Document 15vrawvX94bKL4myZVq2vEzba

received __________________ __________ DuPont-7473 TRADE SECRET Study Title Perfluorooctanoic Acid: Toxicokinetics in the Rat Laboratory Project ID: DuPont-7473 r-o CmO ~XJ i cn o"~--OoH TzxJ ~r. o o F3 o Test Guidelines: U.S. EPA Health Effects Test Guidelines OPPTS 870.7485, Metabolism and Pharmacokinetics (1998) OECD Guidelines for the Testing of Chemicals Section 4: Toxicokinetics, No. 417 (1984) AUTHOR: Raymond A. Kemper, Ph.D. Study Completed on: April 2, 2003 SPONSOR: Association of Plastics Manufacturers of Europe (APME) Fluoropolymers Committee Avenue E. Van Nieuwenhuyse 4 Box 3, B-1160 Brussels Belgium 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 W ork Request Num ber: 13697 Service Code Num ber: 1389 Page 1 of 211 000002 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT This study was conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, which are consistent with the OECD Principles of Good Laboratory Practice (as revised in 1997) published in ENV/MC/CHEM(98)17 and MAFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850) except for the item documented below. The item listed does not impact the validity of the study. The test substance is commercially available and was characterized by the supplier prior to the initiation of this study. A certificate of analysis was provided. Although the characterization was not performed under Good Laboratory Practice Standards, based on our monitoring, we have no reason to doubt the validity of the analysis. Study Director: lond A. Kemper, Ph.D. Research Toxicologist 2 - Q P i -2-00' Date 'iH'M - 2 - 000003 Perfluorooctanoic Acid: Toxicokinetics in the Rat QUALITY ASSURANCE STATEMENT DuPont-7473 Haskell Sample Number(s): 25028,22705-48, 22705-49, 22705-50, 22705-51, 22705-52,22705-53,22705-54, 22705-55 Dates of Inspections: Protocol: August 31, 2001 Conduct: September 28, 2001; October 1, 5, 2001; November 5, 2001; January 7, 2002; February 4, 2002; April 5, 2002; July 17, 25, 2002; August 2, 2002 Records, Reports: February 4-6,10-14,19, 20, 24, 25, 2003 Dates Findings Reported to: Study Director: February 25, 2003 Management: April 1, 2003 Reported by: Joseph C. Hamill Staff Quality Assurance Auditor Date iO?r -3- 000004 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 CERTIFICATION We, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Issued by Study Director: ^ Raymond A. Kemper/Ph.D. Research Toxicologist _______ 2 - / i p r - Z a p '2) Date Approved by: Gary W. Jepson, Ph.D. Principal Research Toxicologist and Manager O j.-'/q - g -Z o o Z Date Approved by: Matthew S. Bogdanff, Ph.D ^D ^B ." Research Manager and Director Date -4- >, 000005 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 TABLE OF CONTENTS Page GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT...................................2 QUALITY ASSURANCE STATEMENT.....................................................................................3 CERTIFICATION.......................................................................................................................... 4 LIST OF TABLES.......................................................................................................................... 7 LIST OF FIGURES..................................................................................................... 10 LIST OF APPENDICES............................................................................................................... 12 STUDY INFORMATION............................................................................................................. 13 STUDY PERSONNEL.................................................................................................................. 14 SUMMARY.................................................................................................................................... 15 INTRODUCTION......................................................................................................................... 17 MATERIALS AND METHODS................................................................................................. 17 A. Regulatory Test Guidelines................................................................................................. 17 B. Test Substance................................................................................................................. ...17 C. Radiolabeled Test Substances............................................................................................. 18 D. Test Species......................................................................................................................... 18 E. Animal Husbandry.............................................................................................................. 19 F. Animal Health Monitoring.................................................................................................. 19 G. Dose Preparation, Analysis and Rates............................... 20 H. Sacrifice............................................................................................................................... 21 I. Pilot Studies........................................................................................................................21 1. Plasma kinetics following Oral Dosing with Non-Radiolabeled PFOA......................21 2. Excretion and Material Balance following Oral Dosing with Radiolabeled PFOA............................................................................................................................. 21 J. Main Study.......................................................................................................................... 22 1. Excretion/Material Balance Experiments (Experiments 1-4)......................................22 2. Pharmacokinetic Experiments (Experiments 5-9)........................................................ 23 3. Distribution Experiments (Experiments 10-12)........................................................... 24 -5 - 000006 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 K. Sample Processing and Quantitation of Radioactive Residues.......................................... 25 1. Whole Blood..................... 26 2. Urine.............................................................................................................................. 26 3. Feces.............................................................................................................................. 26 4. Tissues........................................................................................................................... 26 5. C 0 2................................................................................................................................ 26 6. Volatile Organics...........................................................................................................26 7. Cage Rinse.......................................... 26 8. Residual Feed................................................................................................................ 26 9. Carcass.......................................................................................................................... 27 L. Extraction of PFOA from Plasma for LC/MS Analysis.................................................. .. 27 M. Material Balance............................................................................................................... 27 N. Analytical Methods.............................................................................................................27 1. Instrumentation.............................................................................................................27 2. Chromatographic Methods............................................................................................29 O. Clinical Observations and Mortality...................................................................................34 P. Measurement of Radioactivity............................................................................................34 Q. Data Analyses...................................................................................................................... 34 RESULTS AND DISCUSSION...................................................................................................35 A. Pilot Experiments................................................................................................................ 35 1. Pharmacokinetics in Male and Female Rats following Oral Gavage with 1 mg/kg PFOA...............................................................................................................35 2. Excretion and Material Balance in Male and Female Rats following Oral Gavage with 25 mg/kg 14C-PFOA................................................................................ 35 B. Main Study.......................................................................................................................... 36 1. Excretion/Material Balanceof 14C-PFOA in Male and Female Rats............................ 36 2. Plasma Pharmacokinetics of PFOA in Male and Female R ats....................................39 3. Tissue Distribution of ,4C-PFOA at Plasma Tmax and Tmax/2 in Male and Female Rats...................................................................................................................41 CONCLUSIONS........................................................................................................................... 43 RECORDS AND SAMPLE STORAGE.....................................................................................43 REFERENCES.............................................................................................................................. 44 TABLES......................................................................................................................................... 45 FIGURES..................................................................................................................................... 100 APPENDICES............................................................................................................................. 124 - - 6 - 000007 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 LIST OF TABLES Page 1. Pilot Study - Dosing information for male and female rats....................................................47 2. Pilot Study - Pharmacokinetic parameters in male and female rats following oral gavage with 1 mg/kg PFOA.................................................................................................... 48 3. Pilot Study - Percent of dose recovered in urine in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..................................................................................... 49 4. Pilot Study - Percent of dose recovered in feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..................................................................................... 50 5. Pilot Study - Percent of dose recovered at 168 hours in tissues in male and female rats following oral gavage with 25 mg/kg l4C-PFOA............................................................. 51 6. Pilot Study - Concentration (fig equiv/g) in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..................................................................................... 52 7. Pilot Study - Percent of dose recovered at 168 hours in cage wash and residual feed in male and female rats following oral gavage with 25 mg/kg 14C-PFOA................................. 53 8. Pilot Study - Overall mass balance in male and female rats following oral gavage with 25 mg/kg 14C-PFOA.................................................................................. 54 9. Material Balance - Dosing information for male and female rats......................................... 55 10. Percent of dose recovered in urine in male and female rats following oral gavage with 1 mg/kg ,4C-PFOA....................................................................................................................56 11. Percent of dose recovered in feces in male and female rats following oral gavage with 1 mg/kg I4C-PFOA................................... 57 12. Percent of dose recovered in tissues in male and female rats following oral gavage with 1 mg/kg i4C-PFOA ............................................................................................................ 58 13. Concentration (fig equiv/g) in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA............................................................................................................59 14. Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 1 mg/kg 14C-PFOA....................................................................... 60 15. Overall mass balance in male and female rats following oral gavage with 1 mg/kg 14C-PFOA................................................................................................................................... 61 16. Percent of dose recovered in urine in male and female rats following oral gavage with 5 mg/kg 14C-PFOA....................................................................................................................62 17. Percent of dose recovered in feces in male and female rats following oral gavage with 5 mg/kg 14C-PFOA.................................................................................................................... 63 -7- 000008 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 18. Percent of dose recovered in tissues in male and female rats following oral gavage with 5 mg/kg `"C-PFOA............................................................................................................64 19. Concentration (pig equiv/g) in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA............................................................................................................65 20. Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 5 mg/kg 14C-PFOA.................................................................... 66 21. Overall mass balance in male and female rats following oral gavage with 5 mg/kg 14C-PFOA.............................................. ...................................................................................67 22. Percent of dose recovered in urine in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..................................................................................................................68 23. Percent of dose recovered in feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..................................................................................................................69 24. Percent of dose recovered in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..........................................................................................................70 25. Concentration (pig equiv/g) in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA..........................................................................................................71 26. Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 25 mg/kg 14C-PFOA................................................................... 72 27. Overall mass balance in male and female rats following oral gavage with 25 mg/kg ,4C-PFOA................................................. 73 28. Percent of dose recovered in urine in male and female rats after 14-day repeated administration of 1 mg/kg PFOA, followed by a 1 mg/kg challenge dose of 14C-PFOA (day 15)..................................................................................................................................... 74 29. Percent of dose recovered in feces in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose).............................................................................................. 75 30. Percent of dose recovered in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose)...................................................................................... 76 31. Concentration (pig equiv/g) in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose)...................................................................................... 77 32. Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose)..................................................78 33. Overall mass balance in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose).............................................................................................................79 34. Pharmacokinetics - Dosing information for male and female rats.........................................80 35. Pharmacokinetic parameters in male and female rats following oral gavage with 0.1 mg/kg PFOA.......................................................................................................................81 36. Pharmacokinetic parameters in male and female rats following oral gavage with 1 mg/kg PFOA..........................................................................................................................82 -8- 000009 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 37. Pharmacokinetic parameters in male and female rats following intravenous injection with 1 mg /kg PFOA................................................................................................................83 38. Pharmacokinetic parameters in male and female rats following oral gavage with 5 mg/kg PFOA.......................................................................................................................... 84 39. Pharmacokinetic parameters in male and female rats following oral gavage with 25 mg/kg PFOA................................................................................................... 85 40. Pharmacokinetic parameters in male and female rats following oral gavage with 0.1 mg/kg PFOA (extended time course)................................................................................ 86 41. Tissue Distribution - Dosing information for male and female rats.......................................87 42. Percent of dose recovered at in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA............................................................................................ |. 88 43. Percent of dose recovered at in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA............................................................................................... 89 44. Concentration (pig equiv/g) at in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA............................................................................................... 90 45. Concentration (fig equiv/g) at T , ^ in tissues in male and female rats following oral gavage with 1 mg/kg I4C-PFOA.............................................................................................. 91 46. Percent of dose recovered at Tmmin tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA.............................................................................................. 92 47. Percent of dose recovered at Tmax/2in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA.............................................................................................. 93 48. Concentration (pig equiv/g) at in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA............................................................................................... 94 49. Concentration (fig equiv/g) at in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA............................................................................................... 95 50. Percent of dose recovered at in tissues in male and female rats following oral gavage with 25 mg/kg I4C-PFOA............................................................................................ 96 51. Percent of dose recovered at in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA............................................................................... .............97 52. Concentration (fig equiv/g) at in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA............................................................................................ 98 53. Concentration (fig equiv/g) at in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA............................................................................................. 99 {r / .f ' i i -9 - OOOOIO Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 LIST OF FIGURES Page 1. Pilot Study - PFOA concentration in serial plasma samples from male and female rats following oral gavage with 1 mg/kg PFOA.......................................................................... 102 2. Pilot Study - Cumulative recovery of 14C in urine and feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA........ ..........................................................103 3. Cumulative recovery of 14C in urine and feces in male and female rats following oral I gavage with 1 mg/kg 14C-PFOA............................................................................................ 104 4. Terminal tissue:blood concentration ratio in male and female rats following oral gavage with 1 mg/kg 14C-PFOA............................................................................................ 105 5. Cumulative recovery of 14C in urine and feces in male and female rats following oral gavage with 5 mg/kg 14C-PFOA............................................................................................ 106 6. Terminal tissue:blood concentration ratio in male and female rats following oral gavage with 5 mg/kg 14C-PFOA............................................................................................ 107 7. Cumulative recovery of I4C in urine and feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA.......................................................................................... 108 8. Terminal tissue:blood concentration ratio in male and female rats following oral gavage with 25 mg/kg 14C-PFOA.......................................................................................... 109 9. Cumulative recovery of 14C in urin and feces of male and female rats after 14-day repeated oral gavage with 1 mg/kg PFOA, followed by a 1 mg/kg challenge dose of !4C-PFOA (day 15).................................................................................................................110 10. Terminal tissue:blood concentration ratio in male and female rats following 14-day repeated oral gavage with 1 mg/kg 14C-PFOA...................................................................... I l l 11. PFOA concentration in serial plasma samples from male and female rats following oral gavage with 0.1 mg/kg PFOA........................................................................................ 112 12. PFOA concentration in serial plasma samples from male and female rats following oral gavage with 1 mg/kg PFOA........................................................................................... 113 13. PFOA concentration in serial plasma samples from male and female rats following intravenous injection with 1 mg/kg PFOA............................................................................ 114 14. PFOA concentration in serial plasma samples from male and female rats following oral gavage with 5 mg/kg PFOA........................................................................................... 115 15. PFOA concentration in serial plasma samples from male and female rats following oral gavage with 25 mg/kg PFOA......................................................................................... 116 16. PFOA concentration in serial plasma samples from male and female rats following oral gavage with 0.1 mg/kg PFOA (extended time course).................................................. 