404.1 390.4 378.9 Day 45 427.7 369.0 441.3 411.2 400.0 Day 52 453.0 379.1 472.9 430.9 413.7 Day 59 474.8 393.7 490.6 449.3 441.1 Day 67 496.2 405.6 508.7 455.0 453.0 ANIMAL NUMBER Day 73 665516 665518 665519 665520 665521 517.8 408.0 530.8 477.1 479.0 Day 80 543.6 415.1 545.0 503.5 493.8 Day 87 543.0 422.3 553.8 504.6 499.9 TEST DAY Day 94 564.4 430.7 562.4 514.2 507.2 SD test day 94 SD test day 94 SD test day 94 SD test day 94 SD test day 94 Company Sanitized. Does not contain TSCA CSt -49- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 APPENDIX B Individual Clinical Observations Company Sanitized. Does not contain TSCA CBl Company Sanitized. Does not contain TSCA OBJ )) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal Observation GROUP I Days 646932 General observation, No Abnormality Detected 1 Eye Observations, Exophthalmus, Left Eye Observations, Bled via Orbital for Clin Path, Left 2-9 1,5 Discharge, Eye left, Black 10 646933 Sacrificed by design General observation, No Abnormality Detected 10 1-5 Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Dark, Left 1,5 6-10 Discharge, Eye left, Black 6-7 Swollen Observations, Mouth 10 Sacrificed by design 10 646934 General observation, No Abnormality Detected 1-5,10 Eye Observations, Bled via Orbital for Clin Path, Left 1,5 Discharge, Eye left, Black 6-9 Sacrificed by design 10 -51 - ') DuPont-11558 H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I (Continued) Animal 646935 Observation General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Days 1-10 1.5 Sacrificed by design 10 646936 General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left 1-10 1.5 Sacrificed by design 10 DuPont-11558 Company Sanitized. Does not contain TSCA CBl - 52- Company Sanitized. Does not contain TSCA CBJ ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal 646937 646938 GROUP III Observation General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left Eye Observations, Bledvia Orbital for Clin Path, Right Sacrificed by design General observation, No Abnormality Detected Eye Observations, Enophthalmus, Right Eye Observations, Exophthalmus, Right Eye Observations, Bledvia Orbital for Clin Path, Bilateral Eye Observations, Bledvia Orbital for Clin Path, Right Eye Observations, Corneal Opacity, Right Discharge, Eye right, Black Sacrificed by design Days 1-10,17-94 13 24,52 94 1-10,17-45 80-94 52-73 13,52 24 80-94 59 94 646939 General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left 1-10,17-94 13 Eye Observations, Bledvia Orbital for Clin Path, Right 24,52 Sacrificed by design 94 - 53- DuPont-11558 H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DEIONIZED WATER (NEGATIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP III (Continued) Animal Observation Days 646940 General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Discharge, Nose, Black Sacrificed by design 1-10,17-80,94 13 24,52 87 94 646941 General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design 1-10,17-94 52 13 24 94 ) DuPont-11558 .Company Sanitized. Does not contain TSCA CBI -54- Company Sanitized. Does not contain TSCA CBI )) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Animal Observation H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROIIP I Days 646910 646911 General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design General observation, No Abnormality Detected 1-10 1,5 10 1,10 Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Dark, Left Sacrificed by design 1,5 2-9 10 646912 General observation, No Abnormality Detected 1-10 646913 646914 Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design General observation, No Abnormality Detected 1 5 10 1-10 1,5 10 1-10 Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design 1,5 10 - 55- ) DuPont-11558 .Company Sanitized. Does not contain TSCA CBt I |43: Biopersistence Screening 10-j^ose Oral Gavage Study in Rats ) H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP III Animal Observation Days 646915 General observation, No Abnormality Detected 1,17-38,73 Eye Observations, Bled via Orbital for Clin Path, Left 13,24,52 Discharge, Eye right, Red 2-10 Hair Loss, Forelimb, Right 80 Hair Loss, Forepaw, Right 45-67,87-94 Sacrificed by design 94 646916 General observation, No Abnormality Detected 1-10,17-94 Eye Observations, Bledvia Orbital for Clin Path, Bilateral 24 Eye Observations, Bledvia Orbital for Clin Path, Left 13 Eye Observations, Bledvia Orbital for Clin Path, Right 52 Sacrificed by design 94 646917 General observation, No Abnormality Detected 1-10,17-94 Eye Observations, Bledvia Orbital for Clin Path, Left 13,24 Eye Observations, Bledvia Orbital for Clin Path, Right 52 Sacrificed by design 94 ') DuPont:--1i 1i55558 - 56- )) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________________________________________________________ H-24019 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP III (Continued) Animal Observation Days 646918 General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left 1-10,17-94 13,24 Eye Observations, Bledvia Orbital for Clin Path, Right 52 Sacrificed by design 94 646919 General observation, No Abnormality Detected 1-10,17-94 Eye Observations, Bledvia Orbital for Clin Path, Left 13,24 Eye Observations, Bledvia Orbital for Clin Path, Right 52 Sacrificed by design 94 DuPont-11558 ty Sanitized. Does not contain TSCA CBI - 57- Company Sanitized. Does not contain TSCA CBI ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Animal Observation Days 646921 General observation, No Abnormality Detected 1,5-10 Eye Observations, Bled via Orbital for Clin Path, Left 1,5 Eye Observations, Dark, Left 2-4 Sacrificed by design 10 646922 General observation, No Abnormality Detected 1-10 Eye Observations, Bledvia Orbital for Clin Path, Left 1 Eye Observations, Bledvia Orbital for Clin Path, Right 5 Sacrificed by design 10 646923 General observation, No Abnormality Detected 1,7-10 Eye Observations, Exophthalmus, Left 4-6 Eye Observations, Bledvia Orbital for Clin Path, Left 1 Eye Observations, Bledvia Orbital for Clin Path, Right 5 Eye Observations, Dark, Left 2-6 Sacrificed by design 10 ') DuPont-11558 - 58- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____________________ ) H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I (Continued) Animal Observation Days 646924 General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left 1-10 1 Eye Observations, Bledvia Orbital for Clin Path, Right 5 Sacrificed by design 10 646925 General observation, No Abnormality Detected Eye Observations, Bledvia Orbital for Clin Path, Left 1-10 1 Eye Observations, Bledvia Orbital for Clin Path, Right 5 Sacrificed by design 10 DuPont-11558 Company Sanitized. Does not contain TSCA CBl -59- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal 646926 GROUP III Observation General observation, No Abnormality Detected Days 1-10,17-94 646927 Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design General observation. No Abnormality Detected 52 13,24 94 1-10,17-94 Eye Observations, Bled via Orbital for Clin Path, Bilateral Eye Observations, Bled via Orbital for Clin Path, Left 52 13,24 646928 