117 OOOOll Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 17. Tissue distribution of 14C at and Tmax/2in male and female rats following oral gavage with 1 mg/kg 14C-PFOA............................................................................................ 118 18. Tissue:blood concentration ratio at and Tamn in male and female rats following oral gavage with 1 mg/kg 14C-PFOA..................................................................................... 119 19. Tissue distribution of 14C at T ^ and T ^ in male and female rats following oral gavage with 5 mg/kg l4C-PFOA............................................................................................ 120 20. Tissue:blood concentration ratio at T ^ and T , ^ in male and female rats following oral gavage with 5 mg/kg 14C-PFOA..................................................................................... 121 21. Tissue distribution of 14C at T ^ and TmM/2in male and female rats following oral gavage with 25 mg/kg 14C-PFOA.......................................................................................... 122 22. Tissue:blood concentration ratio at T ^ and TmK/2in male and female rats following oral gavage with 25 mg/kg )4C-PFOA........................... ....................................................... 123 jN M)t ; -11 - OGOOIE Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 LIST OF APPENDICES Page A. Certificates of Analysis...........................................................................................................126 B. Pilot Pharmacokinetics in Male and Female Rats Following Oral Gavage with 1 mg/kg PFOA........................................................................................................................128 C. Pilot Excretion and Material Balance in Male and Female Rats Following Oral Gavage with 25 mg/kg 14C-PFOA...........................................................................................132 D. Material Balance in Male and Female Rats Following Oral Gavage with 1 mg/kg 14C-PFOA................................................................................................ 141 E. Material Balance in Male and Female Rats Following Oral Gavage with 5 mg/kg l4C-PFOA................................................................................................................................. 148 F. Material Balance in Male and Female Rats Following Oral Gavage with 25 mg/kg 14C-PFOA...................................................................................... 155 G. Material Balance in Male and Female Rats Following Oral Gavage with 1 mg/kg ,4C-PFOA (14 day-repeated dose)...........................................................................................162 H. Pharmacokinetics in Male and Female Rats Following Oral Gavage with 0.1 mg/kg PFOA....................................................................................................................................... 169 I. Pharmacokinetics in Male and Female Rats Following Oral Gavage with 1 mg/kg PFOA............................................................................... 173 J. Pharmacokinetics in Male and Female Rats Following Intravenous Injection with 1 mg/kg PFOA........................................................................................................................177 K. Pharmacokinetics in Male and Female Rats Following Oral Gavage with 5 mg/kg PFOA....................................................................................................................................... 181 L. Pharmacokinetics in Male and Female Rats Following Oral Gavage with 25 mg/kg PFOA....................................................................................................................................... 185 M. Pharmacokinetics in Male and Female Rats Following Oral Gavage with 0.1 mg/kg PFOA (extended time course).................................................................................................189 N. Tissue Distribution at T ^ and T ^ in Male and Female Rats Following Oral Gavage with 1 mg/kg 14C-PFOA..........................................................................................................194 O. Tissue Distribution at and in Male and Female Rats Following Oral Gavage with 5 mg/kg 14C-PFOA..........................................................................................................200 P. Tissue Distribution at and T ^ in Male and Female Rats Following Oral Gavage with 25 mg/kg 14C-PFOA........................................................................................................206 ( - 12- 000013 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 STUDY INFORMATION 9th Collective Nomenclature: Pentadecafluorooctanoic acid Synonvms/Codes: Pentadecafluoro-n-octanoic acid Perfluorocaprylic acid Perfluoroheptanecarboxylic acid Perfluorooctanoic acid PFOA 08316DO (Aldrich Chemical Co., Inc) (Lot No.) Haskell Number: 25028 i CAS Registry Number: 335-67-1 Purity: 100.4% (titration with NaOH) 99.9% (gas liquid chromatography) Known Impurities: None identified Physical Characteristics: White powder Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed. Sponsor: Association of Plastics Manufacturers of Europe (APME) Fluoropolymers Committee Avenue E. Van Nieuwenhuyse 4 Box 3, B-1160 Brussels Belgium Study Initiated/Completed: August 3, 2001 / (see report cover page) - 13- 000014 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 STUDY PERSONNEL Study Director: Raymond A. Kemper, Ph.D. Management: Matthew S. Bogdanffy, Ph.D., D.A.B.T. Primary Technician: Benjamin Robinson, B.A. Analytical Associate: Julia N. Wang, Ph.D. Toxicology Report Preparation: Lisa G. Burchfield, A.A. Management: Nancy S. Selzer, M.S. Laboratory Veterinarian: Thomas W. Mayer, D.V.M., A.C.L.A.M. William Singleton, D.V.M., A.C.L.A.M. -14- 000015 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 SUMMARY Perfluorooctanoic acid (PFOA) is a fluorinated fatty acid analogue used as a surfactant in industrial applications. The purpose of the current study was to characterize the absorption, distribution, elimination and pharmacokinetic behavior of PFOA in male and female rats. Male and female CD rats were administered 14C-PFOA (carbonyl label) by oral gavage at dose levels of 1, 5, and 25 mg/kg. Urine and feces were collected for 28 days in males and 7 days in females, and tissues were analyzed for residual radioactivity at sacrifice. Urine was the primary route of elimination for 14C in both sexes. In males, urinary excretion accounted for approximately 43-62% of the administered dose. In females, the extent of urinary elimination was greater, accounting for approximately 76-84% of the administered dose. In males, fecal excretion of 14C accounted for approximately 6-14% of the administered dose, while in females, approximately 2-6% of the dose was recovered in feces. Pilot experiments demonstrated that 14C was not eliminated as either ,4CC>2 or volatile organic compounds in 14C-PFOA-treated rats. At sacrifice, approximately 13-24% of the dose remained in the tissues of male rats, while females retained less than 0.25% of the dose at any dose level. The highest concentrations of residual 14C were found in the blood, liver and kidney. Overall mean recovery of radioactivity ranged from 83.8 to 91.6% in males and 81.3 to 93.3% in females. Pretreatment of rats with 1 mg/kg PFOA for 14 days had little or no effect on excretion or terminal tissue distribution of a challenge dose of 1 mg/kg 14C. The pharmacokinetic behavior of PFOA was investigated following oral administration of PFOA at dose rates of 0.1,1,5, and 25 mg/kg PFOA, and after intravenous administration of 1 mg/kg PFOA. Plasma was collected for 22 days after dosing in males and 5 days in females. Concentration vs. time data were analyzed by non-compartmental pharmacokinetic methods. Comparison of AUC for the oral and intravenous 1 mg/kg doses indicated that oral bioavailability of PFOA was approximately 100%. Plasma elimination curves were linear with respect to time in male rats at all dose levels, while elimination kinetics were biphasic in females at the 5 mg/kg and 25 mg/kg dose levels. In males, plasma elimination half-lives were independent of dose level and ranged from approximately 138 hours to 202 hours. In females, terminal elimination half-lives ranged from approximately 2.8 hours at the lowest dose to approximately 16 hours at the high dose. To further characterize plasma elimination kinetics, particularly in male rats, animals were given oral PFOA at a rate of 0.1 mg/kg, and plasma samples were collected until PFOA concentrations fell below quantitation limits (24 hours and 2016 hours in females and males, respectively). Estimated plasma elimination half-lives in this experiment were approximately 277 hours in males and 3.4 hours in females. The tissue distribution of 14C at the time corresponding to maximum plasma concentrations (Tmax) and at the time corresponding to a return to 50% of maximum plasma concentration following attainment of maximum concentrations (Tmax/2) was investigated following administration of 1, 5, and 25 mg/kg 14C-PFOA. Tissues were collected at 10.5 and 171 hours after dosing in male rats and at 1.25 and 4 hours after dosing in female rats. For both sexes and at both sampling times, the major tissues for distribution of 14C were liver, kidney and blood. In i t - 15- 000016 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 males, the fraction of the dose found in liver increased from Tmaxto Tmax/2, but remained constant or decreased in other tissues. In females, the fraction of the dose present in all tissues remained constant or decreased between Tmax and Tmax/2. Liver:blood concentration ratios for I4C at Tmax in males were greater than 1, and increased between Tmax and Tmax/2- Kidney:blood concentration ratios at Tmax in females were approximately 2 at all dose levels and remained relatively constant between Tmaxand Tmax/2* Overall, the results of this study demonstrated significant sex differences in excretion, pharmacokinetics, and tissue distribution of PFOA in rats. - 16- 000017 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 INTRODUCTION Perfluorooctanoic acid (PFOA) is a synthetic perfluorinated fatty acid analogue, used as a surfactant in industrial applications. In rat metabolism studies with PFOA, elimination following oral administration was found to be much more rapid in females compared to males.(1"2) The dramatic gender differences elimination rate observed for PFOA may be unique to rats. For example, in rabbits and mice treated with 14C-ammonium perfluorooctanoate, no sex differences in elimination rates of 14C were observed, while in hamsters, elimination was more rapid in males compared to females(3). In dogs, a relatively minor (2-fold) sex difference in urinary elimination of PFOA was reported(4). A comprehensive comparison of PFOA elimination rates and tissue distribution between male and female rats will provide a foundation for conducting mechanistic studies to elucidate the basis for the observed sex differences in PFOA elimination kinetics. The objectives of this study were determine the absorption, distribution, excretion and metabolism of PFOA in male and female rats following oral dosing. MATERIALS AND METHODS A. Regulatory Test Guidelines This study was designed to meet the testing requirements of OECD Guidelines for Testing of Chemicals, Toxicokinetics, 417 (April 1984), and U.S. EPA Health Effects Test Guidelines, OPPTS 870.7485, Metabolism and Pharmacokinetics. B. Test Substance Industrial grade PFOA contains various proportions of branched chain isomers in addition to the linear form. For consistency, the current study was conducted using only the linear isomer. Unlabeled PFOA was obtained from Aldrich Chemicals (Milwaukee, Wisconsin). CAS Registry Number: 335-67-1 Molecular Weight: 414.1 Molecular Formula: C8F 15O2H Haskell Number H-25028 r * T' f'\f::\ ,? -17- 000018 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Structure: Purity: F F FO F FFFF 100.4% Titration 99.9% Gas Liquid Chromatography C. Radiolabeled Test Substances [Carbonyl-14C]Ammonium Perfluorooctanoate (14C-PFOA) was synthesized and characterized by Amersham Pharmacia Biotech (Piscataway, NJ). 14C-PFOA, and supplied as an aqueous solution with a radiochemical concentration of approximately 506 pCi/g and chemical concentration of approximately 4.0 mg/g. Specific Activity: Molecular Weight Haskell Numbers Radiochemical Purity: [C14] Ammonium Perfluorooctanoate 125.4 pCi/mg 432.9 22705-48, 22705-49, 22705-50, 22705-51, 22705-52, 22705-53, 22705-54,22705-55 >95% [*] denotes the position of the radiolabel D. Test Species Male and female Crl:CD(SD)IGS BR rats were obtained from Charles River Laboratories, Inc., Raleigh, North Carolina. The Sprague-Dawley rat was chosen for this study because of the extensive experience with this strain and its suitability with respect to longevity, sensitivity, and low incidence of spontaneous diseases. Furthermore, the Sprague-Dawley rat has been used previously for pharmacokinetic and toxicity testing of PFOA and other fluorinated test materials/1'2* At the time of dosing, male rats weighed 193.3-280.0 g and female rats weighed 155.1-231.5 g. At the time of dosing, rats were sexually mature and the weight variation did not exceed 20% of the mean weight by dose group. Each animal was assigned a unique identification number vi!. i 1* Vi - 18- 000019 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 that was used throughout the study. The last 3 digits of the animal identification number were marked on the tail of each animal in indelible ink. . Animal Husbandry Upon arrival at Haskell Laboratory, rats were removed from shipping cartons and housed in appropriate cages, according to Standard Operating Procedures LA003-P-006 unless specified in the study records. Animals were maintained under quarantine for at least six days unless approved otherwise by the site veterinarian, had three recorded weight gains and no abnormalities detected. After the quarantine period, rats were selected for study. Animal rooms were targeted at a temperature of 231C and a relative humidity of 40-60%. j Animal rooms are artificially illuminated (fluorescent light) on an approximate 12-hour light/dark cycle. Throughout the dosing period with test compound, the rats were housed individually in appropriate cages according to the SOP. All animals were provided tap water and fed PMI Nutrition International, LLC Certified Rodent LabDiet 5002 ad libitum. Animals were fasted overnight prior to dosing and were allowed access to food approximately 24 hours after dosing. The actual duration of fasting is documented in the study records. F. Animal Health Monitoring 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 of the study: Water samples are analyzed for total bacterial counts, and the presence of 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 of key contaminants, including specified heavy metals, aflatoxin, chlorinated hydrocarbons, and organophosphates. The presence of these contaminants below the maximum concentration stated by the manufacturer would not be expected to impact the integrity of the study. irr r r r -19- OOZ Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 The animal health and environmental monitoring program is administered by the attending laboratory animal veterinarian. Evaluation of these data did not indicate any conditions that affected the validity of the study. G. Dose Preparation, Analysis and Rates The primary route of test substance administration was oral gavage. This route was chosen at the request of the sponsor. For one experiment, PFOA was administered by intravenous injection to allow estimation of oral bioavailability. PFOA and/or 14C-PFOA was mixed with water as required to achieve the target dose concentrations. The pH of the dose solutions was adjusted to approximately 6.0 with ammonium hydroxide to ensure complete dissolution of the test material. 14C-PFOA was diluted with PFOA to the appropriate specific activity for each dose level. For excretion material balance and tissue distribution experiments, dose solutions were targeted to deliver approximately 20-30 f i d 14C per animal. For oral dosing, a dose volume of approximately 4 mL/kg body weight was used. For intravenous dosing, a dose volume of approximately 2 mL/kg was used. Dose solutions were prepared as follows: Absorption/Plasma Pharmacokinetic Experiments Dose level Dose Volume Chemical Concentration (mg/kg) (mL/kg) (mg/mL) 0.1 1.0 1.0 (iv) 5.0 25.0 4 4 2 4 4 0.025 0.250 0.500 1.250 6.250 Material Balance/Tissue Distribution Experiments Dose Dose Radiochemical Specific Chemical Level Volume Dose Activity Concentration (mg/kg) (mL/kg) (itCi/kg) (ftCi/mg) (mg/mL) 1.0 4 5.0 4 25.0 4 120 120.0 120 24.0 120 4.8 0.25 1.25 6.25 Radiochemical Concentration (nCi/mL) 30 30 30 Dose levels were determined based on previous toxicokinetic and toxicity studies with PFOA.(1) Preparation and data management for radioactive dose solutions was carried out using the DEBRA5 laboratory information management system (Lablogic LTD., Sheffield, UK). Dose preparations for the single dose experiments were prepared prior to the day of use and stored at approximately 4C until used. Dose solution for the repeat dose experiments was prepared in bulk, stored at approximately 4C, and used as needed. -20- '" k 000021 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 HPLC-MS was used to analyze dose solutions for chemical concentration. Dose solutions containing J4C-PFOA were assayed in triplicate for radioactive concentration by liquid scintillation counting (LSC), and radiochemical purity was determined by HPLC using radiometric detection. H. Sacrifice Rats were sacrificed by CO2 asphyxiation and exsanguinated by cardiac puncture. I. Pilot Studies The following pilot studies were conducted to determine the plasma kinetics of PFOA and to characterize PFOA elimination rates following oral administration. The data from these experiments were used in setting the whole blood serial collection time-points and determining the principal routes and rates of elimination. 1. Plasma kinetics following Oral Dosing with Non-Radiolabeled PFOA Two male and two female rats with surgically implanted jugular vein cannulas were obtained from the vendor and allowed to recover for at least one day after surgery before dosing. Rats were administered PFOA at a dose level of 1 mg/kg by the oral route and housed in metabolism cages. Following dosing, serial blood samples were removed from the jugular vein cannula and placed into tubes containing EDTA. In cases where patency of the cannulas became compromised, whole blood was collected from the tail vein. Samples were collected predose, and at 0.5,2, 4, 8, 12,16, and 24 hours post dose, and at 24-hour intervals thereafter up to 168 hours. Whole blood samples were maintained on wet ice. Plasma was separated from red cells by centrifugation. Plasma was stored frozen at < -10C until processed for analysis. 2. Excretion and Material Balance following Oral Dosing with Radiolabeled PFOA Two male and two female rats were administered the test material formulated with ,4C-PFOA at the 25 mg/kg dose level by oral gavage. Following dosing, rats were housed individually in glass metabolism units suitable for the collection of urine, feces, expired air and volatile organics. Urine and feces were collected on dry ice at approximately 4, 8,12, and 24 hours post-dose, and every 24 hours thereafter until sacrifice. Expired air was passed through two scrubbing towers, one containing 2 N NaOH to trap expired 14C 02 and the second containing ethylene glycol to trap volatile organic compounds. Scrubbing towers were replaced every 24 hours post-dose until sacrifice. Animals were sacrificed after 168 hours post-dosing. -21 - 000022 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 At the conclusion of the radiolabeled dose experiments, cages were rinsed with detergent and water (50:50 v/v), followed by water, and finally with acetone. The following tissues were collected at sacrifice: Blood Kidney Lung Uterus Spleen G.I. tract and contents Fat Muscle Testes Bone Adrenals Prostate Liver Heart Ovaries Brain Skin sample Thymus After collection, tissue samples were stored at <-10C until processing and analysis. The remaining carcass was homogenized and assayed so that total material balance could be determined. J. Main Study 1. Excretion/Material Balance Experiments (Experiments 1-4) The objectives of the excretion/material balance experiments were to: determine the rates and routes for elimination of PFOA. determine the material balance for total radioactivity administered. determine if repeated (daily) low oral dose exposure alters the excretion kinetics of PFOA. Dose Routes and Frequency The excretion/material balance experiments were comprised of four dose groups. Experiment 1 2 3 4 Dose Group 1 mg/kg, single oral dose 5 mg/kg, single oral dose 25 mg/kg, single oral dose 1 mg/kg, 14-day repeated dose (unlabeled) 1 mg/kg 14C-PFOA challenge dose (day 15) Each dose group (single dose) was composed of 4 male rats and 4 female rats. Each dose group received dose formulated with 14C-PFOA. For the daily (repeat) low-dose oral gavage experiment, four male and four female rats were administered non-radiolabeled test compound at the low dose rate once per day for -22- 000023 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 14 consecutive days. Rats were administered a challenge dose of the test material formulated with 14C-PFOA on day 15. Experimental Design Following administration of the radiolabeled dose, urine and feces of male rats were collected on dry ice at approximately 4, 8,12, and 24 hdurs following dosing, and at 24-hour intervals thereafter through 336 hours (14 days). Excreta were collected at 48-hour intervals from 336 hours until sacrifice at 672 hours (28 days). For female rats, excreta was collected on dry ice at 4, 8,12 and 24 hours following dosing and at 24-hour intervals thereafter until sacrifice at 168 hours (7 days). As specified in OECD and OPPTS guidelines, since less than 1% of the administered dose was eliminated as expired CO2 and volatile organic compounds in the pilot study, CO2 and volatile organics were not collected in the main study experiments. Moreover, since less than 20% of the administered dose was recovered in the feces in the pilot experiment, bile was not collected in the main study experiments. The following tissues were collected at sacrifice: Blood Kidney Lung Uterus Spleen G.I. tract and contents Fat Muscle Testes Bone Adrenals Prostate Liver Heart Ovaries Brain Skin sample Thymus Thyroid After collection, tissue samples were stored at <-10C until processing and analysis. The remaining carcass was homogenized and assayed so that total material balance could be determined. At the conclusion of the radiolabeled dose experiments, cages were rinsed with detergent and water (50:50 v/v), followed by water, and finally with acetone. Cage wash was stored at room temperature. 2. Pharmacokinetic Experiments (Experiments 5-9) The principal objective of the pharmacokinetic experiments was to determine the extent of absorption of PFOA following oral dosing and to characterize rates of elimination of PFOA from plasma using plasma pharmacokinetic methods. -23- 000024 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dose Routes and Frequency The pharmacokinetic experiments were comprised of five dose groups. DuPont-7473 Experiment 5 6 7 8 9 Dose Group 0.1 mg/kg, oral 1 mg/kg, oral 1 mg/kg, intravenous 5 mg/kg, oral 25 mg/kg, oral 0.1 mg/kg, oral, extended time course Each dose group was composed of 4 male and 4 female rats. Each dose group received nonradiolabeled test compound. The intravenous dose provided a reference for estimating bioavailability of PFOA by the oral exposure routes. Experimental Design Rats with surgically implanted jugular vein cannulas were obtained from the vendor and allowed to recover for at least additional one day after arrival before dosing. Following dosing, serial whole blood samples were drawn from the cannula and placed into Microtainer tubes containing EDTA. In some cases, whole blood was collected from the tail vein. For male rats, blood samples were collected pre-dose, and at 0.25,0.5, 1, 2,4, 8, 12, 16, and 24 hours post-dose, at 24-hour intervals through 192 hours, then at 48-hour intervals from 192 hours through 528 hours. For female rats, blood samples were collected pre-dose, and at 0.25,0.5,1,2,4, 8,12,16,24, 36, 48,72, and 96 hours post-dose. In some cases, whole blood from donor animals of the same age and sex was infused into test animals via the jugular vein cannula following sample removal to prevent volume depletion, as per Haskell Animal Welfare Committee guidelines. Rats were sacrificed after the final sampling time. Whole blood was placed on wet ice, and plasma was separated by centrifugation and frozen at <-10C until analyzed. To further characterize the potential persistence of PFOA in rat plasma, a satellite group of four rats per sex was administered PFOA at a rate of approximately 0.1 mg/kg by oral gavage, and plasma samples were obtained over a more extended sampling period than used in previous toxicokinetics experiments. In female rats, plasma samples were obtained through 312 hours post dose, while in males, samples were obtained through 2016 hours (84 days). Samples were analyzed by LC-MS using multiple reaction monitoring (MRM, Section N). 