Sacrificed by design General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Sacrificed by design 94 1-10,17-94 13,24,52 94 ) DuPont-11558 Company Sanitized, Does not contain TSCA CBS - 60- ) H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats ) H-24020 (POSITIVE CONTROL) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal 646929 GROUP III (Continued) Observation General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Left Eye Observations, Bled via Orbital for Clin Path, Right Days 1-10,17-31 13,24 52 Hair Loss, Forelimb, Bilateral 52-73 Hair Loss, Neck, Left Hair Loss, Neck, Ventral Wound, Superficial, Face Sacrificed by design 73 38-67,80-94 59 94 646930 General observation, No Abnormality Detected 1-10,17-94 Eye Observations, Bled via Orbital for Clin Path, Left 13,24 Eye Observations, Bled via Orbital for Clin Path, Right Sacrificed by design 52 94 ) DuPont-11558 Company Sanitized Does not contain TSCA CSI -61 - Company Sanitized. Does not contain TSCA CBt )) ) H-24743: Biopersistence Screening 10-Dose ( >ral Gavage Study in Rats______________________________________________________________________________________________DuPont-11558 H-24743 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS GROUP I Animal Observation Days 665511 General observation, No Abnormality Detected 1-10 Eye Observations, Bled via Orbital for Clin Path, Right 1,5 Sacrificed by design 10 665512 General observation, No Abnormality Detected 1-10 Eye Observations, Bled via Orbital for Clin Path, Right 1,5 665513 Sacrificed by design General observation, No Abnormality Detected 10 1-10 Eye Observations, Bled via Orbital for Clin Path, Right 1,5 Sacrificed by design 10 665514 General observation, No Abnormality Detected 1-10 Eye Observations, Bled via Orbital for Clin Path, Right 1,5 Sacrificed by design 10 665515 General observation, No Abnormality Detected 1-10 Eye Observations, Bled via Orbital for Clin Path, Right 1,5 Sacrificed by design 10 - 62- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats H-24743 (TEST SUBSTANCE) INDIVIDUAL CLINICAL OBSERVATIONS IN MALE RATS Animal 665516 GROUP III Observation General observation, No Abnormality Detected Days 1-94 665518 Eye Observations, Bled via Orbital for Clin Path, Right General observation, No Abnormality Detected 24.52 1-45,59-94 Eye Observations, Bled via Orbital for Clin Path, Right 24.52 665519 665520 Hair Loss, Forelimb, Bilateral General observation, No Abnormality Detected Eye Observations, Bled via Orbital for Clin Path, Right General observation, No Abnormality Detected 52 1-94 24.52 1-94 Eye Observations, Bled via Orbital for Clin Path, Right 24-52 665521 General observation, No Abnormality Detected 1-94 Eye Observations, Bled via Orbital for Clin Path, Right 24-52 DuPont-11558 Company Sanitized. Does not contain TSCA CBt - 63- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 APPENDIX C Terms and Calculations Factors Influencing Interpretation of Kinetic Analysis Company Sanitized. Does not contain TSCA CBf 6 4- - H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats___________________________________________________________ DuPont-11558 Terms: Active % Active Mol Wt Active Formulation Dose % F in Active Mol Wt F TERMS AND CALCULATIONS Fluorine containing compound The % o f formulation that is made up o f fluorine containing components The molecular weight of the fluorine containing components (g/mole) 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) The molecular weight o f fluorine g/mol Compound Calculations: Dose Active (mg/kg) The mg of fluorine containing compound administered per kg of animal body weight. = (% active/100) x Formulation Dose Dose Active (mmole/kg) The mmole o f fluorine containing compound administered per kg of animal body weight = dose [mg/kg] / Mol W t Active [mg/mmol] Dose F (mg/kg) The mg Fluorine administered per kg o f animal body weight = (% F in active/100) x Dose Active [mg/kg] Dose F (mmol/kg) The mmole o f fluorine administered per kg o f animal body weight = Dose F [mg/kg] / Mol Wt F [mg/mmol] Molar Ratio (Active/F) The moles o f fluorine containing compound per mole o f fluorine = Dose Active [mmol/kg] / Dose F [mmol/kg] Formulation Dose Normalization Factor The formulation dose that would be required to administer the amount of active needed to achieve the normalized dose = (Normalized dose o f Active [mmol/kg] / Dose Active [mmol/kg]) x Formulation Dose 6 5- - Company Sanitized. Does not contain tsca H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 TERMS AND CALCULATIONS Individual Animal Measurement: ppm F The ppm fluoride measured Individual Animal Calculations: ppm F minus Bkg 0.2 ppm The ppm fluoride measured minus the background fluoride measured in control animal. In this case the value was established at 0.2 ppm. ppm F normalized to 0.1 mmol/kg Dose The ppm fluoride minus background that would be expected if the active dose was 0.1 mmol/kg instead o f the actual active dose. This assumes linearity between administered dose and blood fluorine levels, but is needed because different doses o f active were used in the study. = (0.1 [mmol/kg] / Active dose [mmol/kg]) x (ppm F in blood minus background) [molar equivalents o f active The molar [pmol/L] concentration o f fluorine containing compound based on the ppm fluorine normalized to 0.1 mmol/kg active dose. This assumes that all fluorine is derived from the fluorine-containing component in the formulation. Note: 1 ppm - 1 mg/L = (Normalized ppm [mg/L] fluorine / Mol W t F [mg/mmol]) x molar ratio active/F [mmol active/mmol F] x 1000 pmol/mmol Company Sanitized. Does not contain rssca c w - 66- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 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 o f fluorinated compounds - Different active and formulation doses were used - Different vehicles were used to deliver formulations - Each compound may have very different potency for producing toxicity Assumptions: (May or may not be justified in all cases) - 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 Fluorine is 0.2 ppm - % F data is the % Fluorine o f the active (Fluorine containing component(s) in the formulation) - Molecular weight is the molecular weight o f the active component in the formulation Company Sanitized. Does not contain T SCA o | H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 APPENDIX D Individual Fluorine Levels in Blood Company Sanitized. Does not contain TSCA c w - 68- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24019 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 497 10 100 % F in Active: Mol Wt. F (g/mol): 65 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):_____ 10 0.020 0.342 Molar Ratio (Active/F): 0.059 Dose F (mg/kg): 6.5 Rat Number Test Day Sample ppm F in Blood Group I 646910 646911 646912 646913 646914 1 1 1 1 1 2.6 2.7 2.5 2.6 2.6 ppm F in Blood Minus Bkg 0.2 ppm 2.4 2.5 2.3 2.4 2.4 ppm F in Blood Normalized to 0.1 mmoles/kg Dose 11.93 12.43 11.43 11.93 11.93 pmolar Equivalents of Active in Blood 36.92 38.46 35.38 36.92 36.92 Group I 646910 646911 646912 646913 646914 5 5 5 5 5 30.1 30.3 31.3 32.2 32.7 29.9 