3. Distribution Experiments (Experiments 10-12) The principal objective of the distribution experiments was to determine the concentration and clearance of PFOA in selected tissues and organs at the time corresponding to maximum plasma concentrations (Tmax) and at the time corresponding to a return to 50% of maximum plasma concentration following attainment of maximum concentrations (Tmax/2) for PFOA. Tissues were selected based upon previous toxicology data and the results of the pilot excretion/material M.\-"vc 1, ij--L ' -24- 000025 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 balance experiment. Sampling times were determined based on the results of the plasma pharmacokinetic experiments. Dose Routes and Frequency The distribution experiments were comprised of the following six dose groups. Experiment 10 11 12 Dose Level 1 mg/kg 1 mg/kg 5 mg/kg 5 mg/kg 25 mg/kg 25 mg/kg Sacrifice Time TA max Tm ax/2 Tmax T m ax/2 T1 max Tm ax/2 Each dose group was composed of 4 male and 4 female rats. Each dose group received dose formulated with 14C-PFOA. Experimental Design Male and female rats were administered 14C-PFOA by oral gavage at the nominal dose rates indicated (Section G). Male rats were sacrificed approximately 10.5 hours (T^x) and 171 hours (Tmax/2) after dosing. Females rats were sacrificed approximately 1.25 hours (Tmax) and 4 hours (Tmax/2) after dosing The following tissues were collected at sacrifice: Blood Kidney Lung Uterus Spleen G.I. tract and contents Thyroid Fat Muscle Testes Bone Adrenals Prostate Carcass Liver Heart Ovaries Brain Skin sample Thymus After collection, tissue samples were stored at <-10C until processing and analysis. Total radioactivity in tissues will be determined by oxidation/LSC. K. Sample Processing and Quantitation of Radioactive Residues Solid samples were oxidized using a Packard Model 307 Oxidizer (Perkin-Elmer Life Sciences, Boston, MA). LSC was carried out using a Packard Tricarb liquid scintillation analyzer (PerkinElmer Life Sciences, Boston, MA). Ha-... *. -25- 000026 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 1. Whole Blood Whole blood samples were maintained on wet ice and stored at approximately 4C until analysis. Aliquots of whole blood were applied to combustion cones and allowed to dry at room temperature. Dried samples were combusted and analyzed by LSC. Analyses were carried out in triplicate. 2. Urine Urine samples from each analysis were thawed at room temperature and mixed prior to analysis by vortexing briefly. Aliquots of each urine sample were analyzed in triplicate by LSC. 3. Feces Feces from each collection interval were thawed at room temperature prior to processing. Thawed feces were mixed with an appropriate volume of distilled water and homogenized using a hand held homogenizer. Aliquots of the homogenate were applied to combustion cones and allowed to dry at room temperature. Dried homogenates were combusted, and analyzed by LSC. 4. Tissues Tissues were thawed at room temperature prior to processing. For smaller tissues, the entire tissue was divided into two or three aliquots and applied to combustion cones. Tissue aliquots were combusted and analyzed by LSC. 5. C 0 2 Aliquots of C 0 2trap solution were analyzed in triplicate by LSC. 6. Volatile Organics Aliquots of volatile organics trap solution were analyzed in triplicate by LSC. 7. Cage Rinse Aliquots of cage rinse were analyzed in triplicate by LSC. 8. Residual Feed Residual feed was separated from feces, stored at <-10 C and thawed at room temperature prior to processing. Samples of residual feed were mixed with an appropriate volume of distilled water and homogenized using a hand held homogenizer. Aliquots of the homogenate were applied to combustion cones and allowed to dry at room temperature. Dried homogenates were combusted and analyzed by LSC. -26- 000027 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 9. Carcass Frozen carcasses were chopped into smaller pieces, mixed with an appropriate volume of distilled water, and homogenized using an electric blender. Aliquots of the homogenate were applied to combustion cones and allowed to dry at room temperature. Dried homogenates were combusted, and analyzed by LSC. L. Extraction of PFOA from Plasma for LC/MS Analysis Plasma samples were processed by solid phase extraction (SPE) using 96-well Isolute array protein precipitation columns (Jones Chromatography, Lakewood, CO). In the pilot experiment, perfluoroheptanoic acid (Aldrich Chemicals, Milwaukee, WI) was used as an internal standard for quantitation of PFOA. In all main study experiments, perfluorononanoic acid (Aldrich Chemicals, Milwaukee, WI) was used as the internal standard. Plasma samples were thawed, and a 75 fiL aliquot was applied to the SPE array. Internal standard was added (10 L of a 3 /xg/mL solution in water), followed by 0.215 mL of ACN. The SPE array was eluted slowly under vacuum into a 96-well receiver plate, and the extracts were analyzed by LC/MS. All samples were extracted and analyzed in duplicate. M. Material Balance Total recovery for rats receiving 14C-PFOA was expected to be 100 10% of the amount administered. N. Analytical Methods 1. Instrumentation System 1: Detector: Hewlett-Packard Series 1100 Liquid Chromatograph, equipped with quaternary pump, column heater, and autosampler Packard 500TR Flow Scintillation Analyzer, with CaF2solid flow cell System 2: Detector: Waters 2790 Liquid Chromatograph, equipped with quarternary pump, column heater, and autosampler Quattro Ultima Mass Spectrometer Mode: SIR and Full Scan Analyzer Conditions: Source: LM 1 Resolution: HM 1 Resolution: Ion energy 1: Entrance: Collision: Negative Electrospray 10.0 10.0 1.0 50 0 -27- 000023 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Exit: LM 2 resolution: HM 2 resolution Ion energy 2: Multiplier (V): SIR dwell time(s): 50 12.0 12.0 2.0 650 0.25 Mode: MRM Analyzer Conditions: Source: LM 1 Resolution: HM 1 Resolution: Ion energy 1: Entrance: Collision: Exit: LM 2 resolution: HM 2 resolution Ion energy 2: Multiplier (V): Negative Electrospray 14.0 14.0 0.5-1.0 -2-50 10-13 5-50 15.0 15.0 1.0 650 MS Method 1: Mode: Time: Chi: Ch2: SIR, 2 masses 0-10.0 min 413.00 463.00 MS Method 2: Mode: Time: Chi: SER, 1 mass 0-10.0 min 413.00 MS Method 3: Mode: Time: Chi: Ch2: SIR, 2 masses 0-7.0 min 413.00 363.00 MS Method 4: Mode: Time: Mass Range: Scan Time: Full Scan 0-10.0 min 300-700 Da 0.8 s MS Method 5: Mode: Time: Chi: Ch2: Dwell Time: MRM, 2 mass pairs 0-12.0 min (413.00-368.60) (462.90 - 418.90) 0.25 s System 3: Hewlett-Packard Series 1100 Liquid Chromatograph, equipped with quaternary pump, column heater, and autosampler Detector: Quattro LC Mass Spectrometer Mode: Full Scan Analyzer Conditions: tf 'jrrV - 28- 000023 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Source: LM 1Resolution: HM 1 Resolution: Ion energy 1: Entrance: Exit: LM 2 resolution: HM 2 resolution Ion energy 2: Multiplier (V): Negative Electrospray 15.0 15.0 0 50 50 12.0 12.0 1.0 650 MS Method 6: Mode: Time: Mass Range: Scan Time: Full Scan 0-10.0 min 170-500 Da 1.0 s System 4: Hewlett-Packard Series 1100 Liquid Chromatograph, equipped with quaternary pump, column heater, and autosampler Detector: Finnigan LCQ Mass Spectrometer Mode: MRM MS Method 7: Analyzer Conditions: Source: Scan Events Scan 1: Negative Electrospray 1 412.8 - (368-370) [12C-PFOA only] 414.8 - (368-370) [14C-PFOA only] (413.5-413.8) - (368-370) [12C/14C Mixture] MS Method 8: Source: Scan Events Scan 1: Scan 2: Negative Electrospray 2 412.8 - (368-370) 463.0-(418-420) Mode: SIR MS Method 9: Source: Scan Events Scan 1: Negative Electrospray 1 SIR (411.9-415.9) 2. Chromatographic Methods Method 1: Instrument Column: Column temperature: Mobile phases: Timetable: Dose radiochemical purity analysis (all 14C experiments) System 1 Waters Symmetry C18,2.1 x 30 mm, 3.5 pm Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Time (min) 0.00 %B 20.0 -29- 000030 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 2.50 20.0 7.00 90.0 10.0 90.0 Flow rate: Stop time: Injection volume: 0.25 mL/min 15.00 min 10 pL Method 2: Dose concentration and plasma concentration analysis (pilot pharmacokinetics experiment) Instrument MS: Column: Column temperature: Mobile phases: System 2 MS Method 3 Waters x Terra MS C l8, 2.1 x 30 mm, 3.5 pm Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 5.00 5.10 7.00 %B 20.0 100.0 20.0 20.0 Flow rate: Stop time: Injection volume: 0.30 mL/min 7.00 min 5 pL Method 3: Dose concentration analysis (pilot material balance exp Instrument MS: Column: Column temperature: Mobile phases: System 3 MS Method 6 Phenomenex Synergi Max-RP, 2.0 x 20 mm, 2 pm Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 0.50 2.00 3.00 3.10 10.0 %B 20.0 20.0 100.0 100.0 20.0 20.0 Flow rate: Stop time: Injection volume: 0.30 mL/min 10.00 min 5 pL Method 4: Dose concentration analysis (main study experiments 1, 4, 6,9, and 10) Instrument MS: Column: System 4 MS Method 7 Zorbax SB-C18 Rapid Resolution, 4.6 x 75 mm, 3.5 pm -30- 000031 Perfluorooctanoic Acid: Toxicokinetics in the Rat Column temperature: Mobile phases: Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 2.00 4.00 4.01 7.00 %B 5.0 5.0 100.0 5.00 5.0 Flow rate: Stop time: Injection volume: 1.0 mL/min 7.00 min 5 fiL Method 5: Dose concentration analysis (main study experiment 8) Instrument MS: Column: Column temperature: Mobile phases: System 4 MS Method 7 Zorbax SB-C18 Rapid Resolution, 4.6 x 75 mm, 3.5 fim Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 2.00 8.00 8.01 10.0 %B 5.0 5.0 100.0 5.00 5.0 Flow rate: Stop time: Injection volume: 1.0 mL/min 10.00 min 5 /iL Method 6: Plasma concentration analysis (main study experiment Instrument MS: Column: Column temperature: Mobile phases: System 4 MS Method 8 Zorbax SB-C18 Rapid Resolution, 4.6 x 75 mm, 3.5 /im Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 2.00 8.00 8.01 10.0 %B 5.0 5.0 90.0 5.00 5.0 Flow rate: Stop time: Injection volume: 1.0 mL/min 10.00 min 5 pL **' . .v -31 - DuPont-7473 000032 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Method 7: Instrument MS: Column: Column temperature: Mobile phases: Timetable: Flow rate: Stop time: Injection volume: Method 8: Instrument MS: Column: Column temperature: Mobile phases: Timetable: Flow rate: Stop time: Injection volume: Method 9: Instrument MS: Column: Column temperature: Mobile phases: Timetable: Dose concentration analysis (main study experiment 12) System 4 MS Method 9 Zorbax SB-C18 Rapid Resolution, 4.6 x 75 mm, 3.5 /xm Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Time (min) 0.00 2.00 8.00 10.0 %B 5.0 5.0 95.0 95.0 1.0 mlVmin 10.00 min 5 fiL Dose concentration analysis (main study experiment 5) System 4 MS Method 7 Waters x Terra MS C18 2.0 x 30 mm, 2.5 xm Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Time (min) 0.00 9.00 %B 25.0 100.0 0.3 mL/min 10.00 min 5 iiL Dose concentration analysis (main study experiment 7) and plasma concentration analysis (main study experiments 5 [males], and 6-8) System 2 MS Method 1 (plasma analysis) MS Method 2 (dose analysis) Phenomenex Synergi Max-RP, 2.0 x 20 mm, 2 /xm Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Time (min) 0.00 0.50 10.0 %B 10.0 10.0 80.0 000033 Perfluorooctanoic Acid: Toxicokinetics in the Rat Flow rate: Stop time: Injection volume: 1.0 mL/min (0.0-0.5 min) 0.25 mL/min (0.5-10.0 min) 10.00 min 5 iL Method 10: Dose concentration analysis (main study experiments 2 and 11) Instrument MS: Column: Column temperature: Mobile phases: System 2 MS Method 4 Phenomenex Synergi Max-RP, 2.0 x 20 mm, 2 fim Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 0.50 10.0 %B 10.0 10.0 80.0 How rate: Stop time: Injection volume: 1.0 mL/min (0.0-0.5 min) 0.25 mL/min (0.5-10.0 min) 10.00 min 5 ixL Method 11: Plasma concentration analysis (main study experiment 9) Instrument MS: Column: Column temperature: Mobile phases: System 2 MS Method 5 Phenomenex Synergi Max-RP, 2.0 x 20 mm, 2 jam Ambient A: 50 mM Ammonium Acetate B: Acetonitrile Timetable: Time (min) 0.00 0.50 10.0 10.01 11.00 11.01 %B 10.0 10.0 80.0 100.0 100.0 10 How rate: Stop time: Injection volume: 1.0 mL/min (0.0-0.5 min) 0.25 mL/min (0.5-12.0 min) 12.00 min 5 fiL Method 12: Dose concentration analysis (main study experiment 3) Instrument MS: Column: Column temperature: Mobile phases: System 4 MS Method 9 Zorbax SB-C18 Rapid Resolution, 4.6 x 75 mm, 3.5 /im Ambient A: 50 mM Ammonium Acetate B: Acetonitrile DuPont-7473 -33- 000034 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Timetable: Flow rate: Stop time: Injection volume: Time (min) 0.00 2.00 4.00 5.00 5.01 8.00 1.0 mL/min 10.00 min 5 fiL %B 5.0 5.0 100.0 100.0 5.0 5.0 O. Clinical Observations and Mortality Cage-site examinations to detect moribund or dead rats and abnormal behavior and/or appearance among rats was conducted at least once daily throughout the study. Moribund rats were sacrificed. P. Measurement of Radioactivity Radioactivity in selected liquid samples was assayed by LSC. Total radioactive residues in solid samples were determined by combustion, trapping liberated 14C02 followed by LSC. Samples for LSC were analyzed for 10 minutes, or until 160,000 disintegrations (0.5% 2c) was accumulated, whichever came first. Q. Data Analyses Group data is represented as a mean SD. Non-compartmental pharmacokinetic analysis of plasma PFOA time course data was accomplished using WinNonlin (Pharsight, Mountain View, CA) using models 200 (single oral bolus) and 201 (single IV bolus). Values for Tmax and Tmax/2 across dose levels were analyzed by one-way ANOVA using Microsoft Excel. -34- 000035 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 RESULTS AND DISCUSSION A. Pilot Experiments 1. Pharmacokinetics in Male and Female Rats following Oral Gavage with 1 mg/kg PFOA (Appendix B, Tables 1-2, Figure 1). Analysis of the dose solution indicated an actual concentration of 0.206 mg/mL, compared to a target concentration of 0.25 mg/mL. Thus, the actual dose rates for male and female rats were 0.78 0.05 mg/kg and 0.80 0.01 mg/kg, respectively. , In male rats, mean plasma concentrations reached a peak of approximately 26 pg/mL after approximately 7 hours (Tmax), and declined slowly thereafter throughout the observation period (168 hours). The plasma half-life for male rats observed in this experiment was approximately 103 hours. The peak plasma concentrations of PFOA observed in females rats (~25 jug/mL) was similar to that seen in male rats. However, peak plasma concentrations of PFOA were achieved within 0.5 hours of dosing in female rats, and plasma concentrations dropped rapidly thereafter. In female rats, PFOA was no longer detectable by 96 hours post-dose. The apparent half-life of PFOA in female rats in this experiment was approximately 3 hours. Furthermore, plasma clearance of PFOA was approximately 32 times greater in female rats compared to male rats. Based on these data, the observation periods for toxicokinetic experiments in the main study were set at 528 hours and 96 hours in male and female rats, respectively. Furthermore, an additional time point was added at 0.25 hours to more accurately define T^x for PFOA in female rats. 2. Excretion and Material Balance in Male and Female Rats following Oral Gavage with 25 mg/kg 14C-PFOA (Appendix C, Tables 1,3-8, Figure 2) Upon analysis, the concentration of test substance in the dose solution was determined to be approximately 6.37 mg/g, compared with a target value of 6.25 mg/g. Based on the volume of dose solution administered, male and female rats received PFOA doses of approximately 18.1 and 18.3 mg/kg, respectively. The radiochemical dose rate was approximately 77 fiCi/kg for both males and females. For both sexes, urinary excretion was the primary route for elimination of 14C-PFOA, with males and females eliminating approximately 30 and 92% of the administered dose by this route, respectively. Urinary excretion of 14C-PFOA in females was rapid, and virtually complete within 24 hours. In contrast, urinary elimination of radioactivity in males was gradual and essentially linear throughout the observation period. The other significant route of elimination for 14C-PFOA was in the feces. Males excreted approximately 8.5% of the administered radioactivity by this route, while females only eliminated approximately 0.5% of the dose in the feces. No radioactivity was detected in either the CO2 trap or the volatile organics trap at any timepoint, confirming that exhalation is not a significant route of elimination for PFOA (Table 8). -35000036 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 At sacrifice (168 hours) approximately 54% of the administered radioactivity remained in the body of male rats, while less than 0.2% of the dose remained in females (Table 5). For both sexes, the highest amounts of residual radioactivity were found in the liver, followed by the kidney. In males, significant amounts of radioactivity were also found in the carcass and whole blood. Mean terminal tissue concentration of PFOA in males ranged from 0.57 fig equiv/g in brain to 57.38 fig equiv/g in liver (Table 6). In female rats, mean terminal liver and kidney concentrations of PFOA were 0.030 fig equiv/g and 0.023 fig equiv/g, respectively. Overall, mean recovery of radioactivity in male and female rats was 93.93% and 93.90%, respectively (Table 8). Since no radioactivity was recovered in either the CO2 or the volatile organic fraction, these ^ fractions were not collected in the main study experiments. Furthermore, since less than 20% of the administered dose was recovered in the feces of either sex, bile was not collected in main study experiments. B. Main Study 1. Excretion/Material Balance of 14C-PFOA in Male and Female Rats (Tables 9-33, Figures 3-10, Appendices D-G) The objective of the excretion/material balance experiments was to characterize the rates and dose dependence of fecal and urinary excretion of 14C-PFOA, and to determine the terminal tissue distribution of residual 14C-PFOA in male and female rats. In addition, experiments were performed to determine potential effects of repeated exposure to PFOA on the excretion and disposition of 14C-PFOA. Dosing (Table 9, Appendices D-G) Excretion and material balance of 14C-PFOA following oral administration were investigated in male and female rats at nominal dose rates of 1, 5 and 25 mg/kg body weight. Actual dose rates were calculated based on analysis of the dose solutions and are presented in Table 9. For all experiments, actual dose rates fell within 15% of target dose rates. Animals received approximately 21-28 f i d of 14C-PFOA in the single dose experiments and approximately 2940 f i d of 14C-PFOA for the repeated dose experiment (Table 9). The actual dose rate of unlabeled PFOA used for the 14 day repeated dosing was approximately 0.93 mg/kg, based on analysis of the dose solution and the volume administered. Experiment 1: Excretion and terminal disposition o f14Cfollowing administration of 1 mg/kg 14C-PFOA by oral gavage (Tables 10-15, Figures 3-4, Appendix D). Urinary excretion was the primary route of elimination for 14C in both males and females (Table 10). Within 28 days, male rats excreted approximately 43% of the administered dose in the urine. Urinary excretion of 14C was gradual in males throughout the observation period, and cumulative recovery of 14C in the urine was still increasing at 28 days (Figure 3). Female rats excreted approximately 76% of the administered dose of 14C in the urine over 7 days. In female rats, urinary excretion of 14C was rapid, and nearly complete within 72 hours after dosing. iron- -36- 000037 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Significant amounts of 14C were also detected in feces of both male and female rats over their respective observation periods (Table 11). The extent of fecal excretion of 14C was greater in males than in females. In both sexes, the majority of fecal elimination of 14C occurred within the first 72 hours after dosing (Figure 3). At the termination of the experiment, approximately 24% of the dose remained in the tissues in male rats, compared to only 0.23% in females (Table 12). In both sexes, liver contained the highest concentrations of 14C (Table 13). In males, 14C was detected in all tissues examined, and ranged from 0.008 fig equiv/g in brain to approximately 2 fig equiv/g in liver. In females, 14C was detected in liver, kidney, whole blood, and skin, and mean concentrations of 14C were less than 0.003 fig equiv/g for all tissues. For male rats, the terminal tissue blood concentration ratio for 14C in liver was nearly 6 (Figure 4). Tissue:blood concentration ratios were approximately 1 or less in all other tissues in males. In females, tissue:blood concentration ratios greater than 1 were observed in liver and kidney. Approximately 3% of the dose was recovered in the cage wash and residual feed fractions (Table 14). Total radioactivity recovered from excreta, tissues, cage wash and residual feed amounted to approximately 84% and 81% of the administered dose in male and female rats, respectively (Table 15). Experiment 2: Excretion and terminal disposition o f 14Cfollowing administration of 5 mg/kg 14C-PFOA by oral gavage (Tables 9,16-21, Figures 5-6, Appendix E). Over the course of the experiment, urinary excretion accounted for approximately 62% and 78% of the administered dose of PFOA in male and female rats, respectively (Table 16). Cumulative recovery of 14C in the urine increased gradually throughout the experiment in male rats, but was essentially complete in females within the first 72 hours after dosing (Figure 5). In this experiment, fecal excretion of 14C accounted for approximately 5-6% of the administered dose in both male and female rats (Table 17). Cumulative fecal excretion of radioactivity was more gradual at this dose level compared to the 1 mg/kg dose level, and continued to increase throughout the observation period in both sexes (Figure 5). At the conclusion of the experiment, approximately 20% and 0.1% of the administered dose was recovered from the tissues of male and female rats, respectively (Table 18). In male rats, highest concentrations of 14C were found in liver, kidney and whole blood (Table 19). In males, tissue concentrations of 14C ranged from 0.027 fig equiv/g in brain to 5.33 fig equiv/g in liver. Tissue:blood concentration ratios were 1 or less in all tissues of male rats except liver, for which a tissue:blood ratio of approximately 5 was observed (Figure 6). In females, only liver, kidney and carcass contained detectable concentrations of 14C. Tissue concentrations of 14C in female rats were less than 0.02 fig equiv/g in all tissues. Since 14C was not detected in whole blood of female rats, tissue:blood concentration ratios could not be calculated. Approximately 3-4% of the dose was recovered in the cage wash and residual feed for both sexes (Table 20). The overall material balance for 14C-PFOA was 91.6% and 86.8% in male and female rats respectively (Table 21). h -37- 000038 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Experiment 3: Excretion and terminal disposition o f 14Cfollowing administration o f 25 mg/kg 14C-PFOA by oral gavage (Tables 9, 22-27, Figures 7-8, Appendix F). As seen for the 1 and 5 mg/kg dose levels, urinary excretion was the primary route of elimination of 14C following oral administration of 14C-PFOA at the 25 mg/kg dose rate (Table 22). Male rats eliminated approximately 53% of the administered dose in the urine. Cumulative urinary elimination in males was more rapid over the first 96 hours following dose administration, but slowed thereafter and appeared to be approaching a plateau after 10-11 days (Figure 7). Female rats eliminated approximately 84% of the dose in the urine, primarily in the first 72 hours after dosing. Fecal excretion accounted for elimination of approximately 12.5 and 1.9% of the administered dose in male and female rats respectively (Table 23). In females, cumulative fecal excretion of 14C was essentially complete within 48 hours after dose administration (Figure 7). In males, fecal excretion of 14C continued throughout the observation period. At sacrifice, approximately 13% of the administered dose was recovered from tissues of male rats (Table 24). In females, approximately 0.2% of the dose was recovered from the tissues. As seen for the two lower dose groups, the highest concentrations of 14C were found in the liver, kidney and whole blood of male rats, and in the liver and kidney of female rats (Table 25). For both sexes, the highest tissue concentrations were observed in the liver. In males, a tissue:blood ratio of approximately 3 was observed in the liver, while this ratio was close to or less than 1 in all other tissues (Figure 8). Radioactivity was not detected in the whole blood of female rats in this experiment, so tissue:blood ratios could not be calculated. In this experiment, the cage wash and residual feed contained approximately 5-7% of the administered radioactivity (Table 26).The overall material balance of 14C in this experiment was 83.9% for male rats and 93.3% for female rats (Table 27). Experiment 4: Excretion and terminal disposition o f 14Cfollowing administration o f 1 mg/kg 14C-PFOA by oral gavagefollowing 14-day repeated dosing with 1 mg/kg PFOA (Tables 9, 2833, Figures 9-10, Appendix G). Overall, approximately 52% and 68% of the administered radioactivity was recovered in the urine of male and female rats, respectively (Table 28). Similar to the single gavage dose experiments, urinary excretion of ,4C continued gradually throughout the observation period in male rats, but was nearly complete within 72 hours for female rats (Figure 9). Thus, repeated administration of PFOA for 14 days prior to challenge with 14C-PFOA did not have significant effects on either the rate or extent of urinary 14C excretion in male or female rats. Fecal excretion accounted for approximately 20% and 12% of the administered dose in male and female rats respectively (Table 29). However, the variability associated with the female feces is very high, possibly due to contamination of the fecal samples with urine. Thus, the high mean value for recovery of 14C in female feces may be artifactual. Apparent rates of fecal 14C excretion in male and female rats were similar in this experiment (Figure 9). - 38- 000039 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Approximately 13% and 0.31% of the dose was recovered in the tissues of male and female rats respectively (Table 30). For both males and females, the highest concentrations of 14C were detected in liver and kidney (Table 31). In males, the concentration of 14C retained in liver after repeated exposure is lower than after single exposure (0.94 /xg equiv/g vs. 1.99 pig equiv/g). In other tissues in males, 14C concentrations were similar. In female tissues, 14C concentrations were less than 0.01 pig equiv/g for both single and repeated exposures (Tables 12 and 31). Tissue:blood concentration ratios were greater than 1 in the liver of male rats (~3.5), and in both liver (~ 7.5) and kidney (~ 6) of female rats (Figure 10). Combined, the cage wash and residual feed accounted for approximately 2.5% and 8.4% of the administered radioactivity in male and female rats respectively (Table 32). In this experiment, the mean overall recovery of radioactivity was approximately 88.0% in males and 89.6% in females (Table 33). 2. Plasma Pharmacokinetics of PFOA in Male and Female Rats (Tables 34-40, Figures 11-16, Appendices H-M) The objectives of the pharmacokinetic experiments were to characterize the plasma pharmacokinetic parameters of PFOA in male and female rats following single oral and intravenous administration and to estimate oral bioavailability of PFOA. Dosing (Table 34, Appendices H-M) Non-compartmental pharmacokinetic parameters were estimated after oral dosing at four nominal dose rates (0.1,1.0, 5.0 and 25.0 mg/kg) and after intravenous administration at a nominal dose rate of 1 mg/kg. Dose solutions were prepared and analyzed by LC/MS as described under Materials and Methods. Actual dose rates for all pharmacokinetic experiments are summarized in Table 34. Actual dose rates fell within 15% of target dose rates for all experiments. Experiment 5: Plasma pharmacokinetics of PFOA in male andfemale rats following administration of 0.1 mg/kg PFOA by oral gavage (Table 35, Figure 11, Appendix H). Non-compartmental pharmacokinetic parameters for 0.1 mg/kg PFOA are presented in Table 35. While similar peak plasma concentrations were observed in males and females, the time required to achieve the plasma peak was much longer in males than in female rats. In both sexes, elimination of PFOA was essentially linear with respect to time following achievement of peak plasma levels (Figure 11). However, the elimination half life (T1/2) was nearly 63 times longer in male rats compared to females. Similarly, plasma clearance was much more rapid (~ 34X) in female rats compared to males. Experiment 6: Plasma pharmacokinetics of PFOA in male andfemale rats following administration o f l mg/kg PFOA by oral gavage and by intravenous injection (Tables 36-37, Figures 12-13, Appendix I-J). Non-compartmental pharmacokinetic parameters for 1 mg/kg PFOA following oral administration are presented in Table 36. Peak plasma concentrations of PFOA in male and h -39- 000040 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 female rats were comparable, though somewhat higher in males. Significant sex differences in other pharmacokinetic parameters are evident, and are similar in magnitude to those observed at the 0.1 mg/kg dose rate. Plasma elimination curves were still essentially linear in both sexes, though some there is some evidence for biphasic behavior in females at later time points (Figure 12). One male rat was found dead on day 9 of this experiment. This event does not appear to be compound-related in that no signs of morbidity were observed in other animals in this experiment or in the other pharmacokinetic experiments. Pharmacokinetic parameters for intravenous administration of PFOA are summarized in Table 37, and plasma elimination curves are presented in Figure 13. Values for common pharmacokinetic parameters were similar between oral and intravenous routes of administration within each sex, and similar sex differences in plasma elimination rates were observed for both routes. Oral bioavailability can be estimated as the ratio of the AUC for oral administration to the AUC for intravenous administration. Comparison of values for oral and intravenous AUC suggests that oral bioavailability of PFOA was essentially 100% (95.5-114.9%). The origin of the spike in plasma PFOA levels in female rats at 12 hours is unknown, and may be artifactual, since it was not observed in either sex dosed orally or in males dosed intravenously. Experiment 7: Plasma pharmacokinetics o f PFOA in male andfemale rats following administration of 5 mg/kg PFOA by oral gavage (Table 38, Figure 14, Appendix K). Non-compartmental pharmacokinetic parameters for 5 mg/kg PFOA are presented in Table 38. At the 5 mg/kg dose level, significant sex differences were observed for all pharmacokinetic parameters calculated. Males exhibited higher peak plasma PFOA concentrations than females, and as with lower dose levels, elimination of PFOA from plasma is much more rapid in females compared to males. In males, the plasma elimination curve is linear at this dose level (Figure 14) and kinetic parameters are similar to those observed at lower doses. However, the plasma elimination curve is biphasic in females, resulting in an increased elimination half-life compared to lower doses. Experiment 8: Plasma pharmacokinetics o f PFOA in male andfemale ratsfollowing administration of 25 mg/kg PFOA by oral gavage (Table 39, Figure 15, Appendix L). Non-compartmental pharmacokinetic parameters for 25 mg/kg PFOA are presented in Table 39. With the exception of the peak plasma concentration, the marked sex differences in pharmacokinetic parameters observed at lower doses are also seen at the highest dose level. While plasma elimination was linear with respect to time in male rats at 25 mg/kg, plasma elimination of PFOA in female rats was clearly biphasic (Figure 15), with a consequent increase in terminal plasma elimination half-life compared to lower dose levels. Experiment 9: Plasma pharmacokinetics of PFOA in male andfemale rats following administration of 0.1 mg/kg PFOA by oral gavage with extended observation interval (Table 40, Figure 16, Appendix M). To further characterize plasma elimination kinetics of PFOA in male and female rats, a second experiment was carried out at the 0.1 mg/kg dose levels using a more sensitive analytical method. -40- 000041 > Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Plasma samples were collected and analyzed until the PFOA concentration dropped below quantifiable levels. In this experiment, PFOA was detected in female rat plasma through 24 hours after dosing, while male rat plasma contained quantifiable levels of PFOA through 2016 hours (84 days). Analysis of the plasma elimination data from this experiment yielded results similar to those obtained in the initial 0.1 mg/kg pharmacokinetic experiment (Table 40). The primary differences between experiments were seen with males, for which the estimates for elimination half-life were somewhat greater, and the values for plasma clearance consequently lower. As in the first 0.1 mg/kg experiment, plasma elimination curves were linear with respect to time for both sexes (Figure 16). 3. Tissue Distribution of 14C-PFOA at Plasma Tma, and Tmav/? in Male and Female Rats (Tables 41-53, Figures 17-22, Appendices N-P) The tissue distribution experiments were carried out to characterize the tissue burden of 14C-PFOA and to investigate the kinetics of elimination and accumulation of PFOA in various tissues of male and female rats. Dosing and Sampling Times (Table 41, Appendices N-P) Tissue distribution of 14C at Tmax and Tmax/2 following oral administration was investigated in male and female rats at nominal dose rates of 1, 5 and 25 mg/kg body weight. Actual dose rates were calculated based on analysis of the dose solutions and are presented in Table 41. For all experiments, actual dose rates fell within 15% of target dose rates. Animals received approximately 21-28 fiCi of 14C-PFOA in these experiments (Table 41). Values for Tmax and Tmax/2 for male and female rats were determined from pharmacokinetic experiments. Tmax/2 was calculated as Tmax + T1/2. For cases in which biphasic elimination was evident, the rapid phase T1/2 was used for calculation of Tmax/2- Values for Tmax and Tmax/2 were analyzed by one-way ANOVA to establish whether different sampling times were required for each dose level. ANOVA analysis indicated that there were no significant differences in TmaXor Tmax/2 across dose levels within each sex. For this reason, mean values for Tmax and Tmax/2 were calculated across all dose levels to establish sampling times for tissue distribution. The resultant sampling times for male rats were 10.5 hours and 171 hours. Sampling times for female rats were 1.25 hours and 4 hours. Experiment 10: Tissue distribution o f I4C at plasma Tmaxand Tmaxa in male andfemale rats following oral administration o f 1 mg/kg u C-PFOA (Tables 42-45, Figures 17-18, Appendix N) The overall body burden of 14C following administration of 1 mg/kg 14C-PFOA was approximately 85% and 51% of the administered dose in male and female rats, respectively (Table 42). In male rats, the highest percentages of the administered dose at Tmax were present in whole blood, liver, skin and muscle (Figure 17). In females, the largest fractions of the dose were found in the GI contents, GI tract, liver and whole blood. The total amount of radioactivity recovered in the body at Tmax/2 decreased to approximately 57% and 29% of the administered -41 - 000042 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 dose in males and females, respectively (Table 43). In male rats, the percentage of the dose found in the liver increased at Tmax/2 relative to Tmax, but percentages in other tissues decreased. In females, the tissue burden of 14C at Tmax/2either decreased or remained constant relative to Tmax- In male rats at Tmax, the highest tissue concentrations of 14C were found in liver, followed by blood and kidney (Table 44). In females at Tmax, the GI contents and tract contained the highest concentrations of 14C, followed by kidney, liver and blood. In males at Tmax/2. tissue concentrations of ,4C decreased relative to Tmax, except in liver where the concentrations were similar at both time points (Table 45). In female rats, tissues concentrations were similar between sampling times for all tissues, except GI contents, where the concentration of 14C decreased. To further characterize tissue distribution of 14C over time, tissue:blood concentration ratios were calculated and are presented in Figure 18. In male rats, tissue blood ratios greater than 1 were observed only in liver, and the liver:blood concentration ratio increased significantly between Tmax and Tmax/2- In female rats, tissuerblood ratios greater than 1 were observed in the GI contents and kidney. In other tissues, ratios were close to 1, and in all tissues, concentration ratios either decreased or remained constant between Tmax and Tmax/2- Experiment 11: Tissue distribution o f14C at plasma Tmaxand Tmaxn in male andfemale rats following oral administration o f 5 mg/kg 14C-PFOA (Tables 46-49, Figures 19-20, Appendix O) The body burden of 14C at Tmaxfollowing oral administration of 5 mg/kg 14C-PFOA was approximately 88% and 37% of the administered dose in male and female rats, respectively (Table 46). The tissue distribution of 14C at Tmax in male and female rats was similar to that observed at the 1 mg/kg dose level, except that the proportion of the dose remaining in the GI contents in females was somewhat lower (Figure 19). A decrease in total body radioactivity similar to that observed following 1 mg/kg 14C-PFOA was observed at Tmax/2 (Table 47). Changes in the tissue distribution of ,4C at Tmax/2 were also similar to those observed at the 1 mg/kg dose level, except that a clear decrease in liver 14C burden was evident in female rats in this experiment. For both male and female rats, the highest concentrations of 14C were found in liver, kidney and blood at Tmax and Tmax/2 (Tables 48-49). Liverblood 14C concentration ratios (Figure 20) in male rats at Tmax were greater than 1, and increased between Tmax and Tmax/2- Tissue:blood ratios for other tissues in males remained constant over this interval. Liver.blood ratios in male rats following 5 mg/kg PFOA were generally lower than were observed at the 1 mg/kg dose level. In females, tissueiblood 14C concentration ratios were similar to those observed at 1 mg/kg, with the exception of the GI contents. -42- 000043 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Experiment 12: Tissue distribution o f 14C at plasma Tmaxand Tm ax/2 in male andfemale rats following oral administration o f 25 mg/kg 14C-PFOA (Tables 50-53, Figures 21-22, Appendix P) In general, body 14C burdens in male and female rats at T^x and Tmax/2 following administration of 25 mg/kg 14C-PFOA were similar to those observed at lower dose levels (Tables 50-51). Moreover, the tissue distribution of the remaining dose at both sampling times was similar to lower dose groups for both sexes (Figure 21). For both male and female rats, the highest concentrations of 14C were found in liver, kidney and blood at Tmax and Tmax/2 (Tables 52-53). The decrease in tissue concentration in female rats between Tmaxand Tmax/2 was somewhat lower than expected. This may have been the due to the use of Tmaxand Tmax/2 to estimate Tmax- This method underestimated Tmax/2 for female rats at the 25 mg/kg dose level. Livenblood 14C concentration ratios (Figure 22) in male rats at Tmax were significantly greater than 1, and increased between Tmax and Tmax/2- Tissue:blood ratios for other tissues in males were less than 1 and remained constant over this interval. In females, the kidney:blood ratio was significantly greater than 1, and remained relatively constant between Tmax and Tmax/2-For other tissues, the ratio was approximately 1 or less and did not change appreciably between sampling times. CONCLUSIONS The results of these studies demonstrate significant sex differences in tissues distribution, elimination, and pharmacokinetic behavior of PFOA in rats. Urinary excretion was the primary route of elimination of PFOA in both males and female, while fecal excretion played a relatively minor role. Elimination of PFOA by both routes was rapid and nearly complete in females. In males, elimination was much slower than in females, and up to 20% of the dose remained in the body after 28 days. PFOA-derived 14C was not eliminated as 14C 02or as volatile organic compounds. Sex differences in excretion of PFOA were reflected in plasma pharmacokinetic parameters in male and female rats. Oral bioavailability of PFOA was approximately 100%. Plasma elimination was slow in male rats, and no evidence of dose-dependent elimination was observed. In females, plasma elimination was rapid and displayed biphasic behavior at the higher dose levels. In both males and females, the main tissues for distribution of PFOA were the blood, liver, and kidney. The blood:liver concentration ratio of PFOA in males increased over time, suggesting that this tissue may serve as a depot for PFOA in male rats. The results of these studies constitute a comprehensive description of PFOA toxicokinetics in male and female rats. 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. -43 - 000044 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 REFERENCES 1. DuPont Haskell Laboratoiy.(1997). Hazard Characterization for Human Health C8 Exposure. Unpublished report. 2. Vanden Heuval, J.P., Kuslikis, B.I., Van Rafelghem, M J. and Peterson, R.E. (1991) Tissue Distribution, Metabolism, and Elimination of Perfluorooctanoic Acid in Male and Female Rats. J. Biochem. Toxicol. 6(2): 83-92. 3. DuPont Haskell Laboratory (1982). Excretion and disposition of 14C-ammonium perfluorooctanoate in male and female rats, mice, hamsters and rabbits. Unpublished report, HLR 62-82. 4. Hanhijarvi, H. (1988) A proposed species differences in the renal excretion of perfluorooctanoic acid in the beagle dog and rat. In: Beynen, A.C and Solleveld, H.A. (Eds) New Developments in Biosciences: Their Implication for Laboratory Animals Science. Martinus Nijhoff Publishers. Dordrecht, Netherlands. -44- 000045 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 TABLES -45 - 000046 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 TABLES ABBREVIATIONS: EXPLANATORY NOTES AUCjnf AUCinf/D Aux Cip Cmax equiv hr or h Lambda-Z fid m NA ND SD Ti/2 Tmax Tmax/2 area under the plasma concentration vs. time curve, extrapolated to infinity AUCinf, normalized to dose auxiliary plasma clearance maximum plasma concentration equivalent hour(s) terminal elimination constant microcurie, 2.22xl06 dpm minute(s) not applicable not detected standard deviation terminal elimination half-life time to Cmax time to Vi Cmax following attainment of maximum concentrations - 46- 000047 O- Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 1: Pilot Study - Dosing information for male and female rats Test Substance Nominal Dose Animal Weight Dose Volume Actual Dose Radioactivity _______ (g)_______ _______ (mL)_______ Rate (mg/kg) Received (gCi) Sex Mean S D ________Mean______SD________ Mean_____ SD_________Mean_____ SD Pharmacokinetics PFOA 1 mg/kg Male 253.7 Female 228.7 20.7 4.0 0.96 0.02 0.89 0.01 0.78 0.05 0.80 0.01 NA NA NA NA Material Balance l4C-PFOA 25 mg/kg Male 250.1 Female 157.2 12.4 2.9 0.71 0.05 0.45 0.01 18.11 18.26 0.38 0.15 19.284 1.361 12.210 Q.326 on - 4 7 - 000048 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 2: Pilot Study - Pharmacokinetic parameters in male and female rats following oral gavage with 1 mg/kg PFOA P aram eter Units T*max hr c'-'max Lambda z Otg/mL) (1/hr) T,a (hr) AUCinf (hr-ng/mL) ATJCjNp/D (hr-^g/mL/mg-kg) Clp (mL-kg/hr) Males Mean SD 7.00 7.07 25.98 0.0068 5.37 0.0006 103.30 13.30 3647.75 671.71 4688.66 567.27 0.21 0.03 Females Mean SD 0.50 0.00 25.30 0.24 11.46 0.02 2.96 0.33 123.35 30.66 154.99 37.19 6.64 1.59 f . -48 - 000049 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 3: Pilot Study - Percent of dose recovered in urine in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total* Male Mean SD 0.217 1.176 1.135 2.570 3.916 4.412 4.580 4.421 3.691 4.194 0.104 0.285 0.300 0.644 0.320 1.278 0.083 0.008 0.591 0.106 30.310 2.539 Female Mean SD 28.493 32.162 17.559 9.968 2.560 0.546 0.351 0.206 0.149 0.147 0.993 9.208 5.326 2.291 0.613 0.087 0.013 0.016 0.000 0.037 92.140 1.849 aTotals are calculated from individual animal data. n=2 ^777 -49- 000050 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 4: Pilot Study - Percent of dose recovered in feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h 0.028 0.045 0.507 4.247 1.102 0.434 0.501 0.477 0.846 0.313 NA NA 0.705 4.183 0.739 0.088 0.380 0.394 1.012 0.298 NA NA 0.123 0.217 0.113 0.030 0.014 0.025 0.012 0.028 NA NA 0.109 0.162 0.086 NA 0.006 NA NA 0.008 Total8 8.462 2.057 0.528 0.419 Totals are calculated from individual animal data. n=2 ^ *"? m f , -50- 000051 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 5: Pilot Study - Percent of dose recovered at 168 hours in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Mean SD Female Mean SD prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thymus testes ovaries adrenals uterus muscle bone Total' 0.034 18.619 0.525 5.761 0.025 0.065 0.201 0.415 0.046 25.628 1.189 0.988 0.228 0.046 0.263 NA 0.008 NA 0.223 0.078 54.338 0.006 2.603 0.211 1.194 0.002 0.000 0.004 0.065 0.006 0.259 0.013 0.132 0.013 0.008 0.033 NA 0.001 NA 0.044 0.011 3.630 NA 0.288 NA NA NA NA NA NA NA 0.008 0.002 NA NA NA NA NA NA NA NA NA 0.154 NA NA NA NA NA NA NA NA NA 0.001 0.001 NA NA NA NA NA NA NA NA NA 0.202 " Totals are calculated from individual animal data. n=2 T -51 - 000052 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 6: Pilot Study - Concentration (jig equiv/g) in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thymus testes ovaries adrenals uterus muscle bone n=4 Male Mean SD 3.009 3.755 7.149 25.548 0.570 1.494 7.890 11.657 3.049 57.379 17.494 3.771 0.543 3.069 3.855 NA 4.348 NA 2.115 3.394 0.137 0.393 0.559 0.679 0.007 0.684 0.222 0.181 0.210 3.746 0.129 0.345 0.004 0.000 0.166 NA 0.354 NA 0.351 1.058 ______ Female Mean SD NA 0.060 NA NA NA NA NA NA NA 0.030 0.023 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 0.003 0.011 NA NA NA NA NA NA NA NA NA -52- 000053 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 7: Pilot Study - Percent o f dose recovered at 168 hours in cage wash and residual feed in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Mean SD Female Mean SD cage wash residua] feed 0.820 NA 0.274 NA 0.605 0.451 0.306 0.503 TotaT 0.820 0.274 1.055 0.809 aTotals are calculated from individual animal data. n=2 -53- 1)1)0054 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 8: Pilot Study - Overall mass balance in male and female rats following oral gavage with 25 mg/kg 14C-PFOA urine feces tissues cage wash & residual feed C 02trap A volatile organics Total Percent of Administered Dose Male Female Mean SD Mean SD 30.310 8.462 54.338 0.820 ND ND 2.539 2.057 3.630 0.274 ND ND 92.140 0.528 0.154 1.055 ND ND 1.849 0.419 0.202 0.809 ND ND 93.929 3.422 93.902 2.581 - 5 4 - 000055 Perfluorooctanoic Acid: Toxicokinetics in the Rat Table 9: Material Balance - Dosing information for male and female rats DuPont-7473 Test Substance Nominal Dose Animal Weight (E) Sex Mean SD Dose Administered (E) Mean SD Material Balance l4C-PFOA 1 mg/kg Male 233.4 Female 174.6 6.2 8.9 0.93 0.02 0.68 0.05 u C-PFOA 5 mg/kg Male 197.6 Female 200.3 3.2 7.4 0.77 0.04 0.80 0.03 u C-PFOA 25 mg/kg Male 209.8 Female 188.5 6.3 5.2 0.85 0.03 0.75 0.02 14C-PFOA 1mg/kg* Male 343.2 Female 243.5 11.1 11.0 1.34 0.06 0.97 0.03 *14-day repeated dose with PFOA, followed by a single dose with uC-PFOA. Actual Dose Rate (mg/kg) Mean SD 1.15 0.85 4.72 4.83 26.07 25.57 0.92 0.94 0.02 0.01 0.15 0.03 0.89 0.20 0.02 0.01 Radioactivity Received (gCi) Mean SD 28.180 22.205 22.125 22.936 24.320 21.438 39.994 29.043 0.579 1.175 1.025 0.975 0.964 0.519 1.935 0.966 <? j H i -55- 000056 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 10: Percent of dose recovered in urine in male and female rats following oral gavage with 1 mg/kg wC-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 0.048 0.408 0.896 2.114 3.370 2.978 2.320 2.101 2.155 1.914 1.735 1.626 1.750 1.496 1.472 1.086 1.191 2.340 2.457 2.377 2.272 2.062 1.499 1.619 0.042 0.230 0.199 0.298 0.481 0.240 0.163 0.186 0.395 0.408 0.338 0.348 0.363 0.149 0.275 0.431 0.323 0.585 0.248 0.534 0.718 0.846 0.140 0.327 11.505 30.930 16.002 10.713 3.119 1.538 0.826 0.432 0.329 0.480 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 5.954 . 