30.1 31.1 32.0 32.5 148.60 149.60 154.57 159.04 161.53 460.00 463.08 478.46 492.31 500.00 Group I 646910 10 71.5 646911 10 70.5 646912 10 66.9 646913 10 62.5 646914 10 68.6 71.3 70.3 66.7 62.3 68.4 354.36 349.39 331.50 309.63 339.95 1096.92 1081.54 1026.15 958.46 1052.31 Company Sanitized. Does not contain TSCA C8I -69- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats___________________________________________________________DuPont-11558 Rat Number Test Day Sample PPm F in Blood ppm F in Blood Minus Bkg 0.2 ppm ppm F in Blood Normalized to 0.1 mmoles/kg Dose pmolar Equivalents of Active in Blood Group III 646915 646916 646917 646918 646919 13 13 13 13 13 55.0 55.3 52.2 53.8 53.6 54.8 55.1 52.0 53.6 53.4 272.36 273.85 258.44 266.39 265.40 843.08 847.69 800.00 824.62 821.54 Group III 646915 646916 646917 646918 646919 Group III 646915* 646916 646917 646918 646919 24 24 24 24 24 52 52 52 52 52 38.9 43.8 37.6 35.4 42.4 23.2 20.3 25.9 24.9 23.5 38.7 43.6 37.4 35.2 42.2 23.0 20.1 25.7 24.7 23.3 192.34 216.69 185.88 174.94 209.73 114.31 99.90 127.73 122.76 115.80 595.38 670.77 575.38 541.54 649.23 353.85 309.23 395.38 380.00 358.46 Group III 646915 646916 646917 646918 646919 94 94 94 94 94 13.3 11.7 12.9 11.0 14.1 13.1 11.5 12.7 10.8 13.9 65.11 57.16 63.12 53.68 69.08 201.54 176.92 195.38 166.15 213.85 Company Sanitized. Does not contain T SC A CBS / -- N H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24020 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 426 20 100 % F in Active: Mol Wt. F (g/mol): 69 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg): 20 0.047 0.726 Molar Ratio (Active/F): 0.065 Dose F (mg/kg): 13.8 (/***\ Rat Number Test Day Sample ppm F in Blood Group I 646921 1 10.6 646922 1 10.7 646923 1 9.2 646924 1 5.3 646925 1 11.2 ppm F in Blood Minus Bkg 0.2 ppm 10.4 10.5 9.0 5.1 11.0 ppm F in Blood Normalized to 0.1 mmoles/kg Dose 22.15 22.37 19.17 10.86 23.43 pmolar Equivalents of Active in Blood 75.36 76.09 65.22 36.96 79.71 Group I 646921 646922 646923 646924 646925 5 5 5 5 5 78.7 83.2 77.0 66.1 69.6 78.5 83.0 76.8 65.9 69.4 167.21 176.79 163.58 140.37 147.82 568.84 601.45 556.52 477.54 502.90 Group I 646921 10 63.0 646922 10 69.1 646923 10 60.3 646924 10 62.5 646925 10 53.9 62.8 68.9 60.1 62.3 53.7 133.76 146.76 128.01 132.70 114.38 455.07 499.28 435.51 451.45 389.13 Company Sanitized. Does not contain TSCA CBI -71 - H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats_____ __________________________________ Rat Number 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 Group III 646926 646927 646928 646929 646930 13 13 13 13 13 34.8 30.0 23.7 33.7 25.4 34.6 29.8 23.5 33.5 25.2 73.70 63.47 50.06 71.36 53.68 DuPont-11558 pmolar Equivalents of Active in Blood 250.72 215.94 170.29 242.75 182.61 Group III 646926 646927 646928 646929 646930 24 24 24 24 24 11.9 10.4 8.1 15.7 9.8 11.7 10.2 7.9 15.5 9.6 24.92 21.73 16.83 33.02 20.45 84.78 73.91 57.25 112.32 69.57 Group III / '" 'N 646926 52 2.5 646927 52 1.7 646928 52 1.3 646929 52 4.0 646930 52 1.8 2.3 1.5 1.1 3.8 1.6 4.90 16.67 3.20 10.87 2.34 7.97 8.09 27.54 3.41 11.59 Group III 646926 646927 646928 646929 646930 94 94 94 94 94 0.8 <0.5 <0.5 0.9 <0.5 0.6 * * 0.7 * 1.28 4.35 ** ** 1.49 5.07 ** * Below LOQ (Limit o f Quantification) /""""N, Company Sanitized. Does not contain TSC CBI - 72- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24743 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 364.1 300 100 % F in Active: Mol Wt. F (g/mol): 67.9 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg): 300 0.824 10.721 