5.092 3.043 3.290 1.428 0.634 0.153 0.086 0.095 0.220 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total* 43.283 3.015 75.872 4.066 *Totals are calculated from individual animal data. n=4 - 5 6 - 000057 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 11: Percent of dose recovered in feces in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 NA 2.269 2.433 5.554 0.838 0.292 0.238 0.126 0.160 0.190 0.255 0.299 0.190 0.204 0.098 0.256 0.227 0.387 0.232 0.284 0.309 0.457 0.262 0.198 NA NA 2.625 3.577 0.517 0.225 0.245 0.046 0.039 0.124 0.268 0.297 0.177 0.204 0.035 0.165 0.159 0.235 0.128 0.158 0.193 0.313 0.195 0.096 NA NA 1.239 0.580 0.108 0.028 0.106 0.046 0.033 0.030 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 1.944 0.898 0.092 0.005 0.092 0.028 0.019 0.017 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total3 14.055 4.003 2.169 2.923 *Totals are calculated from individual animal data. n=4 < ' - 5 7 - 000058 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 12: Percent of dose recovered in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male (Day 28) Mean SD Female (168 Hours) Mean SD prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone 0.020 7.388 0.090 1.265 0.006 0.017 0.064 0.108 0.015 13.400 0.522 0.344 0.061 0.002 0.011 0.103 NA 0.002 NA 0.095 0.032 0.009 1.440 0.030 0.387 0.002 0.007 0.014 0.016 0.004 2.467 0.132 0.070 0.012 0.001 0.004 0.029 NA 0.001 NA 0.025 0.011 NA 0.215 0.001 0.004 NA NA NA NA NA 0.013 0.002 NA 0.003 NA NA NA NA NA NA NA NA NA 0.125 NA 0.002 NA NA NA NA NA 0.003 0.001 NA NA NA NA NA NA NA NA NA NA Total11 23.545 4.062 0.232 0.125 aTotals are calculated from individual animal data. n=4 -58- 000059 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 13: Concentration (jig equiv/g) in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone n=4 Male Mean SD 0.050 0.061 0.115 0.357 0.008 0.014 0.121 0.171 0.054 1.987 0.357 0.051 0.008 0.066 0.047 0.092 NA 0.056 NA 0.035 0.054 0.016 0.015 0.029 0.099 0.002 0.007 0.035 0.046 0.014 0.427 0.054 0.016 0.003 0.034 0.014 0.035 NA 0.024 NA 0.008 0.011 Female Mean SD NA 0.002 0.001 0.001 NA NA NA NA NA 0.002 0.002 NA 0.000 NA NA NA NA NA NA NA NA NA 0.001 NA 0.000 NA NA NA NA NA 0.000 0.001 NA NA NA NA NA NA NA NA NA NA M **:i t , ' -59- 000060 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 14: Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male (Day 28)________ Mean SD Female _______(168 Hours) Mean SD cage wash residual feed 2.212 0.736 0.528 0.450 2.854 0.198 1.287 0.031 Total" 2.948 0.732 3.052 1.311 "Totals are calculated from individual animal data. n=4 P# - 60- 0U061 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 15: Overall mass balance in male and female rats following oral gavage with 1 mg/kg 14C-PFOA urine feces tissues cage wash & residual feed Total Male Mean SD 43.238 14.055 23.545 2.948 3.015 4.003 4.062 0.732 83.830 4.006 Female Mean SD 75.872 2.169 0.232 3.052 4.066 2.923 0.125 1.311 81.325 4.546 L . TV -61 - 000062 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 16: Percent of dose recovered in urine in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 0.102 0.607 1.043 2.575 4.445 3.931 3.473 3.748 3.346 3.454 2.925 2.634 2.674 2.379 2.241 2.045 1.869 3.214 3.379 3.003 2.788 2.314 2.258 1.935 0.081 0.319 0.468 0.441 0.584 0.784 0.675 0.684 0.151 0.779 0.212 0.244 0.248 0.193 0.284 0.289 0.298 0.306 0.336 0.268 0.314 0.285 0.151 0.249 11.696 35.213 17.189 13.565 3.026 1.008 0.622 0.642 0.441 0.314 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 10.464 3.547 3.358 6.189 0.831 0.579 0.209 0.480 0.319 0.132 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total" 62.201 3.656 77.867 6.034 *Totals are calculated from individual animal data. n= 4 e JT - -62- 000063 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 17: Percent of dose recovered in feces in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 NA NA 1.027 0.740 0.536 0.580 0.355 0.197 0.176 0.140 0.128 0.111 0.149 0.145 0.119 0.088 0.101 0.204 0.161 0.124 0.107 0.120 0.115 0.148 NA NA 1.989 0.249 0.381 0.489 0.314 0.094 0.078 0.039 0.035 0.027 0.082 0.051 0.073 0.030 0.032 0.082 0.071 0.027 0.031 0.049 0.027 0.091 0.003 NA 1.134 3.934 0.560 0.341 0.146 0.119 0.106 0.114 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 0.069 5.118 0.630 0.357 0.169 0.141 0.131 0.112 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total* 5.568 1.779 5.886 5.387 *Totals are calculated from individual animal data. n=4 fi0<. 000064 -63- Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 18: Percent of dose recovered in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male (Day 28) Mean SD Female (168 Hours) Mean SD prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone 0.015 6.677 0.074 1.183 0.006 0.018 0.056 0.096 0.014 10.904 0.388 0.259 0.055 0.002 0.014 0.093 NA 0.006 NA 0.065 0.028 0.003 1.214 0.016 0.201 0.001 0.002 0.012 0.030 0.003 1.390 0.031 0.045 0.015 0.001 0.003 0.014 NA 0.006 NA 0.008 0.005 NA 0.121 NA NA NA NA NA NA NA 0.016 0.002 NA NA NA NA NA NA NA NA NA NA NA 0.045 NA NA NA NA NA NA NA 0.002 0.000 NA NA NA NA NA NA NA NA NA NA Total* 19.949 2.755 0.109 0.072 "Totals are calculated from individual animal data. -64- 00006S Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 19: Concentration (jig equiv/g) in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone n=4 Male Mean SD 0.142 0.179 0.345 1.103 0.027 0.052 0.349 0.492 0.159 5.331 0.935 0.165 0.028 0.176 0.145 0.264 NA 0.468 NA 0.102 0.174 0.043 0.025 0.073 0.161 0.005 0.013 0.061 0.081 0.013 0.670 0.078 0.031 0.011 0.049 0.012 0.042 NA 0.574 NA 0.011 0.023 Female Mean SD NA 0.007 NA NA NA NA NA NA NA 0.016 0.010 NA NA NA NA NA NA NA NA NA NA NA 0.003 NA NA NA NA NA NA NA 0.003 0.001 NA NA NA NA NA NA NA NA NA NA -65/\; 000066 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 20: Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male (Day 28) Mean SD Female (168 Hours) Mean SD cage wash residual feed 3.652 0.377 1.966 0.434 2.417 0.541 0.864 0.450 Total* 3.935 2.362 2.958 0.845 aTotals are calculated from individual animal data. n=4 - 66- O67 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 21: Overall mass balance in male and female rats following oral gavage with 5 mg/kg 14C-PFOA urine feces tissues cage wash & residual feed Total Male Mean SD 62.201 5.568 19.949 3.935 3.656 1.779 2.755 2.362 91.652 2.577 Female Mean SD 77.867 5.886 0.109 2.958 6.034 5.387 0.072 0.845 86.819 5.169 A ? . -67 - 000068 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 22: Percent of dose recovered in urine in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 0.251 0.771 1.350 3.258 6.803 4.609 4.026 3.779 3.238 2.567 2.066 1.656 1.856 1.715 1.539 1.182 1.124 1.495 1.151 2.249 1.953 1.819 1.522 1.287 0.190 0.400 0.568 0.921 1.914 1.811 1.361 0.741 0.635 0.579 0.527 1.088 0.362 0.442 0.489 0.321 0.381 0.557 0.435 0.394 0.256 0.106 0.339 0.591 23.794 28.586 14.200 9.855 3.714 1.539 1.080 0.706 0.434 0.474 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 8.041 11.300 3.042 4.783 0.966 0.428 0.552 0.313 0.196 0.286 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total" 53.265 8.490 84.381 12.023 *Totals are calculated from individual animal data. n=4 ft yj_ ; - 6 8 - 000069 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 23: Percent of dose recovered in feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 0.020 0.181 0.125 1.667 0.986 0.338 0.436 0.522 0.507 0.495 0.722 0.678 0.933 0.325 0.319 0.459 0.421 0.798 1.320 0.191 0.307 0.297 0.322 0.260 NA 0.249 0.139 1.677 1.018 0.073 0.221 0.289 0.138 0.274 0.414 0.496 0.418 0.113 0.247 0.421 0.332 0.388 0.335 0.053 0.184 0.202 0.073 0.139 0.017 0.002 0.560 0.749 0.280 0.171 0.144 0.042 0.069 0.130 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 0.783 1.187 0.354 0.208 0.111 0.032 0.063 0.166 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total" 12.490 4.153 1.868 2.546 aTotals are calculated from individual animal data. n=4 -69- ' i t 000070 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 24: Percent of dose recovered in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male (Day 28) Mean SD Female (168 Hours) Mean SD prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone Total8 0.010 5.355 0.051 0.769 0.005 0.014 0.045 0.077 0.016 5.521 0.389 0.357 0.056 0.002 0.009 0.068 NA 0.003 NA 0.041 0.018 12.803 0.004 2.637 0.029 0.366 0.002 0.007 0.028 0.035 0.010 1.636 0.183 0.187 0.034 0.001 0.006 0.031 NA 0.002 NA 0.016 0.006 5.005 NA 0.188 0.001 NA NA NA NA NA NA 0.015 0.003 NA NA NA NA NA NA NA NA NA NA 0.160 NA 0.068 0.001 NA NA NA NA NA NA 0.008 0.000 NA NA NA NA NA NA NA NA NA NA 0.102 8Totals are calculated from individual animal data. n=4 -, i t f. v - 70 - 000071 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 25: Concentration (/ig equiv/g) in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone n=4 Male ______ Mean SD 0.798 0.962 1.790 5.771 0.139 0.291 1.863 2.658 1.248 16.609 6.229 1.166 0.156 1.164 0.854 1.227 NA 1.150 NA 0.476 0.707 0.493 0.529 0.905 2.828 0.068 0.156 1.029 1.375 0.903 5.892 2.745 0.755 0.100 0.671 0.577 0.590 NA 0.589 NA 0.225 0.274 _________Female Mean SD NA 0.056 0.024 NA NA NA NA NA NA 0.078 0.074 NA NA NA NA NA NA NA NA NA NA NA 0.022 0.016 NA NA NA NA NA NA 0.041 0.013 NA NA NA NA NA NA NA NA NA NA r ; .< -71 - 000072 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 26: Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male (Day 28) Mean SD Female (168 Hours) Mean SD cage wash residual feed 3.997 1.347 1.614 1.850 1.856 4.996 1.316 7.569 Total8 5.344 3.336 6.852 8.360 8Totals are calculated from individual animal data. n=4 kf i -72- 000073 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 27: Overall mass balance in male and female rats following oral gavage with 25 mg/kg 14C-PFOA urine feces tissues cage wash & residual feed Total Male Mean SD 53.265 12.490 12.803 5.344 8.490 4.153 5.005 3.336 83.902 3.433 Female Mean SD 84.381 1.868 0.160 6.852 12.023 2.546 0.102 8.360 93.260 . 3.554 K m - 73- 000074 Perfluorooctanoic Acid: Toxicokinetics in die Rat DuPont-7473 Table 28: Percent of dose recovered in urine in male and female rats after 14-day repeated administration of 1 mg/kg PFOA, followed by a 1 mg/kg challenge dose of 14CPFOA (day 15) Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 0.155 0.704 0.929 2.832 5.107 3.788 2.659 2.488 2.681 2.298 1.784 1.918 1.561 1.810 1.871 1.998 2.279 3.213 2.905 2.381 2.035 1.850 1.744 1.444 0.143 0.224 0.451 0.631 1.731 1.044 1.419 1.273 1.162 0.403 0.629 0.422 0.637 0.483 0.620 0.689 0.434 0.238 0.344 0.465 0.531 0.485 0.264 0.465 7.547 32.922 9.853 10.020 5.984 1.375 1.281 0.556 0.558 0.329 NA NA NA NA NA NA NA NA NA NA NA NA NA NA 5.777 14.134 2.887 4.319 2.579 0.854 0.894 . 0.239 0.462 0.253 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total* 52.430 7.959 68.537 16.631 *Totals are calculated from individual animal data. n=4 - 74- 000075 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 29: Percent of dose recovered in feces in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose) Timepoint Male Mean SD Female Mean SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day-8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 NA 0.203 1.072 6.752 1.254 0.817 1.044 0.376 0.769 1.074 1.150 1.113 1.060 0.712 0.465 0.345 0.096 0.342 0.202 0.275 0.319 0.220 0.076 0.108 NA 0.318 1.745 3.460 0.458 0.473 0.774 0.207 0.315 0.578 0.714 0.427 1.170 0.815 0.433 0.400 0.036 0.295 0.053 0.250 0.242 0.202 0.021 0.091 NA 3.599 0.172 4.539 2.883 0.764 0.121 0.160 0.143 0.047 NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA 4.530 0.154 7.406 3.972 0.824 0.093 0.124 0.072 0.019 NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total3 19.841 6.620 12.384 15.775 3Totals are calculated from individual animal data. n=4 - 75 - 000076 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 30: Percent of dose recovered in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose) Male (Day 28) Mean SD Female (168 Hours) Mean SD prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone 0.008 5.226 0.078 0.977 0.004 0.019 0.040 0.074 0.012 6.113 0.346 0.177 0.041 0.001 0.012 0.054 NA 0.002 NA 0.052 0.019 0.004 1.886 0.035 0.573 0.002 0.006 0.016 0.030 0.006 1.684 0.164 0.083 0.021 0.001 0.006 0.021 NA 0.001 NA 0.023 0.007 NA 0.262 0.001 0.004 NA NA NA 0.000 NA 0.037 0.005 NA NA NA NA NA NA NA NA NA NA NA 0.075 0.000 0.001 NA NA NA NA NA 0.006 0.002 NA NA NA NA NA NA NA NA NA NA Total3 13.251 4.430 0.309 0.083 "Totals are calculated from individual animal data. n=4 -76- 00007? Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 31: Concentration (fig equiv/g) in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose) prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone n=4 Male Mean SD 0.022 0.042 0.083 0.293 0.007 0.009 0.084 0.121 0.036 0.940 0.272 0.038 0.006 0.034 0.042 0.052 NA 0.044 NA 0.025 0.032 0.011 0.016 0.032 0.147 0.003 0.003 0.034 0.052 0.015 0.243 0.111 0.018 0.003 0.018 0.021 0.024 NA 0.027 NA 0.010 0.011 ___ Female Mean SD NA 0.003 0.002 0.001 NA NA NA 0.001 NA 0.008 0.006 NA NA NA NA NA NA NA NA NA NA NA 0.001 0.001 0.000 NA NA NA NA NA 0.001 0.001 NA NA NA NA NA NA NA NA NA NA -77- 000078 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 32: Percent of dose recovered in cage wash and residual feed in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose) Male (Day 28)________ Mean SD Female (168 Hours) Mean SD cage wash residual feed 2.265 0.263 1.170 0.349 3.643 4.766 1.982 8.615 Total" 2.528 1.465 8.409 8.329 aTotals are calculated from individual animal data. n=4 -78- 000079 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 33: Overall mass balance in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeat dose) urine feces tissues cage wash & residual feed Total Percent of Administered Dose Male Female Mean SD Mean SD 52.430 19.841 13.251 2.528 7.959 6.620 4.430 1.465 68.537 12.384 0.309 8.409 16.631 15.775 0.083 8.329 88.049 1.651 89.638 3.348 -79- 000060 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 34: Pharmacokinetics - Dosing information for male and female rats Test Substance Nominal Dose Animal Weight (g) Sex Mean SD Pharmacokinetics PFOA 0.1 mg/kg Male 216.3 Female 176.4 11.2 8.6 PFOA 1 mg/kga Male 248.7 Female 197.0 1.6 9.7 PFOA 1 mg/kgb Male 248.3 Female 200.6 9.3 2.7 PFOA PFOA 5 mg/kg 25 mg/kg Male 218.0 Female 187.3 Male 225.0 Female 194.8 3.4 3.6 3.7 14.4 PFOA 0.1 mg/kgc Male 272.5 Female 201.0 5.8 11.8 "Oral gavage ^Intravenous injection 'Extended time course Dose Volume (mL) Mean SD 0.86 0.04 0.71 0.03 1.00 0.01 0.79 0.04 0.50 0.02 0.40 0.01 0.87 0.01 0.75 0.01 0.90 0.01 0.78 0.06 1.10 0.00 0.81 0.05 Actual Dose Rate (mg/kg) Mean SD 0.112 0.000 0.113 0.000 1.01 0.00 1.01 0.01 1.11 0.01 1.11 0.01 5.51 0.02 5.53 0.02 26.76 26.94 0.31 0.09 0.097 0.002 0.097 0.000 -80- 000081 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 35: Pharmacokinetic parameters in male and female rats following oral gavage with 0.1 mg/kg PFOA P aram eter Units Maies Mean SD Tmax c'"'max Lambda z Tm AUCjnf AUCinf/D Clp hr (ig/mL) (1/hr) (hr) (hr-/ig/mL) (hr-/xg/mLVmg/kg (mL/kg-hr) 10.25 0.598 0.004 201.774 123.224 1096.811 0.962 6.45 0.127 0.001 37.489 35.476 310.491 0.240 Females Mean SD 0.56 0.67 0.231 3.206 3.584 31.721 32.359 0.31 0.07 0.066 0.905 0.666 5.880 6.025 H) -81 000052 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 36: Pharmacokinetic parameters in male and female rats following oral gavage with 1 mg/kg PFOA Parameter Units Males Mean SD T1max Cmax Lambda z T,fl AUCinf AUCinf/D Clp hr (ftg/mL) (1/hr) (hr) (hr/tg/mL) (hr-/ig/mL/mg/kg (mL/kg-hr) 9.00 8.431 0.005 138.343 1194.463 1176.009 0.871 3.83 1.161 0.001 31.972 215.578 206.316 0.158 Females Mean SD 1.13 4.782 0.213 3.457 39.072 38.635 27.286 0.63 1.149 0.053 1.111 10.172 10.093 7.159 - 82- 000083 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 37: Pharmacokinetic parameters in male and female rats following intravenous injection with 1 mg /kg PFOA Parameter Units Males Mean SD Lambda z T,fl AUCinf AUCinf^D Clp (1/hr) (hr) (hr/tg/mL) (hr/ig/mL/mg/kg (mL/kg-hr) 0.004 185.584 1249.817 1123.384 0.896 0.000 19.558 113.167 100.488 0.082 Females Mean SD 0.250 2.844 33.998 30.747 34.040 0.047 0.514 7.601 6.759 9.230 - 83 - 000084 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 38: Pharmacokinetic parameters in male and female rats following oral gavage with 5 mg/kg PFOA Parameter Units Males Mean SD Tmax c'-'max Lambda z Tlfl AUCjnf AUCfNp/D Clp hr (/xg/mL) (1/hr) (hr) (hr/xg/mL) (hr/xg/mL/mg/kg (mL/kg-hr) 15.0 44.75 0.0041 174.19 6733.70 1221.89 0.85 10.5 6.14 0.0007 28.92 1392.83 250.28 0.21 Females Mean SD 1.50 20.36 0.15 4.60 114.90 20.78 48.48 0.58 1.58 0.02 0.64 11.23 2.01 4.86 - 84- 0000S5 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 39: Pharmacokinetic parameters in male and female rats following oral gavage with 25 mg/kg PFOA Parameter Units Males Mean SD T1max c'--max Lambda z Tin AUCinf AUCinf/D Clp hr (/ig/mL) (1/hr) (hr) (hr-jig/mL) (hr/ig/mL/mg/kg (mL/kg-hr) 7.5 160.00 0.0046 157.47 25155.61 942.65 1.13 6.2 11.95 0.0012 38.39 7276.96 284.67 0.31 Females Mean SD 1.25 132.65 0.059 16.22 795.76 29.54 35.06 0.87 45.95 0.037 9.90 187.51 6.92 6.88 A -85- 000086 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 40: Pharmacokinetic parameters in male and female rats following oral gavage with 0.1 mg/kg PFOA (extended time course) Parameter Units Males Mean SD T1max c'-'max Lambda z Tjfl AUCinf AUCjnf'D Clp hr (/rg/mL) (1/hr) (hr) (hr/rg/mL) (hr/ig/mL/mg/kg (mL/kg'hr) 5.5 1.08 0.0026 277.10 206.38 2111.28 0.51 7.0 0.42 0.0007 56.62 59.03 586.77 0.17 Females Mean SD 1.25 0.52 0.22 3.44 3.34 34.39 29.30 0.50 0.08 0.07 1.26 0.32 3.29 3.06 -86- 000087 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 41: Tissue Distribution - Dosing information for male and female rats Test Substance Tom* I4c -p f o a 14c -p f o a I4c -p f o a Ttraodi I4c -p f o a I4c -pf o a 14c -p f o a Nominal Dose Animal Weight ____ (g) Sex Mean SD 1 mg/kg 5 mg/kg 25 mg/kg Male 202.2 Female 192.4 Male 229.1 Female 193.2 Male 216.2 Female 190.3 3.4 2.2 6.1 5.4 6.8 3.0 1 mg/kg 5 mg/kg 25 mg/kg Male 203.2 Female 189.5 Male 225.1 Female 192.3 Male 218.3 Female 193.2 2.4 6.4 7.6 9.6 7.6 4.1 Dose Volume (inL) Mean SD 0.80 0.01 0.79 0.02 0.92 0.03 0.77 0.02 0.88 0.03 0.77 0.01 0.81 0.02 0.76 0.02 0.90 0.03 0.78 0.04 0.88 0.03 0.78 0.02 Actual Dose Rate (mg/kg) Mean SD 0.87 0.88 4.72 4.68 25.02 24.90 0.02 0.01 0.07 0.03 0.08 0.28 0.88 0.89 4.71 4.72 24.95 24.90 0.01 0.01 0.02 0.05 0.12 0.09 Radioactivity Received (gCi) Mean SD 23.463 22.725 28.391 23.754 23.792 20.837 0.384 0.171 0.985 0.601 0.790 0.338 23.942 22.548 27.822 23.838 23.948 21.149 0.517 0.620 0.922 1.214 0.900 0.444 V -87- ooooss Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 42: Percent of dose recovered at Tinaxin tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male (10 h 30 m) Mean SD Female (1 h 15 m) Mean SD prostate skin2 whole blood2 brain fat2 heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle2 uterus bone2 0.083 14.772 22.148 0.071 2.281 0.451 0.740 0.086 21.708 1.949 2.930 2.083 0.008 0.085 0.755 NA 0.019 12.025 NA 3.273 0.039 2.135 0.692 0.018 0.467 0.119 0.147 0.011 5.627 0.402 0.929 0.625 0.005 0.008 0.079 NA 0.004 0.648 NA 0.538 NA 0.434 5.740 0.037 0.134 0.198 0.454 0.063 7.060 3.288 10.699 21.956 0.010 0.052 NA 0.047 0.014 0.170 0.243 0.101 NA 0.162 1.507 0.009 0.032 0.079 0.148 0.027 1.266 0.948 9.066 13.477 0.003 0.017 NA 0.019 0.005 0.051 0.091 0.017 Totalb 85.465 6.426 50.698 16.485 2 Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. b Totals are calculated from individual animal data, n=4 -88- 000089 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 43: Percent of dose recovered at Tmax/2 in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male (171 h) Mean SD Female (4 h) Mean SD prostate skin8 whole blood8 brain fat8 heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle8 uterus bone8 Total" 0.030 6.061 8.232 0.022 0.593 0.195 0.341 0.045 32.627 1.140 0.980 0.239 0.004 0.051 0.356 NA 0.010 4.984 NA 1.120 57.026 0.002 0.274 1.218 0.002 0.136 0.024 0.043 0.006 3.601 0.215 0.300 0.025 0.003 0.018 0.037 NA 0.001 0.745 NA 0.094 3.379 NA 0.403 4.438 0.047 0.164 0.253 0.546 0.058 6.817 2.769 8.462 3.891 0.016 0.058 NA 0.048 0.018 0.258 0.374 0.153 28.772 NA 0.096 1.625 0.008 0.079 0.055 0.082 0.006 1.537 0.784 6.519 2.395 0.021 0.024 NA 0.006 0.004 0.089 0.247 0.052 10.976 8 Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. b Totals are calculated from individual animal data, n=4 -89- 001X090 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 44: Concentration (/tg equiv/g) at Tma*in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone n=4 Male (10 h 30 m) Mean SD 0.454 0.672 2.587 0.066 0.281 0.973 1.157 0.339 4.627 1.710 0.528 0.218 0.399 0.277 0.540 NA 0.531 NA 0.257 0.386 0.110 0.097 0.024 0.012 0.050 0.161 0.151 0.058 0.729 0.302 0.151 0.025 0.088 0.039 0.051 NA 0.130 NA 0.008 0.053 Female (1 h 15 m) Mean SD NA 0.298 1.717 0.033 0.142 0.440 0.701 0.201 1.938 3.073 2.184 9.170 0.385 0.175 NA 0.422 0.238 0.410 0.119 0.171 NA 0.114 0.569 0.008 0.064 0.151 0.192 0.065 0.260 0.852 2.185 5.910 0.117 0.055 NA 0.167 0.076 0.231 0.035 0.053 -90- 000091 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 45: Concentration (fig equiv/g) at Tma* in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals uterus muscle bone n=4 Male (171 h) Mean SD 0.153 0.280 0.976 0.021 0.074 0.339 0.416 0.125 4.180 0.773 0.140 0.025 0.298 0.141 0.231 NA 0.175 NA 0.108 0.135 0.035 0.011 0.143 0.001 0.017 0.027 0.025 0.008 0.628 0.151 0.010 0.002 0.187 0.033 0.029 NA 0.021 NA 0.016 0.012 Female (4 h) Mean SD NA 0.415 1.753 0.041 0.157 0.533 0.853 0.227 2.064 2.897 1.567 1.639 1.289 0.219 NA 0.465 0.308 0.565 0.187 0.188 NA 0.073 0.212 0.005 0.020 0.067 0.098 0.028 0.503 0.800 1.154 1.039 2.094 0.109 NA 0.063 0.056 0.228 0.083 0.015 -91 - 000092 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 46: Percent of dose recovered at Tm in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male (10 h 30 m) Mean SD Female (1 h 15 m) Mean SD prostate skin" whole blood" brain fat" heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle" uterus bone" 0.071 15.565 24.919 0.051 2.815 0.443 0.593 0.096 18.750 2.170 2.508 2.632 0.011 0.085 0.693 NA 0.022 13.565 NA 3.047 0.045 0.899 1.942 0.021 0.225 0.037 0.376 0.017 2.434 0.354 0.713 0.934 0.006 0.012 0.180 NA 0.004 0.576 NA 0.544 NA 0.624 8.089 0.066 0.220 0.388 0.827 0.101 11.190 4.293 7.142 2.896 0.008 0.105 NA 0.071 0.026 0.325 0.354 0.174 NA 0.142 2.080 0.019 0.111 0.057 0.102 0.021 2.192 0.771 2.594 2.305 0.002 0.030 NA 0.012 0.005 0.010 0.046 0.057 Totalb 88.033 1.420 36.897 3.187 " Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. b Totals are calculated from individual animal data. n= 4 -92- 000093 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 47: Percent of dose recovered at Tmax/2 in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male (171 h) Mean SD Female (4h) Mean SD prostate skina whole blood" brain fata heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle* uterus bonea 0.057 7.233 11.140 0.023 0.916 0.252 0.344 0.060 25.231 1.212 1.052 0.270 0.004 0.053 0.372 NA 0.009 6.429 NA 1.375 0.020 0.430 0.624 0.008 0.205 0.030 0.194 0.007 1.289 0.115 0.202 0.028 0.002 0.003 0.062 NA 0.001 0.648 NA 0.169 NA 0.307 5.411 0.045 0.110 0.236 0.570 0.060 7.176 2.685 8.255 5.601 0.006 0.068 NA 0.041 0.015 0.229 0.247 0.142 NA 0.121 1.466 0.010 0.069 0.051 0.179 0.012 0.982 0.736 8.967 6.165 0.002 0.021 NA 0.012 0.004 0.031 0.068 0.078 Totalb 56.031 1.025 31.201 12.626 a Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. b Totals are calculated from individual animal data. -93- 000094 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 48: Concentration (fig equiv/g) at Tinax in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle uterus bone n=4 Male (10 h 30 m) Mean SD 2.012 3.865 15.886 0.310 1.896 5.386 6.996 2.014 23.456 11.018 2.789 1.750 3.042 1.922 2.754 NA 3.449 1.584 NA 1.966 1.278 0.224 1.227 0.028 0.138 0.369 0.744 0.146 3.178 2.089 0.609 0.542 0.836 0.286 0.154 NA 0.714 0.069 NA 0.333 Female (1 h 15 m) Mean SD NA 2.778 13.889 0.330 1.270 4.501 6.992 1.725 16.934 26.042 7.938 4.861 2.389 1.689 NA 3.708 2.810 4.955 1.067 1.360 NA 0.474 1.683 0.073 0.271 0.783 0.873 0.269 3.409 5.457 2.405 4.149 0.256 0.127 NA 0.442 0.605 1.549 0.179 0.259 -94- 000095 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 49: Concentration (jig equiv/g) at T.W2 in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle uterus bone n=4 Male (171 h) Mean SD 1.032 1.792 7.085 0.155 0.616 2.442 3.101 0.868 18.242 4.459 0.963 0.175 1.652 1.142 1.244 NA 1.115 0.749 NA 0.886 0.157 0.116 0.393 0.021 0.138 0.213 0.534 0.061 1.611 0.564 0.124 0.027 0.445 0.236 0.211 NA 0.183 0.077 NA 0.110 Female (4h) Mean SD NA 2.532 9.065 0.241 0.868 2.889 4.819 1.199 11.295 15.737 9.904 12.202 1.990 1.234 NA 2.880 2.053 4.578 0.767 1.070 NA 0.786 1.406 0.078 0.200 0.583 1.265 0.221 1.064 2.948 11.648 12.659 0.387 0.288 NA 0.756 0.330 1.065 0.242 0.360 -95- 000096 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 50: Percent of dose recovered at in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male (10 h 30 m) Mean SD Female (1 h 15 m) Mean SD prostate skin8 whole blood8 brain fat8 heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle8 uterus bone8 0.067 13.836 22.905 0.063 2.153 0.461 0.863 0.106 17.528 2.293 2.784 4.186 0.009 0.120 0.623 NA 0.026 12.855 NA 3.062 0.018 0.969 1.177 0.007 0.430 0.053 0.103 0.015 0.900 0.286 0.608 1.349 0.002 0.025 0.098 NA 0.004 0.841 NA 0.438 NA 0.380 7.158 0.058 0.147 0.317 0.678 0.091 10.538 5.867 6.923 2.491 0.009 0.091 NA 0.071 0.031 0.441 0.358 0.157 NA 0.166 2.232 0.008 0.053 0.035 0.067 0.007 1.723 0.946 1.846 1.548 0.003 0.032 NA 0.012 0.005 0.116 0.124 0.072 Totalb 83.937 3.680 35.803 2.554 Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. b Totals are calculated from individual animal data. -96- 000097 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 51: Percent of dose recovered at Tma, n in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male (171 h) Mean SD Female (4 h) Mean SD prostate skin* whole blood8 brain fat" heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle" uterus bone" 0.028 5.419 7.904 0.019 0.628 0.164 0.303 0.042 20.145 1.003 0.808 0.210 0.005 0.045 0.224 NA 0.009 4.253 NA 0.906 0.012 0.237 1.032 0.002 0.110 0.032 0.057 0.005 3.098 0.122 0.189 0.084 0.001 0.010 0.031 NA 0.003 0.358 NA 0.100 NA 0.415 6.407 0.058 0.148 0.287 0.775 0.070 9.080 4.749 3.547 1.121 0.007 0.077 NA 0.070 0.021 0.304 0.365 0.181 NA 0.175 1.406 0.018 0.065 0.069 0.204 0.002 0.895 0.393 1.306 1.010 0.002 0.020 NA 0.012 0.001 0.099 0.029 0.090 Total" 42.112 4.740 27.680 2.569 " Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. b Totals are calculated from individual animal data. "777" - 97- 000098 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 52: Concentration (/xg equiv/g) at Tma, in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle uterus bone n=4 Male (10 h 30 m) Mean SD 12.048 18.222 77.460 1.807 7.700 27.064 37.162 10.237 104.841 58.898 16.435 14.361 13.121 9.882 14.333 NA 17.271 7.964 NA 10.498 3.283 1.288 4.180 0.147 1.560 1.522 2.431 0.612 6.548 6.203 2.346 3.941 3.797 1.762 1.314 NA 1.927 0.546 NA 1.534 Female (1 h 15 m) Mean SD NA 17.036 73.097 1.475 8.428 21.274 34.065 9.835 92.592 159.165 39.253 33.918 15.543 11.813 NA 19.130 15.727 23.816 5.439 6.546 NA 2.814 4.272 0.179 2.738 1.709 3.029 0.888 10.198 15.051 12.486 19.210 5.794 2.405 NA 2.121 2.063 6.137 0.611 0.478 -98- 000099 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Table 53: Concentration (fig equiv/g) at T ^ x r: in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes ovaries adrenals muscle uterus bone n=4 Male (171 h) Mean SD 4.010 7.114 26.647 0.536 2.238 8.192 10.614 3.007 54.704 19.153 3.546 0.650 7.776 3.421 4.022 NA 4.661 2.626 NA 3.095 0.880 0.302 3.490 0.064 0.390 1.192 1.505 0.234 5.989 2.090 0.697 0.186 3.364 0.463 0.663 NA 0.578 0.216 NA 0.341 Female (4 h) Mean SD NA 16.653 60.145 1.551 6.157 18.171 32.621 8.385 72.211 134.318 25.265 12.611 17.409 7.423 NA 18.721 12.419 21.531 5.297 6.893 NA 2.934 5.361 0.457 0.608 3.540 5.884 0.469 6.062 8.888 19.676 12.463 5.319 0.691 NA 2.802 1.449 3.854 0.637 1.631 -99- O O O IO O Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 FIGURES - 100- 000101 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 FIGURES ABBREVIATIONS: EXPLANATORY NOTES %AD Cmax F hr Tina* Tmax/2 percent of administered dose maximum plasma concentration female hour(s) time tO Cjiiax time to V2 Cmax following attainment of maximum concentrations -101 - 000102 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 1: Pilot Study - PFOA concentration in serial plasma samples from male and female rats following oral gavage with 1 mg/kg PFOA *A.' - 102- 000103 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 2: Pilot Study - Cumulative recovery of 14C in urine and feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Urine Feces - 103- 000104 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 3: Cumulative recovery of 14C in urine and feces in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Urine Feces - 104- OOOIOS Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 4: Terminal tissue:blood concentration ratio in male and female rats following oral gavage with 1 mg/kg 14C-PFOA TissueiBlood Concentration Ratio - 105- 000106 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 5: Cumulative recovery of 14C in urine and feces in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Urine Feces 8-1 7- o - o-f--i--i--i--i-->--i----i----i-->--i-- I 0 100 200 300 400 500 600 700 Time (hr) "Animal 104F was omitted because of an extreme value for 24 hour fecal excreon, possibly due to contamination with urine. ? - 106- 000107 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 6: Terminal tissue:blood concentration ratio in male and female rats following oral gavage with 5 mg/kg 14C-PFOA booe muscle uterus adrenals testes ovaries thymus thyroid G.l.contents G.l. tract kidney liver spleen lungs heart fat brain wholeblood skin prostate 0 Males Females' 1T ' 1' 2 I I-- --"1 r-- -- I r-- I 3456 TissuerBlood Concentration Ratio ' Whole blood concentration in females was below the detection limits for the assay. See Table 19 for tissue concentrations. - 107- 000108 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 7: Cumulative recovery of 14C in urine and feces in male and female rats following oral gavage with 25 mg/kg 14C*PFOA Urine Feces 4? -108- 000109 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 8: Terminal tissue:blood concentration ratio in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Tissue:Blood Concentration Ratio aWhole blood concentration in females was below the detection limits for the assay. - 109- OOOllO Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 9: Cumulative recovery of 14C in urine and feces of male and female rats after 14day repeated oral gavage with 1 mg/kg PFOA, followed by a 1 mg/kg challenge dose of 14C-PFOA (day 15) Urine Feces Time (hr) -110 - O O O lll Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 10: Terminal tissue:blood concentration ratio in male and female rats following 14day repeated oral gavage with 1 mg/kg 14C-PFOA bone muscle 1 H Males uterus 1 I IFemales adrenals 1 testes 1 ovaries * thymus 1 thyroid * G.I.cootents ' G.l.tract 1 kidney liver spleen lungs heart fat brain wholeblood skin prostate 0 1 t--'--i--'--i--1--i--1--r 234 56 i ' "i--' i '--i 7 8 9 10 Tissue.Blood Concentration Ratio - Ill- 000112 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 11: PFOA concentration in serial plasma samples from male and female rats following oral gavage with 0.1 mg/kg PFOA Males Females - 112- 000113 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 12: PFOA concentration in serial plasma samples from male and female rats following oral gavage with 1 mg/kg PFOA Males Females 113 - 000114 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 13: PFOA concentration in serial plasma samples from male and female rats following intravenous injection with 1 mg/kg PFOA Males Females - 114- 000115 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 14: PFOA concentration in serial plasma samples from male and female rats following oral gavage with 5 mg/kg PFOA Males Females - 115- , f ' 000116 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 15: PFOA concentration in serial plasma samples from male and female rats following oral gavage with 25 mg/kg PFOA Males Females - 116- 000117 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 16: PFOA concentration in serial plasma samples from male and female rats following oral gavage with 0.1 mg/kg PFOA (extended time course) Males Females - 117- oooH s Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 17: Tissue distribution of 14C at Tm and Tma*^ in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Males Females T t - 118- 000119 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 18: Tissue:blood concentration ratio at Tm,, and Tma,,n in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Males bone muscle 5 3 adrenals testes 3* B thymus f thyroid G.l.contents 1 G.I.tract T kidney 1---- IT ,, spleen lungs a heart fat brain f wholeblood skin prostate s 9i -i ( r- i |i 1 ii 2 i | i i i i |i 3 i i i |i 4 i i i 1' 5 ' n6 Tissue:Blood Concentration Ratio Females bate muscle 1 uterus H adrenals ovaries 3 thymus thyroid k G.l. tract kidney liver spleen 1 lungs heart h 3 fat brain wholeblood skin 3 I-' T ' I 0 12 T3 TT 45 I "I 1 ) I '*I 1 l~' I ''I 6 7 8 9 10 11 12 13 Tissue:Blood Concentration Ratio tf f t - 119- 000120 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 19: Tissue distribution of 14C at Tmax and Tmax/2 in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Males Females % Administered Dose - 120- 000121 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 20: Tissuezblood concentration ratio at T^x and Tmax/2 in male and female rats following oral garage with 5 mg/kg 14C-PFOA Males Females - 121- 000122 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 2 1 : Tissue distribution of 14C at Tma, and Tnx/2 in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Males Females bone muscle uterus adrenals v ovaries thymus thyroid GXcontents GXtract tddney liver spleen lungs heart fat brain wholeblood skin o I I1/2T T5 10 % Administered Dose 15 - 122- OOOlM 3 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Figure 22: Tissue:blood concentration ratio at T^ax and Tmax/2 in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Males .. bone muscle adrenals m a testes Jf thymus thyroid B GJ.contents G.l. tract kidney liver spleen lungs bean -- fat brain S wholeblood skin prostate 0I---- ,---- ,---- .---- 1---- 11---- ----- ----- ----- ----- 2r CHIT Tissue:Blood Concentration Ratio 3 Females t. CUT ------1---- 1----1--------1---- ----1----1----1-- 0.0 0.3 1.0 1.5 2.0 2.5 Tissue.'Blood Concentration Ratio -i 3.0 -123- 000124 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 APPENDICES - 124- 000125 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 APPENDICES EXPLANATORY NOTES ABBREVIATIONS: AUCinf A U C inf/D C ip Cmax equiv F G.I. hr orh Lambda-Z LOD LOQ M Ci m NA NS SD T* Tmax Tmax/2 area under the plasma concentration vs. time curve, extrapolated to infinity AUCinf. normalized to dose plasma clearance maximum plasma concentration equivalent female gastrointestinal hour(s) terminal elimination constant limit of detection limit of quantitation male microcurie, 2.22x106dpm minute(s) not applicable no sample standard deviation terminal elimination half-life time tO Cmax time to */2 Cmax following attainment of maximum concentrations - 125- 000126 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix A Certificates of Analysis - 126- 000127 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 S ignaa-A ldrich C e rtifica te o f A nalysis Page 1 of I Si ALDRICH*______________________________ CertificateoiAnalysis TEST Produci Name Product Number CAS Number Formula Formule Weight APPEARANCE NFRARED SPECTRUM TITRATION GAS LIQUID CHROMATOGRAPHY TITRATION DUALITY CONTROL ACCEPTANCE DATE SPECIFICATION PenladecaHuorooctanolc acid, 96% 171468 33S671 C FgfC F^C O jH 414.06 WHITE TO OFF-WHITE POWDER CONFORMS TO STRUCTURE AND STANDARD AS ILLUSTRATED 95.5% -104.6% (WITH NAOH) 95.5% (MINIMUM) 2 % H20 (MAXIMUM) LOT {08316DO} RESULTS WHITE POWDER CONFORMS TO STRUCTURE AND STANDARD. 100.4% (WITH NAOH) 99.9% 1.48% H20 (WITH `KARL FISCHER' REAGENT) JUNE 2001 David Swessel, Supervisor Q uality Control http://info.sial.coin/cgi-bin/gx.cgi/Applogic+COAInfo.RetumCOfA - 127 - 0 8 /0 1 /2 0 0 1 o o o i s Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix B Pilot Pharmacokinetics in Male and Female Rats Following Oral Gavage with 1 mg/kg PFOA - 128 - 000129 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats DuPont-7473 Dose Concentration (mg/mL): 0.206 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 1.0 Male Animal Weight Dose Volume Actual Dose Rate Animal_____________________________ (g)____________________(mL)_________________ (mg/kg) 651159 651161 239.0 0.94 0.810 268.3 0.97 0.745 Mean 253.7 0.96 0.778 Standard Deviation 20.7 0.02 0.046 Female Animal 651162 651163 Mean Standard Deviation Animal Weight (r) 231.5 225.8 228.7 4.0 Dose Volume (mL) 0.89 0.88 0.89 0.01 Actual Dose Rate _________ (mg/kg)________ 0.792 0.803 0.798 0.008 - 129- 000130 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/xg/mL) in serial plasma in male and female rats following oral gavage with 1 mg/kg PFOA Male Time (h) 0.5 2 4 8 12 16 24 48 72 96 120 144 168 Animal Number 651159 651161 24.64 29.78 22.40 23.74 24.18 21.86 21.40 23.56 16.46 12.58 11.14 13.34 8.39 8.88 17.44 19.00 20.88 22.18 20.90 20.68 14.46 12.78 12.56 9.04 7.40 7.44 Mean 16.76 23.61 20.70 22.31 23.18 21.38 21.04 19.01 14.62 12.57 10.09 10.37 7.92 SD 11.1 8.7 2.4 2.0 1.4 0.7 0.5 6.4 2.6 0.0 1.5 4.2 0.7 Female Time (h) 0.5 2 4 8 12 16 24 48 72 96 120 144 168 Animal Number 651162 651163 17.20 14.32 9.06 4.05 1.76 0.67 0.12 0.05 0.07 0.00 0.00 0.01 0.00 33.40 25.08 13.16 4.71 1.31 0.39 0.09 0.01 0.01 0.00 0.00 0.00 0.00 Mean 25.30 19.70 11.11 4.38 1.53 0.53 0.10 0.03 0.04 0.00 0.00 0.00 0.00 SD 11.5 7.6 2.9 0.5 0.3 0.2 0.0 0.0 0.0 0.0 0.0 0.0 0.0 - 130- 00013t Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following oral gavage with 1 mg/kg PFOA Male Lambda z AUCint AUCinf/D Cip Animal # Tm, (hr) (fig/mL) (1/hr) Tiflihr) (hrue/m L) (hr'Mg/mL/mg'kg) (mL'kg/hr) 651159 651161 2.00 12.00 29.78 22.18 0.0062 0.0074 112.60 4122.72 93.99 3172.78 5089.78 4287.54 0.20 0.23 Mean SD 7.00 25.98 0.0068 103.30 3647.75 7.07 5.37 0.0006 13.30 671.71 4688.66 567.27 0.21 0.03 Female Cnaj Lambda z AUCdjf AUCinf/D Cip Animal # Timi (hr) (ME/mL) (1/hr) Tm (hr) (hr-M/mL) (hr'/i/mL/mg'ke) (mL*kg/hr) 651162 651163 0.50 17.20 0.22 0.50 33.40 0.25 3.19 101.67 2.72 145.03 128.69 181.29 7.77 5.52 Mean SD 0.50 25.30 0.24 2.96 123.35 0.00 11.46 0.02 0.33 30.66 154.99 37.19 6.64 1.59 ihr- - 131 - 000132 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix C Pilot Excretion and Material Balance in Male and Female Rats Following Oral Gavage with 25 mg/kg 14C-PFOA - 132- 000133 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats DuPont-7473 Dose Concentration (mg/g): 6.369 Nominal Dose Volume (mlAg): 4.0 Nominal Dose Rate (mg/kg): 25.0 Male Animal 101M 102M Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (E) (g) (mg/kg) 258.8 241.3 0.75 0.68 18.384 17.843 250.1 12.4 0.71 0.05 18.114 0.382 Radioactivity Received (fiCi) 20.246 18.322 19.284 1.361 Female Animal 101F 102F Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (g) (g) (mg/kg) 159.2 155.1 0.46 0.44 18.364 18.151 157.2 2.9 0.45 0.01 18.257 0.150 Radioactivity Received OiCi) 12.440 11.980 12.210 0.326 - 133- 000134 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in urine in male and female rats following oral gavagfe with 25 mg/kg 14C-PFOA Male urine urine urine urine urine urine urine urine urine urine Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101M 0.290 1.377 1-347 3.025 4.142 5.316 4.639 4.427 3.273 4.269 32.105 102M 0.143 0.974 0.923 2.114 3.689 3.508 4.521 4.415 4.109 4.119 28.515 Mean 0.217 1.176 1.135 2.570 3.916 4.412 4.580 4.421 3.691 4.194 30.310 SD 0.104 0.285 0.300 0.644 0.320 1.278 0.083 0.008 0.591 0.106 2.539 Female urine urine urine urine urine urine urine urine urine urine Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101F 29.195 38.673 13.793 8.348 2.126 0.484 0.341 0.217 0.149 0.121 93.447 102F 27.791 25.651 21.325 11.588 2.993 0.607 0.360 0.195 0.149 0.173 90.832 Mean 28.493 32.162 17.559 9.968 2.560 0.546 0.351 0.206 0.149 0.147 92.140 SD 0.993 9.208 5.326 2.291 0.613 0.087 0.013 0.016 0.000 0.037 1.849 - 134- 00013S Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101M 0.028 NS 1.005 1.289 0.579 0.496 0.770 0.755 1.561 0.524 7.007 102M NS 0.045 0.008 7.205 1.624 0.372 0.232 0.198 0.130 0.102 9.916 Mean 0.028 0.045 0.507 4.247 1.102 0.434 0.501 0.477 0.846 0.313 8.462 SD NA NA 0.705 4.183 0.739 0.088 0.380 0.394 1.012 0.298 2.057 Female feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101F NS NS 0.200 0.331 0.174 0.030 0.018 0.025 0.012 0.034 0.824 102F NS NS 0.046 0.102 0.052 <LOQ 0.009 <LOQ <LOQ 0.022 0.231 Mean NA NA 0.123 0.217 0.113 0.030 0.014 0.025 0.012 0.028 0.528 SD NA NA 0.109 0.162 0.086 NA 0.006 NA NA 0.008 0.419 - 135- 000136 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.l. contents thymus testes adrenals muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h Total 101M 0.038 16.778 0.674 4.917 0.023 0.065 0.198 0.369 0.041 25.811 1.180 0.895 0.219 0.040 0.239 0.007 0.192 0.085 51.771 102M 0.029 20.459 0.375 6.605 0.026 0.065 0.204 0.461 0.050 25.445 1.198 1.081 0.237 0.051 0.286 0.008 0.254 0.070 56.904 Mean 0.034 18.619 0.525 5.761 0.025 0.065 0.201 0.415 0.046 25.628 1.189 0.988 0.228 0.046 0.263 0.008 0.223 0.078 54.338 SD 0.006 2.603 0.211 1.194 0.002 0.000 0.004 0.065 0.006 0.259 0.013 0.132 0.013 0.008 0.033 0.001 0.044 0.011 3.630 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thymus ovaries adrenals uterus muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h Total 101F <LOD <LOQ <LOQ <LOD <LOD <LOD cLOD cLOD 0.009 0.002 cLOD cLOD cLOD cLOD cLOD cLOD cLOD cLOD 0.011 102F 0.288 cLOD cLOD cLOD cLOD cLOD cLOD cLOD 0.007 0.001 cLOD cLOD cLOD cLOD cLOD cLOD cLOD cLOD 0.296 Mean 0.288 NA NA NA NA NA NA NA 0.008 0.002 NA NA NA NA NA NA NA NA 0.154 SD NA NA NA NA NA NA NA NA 0.001 0.001 NA NA NA NA NA NA NA NA 0.202 - 136- 000137 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in cage wash and residual feed in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Timepoint cage wash residua] feed 168 h 168 h Total 101M 0.626 NS 0.626 102M 1.014 NS 1.014 Mean 0.820 NA 0.820 SD 0.274 NA 0.274 Female Timepoint cage wash residual feed 168 h 168 h Total 101F 0.821 0.806 1.627 102F 0.388 0.095 0.483 Mean 0.605 0.451 1.055 SD 0.306 0.503 0.809 - 137- 000138 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in C 02trap in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male CO2 trap A CO2 trap A C 02trap A C 0 2trap A C 0 2trap A CO2 trap A C 02trap A Timepoint 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101M_______ 102M_______Mean________ SD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA Female C 0 2trap A C 02trap A C 02trap A C 02trap A C 02trap A C 02trap A C 02trap A Timepoint_____101F_______ 102F_______ Mean________ SD 24 h 48 h 72 h 96 h 120 h 144 h 168 h <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD NA NA NA NA NA NA NA NA NA NA NA NA NA NA Total NA NA NA NA - 138- 000139 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity as volatile organics (VO) in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male VO VO VO VO VO VO VO Timepoint 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101M <L0D <LOD <LOD <LOD <LOD <LOD <LOD NA 102M <LOD <LOD <LOD <LOD <LOD <LOD <LOD NA Mean NA NA NA NA NA NA NA NA SD NA NA NA NA NA NA NA NA Female VO VO VO VO VO VO VO Timepoint 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 101F <LOD <LOD <LOD <LOD <LOD <LOD <LOD NA 102F <LOD <LOD <LOD <LOD <LOD <LOD <LOD NA Mean NA NA NA NA NA NA NA NA SD NA NA NA NA NA NA NA NA - 139 - 000140 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/tg equiv/g) in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Timepoint prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.l. contents thymus testes adrenals muscle bone 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 101M 3.106 3.478 6.753 25.068 0.575 1.010 7.733 11.530 3.197 60.028 17.585 3.527 0.546 3.069 3.738 4.598 1.867 4.142 102M 2.912 4.033 7.544 26.028 0.565 1.978 8.047 11.785 2.900 54.730 17.402 4.015 0.540 3.070 3.972 4.098 2.364 2.645 Mean 3.009 3.755 7.149 25.548 0.570 1.494 7.890 11.657 3.049 57.379 17.494 3.771 0.543 3.069 3.855 4.348 2.115 3.394 SD 0.137 0.393 0.559 0.679 0.007 0.684 0.222 0.181 0.210 3.746 0.129 0.345 0.004 0.000 0.166 0.354 0.351 1.058 Female Timepoint carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thymus ovaries adrenals uterus muscle bone 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 101F <LOD <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.032 0.030 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 102F 0.060 <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.028 0.015 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD Mean 0.060 NA NA NA NA NA NA NA 0.030 0.023 NA NA NA NA NA NA NA NA SD NA NA NA NA NA NA NA NA 0.003 0.011 NA NA NA NA NA NA NA NA - 140- 000141 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix D Materia] Balance in Male and Female Rats Following Oral Gavage with 1 mg/kg 14C-PFOA TT" - 141 - 0001*2 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats DuPont-7473 Male Dose Concentration (mg/g): 0.289 Female Dose Concentration (mg/g): 0.214 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 1.0 Male Animal 101M 102M 103M 104M Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (8) (s) (mg/kg) 240.9 225.9 232.6 234.3 0.93 0.90 0.93 0.95 1.119 1.155 1.157 1.170 233.4 6.2 0.93 0.02 1.150 0.022 Radioactivity Received OxCi) 28.301 27.392 28.240 28.786 28.180 0.579 Female Animal 101F 102F 103F 104F Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (g) (g) (mg/kg) 172.0 163.0 182.2 181.0 0.70 0.60 0.70 0.70 0.852 0.846 0.858 0.837 174.6 8.9 0.68 0.05 0.848 0.009 Radioactivity Received (iiCi) 21.973 20.692 23.446 22.710 22.205 1.175 - 142- 000143 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in urine in male and female rats following oral gavage with 1 mg/kg ,4C-PFOA Male urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine Tiroepoint_______ 101M_________ 102M_________ I03M_________ 104M_________ Mean__________ SD 4b 8b 12 b 24 h 48 h 72 h 96 b 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 0.038 0.619 0.626 2.254 2.915 2.773 2.174 2.364 2.133 2.061 1.296 1.187 1.497 1.311 1.612 1.207 1.511 2.147 2.483 2.400 1.953 1.869 1.644 1.342 0.024 0.588 1.040 1.808 3.001 3.175 2.278 2.056 1.726 1.801 1.649 1.523 1.395 1.441 1.343 1.303 1.270 2.728 2.203 1.704 1.895 1.210 1.368 1.553 0.110 0.268 0.867 1.932 3.705 2.767 2.272 2.060 2.682 2.378 2.047 1.816 1.952 1.636 1.775 1.386 1.242 2.882 2.356 2.391 1.890 1.935 1.389 1.489 0.020 0.158 1.051 2.461 3.858 3.197 2.554 1.925 2.079 1.414 1.949 1.978 2.155 1.594 1.157 0.449 0.742 1.601 2.787 3.012 3.348 3.232 1.593 2.092 0.048 0.408 0.896 2.114 3.370 2.978 2.320 2.101 2.155 1.914 1.735 1.626 1.750 1.496 1.472 1.086 1.191 2.340 2.457 2.377 2.272 2.062 1.499 1.619 0.042 0.230 0.199 0.298 0.481 0.240 0.163 0.186 0.395 0.408 0.338 0.348 0.363 0.149 0.275 0.431 0.323 0.585 0.248 0.534 0.718 0.846 0.140 0.327 Total 41.416 40.082 45.227 46.406 43.283 3.015 Female urine urine urine urine urine urine urine urine urine urine Tiroepoint________3OIF__________ 302F__________ 103F__________ 104F_________ Mean__________ SD 4b 8h 12 h 24 b 48 h 72 h 96 h 120 h 144 h 168 h 20.276 27.003 16.385 8.052 1.858 0.875 0.627 0.314 0.200 