MDooslaerFR(amtigo/k(Ag)c:tive/F): 0.077 203.7 Rat Number Test Day Sample ppm F in Blood Group I 665511 1 8.74 665512 1 4.27 665513 1 7.19 665514 1 15.77 665515 1 9.38 ppm F in Blood Minus Bkg 0.2 ppm 8.5 4.1 7.0 15.6 9.2 ppm F in Blood Normalized to 0.1 mmoles/kg Dose 1.04 0.49 0.85 1.89 1.11 pmolar Equivalents of Active in Blood 4.19 2.00 3.43 7.64 4.51 Group I 665511 665512 665513 665514 665515 5 5 5 5 5 4.38 3.36 3.06 5.26 29.7 4.2 3.2 2.9 5.1 29.5 0.51 2.05 0.38 1.55 0.35 1.40 0.61 2.48 3.58 14.48 Group I 665511 10 6.87 6.7 0.81 3.27 665512 10 5.31 5.1 0.62 2.51 665513 10 6.62 6.4 0.78 3.15 665514 10 15.05 14.9 1.80 7.29 665515 10 11.65 11.5 1.39 5.62 Company Sanitized. Does not confai t r / 'a cwt H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats _______ ____________________________________________ DuPont-11558 Rat Number 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 pmolar Equivalents o f Active in Blood Group III 665516 665518 665519 665520 665521 13 13 13 13 13 5.22 6.12 5.03 7.97 7.66 5.0 5.9 4.8 7.8 7.5 0.61 2.46 0.72 2.91 0.59 2.37 0.94 3.81 0.91 3.66 Group III 665516 24 4.16 4.0 0.48 1.94 665518 24 3.76 3.6 0.43 1.75 665519 24 2.76 2.6 0.31 1.26 665520 24 3.75 3.6 0.43 1.74 665521 24 3.25 3.1 0.37 1.50 Group III 665516 52 <.5 * * * 665518 52 0.58 665519 52 <.5 0.4 * 0.05 0.19 ** 665520 52 <.5 * * * 665521 52 <.5 * * * Group III 665516 94 <.5 * * * 665518 94 <.5 * * * 665519 94 <.5 * * * 665520 94 <.5 * * * 665521 94 <.5 * * * * Below LOQ (Limit o f Quantification) Company Sanitized. Does not contain TSCA CBl -74- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 APPENDIX E Individual Fluorine Levels in Liver Company Sanitized. Does not contain TSCA c a l - 75- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24019 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 497 10 100 % F in Active: Mol Wt. F (g/mol): 65 19 Calculated Values: Dose Active (mg/kg): 10 Dose Active (mmole/kg): 0.020 Dose F (mmol/kg):____________________ 0.342 Molar Ratio (Active/F): 0.059 Dose F (mg/kg): 6.5 Rat Number Test Day Sample ppm F in Liver Group I 646910 10 320.9 646911 10 334.0 646912 10 318.6 646913 10 281.6 646914 10 306.6 ppm F in Liver Minus Bkg 0.2 ppm 320.7 333.8 318.4 281.4 306.4 ppm F in Liver Normalized to 0.1 mmoles/kg Dose pmolar Equivalents of Active in Liver 1593.88 1658.99 1582.45 1398.56 1522.81 4933.85 5135.38 4898.46 4329.23 4713.85 Group III 646915 646916 646917 646918 646919 94 94 94 94 94 85.6 75.9 87.5 78.0 94.2 85.4 75.7 87.3 77.8 94.0 424.44 376.23 433.88 386.67 467.18 1313.85 1164.62 1343.08 1196.92 1446.15 Company Sanitized. Does not contain T SC A CBl H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24020 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 426 20 100 % F in Active: Mol Wt. F (g/mol): 69 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg):_____ 20 0.047 0.726 Molar Ratio (Active/F): 0.065 Dose F (mg/kg): 13.8 Rat Number Test Day Sample ppm F in Liver Group I 646921 10 118.3 646922 10 122.7 646923 10 114.3 646924 10 121.6 646925 10 122.1 ppm F in Liver Minus Bkg 0.2 ppm 118.1 122.5 114.1 121.4 121.9 ppm F in Liver Normalized to 0.1 mmoles/kg Dose pmolar Equivalents o f Active in Liver 251.55 260.93 243.03 258.58 259.65 855.80 887.68 826.81 879.71 883.33 Group III 646926 646927 646928 646929 646930 94 94 94 94 94 1.8 1.2 2.2 4.7 2.9 1.6 1.0 2.0 4.5 2.7 3.41 11.59 2.13 7.25 4.26 14.49 9.59 32.61 5.75 19.57 Company Sanitized. Does not contain T SC A C&l H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24743 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 364.1 300 