0.205 7.612 27.422 16.430 11.912 5.041 1.285 0.999 0.498 0.331 0.425 10.156 37.996 11.913 8.060 2.233 1.618 0.846 0.495 0.357 0.569 7.975 31.297 19.279 14.826 3.344 2.373 0.831 0.420 0.427 0.721 11.505 30.930 16.002 10.713 3.119 1-538 0.826 0.432 0.329 0.480 5.954 5.092 3.043 3.290 1.428 0.634 0.153 0.086 0.095 0.220 Total 75.795 71.955 74.243 81.493 75.872 4.066 - 143- 000144 Periluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in feces in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8h 12 h 24 b 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 Total 101M NS NS 0.203 10.313 1.391 0.627 0.600 0.178 0.213 0.212 0.653 0.694 0.455 0.506 0.147 0.378 0.217 0.609 0.363 0.485 0.578 0.541 0.494 0.165 20.022 702M NS 2.269 6.016 1.634 0.147 0.181 0.166 0.087 0.158 0.069 0.174 0.363 0.123 0.099 0.073 0.050 0.066 0.122 0.112 0.098 0.204 0.212 0.119 0.144 12.686 103M NS NS 2.743 5.019 0.840 0.217 0.117 0.151 0.118 0.125 0.109 0.070 0.086 0.066 0.073 0.196 0.180 0.259 0.319 0.284 0.315 0.211 0.083 0.141 11.722 104M NS NS 0.771 5.249 0.972 0.143 0.067 0.086 0.150 0.353 0.084 0.070 0.096 0.145 0.097 0.401 0.446 0.558 0.133 0.270 0.140 0.865 0.351 0.341 11.788 Mean NA 2.269 2.433 5.554 0.838 0.292 0.238 0.126 0.160 0.190 0.255 0.299 0.190 0.204 0.098 0.256 0.227 0.387 0.232 0.284 0.309 0.457 0.262 0.198 14.055 SD NA NA 2.625 3.577 0.517 0.225 0.245 0.046 0.039 0.124 0.268 0.297 0.177 0.204 0.035 0.165 0.159 0.235 0.128 0.158 0.193 0.313 0.195 0.096 4.003 Female feces feces feces feces feces feces feces feces feces feces Timepoint________ 101F__________ 102F__________ 103F__________ 104F__________ Mean___________ SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total NS NS 0.275 0.220 0.095 0.029 0.071 0.064 0.052 0.026 0.832 NS NS 0.265 0.118 0.075 0.020 0.233 0.074 0.026 0.008 0.819 NS NS 4.155 1.924 0.237 0.030 0.102 0.017 0.044 0.037 6.546 NS NS 0.261 0.059 0.023 0.032 0.017 0.027 0.010 0.049 0.478 NA NA 1.239 0.580 0.108 0.028 0.106 0.046 0.033 0.030 2.169 NA NA 1.944 0.898 0.092 0.005 0.092 0.028 0.019 0.017 2.923 ,s - 144- 00014S Perfluorooctanoic Acid: Toxicokinetics in the Rat _____ _____________________________DuPont-7473 Percent recovery of radioactivity in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Total 101M 0.011 6.129 0.065 0.878 0.004 0.008 0.049 0.093 0.011 14.379 0.473 0.302 0.047 0.001 0.008 0.079 0.001 0.080 0.024 22.642 102M 0.020 8.078 0.105 1.614 0.008 0.017 0.075 0.109 0.016 12.553 0.445 0.354 0.072 0.004 0.014 0.138 0.003 0.128 0.047 23.800 103M 0.032 9.086 0.125 1.582 0.008 0.024 0.076 0.130 0.020 16.212 0.719 0.438 0.069 0.002 0.013 0.114 0.003 0.101 0.034 28.788 104M 0.017 6.260 0.066 0.986 0.005 0.020 0.055 0.101 0.011 10.455 0.451 0.282 0.054 0.001 0.007 0.080 0.002 0.072 0.024 18.949 Mean 0.020 7.388 0.090 1.265 0.006 0.017 0.064 0.108 0.015 13.400 0.522 0.344 0.061 0.002 0.011 0.103 0.002 0.095 0.032 23.545 SD 0.009 1.440 0.030 0.387 0.002 0.007 0.014 0.016 0.004 2.467 0.132 0.070 0.012 0.001 0.004 0.029 0.001 0.025 0.011 4.062 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h Total 101F 0.105 <LOQ <LOQ <LOD cLOD <LOD cLOD <LOD 0.008 0.001 cLOD <LOD <LOQ <LOD <LOD <LOD <LOD <LOD <LOQ 0.114 102F 0.388 <LOQ 0.002 <LOD <LOD <LOD <LOD <LOD 0.012 0.002 <LOD <LOQ <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 0.404 103F 0.148 0.001 0.005 <LOD <LOD <LOD <LOQ <LOD 0.016 0.004 <LOQ 0.003 <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 0.177 104F 0.217 <LOQ <LOQ <LOD <LOD <1X>D <LOD <LOD 0.014 0.002 <LOD <LOD cLOD cLOD cLOD cLOD cLOD cLOD cLOQ 0.233 Mean 0.215 0.001 0.004 NA NA NA NA NA 0.013 0.002 NA 0.003 NA NA NA NA NA NA NA 0.232 SD 0.125 NA 0.002 NA NA NA NA NA 0.003 0.001 NA NA NA NA NA NA NA NA NA 0.125 - 145 - 000146 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in cage wash and residual feed in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male Timepoint cage wash residual feed Day 28 Day 28 Total 101M 1.813 0.513 2.326 102M 2.912 0.490 3.402 103M 1.794 0.531 2.325 104M 2.328 1.411 3.739 Mean 2.212 0.736 2.948 SD 0.528 0.450 0.732 Female Timepoint cage wash residual feed 168 h 168 h Total 101F 3.183 0.182 3.365 102F 2.131 0.176 2.307 103F 4.510 0.243 4.753 104F 1.592 0.192 1.784 Mean 2.854 0.198 3.052 SD 1.287 0.031 1.311 - 146- V 000147 Perfluorooctanoic Acid: Toxicokinetics in the Rat _______ DuPont-7473 Concentration (fig equiv/g) in tissues in male and female rats following oral gavage with 1 mg/kg 14c -p f o a Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 101M 0.033 0.043 0.084 0.259 0.006 0.005 0.084 0.117 0.044 1.892 0.324 0.033 0.006 0.036 0.034 0.061 0.034 0.027 0.048 102M 0.057 0.074 0.146 0.442 0.010 0.021 0.152 0.220 0.069 2.325 0.377 0.065 0.012 0.098 0.055 0.105 0.083 0.044 0.070 103M 0.070 0.074 0.132 0.441 0.009 0.017 0.150 0.194 0.062 2.310 0.423 0.063 0.010 0.093 0.062 0.134 0.069 0.039 0.055 104M 0.041 0.055 0.097 0.283 0.007 0.013 0.099 0.151 0.041 1.423 0.304 0.042 0.006 0.039 0.036 0.066 0.037 0.029 0.046 Mean 0.050 0.061 0.115 0.357 0.008 0.014 0.121 0.171 0.054 1.987 0.357 0.051 0.008 0.066 0.047 0.092 0.056 0.035 0.054 SD 0.016 0.015 0.029 0.099 0.002 0.007 0.035 0.046 0.014 0.427 0.054 0.016 0.003 0.034 0.014 0.035 0.024 0.008 0.011 Female Timepoint carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h `Value above LOD, but <0.001 101F 0.001 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.001 0.001 <LOD <LOD <LOQ <LOD <LOD <LOD <LOD <LOD <LOQ 102F 0.003 <LOQ 0.000* <LOD <LOD <UDD <LOD <LOD 0.002 0.001 <LOD <LOQ <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 103F 0.001 0.001 0.001 <LOD <LOD <LOD <LOQ <LOD 0.002 0.003 <LOQ 0.000' <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 104F 0.002 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.002 0.001 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOQ Mean 0.002 0.001 0.001 NA NA NA NA NA 0.002 0.002 NA 0.000` NA NA NA NA NA NA NA SD 0.001 NA 0.000 NA NA NA NA NA 0.000 0.001 NA NA NA NA NA NA NA NA NA -147- 000148 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix E Material Balance in Male and Female Rats Following Oral Gavage with 5 mg/kg 14C-PFOA - 148- 000149 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats DuPont-7473 Dose Concentration (mg/g): 1.211 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 5.0 Male Animal 101M 102M 103M 104M Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (g) (g) (mg/kg) 198.0 197.7 201.2 193.3 0.78 0.79 0.80 0.72 4.787 4.807 4.795 4.497 197.6 3.2 0.77 0.04 4.722 0.150 Radioactivity Received OiCi) 22.476 22.534 22.878 20.611 22.125 1.025 Female Animal 101F 102F 103F 104F Mean Standard Deviation Animal Weight (g) 197.8 192.9 210.5 200.1 Dose Administered (g) 0.79 0.77 0.85 0.80 Actual Dose Rate (mg/kg) 4.835 4.801 4.865 4.810 200.3 7.4 0.80 0.03 4.828 0.029 Radioactivity Received (ttCi) 22.677 21.960 24.285 22.821 22.936 0.975 - 149- 000150 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in urine in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine Timepoint 4h 8h 12 h 24 h 48 h 72 b 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 Total 101M 0.195 0.817 0.599 2.231 4.907 4.188 3.558 3.357 3.411 3.434 3.160 2.763 2.866 2.403 2.359 2.197 1.987 3.555 3.608 3.352 3.248 2.587 2.257 2.060 65.099 102M 0.066 0.765 0.737 2.179 3.609 2.764 2.588 4.265 3.120 4.547 2.646 2.298 2.357 2.134 1.816 1.657 1.435 3.300 3.691 2.835 2.544 1.978 2.273 1.680 57.284 103M 0.045 0.240 1.210 2.813 4.774 4.340 3.513 2.985 3.442 2.759 2.972 2.619 2.597 2.372 2.414 2.004 1.941 3.181 3.255 3.069 2.651 2.510 2.067 1.781 61.554 104M <LOD NS 1.625 3.076 4.490 4.432 4.231 4.383 3.410 3.075 2.920 2.856 2.877 2.605 2.373 2.320 2.113 2.818 2.963 2.755 2.710 2.179 2.436 2.220 64.867 Mean 0.102 0.607 1.043 2.575 4.445 3.931 3.473 3.748 3.346 3.454 2.925 2.634 2.674 2.379 2.241 2.045 1.869 3.214 3.379 3.003 2.788 2.314 2.258 1.935 62.201 SD 0.081 0.319 0.468 0.441 0.584 0.784 0.675 0.684 0.151 0.779 0.212 0.244 0.248 0.193 0.284 0.289 0.298 0.306 0.336 0.268 0.314 0.285 0.151 0.249 3.656 Female urine urine urine urine urine urine urine urine urine urine Tiroepoint________ 1Q1F__________ 102F__________ 103F__________ 104F__________ Mean___________ SD 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 b 4.296 37.676 16.617 21.722 3.828 0.737 0.807 0.461 0.219 0.248 NS 33.762 19.676 14.401 3.616 1.833 0.696 1.348 0.909 0.467 19.095 30.895 12.663 7.168 2.564 0.952 0.662 0.483 0.375 0.370 <LOD 38.518 19.801 10.969 2.095 0.510 0.323 0.274 0.261 0.169 11.696 35.213 17.189 13.565 3.026 1.008 0.622 0.642 0.441 0.314 10.464 3.547 3.358 6.189 0.831 0.579 0.209 0.480 0.319 0.132 Total 86.611 76.708 75.227 72.920 77.867 6.034 - 150- 000151 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in feces in male and female rats following oral gavage with 5 mg/kg 14C-PFOA M ale feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces Timepoint________101M__________102M__________ 103M__________ 0 4 M __________Mean 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 b Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 NS NS 4.010 1.027 0.336 0.204 0.209 0.120 0.114 0.090 0.098 0.085 0.084 0.100 0.070 0.058 0.061 0.221 0.084 0.095 0.079 0.062 0.078 0.065 NS NS 0.032 0.742 1.107 1.228 0.825 0.328 0.279 0.135 0.112 0.117 0.265 0.218 0.228 0.129 0.140 0.129 0.175 0.137 0.089 0.129 0.131 0.154 NS NS 0.063 0.770 0.365 0.200 0.216 0.137 0.116 0.151 0.124 0.095 0.099 0.131 0.083 0.077 0.103 0.152 0.133 0.107 0.109 0.180 0.137 0.273 NS NS 0.002 0.420 0.334 0.688 0.170 0.204 0.196 0.185 0.177 0.146 0.146 0.131 0.094 0.089 0.101 0.312 0.251 0.155 0.149 0.108 0.113 0.101 NA NA 1.027 0.740 0.536 0.580 0.355 0.197 0.176 0.140 0.128 0.111 0.149 0.145 0.119 0.088 0.101 0.204 0.161 0.124 0.107 0.120 0.115 0.148 Total 7.350 6.829 3.821 4.272 5.568 NA NA 1.989 0.249 0.381 0.489 0.314 0.094 0.078 0.039 0.035 0.027 0.082 0.051 0.073 0.030 0.032 0.082 0.071 0.027 0.031 0.049 0.027 0.091 1.779 Female feces feces feces feces feces feces feces feces feces feces Timepoint________ 101F__________ 102F 4h 8h 12 h 24 h 48 h 72 h 96 h 120 b 144 b 168 b NS NS NS 3.262 0.319 0.056 0.026 0.069 0.047 0.039 0.003 NS 1.085 0.396 0.145 0.165 0.029 0.059 0.072 0.245 Total 3.818 2.199 J03F NS NS 1.182 0.712 0.277 0.284 0.386 0.328 0.297 0.168 3.634 J04F NS NS NS 11.365 1.499 0.857 0.142 0.021 0.006 0.004 13.894 Mean 0.003 NA 1.134 3.934 0.560 0.341 0.146 0.119 0.106 0.114 5.886 SD NA NA 0.069 5.118 0.630 0.357 0.169 0.141 0.131 0.112 5.387 - 151 - 000152 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male prostate carcass skin whole blood brain fai heart lungs spleen liver kidney G.I. tract G.l. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Total 101M 0.010 5.379 0.055 0.999 0.004 0.018 0.042 0.055 0.011 10.637 0.381 0.201 0.054 0.001 0.010 0.072 0.015 0.056 0.021 18.021 102M 0.017 5.954 0.068 1.043 0.005 0.020 0.049 0.092 0.015 9.392 0.397 0.250 0.039 0.002 0.014 0.098 0.002 0.060 0.028 17.545 103M 0.016 7.394 0.089 1.254 0.007 0.016 0.068 0.121 0.018 10.830 0.350 0.275 0.051 0.003 0.013 0.105 0.003 0.073 0.032 20.718 104M 0.015 7.981 0.084 1.434 0.006 0.016 0.064 0.116 0.013 12.756 0.423 0.308 0.076 0.002 0.017 0.097 0.002 0.070 0.030 23.510 Mean 0.015 6.677 0.074 1.183 0.006 0.018 0.056 0.096 0.014 10.904 0.388 0.259 0.055 0.002 0.014 0.093 0.006 0.065 0.028 19.949 SD 0.003 1.214 0.016 0.201 0.001 0.002 0.012 0.030 0.003 1.390 0.031 0.045 0.015 0.001 0.003 0.014 0.006 0.008 0.005 2.755 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone Timepoint_______ 101F__________ IMF__________ 103F__________ IMF_________ Mean__________ SD 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 0.109 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.016 0.002 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.170 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.018 0.002 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.083 <LOD <LOQ <LOD <LOD <LOD <LOD <LOD 0.018 0.002 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <U)D <LOD <LOD <LOD <LOD 0.013 0.002 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.121 NA NA NA NA NA NA NA 0.016 0.002 NA NA NA NA NA NA NA NA NA 0.045 NA NA NA NA NA NA NA 0.002 0.000 NA NA NA NA NA NA NA NA NA Total 0.127 0.190 0.103 0.015 0.109 0.072 ,i - 1 5 2 - 000153 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in cage wash and residual feed in male and female rats following oral gavage with 5 mg/kg 14C-PFOA M ale Timepoint cage wash residual feed Day 28 Day 28 Total 101M 2.318 0.019 2.337 102M 6.263 0.860 7.123 103M 4.058 0.253 4.311 104M 1.967 <LOD 1.967 Mean 3.652 0.377 3.935 SD 1.966 0.434 2.362 Female Timepoint cage wash residual feed 168 h 168 h Total 101F 1.578 1.159 2.737 102F 3.102 0.510 3.612 103F 3.219 0.409 3.628 104F 1.768 0.086 1.854 Mean 2.417 0.541 2.958 SD 0.864 0.450 0.845 - 153- 000154 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (fig equiv/g) in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA M ale prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 101M 0.098 0.151 0.245 0.941 0.022 0.037 0.289 0.415 0.142 5.504 0.977 0.135 0.022 0.127 0.128 0.211 1.328 0.090 0.142 102M 0.129 0.166 0.342 0.987 0.023 0.047 0.303 0.452 0.160 4.653 1.014 0.147 0.022 0.165 0.155 0.260 0.131 0.096 0.171 103M 0.201 0.203 0.416 1.241 0.032 0.068 0.409 0.603 0.173 6.187 0.837 0.206 0.044 0.244 0.146 0.313 0.221 0.112 0.192 104M 0.141 0.197 0.377 1.244 0.029 0.057 0.394 0.500 0.162 4.979 0.913 0.172 0.025 0.168 0.152 0.273 0.191 0.112 0.192 Mean 0.142 0.179 0.345 1.103 0.027 0.052 0.349 0.492 0.159 5.331 0.935 0.165 0.028 0.176 0.145 0.264 0.468 0.102 0.174 SD 0.043 0.025 0.073 0.161 0.005 0.013 0.061 0.081 0.013 0.670 0.078 0.031 0.011 0.049 0.012 0.042 0.574 0.011 0.023 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 101F 0.006 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.015 0.008 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 102F 0.010 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD 0.019 0.011 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 103F 0.004 <LOD <LOQ <LOD <LOD <LOD <LOD <LOD 0.016 0.011 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 104F <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.012 0.009 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD Mean 0.007 NA NA NA NA NA NA NA 0.016 0.010 NA NA NA NA NA NA NA NA NA SD 0.003 NA NA NA NA NA NA NA 0.003 0.001 NA NA NA NA NA NA NA NA NA - 154- 000155 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix F Material Balance in Male and Female Rats Following Oral Gavage with 25 mg/kg 14C-PFOA -155 - 000156 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats DuPont-7473 Dose Concentration (mg/g): 6.447 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 25.0 Male Animal 301M 302M 303M 304M Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (fi) (g) (mg/kg) 216.3 201.8 213.0 208.1 0.87 0.81 0.83 0.88 25.903 25.815 25.244 27.326 209.8 6.3 0.85 0.03 26.072 0.885 Radioactivity Received (#iCi) 24.915 23.166 23.911 25.287 24.320 0.964 Female Animal 301F 302F 303F 304F Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate (fi) (g) (mg/kg) 183.7 196.0 187.2 187.2 0.73 0.77 0.75 0.74 25.621 25.395 25.831 25.452 188.5 5.2 0.75 0.02 25.574 0.196 Radioactivity Received (fid) 20.930 22.134 21.503 21.188 21.438 0.519 - 156- 000157 Perfluorooctanoic Acid: Toxicokinetics in the Rat _ ____________ ________ _____ DuPont-7473 Percent recovery of radioactivity in urine in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male unse urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine Timepoint 4b 8h 12 b 24 h 48 b 72 h 96 h 120 h 144 b 168 b Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 Total 301M 0.223 1.056 1.569 3.120 9.291 6.681 5.610 4.489 4.040 3.045 2.668 2.539 2.179 1.879 1.334 1.057 0.662 0.993 0.826 1.927 1.812 1.731 1.229 0.703 60.663 302M 0.218 1.155 1.757 4.265 6.501 5.453 4.712 3.663 3.380 2.511 1.857 1.734 2.109 2.261 2.170 1.036 1.285 1.876 1.717 2.803 2.309 1.814 1.804 1.707 60.097 303M 0.054 0.321 0.509 3.577 6.788 3.688 2.866 4.172 2.992 1.771 1.451 0.102 1.397 1.433 1.022 0.973 1.000 1.042 0.792 2.012 1.730 1.763 1.228 0.860 43.543 304M 0.510 0.552 1.563 2.070 4.632 2.613 2.916 2.791 2.540 2.941 2.288 2.250 1.738 1.288 1.631 1.661 1.548 2.069 1.268 2.255 1.959 1.969 1.826 1.878 48.756 Mean 0.251 0.771 1.350 3.258 6.803 4.609 4.026 3.779 3.238 2.567 2.066 1.656 1.856 1.715 1.539 1.182 1.124 1.495 1.151 2.249 1.953 1.819 1.522 1.287 53.265 SD 0.190 0.400 0.568 0.921 1.914 1.811 1.361 0.741 0.635 0.579 0.527 1.088 0.362 0.442 0.489 0.321 0.381 0.557 0.435 0.394 0.256 0.106 0.339 0.591 8.490 Female urine urine urine urine urine urine urine urine urine urine Timepoint 4b 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 301F 35.531 24.640 9.742 8.623 3.079 1.887 1.833 0.487 0.276 0.202 86.300 302F 19.430 19.922 16.571 3.696 3.371 1.510 0.989 1.022 0.651 0.715 67.877 303F 22.344 24.573 15.024 14.514 5.152 1.812 0.994 0.923 0.548 0.727 86.611 304F 17.871 45.210 15.463 12.587 3.254 0.945 0.503 0.391 0.261 0.251 96.736 Mean 23.794 28.586 14.200 9.855 3.714 1.539 1.080 0.706 0.434 0.474 84.381 SD 8.041 11.300 3.042 4.783 0.966 0.428 0.552 0.313 0.196 0.286 12.023 -157- 000158 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in feces in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8b 12 h 24 h 48 b 72 h 96 h 120 h 144 h 168 b Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 Total 301M NS 0.005 0.072 0.275 0.343 0.266 0.192 0.222 0.333 0.394 0.935 0.411 0.519 0.342 0.621 0.729 0.797 0.979 1.456 0.160 0.083 0.111 0.221 0.386 9.852 302M NS 0.357 NS 2.241 0.445 0.288 0.304 0.430 0.467 0.638 1.141 1.074 1.467 0.387 0.411 0.905 0.601 1.246 1.725 0.157 0.436 0.577 0.375 0.144 15.816 303M 0.020 NS 0.020 3.783 2.500 0.382 0.630 0.912 0.649 0.786 0.620 1.113 1.046 0.161 0.173 0.073 0.104 0.584 0.985 0.269 0.478 0.200 0.375 0.375 16.238 304M NS NS 0.283 0.367 0.657 0.417 0.616 0.523 0.578 0.163 0.190 0.112 0.701 0.411 0.070 0.127 0.182 0.383 1.115 0.177 0.230 0.300 0.317 0.136 8.055 Mean 0.020 0.181 0.125 1.667 0.986 0.338 0.436 0.522 0.507 0.495 0.722 0.678 0.933 0.325 0.319 0.459 0.421 0.798 1.320 0.191 0.307 0.297 0.322 . 0.260 12.490 SD NA 0.249 0.139 1.677 1.018 0.073 0.221 0.289 0.138 0.274 0.414 0.496 0.418 0.113 0.247 0.421 0.332 0.388 0.335 0.053 0.184 0.202 0.073 0.139 4.153 Female feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8h 12 h 24 h 48 b 72 h 96 h 120 h 144 h 168 b Total 301F NS NS NS 0.102 0.123 0.127 0.195 0.059 0.158 0.072 0.836 302F NS NS 1.113 2.529 0.808 0.477 0.275 0.078 0.023 0.376 5.679 303F NS NS NS 0.147 0.129 0.043 0.059 0.009 0.068 0.016 0.471 304F 0.017 0.002 0.006 0.219 0.059 0.037 0.045 0.020 0.026 0.054 0.485 Mean 0.017 0.002 0.560 0.749 0.280 0.171 0.144 0.042 0.069 0.130 1.868 SD NA NA 0.783 1.187 0.354 0.208 0.111 0.032 0.063 0.166 2.546 - 158- 000159 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Total 301M 0.008 3.752 0.026 0.291 0.004 0.012 0.031 0.060 0.008 4.469 0.270 0.195 0.022 0.001 0.006 0.054 0.001 0.033 0.016 9.259 302M 0.006 3.935 0.043 0.746 0.003 0.009 0.030 0.056 0.008 4.553 0.225 0.214 0.033 0.001 0.008 0.039 0.002 0.032 0.014 9.957 303M 0.008 4.446 0.043 0.862 0.005 0.011 0.031 0.064 0.030 5.128 0.434 0.587 0.095 0.003 0.005 0.067 0.002 0.034 0.015 11.870 304M 0.016 9.285 0.093 1.175 0.008 0.025 0.087 0.129 0.016 7.935 0.628 0.432 0.072 0.002 0.018 0.110 0.005 0.064 0.026 20.126 Mean 0.010 5.355 0.051 0.769 0.005 0.014 0.045 0.077 0.016 5.521 0.389 0.357 0.056 0.002 0.009 0.068 0.003 0.041 0.018 12.803 SD 0.004 2.637 0.029 0.366 0.002 0.007 0.028 0.035 0.010 1.636 0.183 0.187 0.034 0.001 0.006 0.031 0.002 0.016 0.006 5.005 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h Total 301F <LOQ 0.001 <LOQ <LOD <LOD <LOD <LOD <LOD 0.020 0.003 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.024 302F 0.172 0.001 <LOQ <LOD <LOD <LOD <LOD <LOD 0.019 0.003 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 0.195 303F 0.263 0.002 <LOQ <LOD <LOD <LOD <LOD <LOD 0.003 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 0.268 304F 0.129 0.001 <LOQ <LOD <LOD <LOD <LOQ <LOD 0.018 0.003 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 0.151 Mean 0.188 0.001 NA NA NA NA NA NA 0.015 0.003 NA NA NA NA NA NA NA NA NA 0.160 SD 0.068 0.001 NA NA NA NA NA NA 0.008 0.000 NA NA NA NA NA NA NA NA NA 0.102 - 159- 000160 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in cage wash and residual feed in male and female rats following oral gavage with 25 mg/kg I4C-PFOA Male Timepoint cage wash residual feed Day 28 Day 28 Total 301M_______ 302M_______303M_______304M_______ Mean________ SD 3.125 0.002 2.412 0.716 6.099 4.082 4.351 0.589 3.997 1.347 1.614 1.850 3.127 3.128 10.181 4.940 5.344 3.336 Female Timepoint cage wash residual feed 168 h 168 h Total 301F 0.694 3.661 4.355 302F 3.145 16.071 19.216 303F 2.835 0.052 2.887 304F 0.749 0.201 0.950 Mean 1.856 4.996 6.852 SD 1.316 7.569 8.360 - 160- 000161 Perfluorooctanoic Acid: Toxicokinetics in the Rat _____________________________ __________DuPont-7473 Concentration (/ig equiv/g) in tissues in male and female rats following oral gavage with 25 mg/kg 14c -p f o a Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 301M 0.714 0.655 1.353 4.108 0.106 0.232 1.343 2.085 0.564 14.266 4.434 0.576 0.075 0.623 0.529 1.020 0.784 0.382 0.566 302M 0.425 0.615 1.136 3.934 0.084 0.139 1.207 1.672 0.522 11.108 3.468 0.516 . 