100 % F in Active: Mol Wt. F (g/mol): 67.9 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg): 300 0.824 10.721 Molar Ratio (Active/F): 0.077 Dose F (mg/kg): 203.7 Rat Number Test ppm F Day in Sample Liver Group I 665511 665512 665513 665514 665515 10 44.68 10 41.46 10 33.84 10 82.31 10 78.98 Group III 665516 665518 665519 665520 665521 94 94 94 94 94 5.43 5.95 3.94 4.75 3.97 ppm F in Liver Minus Bkg 0.2 ppm 44.5 41.3 33.6 82.1 78.8 5.2 5.8 3.7 4.6 3.8 ppm F in Liver Normalized to 0.1 mmoles/kg Dose pmolar Equivalents of Active in Liver 5.40 21.84 5.01 20.26 4.08 16.51 9.97 40.31 9.56 38.67 0.63 2.57 0.70 2.82 0.45 1.84 0.55 2.23 0.46 1.85 Company Sanitized. Does not contain TSCA CBI - 78- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats DuPont-11558 APPENDIX F Individual Fluorine Levels in Fat Company Sanitized. Does not contain TSCA CBI - 79- H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24019 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 497 10 100 % F in Active: Mol Wt. F (g/mol): 65 19 Calculated Values: Dose Active (mg/kg): 10 Dose Active (mmole/kg): 0.020 Dose F (mmol/kg):_____________________0.342 Molar Ratio (Active/F): 0.059 Dose F (mg/kg): 6.5 Rat Number Test Day Sample ppm F in Fat Group I 646910 646911 646912 646913 646914 10 10 10 10 10 14.4 14.9 10.9 11.6 12.4 ppm F in Fat Minus Bkg 0.2 ppm 14.2 14.7 10.7 11.4 12.2 G roup III 646915 646916 646917 646918 646919 94 94 94 94 94 1.3 1.1 1.3 0.8 1.5 1.1 0.9 1.1 0.6 1.3 ppm F in Fat Normalized to 0.1 mmoles/kg Dose pmolar Equivalents of Active in Fat 70.57 73.06 53.18 56.66 60.63 218.46 226.15 164.62 175.38 187.69 5.47 16.92 4.47 13.85 5.47 16.92 2.98 9.23 6.46 20.00 Company Sanitized. Does not contain T SC A CBt H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats Data for H-24020 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 426 20 100 % F in Active: Mol Wt. F (g/mol): 69 19 Calculated Values: Dose Active (mg/kg): 20 Molar Ratio (Active/F): 0.065 Dose Active (mmole/kg): 0.047 Dose F (mg/kg): 13.8 Dose F (mmol/kg):______________________ 0.726_________________________________ Rat Number Test ppm F Day in Sample Fat Group I 646921 646922 646923 646924 646925 10 10 10 10 10 6.2 8.5 8.7 8.8 8.3 ppm F in Fat Minus Bkg 0.2 ppm 6.0 8.3 8.5 8.6 8.1 Group III 646926 646927 646928 646929 646930 94 94 94 94 94 ND ND ND ND ND ND ND ND ND ND ND Non-detectable. ppm F in Fat Normalized to 0.1 mmoles/kg Dose pmolar Equivalents of Active in Fat 12.78 17.68 18.11 18.32 17.25 43.48 60.14 61.59 62.32 58.70 ND ND ND ND ND ND ND ND ND ND Company Sanitized. Does not contain TSCA CB1 -81 - H-24743: Biopersistence Screening 10-Dose Oral Gavage Study in Rats___________________________________ Data for H-24743 DuPont-11558 Given: Mol Wt. Active (g/mole): Formulation Dose (mg/kg): % Active (F Containing) in Formulation: 364.1 300 100 % F in Active: Mol Wt. F (g/mol): 67.9 19 Calculated Values: Dose Active (mg/kg): Dose Active (mmole/kg): Dose F (mmol/kg): 300 0.824 10.721 Molar Ratio (Active/F): 0.077 Dose F (mg/kg): 203.7 Rat Number Test Day Sample ppmF in Fat Group I 665511 665512 665513 665514 665515 10 97.26 10 35.49 10 65.62 10 121.64 10 181.03 Group III 665516 665518 665519 665520 665521 94 94 94 94 94 29.18 34.66 9.72 24.36 13.77 ppm F in Fat Minus Bkg 0.2 ppm 97.1 35.3 65.4 121.4 180.8 29.0 34.5 9.5 24.2 13.6 ppm F in Fat Normalized to 0.1 mmoles/kg Dose pmolar Equivalents of Active in Fat 11.78 4.28 7.94 14.74 21.95 47.65 17.32 32.12 59.62 88.77 3.52 14.23 4.18 16.92 1.16 4.67 2.93 11.86 1.65 6.66 Company Sanitized. Does not contain TSCA CBt