0.076 0.545 0.492 0.649 0.886 0.322 0.537 303M 0.538 0.836 1.548 5.098 0.130 0.289 1.507 2.181 2.432 16.192 7.674 2.077 0.282 1.748 0.686 1.195 0.902 0.390 0.609 304M 1.516 1.742 3.125 9.943 0.238 0.506 3.396 4.693 1.474 24.870 9.338 1.496 0.189 1.742 1.710 2.044 2.030 0.810 1.115 Mean 0.798 0.962 1.790 5.771 0.139 0.291 1.863 2.658 1.248 16.609 6.229 1.166 0.156 1.164 0.854 1.227 1.150 0.476 0.707 SD 0.493 0.529 0.905 2.828 0.068 0.156 1.029 1.375 0.903 5.892 2.745 0.755 0.100 0.671 0.577 0.590 0.589 0.225 0.274 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus m uscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 301F <LOQ 0.019 <LOQ <LOD <LOD <LOD <LOD <LOD 0.092 0.088 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 302F 0.053 0.015 <LOQ <LOD <LOD <LOD <LOD <LOD 0.117 0.067 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 303F 0.080 0.048 <LOQ <LOD <LOD <LOD <LOD <LOD 0.021 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOQ 304F 0.036 0.014 <LOQ <LOD <LOD <LOD <LOQ <LOD 0.079 0.065 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOQ Mean 0.056 0.024 NA NA NA NA NA NA 0.078 0.074 NA NA NA NA NA NA NA NA NA SD 0.022 0.016 NA NA NA NA NA NA 0.041 0.013 NA NA NA NA NA NA NA NA NA -161 - 000162 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix G Material Balance in Male and Female Rats Following Oral Gavage with 1 mg/kg 14C-PFOA (14 day-repeated dose) - 162- 000163 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats DuPont-7473 Dose Concentration (mg/g): Nominal Dose Volume (mL/kg): Nominal Dose Rate (mg/kg): 0.236 4.0 1.0 Male Animal 501M 502M 503M 504M Animal Weight* Dose Administered (r) (g) 332.9 334.9 349.2 355.8 1.28 1.29 1.39 1.40 Actual Dose Rate (mg/kg) 0.906 0.907 0.942 0.925 Radioactivity Received (nCi) 38.186 38.455 41.652 41.682 Mean Standard Deviation 343.2 11.1 1.34 0.06 0.920 0.017 39.994 1.935 aBody weights at time of administration of 14C-PFOA challenge dose. At the initiation of the 14-day repeat dosing, male body weights were 280-294 g. Female Animal 505F 506F 507F 508F Animal Weight* Dose Administered Actual Dose Rate (R) (g) (mg/kg) 232.9 257.5 237.0 246.6 0.94 1.01 0.95 0.99 0.954 0.922 0.944 0.950 Radioactivity Received (lid) 28.117 30.059 28.326 29.671 Mean Standard Deviation 243.5 11.0 0.97 0.03 0.942 0.014 29.043 0.966 aBody weights at time of administration of 14C-PFOA challenge dose. At the initiation of the 14-day repeat dosing, female body weights were 245-255 g. - 363 - 000164 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in urine in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeated dose) Male urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine urine Timepoint 4b 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 Total 501M 0.318 1.024 1.124 2.695 6.099 3.608 1.147 1.592 1.934 2.025 0.980 2.291 2.082 1.956 1.852 2.078 2.619 3.136 3.369 2.751 2.212 2.241 1.936 1.009 52.078 502M 0.231 0.584 0.645 2.046 3.159 2.605 1.893 1.239 1.619 1.887 1.881 1.315 1.096 1.562 1.018 1.092 1.749 2.911 2.832 2.509 2.361 1.848 1.700 1.517 41.299 503M 0.029 0.523 1.467 3.036 4.214 3.794 3.265 3.247 2.984 2.714 2.512 1.977 2.135 2.415 2.157 2.052 2.098 3.378 2.878 2.563 2.322 2.143 1.952 2.065 57.920 504M 0.041 0.683 0.479 3.550 6.954 5.143 4.332 3.874 4.188 2.564 1.764 2.089 0.929 1.308 2.458 2.770 2.648 3.425 2.540 1.701 1.243 1.166 1.388 1.184 58.421 Mean 0.155 0.704 0.929 2.832 5.107 3.788 2.659 2.488 2.681 2.298 1.784 1.918 1.561 1.810 1.871 1.998 2.279 3.213 2.905 2.381 2.035 1.850 1.744 1.444 52.430 SD 0.143 0.224 0.451 0.631 1.731 1.044 1.419 1.273 1.162 0.403 0.629 0.422 0.637 0.483 0.620 0.689 0.434 0.238 0.344 0.465 0.531 0.485 0.264 0.465 7.959 Female urine urine urine urine urine urine urine urine urine urine Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 505F 7.619 50.468 13.253 9.275 2.574 0.737 0.325 0.239 0.054 0.036 84.580 506F 13.288 25.271 11.104 16.269 7.575 2.196 1.729 0.725 0.556 0.532 79.245 507F 1.735 37.566 8.256 6.492 5.454 0.545 0.774 0.755 0.450 0.199 62.226 508F NS 18.383 6.798 8.044 8.331 2.021 2.294 0.504 1.171 0.549 48.095 Mean 7.547 32.922 9.853 10.020 5.984 1.375 1.281 0.556 0.558 0.329 68.537 SD 5.777 14.134 2.887 4.319 2.579 0.854 0.894 0.239 0.462 0.253 16.631 - 164- 000165 DuPont-7473 Percent recovery of radioactivity in feces in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeated dose) Male feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Day 8 Day 9 Day 10 Day 11 Day 12 Day 13 Day 14 Day 16 Day 18 Day 20 Day 22 Day 24 Day 26 Day 28 Total 501M NS 0.023 0.028 6.962 0.820 1.501 2.202 0.554 1.206 1.075 1.402 0.599 0.335 0.343 0.177 0.201 0.148 0.784 0.133 0.101 0.321 0.066 0.050 0.060 19.091 502M NS 0.678 3.681 11.323 1.243 0.754 0.742 0.162 0.656 0.638 0.733 1.368 0.740 0.525 1.082 0.940 0.083 0.212 0.251 0.158 0.154 0.184 0.099 0.239 26.645 503M NS 0.092 0.222 3.029 1.890 0.564 0.601 0.234 0.464 0.693 0.434 0.938 0.372 0.077 0.151 0.154 0.088 0.183 0.235 0.195 0.141 0.115 0.086 0.098 11.056 504M NS 0.019 0.355 5.692 1.062 0.450 0.631 0.553 0.749 1.890 2.032 1.546 2.793 1.902 0.449 0.083 0.065 0.189 0.187 0.646 0.660 0.514 0.067 0.036 22.570 Mean NA 0.203 1.072 6.752 1.254 0.817 1.044 0.376 0.769 1.074 1.150 1.113 1.060 0.712 0.465 0.345 0.096 0.342 0.202 0.275 0.319 0.220 0.076 0.108 19.841 SD NA 0.318 1.745 3.460 0.458 0.473 0.774 0.207 0.315 0.578 0.714 0.427 1.170 0.815 0.433 0.400 0.036 0.295 0.053 0.250 0.242 0.202 0.021 0.091 6.620 Female feces feces feces feces feces feces feces feces feces feces Timepoint 4h 8h 12 h 24 h 48 h 72 h 96 h 120 h 144 h 168 h Total 505F NS 0.005 0.003 0.396 0.234 0.077 0.029 0.076 0.099 0.029 0.948 506F NS 0.110 0.211 0.014 0.823 0.615 0.216 0.163 0.073 0.032 2.257 S07F NS 4.752 0.303 2.188 1.702 1.954 0.055 0.067 0.233 0.063 11.317 508F NS 9.527 <LOD 15.556 8.771 0.409 0.185 0.334 0.167 0.065 35.014 Mean NA 3.599 0.172 4.539 2.883 0.764 0.121 0.160 0.143 0.047 12.384 SD NA 4.530 0.154 7.406 3.972 0.824 0.093 0.124 0.072 0.019 15.775 ----- r-- \ (' ' ^ 1 '-- --------------------------------------------- --' ' 000166 Perfluorooctanoic Acid: Toxicokinetics in the Rat_______________________________ ___________ DuPont-7473 Percent recovery of radioactivity in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeated dose) Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Total 501M 0.008 4.723 0.077 0.660 0.003 0.017 0.033 0.057 0.008 5.932 0.268 0.123 0.032 0.001 0.007 0.044 0.002 0.036 0.013 12.044 502M 0.009 6.354 0.085 1.176 0.006 0.028 0.055 0.092 0.014 7.714 0.400 0.281 0,047 0.001 0.019 0.069 0.001 0.063 0.027 16.441 503M 0.012 7.029 0.117 1.683 0.006 0.016 0.051 0.105 0.019 6.969 0.546 0.205 0.068 0.002 0.014 0.073 0.002 0.078 0.023 17.018 504M 0.003 2.798 0.031 0.387 0.002 0.015 0.020 0.041 0.005 3.837 0.169 0.098 0.018 0.000 0.006 0.028 0.001 0.029 0.014 7.502 Mean 0.008 5.226 0.078 0.977 0.004 0.019 0.040 0.074 0.012 6.113 0.346 0.177 0.041 0.001 0.012 0.054 0.002 0.052 0.019 13.251 SD 0.004 1.886 0.035 0.573 0.002 0.006 0.016 0.030 0.006 1.684 0.164 0.083 0.021 0.001 0.006 0.021 0.001 0.023 0.007 4.430 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 h 168 b 168 h 168 h 168 h 168 h 168 b 168 h 168 h Total 505F 0.240 0.001 0.004 <LOD <LOD <LOD <LOQ <LOD 0.037 0.006 <LOQ <LOQ <LOD <LOD cLOD <LOD <LOD <LOD cLOD 0.288 506F 0.291 <LOQ 0.005 <LOD <LOD <LOD <LOD <LOD 0.043 0.006 cLOD cLOD cLOD cLOD cLOD cLOD cLOD cLOD cLOD 0.345 507F 0.170 cLOQ 0.003 cLOD cLOD cLOD cLOQ cLOD 0.028 0.003 cLOQ cLOQ cLOD cLOD cLOD cLOD cLOD cLOD cLOD 0.204 508F 0.346 0.001 0.005 cLOD cLOD cLOD 0.000 cLOD 0.040 0.006 cLOQ cLOQ cLOD cLOD cLOD cLOD cLOD cLOD cLOD 0.398 Mean 0.262 0.001 0.004 NA NA NA 0.000 NA 0.037 0.005 NA NA NA NA NA NA NA NA NA 0.309 SD 0.075 0.000 0.001 NA NA NA NA NA 0.006 0.002 NA NA NA NA NA NA NA NA NA 0.083 - 166- 000167 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity in cage wash and residual feed in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeated dose) Male Timepoint cage wash residual feed Day 28 Day 28 Total 501M 3.912 0.755 4.667 502M 1.354 0.268 1.622 503M 2.276 0.009 2.285 504M 1.516 0.021 1.537 Mean 2.265 0.263 2.528 SD 1.170 0.349 1.465 Female Timepoint cage wash residual feed 168 h 168 h Total 505F 1.558 0.170 1.728 506F 3.852 1.223 5.075 507F 2.904 17.662 20.566 508F 6.259 0.007 6.266 Mean 3.643 4.766 8.409 SD 1.982 8.615 8.329 - 167- 000168 Perfluorooctanoic Acid: Toxicokinetics in the Rat ____________________________ DuPont-7473 Concentration ( j i g equiv/g) in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA (repeated dose) Male prostate carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle bone Timepoint Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 Day 28 S01M 0.020 0.036 0.057 0.209 0.005 0.008 0.068 0.097 0.028 0.929 0.215 0.029 0.004 0.030 0.034 0.037 0.025 0.018 0.026 502M 0.022 0.052 0.125 0.390 0.009 0.011 0.120 0.169 0.049 1.144 0.325 0.053 0.007 0.026 0.065 0.070 0.049 0.033 0.039 503M 0.037 0.056 0.090 0.441 0.009 0.011 0.103 0.159 0.049 1.084 0.397 0.054 0.010 0.060 0.052 0.075 0.079 0.033 0.044 504M 0.010 0.022 0.058 0.131 0.003 0.004 0.044 0.060 0.019 0.601 0.149 0.017 0.003 0.021 0.018 0.027 0.021 0.015 0.020 Mean 0.022 0.042 0.083 0.293 0.007 0.009 0.084 0.121 0.036 0.940 0.272 0.038 0.006 0.034 0.042 0.052 0.044 0.025 0.032 SD 0.011 0.016 0.032 0.147 0.003 0.003 0.034 0.052 0.015 0.243 0.111 0.018 0.003 0.018 0.021 0.024 0.027 0.010 0.011 Female carcass skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone Timepoint 168 h 168 h 168 h 168 h 168 h 168 b 168 h 168 h 168 h 168 h 168 h 168 h 168 b 168 h 168 h 168 h 168 h 168 h 168 h 505F 0.003 0.001 0.001 <LOD <LOD <LOD <LOQ <LOD 0.009 0.007 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD <LOD <LOD 506F 0.003 <LOQ 0.001 <LOD <LOD <LOD <LOD <LOD 0.008 0.006 <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD <LOD 507F 0.002 <LOQ 0.001 <LOD <LOD <LOD <LOQ <LOD 0.006 0.004 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD <LOD <LOD 508F 0.004 0.002 0.001 <LOD <LOD <LOD 0.001 <LOD 0.008 0.007 <LOQ <LOQ <LOD <LOD <LOD <LOD <LOD <LOD <LOD Mean 0.003 0.002 0.001 NA NA NA 0.001 NA 0.008 0.006 NA NA NA NA NA NA NA NA NA SD 0.001 0.001 0.000 NA NA NA NA NA 0.001 0.001 NA NA NA NA NA NA NA NA NA -168- 000169 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix H Pharmacokinetics in Male and Female Rats Following Oral Gavage with 0.1 mg/kg PFOA - 169 - ) 000170 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/mL): 0.028 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 0.1 Male Animal 652092 652093 652095 652096 Mean Standard Deviation Animal Weight (g) 223.3 228.3 205.9 207.5 216.3 11.2 Dose Volume (mL) 0.89 0.91 0.82 0.83 0.86 0.04 DuPont-7473 Actual Dose Rate (mg/kg) 0.112 0.112 0.112 0.113 0.112 0.000 Female Animal 652100 652101 652102 652105 Mean Standard Deviation Animal Weight (g) 179.8 164.5 184.6 176.5 176.4 8.6 Dose Volume (mL) 0.72 0.66 0.74 0.71 0.71 0.03 Actual Dose Rate (mg/kg) 0.113 0.113 0.113 0.113 0.113 0.000 - 170- 000171 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/tg/mL) in serial plasma in male and female rats following oral gavage with 0.1 mg/kg PFOA Male Time (h) 0 0.25 0.5 1 2 4 8 12 16 24 36 48 72 96 120 144 168 192 240 288 336 384 432 480 528 652092 <LOQ 0.438 0.438 0.376 0.452 0.445 0.468 0.573 0.430 0.383 0.291 0.272 0.255 0.240 0.219 0.189 0.221 0.194 0.136 0.145 0.094 0.083 0.070 0.063 0.040 Animal 652093 652095 <LOQ 0.105 0.125 0.185 0.214 0.239 0.339 0.429 0.387 0.347 0.360 0.279 0.320 0.245 0.233 0.220 0.199 0.177 0.148 0.147 0.108 0.085 0.059 0.057 0.053 <LOQ 0.141 0.198 0.267 0.376 0.425 0.491 0.501 0.714 0.443 0.371 0.322 0.320 0.367 0.304 0.309 0.247 0.241 0.181 0.176 0.117 0.102 0.087 0.081 0.067 652096 <LOQ 0.442 0.448 0.676 0.509 0.506 0.570 0.565 0.512 0.421 0.434 0.441 0.402 0.390 0.290 0.288 0.345 0.287 0.242 0.255 0.182 0.160 0.186 0.148 0.092 Mean NA 0.282 0.302 0.376 0.388 0.404 0.467 0.517 0.511 0.399 0.364 0.328 0.324 0.311 0.262 0.251 0.253 0.225 0.177 0.181 0.125 0.108 0.100 0.087 0.063 SD NA 0.184 0.165 0.215 0.128 0.115 0.096 0.067 0.145 0.042 0.059 0.078 0.060 0.079 0.042 0.056 0.064 0.049 0.047 0.052 0.039 0.036 0.058 0.042 0.022 Female Time (h) 0 0.25 0.50 1 2 4 8 16 24 36 48 72 96 652100 <LOQ 0.651 0.512 0.577 0.504 0.318 0.073 <LOQ <LOQ <LOQ <LOQ <LOQ <IjOQ Animal 652101 652102 <LOQ 0.566 0.648 0.487 0.619 0.422 0.146 0.050 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.591 0.770 0.481 0.564 0.352 <LOQ <LOQ <LOQ <LOQ NS NS <LOQ 652105 <LOQ NA 0.591 0.611 0.425 0.431 0.087 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ Mean NA 0.603 0.630 0.539 0.528 0.381 0.102 0.050 NA NA NA NA NA SD NA 0.043 0.109 0.065 0.083 0.055 0.039 NA NA NA NA NA NA - 171- 000172 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following oral gavage with 0.1 mg/kg PFOA Male Animal 652092 652093 652095 652096 Mean SD Cmu Lambda z AUCjnf AUCjnj/D Cip Tmax (hr) (/ig/mL) (1/hr) Tw (hr) (hr-M/mL) (hrM/mL/m/kg) (mL/kg-hr) 12.0 0.573 0.004 184.559 95.469 12.0 0.429 0.004 177.376 98.863 16.0 0.714 0.004 187.517 126.328 1.0 0.676 0.003 257.643 172.235 852.404 882.701 1127.931 1524.207 1.173 1.133 0.887 0.656 10.25 6.45 0.598 0.127 0.004 0.001 201.774 123.224 37.489 35.476 1096.811 310.491 0.962 0.240 Female Animal 652100 652101 652102 652105 Mean SD Cjoax Lambda z AUCinf AUCwf/D Cip Tmax (hr) (/ig/mL) (1/hr) Tin (hr) (hr-fi/mL) (hr/t/mL/mg/k) (mL/k-hr) 0.25 0.651 0.303 2.291 2.885 0.50 0.648 0.170 4.098 4.316 0.50 0.770 0.180 3.860 3.956 1.00 0.611 0.270 2.576 3.182 25.529 38.193 35.006 28.156 39.171 26.182 28.567 35.517 0.56 0.67 0.231 3.206 3.584 0.31 0.07 0.066 0.905 0.666 31.721 5.880 32.359 6.025 - 172- 000170 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix I Pharmacokinetics in Male and Female Rats Following Oral Gavage with 1 mg/kg PFOA - 173- 000174 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/mL): 0.253 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 1.0 Male Animal 653745 653746 653747 653749 Mean Standard Deviation Animal Weight (g) 250.0 246.4 248.7 249.6 248.7 1.6 Female Animal 653759 653760 653761 653762 Mean Standard Deviation Animal Weight (g) 200.4 201.3 182.6 203.5 197.0 9.7 Dose Volume (mL) 1.00 0.99 0.99 1.00 1.00 0.01 Dose Volume (mL) 0.80 0.81 0.73 0.81 0.79 0.04 DuPont-7473 Actual Dose Rate (mg/kg) 1.014 1.018 1.009 1.015 1.014 0.004 Actual Dose Rate (mg/kg) 1.010 1.018 1.011 1.007 1.012 0.005 - 174- 000175 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/tg/mL) in serial plasma in male and female rats following oral gavage with 1 mg/kg PFOA Male Time (b) 653745 Animal 653746 653747 0 <LOQ <LOQ <LOQ 0.25 2.054 2.198 1.688 0.5 3.096 2.978 2.252 1 4.860 3.920 2.768 2 6.848 5.056 3.952 4 9.128 7.798 3.818 8 8.042 7.910 6.800 12 5.098 8.414 5.372 16 6.228 6.890 6.782 24 5.724 4.764 4.792 36 4.222 4.514 4.132 48 4.230 4.754 3.540 72 2.892 3.960 2.904 96 2.580 3.402 3.006 120 3.048 3.142 3.836 144 2.136 2.930 3.250 168 1.610 3.852 2.746 192 2.152 3.952 3.434 240 NS* 1.636 1.358 288 NS 1.507 0.811 336 NS 1.680 0.801 384 NS 1.425 0.828 432 NS 0.802 0.711 480 NS 1.077 0.579 528 NS 0.561 0.384 "Animal # 653745 was found dead on day nine 653749 <LOQ 5.016 6.580 6.312 6.376 7.418 6.698 9.380 9.014 4.624 5.956 4.656 4.126 4.522 4.368 2.994 2.932 2.444 1.471 1.315 1.135 1.013 0.796 0.681 0.397 Mean NA 2.739 3.727 4.465 5.558 7.041 7.363 7.066 7.229 4.976 4.706 4.295 3.471 3.378 3.599 2.828 2.785 2.996 1.488 1.211 1.205 1.089 0.770 0.779 0.447 SD NA 1.533 1.939 1.499 1.312 2.270 0.712 2.154 1.225 0.504 0.849 0.552 0.665 0.834 0.622 0.481 0.921 0.841 0.140 0.360 0.443 0.305 0.051 0.263 0.098 Female T im e(h) 0 0.25 0.50 1 2 4 8 12 16 24 36 48 72 96 653759 <LOQ 3.263 3.602 4.138 4.307 4.278 1.609 0.645 0.219 0.255 <LOQ <LOQ <LOQ 0.070 Animal 653760 653761 <LOQ 4.356 4.314 4.358 4.245 3.404 1.384 0.494 0.189 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 5.914 6.315 6.487 5.604 4.802 2.025 1.108 0.421 0.063 0.036 0.013 <LOQ NS 653762 <LOQ 2.929 3.977 3.909 3.399 2.719 0.788 0.510 0.115 <LOQ 0.023 0.058 NS 0.080 Mean NA 4.116 4.552 4.723 4.389 3.801 1.452 0.689 0.236 0.159 0.029 0.036 NA 0.075 SD NA 1.345 1.211 1.190 0.910 0.923 0.516 0.287 0.131 0.136 0.009 0.032 NA 0.007 - 175- 000176 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following oral gavage with 1 mg/kg PFOA Male Animal 653745 653746 653747 653749 Mean SD c mi Lambda z AUCinf AUCini'/D Cip Tma, (hr) 0ig/mL) (1/hr) Ti/2 (hr) (hr-M/mL) (hi-tie/mL/mc/ke) (mL/kg*hr) 4.0 9.128 0.007 98.054 941.580 932.258 12.0 8.414 0.004 176.052 1432.625 1404.534 8.0 6.800 0.005 142.647 1105.277 1094.334 12.0 9.380 0.005 136.620 1298.369 1272.911 1.073 0.712 0.914 0.786 9.00 8.431 0.005 138.343 1194.463 1176.009 3.83 1.161 0.001 31.972 215.578 206.316 0.871 0.158 Female Animal f'-'max Lambda z AUCinf AUCjnf/D Clp Tna* (hr) (jig/mL) (1/hr) T m (hr) (hr-Mg/mL) (hrag/mL/mg/kg) (mL/kg-hr) 653759 653760 653761 653762 2.00 4.307 0.246 2.822 47.894 47.420 1.00 4.358 0.242 2.858 31.224 30.672 1.00 6.487 0.135 5.118 47.818 47.298 0.50 3.977 0.229 3.029 29.350 29.149 21.088 32.603 21.143 34.310 Mean SD 1.13 4.782 0.213 3.457 39.072 0.63 1.149 0.053 1.111 10.172 38.635 10.093 27.286 7.159 - 176- 000177 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix J Pharmacokinetics in Male and Female Rats Following Intravenous Injection with 1 mg/kg PFOA - 177 - 000178 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/mL): 0.554 Nominal Dose Volume (mL/kg): 2.0 Nominal Dose Rate (mg/kg): 1.0 Male Animal 653740 653742 653743 653744 Mean Standard Deviation Animal Weight (g) 253.2 256.4 235.2 248.5 248.3 9.3 Female Animal 653752 653754 653758 653763 Mean Standard Deviation Animal Weight (g) 201.8 202.7 201.2 196.7 200.6 2.7 Dose Volume (mL) 0.51 0.51 0.47 0.50 0.50 0.02 Dose Volume (mL) 0.40 0.41 0.40 0.39 0.40 0.01 DuPont-7473 Actual Dose Rate (mg/kg) 1.116 1.102 1.108 1.115 1.110 0.007 Actual Dose Rate (mg/kg) 1.099 1.121 1.102 1.099 1.105 0.011 - 178- 0001/9 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (jig/mL) in serial plasma in male and female rats following intravenous injection with 1 mg/kg PFOA Male Time (h) 0 0.25 0.5 1 2 4 8 12 16 24 36 48 72 96 120 144 168 192 240 288 336 384 432 480 528 653740 <LOQ 20.170 9.380 9.202 6.106 6.702 5.658 5.542 8.656 6.972 3.984 4.104 4.642 3.222 2.818 1.852 1.687 1.741 2.295 1.695 1.403 1.677 1.108 0.864 0.652 Animal 653742 653743 <LOQ 11.158 6.470 9.920 8.212 8.916 6.832 7.942 7.330 6.034 4.450 4.468 3.158 2.892 2.630 2.037 1.752 1.694 1.498 1.381 1.372 1.161 1.153 0.989 0.699 <LOQ 8.144 8.454 5.562 5.016 6.568 8.426 8.230 10.408 5.736 4.224 3.252 4.654 3.200 2.468 1.600 1.466 1.481 1.663 1.122 1.030 0.802 0.669 0.696 0.449 653744 <LOQ 7.884 7.444 12.480 8.606 7.264 7.192 7.852 7.574 4.480 4.090 4.358 3.426 2.550 2.890 1.493 1.964 1.874 1.864 1.481 1.431 1.161 0.843 0.812 0.565 Mean NA 11.839 7.937 9.291 6.985 7.363 7.027 7.392 8.492 5.806 4.187 4.046 3.970 2.966 2.702 1.745 1.717 1.697 1.830 1.420 1.309 1.200 0.943 0.840 0.591 SD NA 5.749 1.258 2.856 1.711 1.079 1,140 1.243 1.401 1.029 0.201 0.551 0.791 0.316 0.190 0.246 0.205 0.163 0.344 0.238 0.188 0.360 0.228 0.121 0.110 Female Time (h) 0 0.25 0.50 1 2 4 8 12 16 24 36 48 72 96 653752 <LOQ 4.431 4.709 3.704 3.261 2.593 0.833 3.940 0.121 <LOQ <LOQ <LOQ <LOQ <LOQ Animal 653754 653758 <LOQ 5.525 4.825 4.728 4.041 3.415 1.271 1.798 0.197 0.017 <LOQ <LOQ 0.065 <LOQ <LOQ 4.274 3.830 3.280 3.610 1.898 0.335 0.953 0.022 <LOQ <LOQ <LOQ 0.019 <LOQ 653763 0.012 5.637 4.751 4.052 3.789 3.050 0.729 2.127 0.091 <LOQ 0.092 <LOQ <LOQ <LOD Mean 0.012 4.967 4.529 3.941 3.675 2.739 0.792 2.204 0.108 0.017 0.092 NA 0.042 NA SD NA 0.714 0.468 0.612 0.328 0.654 0.385 1.258 0.073 NA NA NA 0.032 NA - 179- 0001S0 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following intravenous injection with 1 mg/kg PFOA Male Lambda z AUC^r ALJCinp/D Cip Animal_____ (1/hr) Tm (hr) (hr-flg/mL) (hr-Mg/mL/mg/kg) (mL/kg-hr) 653740 653742 653743 653744 Mean SD 0.004 0.004 0.004 0.004 0.004 0.000 194.339 200.779 156.978 190.240 1374.590 1293.389 1107.407 1223.881 185.584 1249.817 19.558 113.167 1227.313 1175.808 997.664 1092.751 1123.384 100.488 0.815 0.851 1.002 0.915 0.896 0.082 Female Lambda z AUCj^r AUCi^/D Lip Animal_____ (1/hr) Tm (hr) (hr-ftg/mL) (hr-Mg/mL/mg/kg) (mL/kg-hr) 653752 653754 653758 653763 0.216 0.268 0.308 0.208 3.211 2.584 2.251 3.331 38.774 39.558 23.078 34.581 35.249 35.320 20.980 31.437 28.370 28.313 47.665 31.810 Mean SD 0.250 0.047 2.844 0.514 33.998 7.601 30.747 6.759 34.040 9.230 . - 180 - 000161 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix K Pharmacokinetics in Male and Female Rats Following Oral Gavage with 5 mg/kg PFOA - 181 - 000162 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/mL): 1.380 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 5.0 Male Animal 654601 654602 654603 654604 Mean Standard Deviation Animal Weight (g) 216.0 223.0 216.0 217.0 218.0 3.4 Female Animal 654605 654606 654608 654610 Mean Standard Deviation Animal Weight (g) 184.0 188.0 192.0 185.0 187.3 3.6 Dose Volume (mL) 0.86 0.89 0.86 0.87 0.87 0.01 Dose Volume (mL) 0.74 0.75 0.77 0.74 0.75 0.01 DuPont-7473 Actual Dose Rate (mg/kg) 5.496 5.509 5.496 5.535 5.509 0.018 Actual Dose Rate (mg/kg) 5.552 5.507 5.536 5.522 5.529 0.019 - 182- )< 0001&3 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/ig/mL) in serial plasma in male and female rats following oral gavage with 5 mg/kg PFOA Male Time (h) 0 0.25 0.5 1 2 4 8 12 16 24 48 72 96 120 144 168 192 240 288 336 384 432 480 528 654601 <LOQ 6.72 17.456 19.728 24.976 24.74 38.02 35.112 36.472 31.968 16.412 24.088 14.456 9.531 9.916 7.855 5.618 5.779 3.92 3.874 3.792 2.868 2.106 1.403 Animal 654602 654603 <LOQ 7.216 10.136 14.128 27.24 41.148 33.036 26.284 25.492 NS 26.408 25.6 19.632 12.967 12.067 12.846 11.481 9.067 11.542 8.903 6.347 5.454 5.783 3.877 <LOQ 4.808 7.132 14.632 31.612 27.776 31.604 27.404 38.368 50.812 16.788 21.792 15.38 15.676 12.666 10.105 12.127 8.807 7.635 5.868 6.45 3.997 4.686 3.367 654604 <LOQ 8.024 9.712 22.896 32.08 42.204 47.128 33.804 40.98 49.02 33.272 29.348 16.496 20.903 11.497 14.831 12.746 9.07 9.601 6.065 6.473 5.036 4.737 3.102 Mean NA 6.692 11.109 17.846 28.977 33.967 37.447 30.651 35.328 43.933 23.220 25.207 16.491 14.769 11.537 11.409 9.182 8.181 8.175 6.178 5.766 4.339 4.328 2.937 SD NA 1.366 4.435 4.211 3.445 8.998 7.015 4.452 6.813 10.401 8.143 3.174 2.254 4.800 1.181 3.061 5.040 1.606 3.254 2.069 1.317 1.156 1.565 1.072 Female Time (h) 0 0.25 0.50 1 2 4 8 12 16 24 36 48 72 96 654605 <LOQ 16.795 18.910 21.450 17.455 14.195 1.793 0.730 0.385 0.248 0.028 <LOQ 0.012 <LOQ Animal 654606 654608 <LOQ 15.055 14.965 19.780 13.640 10.595 2.178 1.738 0.754 0.242 0.080 0.028 <LOQ <LOQ <LOQ 5.230 9.665 9.935 18.405 15.675 4.910 2.144 0.765 0.122 0.024 0.015 <LOQ <LOQ 654610 <LOQ 6.705 9.730 15.055 21.810 15.465 4.201 2.002 0.604 0.081 0.042 0.017 <LOQ <LOQ Mean NA 10.946 13.318 16.555 17.828 13.983 3.270 1.653 0.627 0.173 0.043 0.020 0.012 NA SD NA 5.824 4.480 5.178 3.360 2.351 1.520 0.638 0.177 0.084 0.026 0.007 NA NA t i t ' -, -183- 0001&4 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following oral gavage with 5 mg/kg PFOA Male Animal 654601 654602 654603 654604 Mean SD C Lambda z AUCW AUCW/D Clp Timi (hr) Oig/mL) (1/hr) Tia (hr) (hr-jig/mL) (hr-/ig/mL/mg/kg) (mL/kg-hr) 8.0 38.02 0.0050 139.61 4722.52 4.0 41.15 0.0034 201.71 7505.93 24.0 50.81 0.0036 194.02 6899.71 24.0 49.02 0.0043 161.42 7806.62 859.26 1362.48 1255.41 1410.41 1.16 0.73 0.80 0.71 15.0 44.75 0.0041 174.19 6733.70 10.5 6.14 0.0007 28.92 1392.83 1221.89 250.28 0.85 0.21 Female Animal 654605 654606 654608 654610 Mean SD ^max Lambda z AUCinf AUCjnf/D Clp TTM, (hr) (lig/mL) (1/hr) Tw (hr) (hr-Mg/mL) (hr-gg/mL/mg/kg) (mL/kg-hr) 1.0 21.450 0.169 4.103 111.998 1.0 19.780 0.126 5.521 100.407 2.0 18.405 0.163 4.261 121.502 2.0 21.810 0.153 4.536 125.685 20.173 18.233 21.948 22.761 49.573 54.847 45.563 43.935 1.50 20.36 0.15 0.58 1.58 0.02 4.60 114.90 0.64 11.23 20.78 2.01 48.48 4.86 - 184- 0001.5 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix L Pharmacokinetics in Male and Female Rats Following Oral Gavage with 25 mg/kg PFOA - 185- 0001S6 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/mL): 6.726 Nomina] Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 25.0 Male Animal 655168 655169 655170 655172 Mean SD Animal Weight (g) 225.0 221.0 224.0 230.0 225.0 3.7 Female Animal 655175 655176 655177 655178 Mean SD Animal Weight (g) 182.0 183.0 210.0 204.0 194.8 14.4 Dose Volume (mL) 0.90 0.88 0.90 0.90 0.90 0.01 Dose Volume (mL) 0.73 0.73 0.84 0.82 0.78 0.06 DuPont-7473 Actual Dose Rate (mg/kg) 26.90 26.78 27.02 26.32 26.76 0.31 Actual Dose Rate (mg/kg) 26.98 26.83 26.90 27.04 26.94 0.09 - 186 - O O O IB V Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/ig/mL) in serial plasma in male and female rats following oral gavage with 25 mg/kg PFOA Male ______________________ Animal ________________ Time (h) 655168 655169 65S17C 655172_______ Mean 0 <LOQ <LOQ <LOQ <LOQ NA 0.25 16.375 70.375 67.725 70.175 56.163 0.5 26.925 97.400 81.575 85.525 72.856 1 56.175 115.475 106.500 121.650 99.950 2 91.575 177.525 131.375 144.000 136.119 4 156.875 153.175 149.225 143.925 150.800 8 120.800 134.550 143.725 154.775 138.463 12 134.500 155.200 136.250 150.975 144.231 16 136.550 151.425 150.825 135.325 143.531 24 103.175 147.225 134.375 122.775 126.888 48 90.425 104.900 91.125 114.775 100.306 72 75.775 85.050 65.600 76.250 75.669 96 61.275 63.250 60.500 84.775 67.450 120 53.025 59.150 57.325 81.750 62.813 144 42.250 54.075 50.325 83.200 57.463 168 47.175 46.825 32.875 75.575 50.613 192 36.875 37.325 23.000 47.475 36.169 240 25.525 27.825 15.975 55.650 31.244 288 21.275 22.675 13.250 43.475 25.169 336 21.475 25.775 11.150 43.500 25.475 384 13.195 21.960 9.225 32.075 19.114 432 22.395 18.525 7.715 27.690 19.081 480 9.385 15.700 6.280 29.410 15.194 528 8.050 10.160 4.445 15.930 9.646 SD NA 26.552 31.351 29.839 35.512 5.547 14.388 10.395 8.786 18.698 11.724 7.953 11.608 12.883 17.854 17.925 10.051 17.061 12.891 13.493 10.147 8.458 10.256 4.808 Female Time (h) 0 0.25 0.50 1 2 4 8 12 16 24 36 48 72 96 655175 <LOQ 68.020 200.270 145.180 125.270 154.950 33.578 11.726 4.671 0.349 0.067 0.056 0.058 0.033 Animal 655176 655177 <LOQ 62.380 101.140 80.530 97.560 88.640 19.192 9.130 2.257 0.381 0.203 0.096 0.013 <LOD <LOQ 79.860 95.090 74.110 122.240 87.100 37.070 6.624 2.014 0.446 0.109 0.044 0.074 <LOD 655178 0.019 106.400 99.930 106.720 106.950 68.240 36.072 7.072 1.803 0.276 0.160 0.046 <UDD <LOD Mean 0.019 79.165 124.108 101.635 113.005 99.733 31.478 8.638 2.686 0.363 0.135 0.060 0.048 0.033 SD NA 19.563 50.842 32.275 13.050 37.962 8.321 2.330 1.336 0.070 0.059 0.024 0.031 NA - 187- 000168 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following oral gavage with 25 mg/kg PFOA Male Animal 655168 655169 655170 655172 Mean SD Cm** Lambda z AUCjnf AUCjnf/D Cip Tm (hr) (M/mL) (1/hr) T,fl (hr) (hr-Mg/mL) (hr/ig/mL/mg/kg) (mL/kg-hr) 4.0 156.875 0.0047 147.100 21698.88 2.0 177.525 0.0042 165.499 25579.15 16.0 150.825 0.0062 112.565 18242.08 8.0 154.775 0.0034 204.712 35102.32 806.65 955.16 675.13 1333.67 1.240 1.047 1.481 0.750 7.5 160.00 0.0046 157.47 25155.61 6.2 11.95 0.0012 38.39 7276.96 942.65 284.67 1.13 0.31 Female Animal 655175 655176 655177 655178 Mean SD Cnax Lambda z AUCinf AUCWD C1p Tfimi (hr) (MR/mL) (1/hr) Tin (hr) (hr-gJmL) (hr-jig/mL/mg/kg) (mL/kg-hr) 0.5 200.270 0.025 28.157 1071.376 0.5 101.140 0.071 9.738 661.311 2.0 122.240 0.034 20.366 752.314 2.0 106.950 0.105 6.625 698.043 39.710 24.648 27.967 25.815 25.183 40.571 35.756 38.737 1.25 132.65 0.059 16.22 795.76 0.87 45.95 0.037 9.90 187.51 29.54 6.92 35.06 6.88 - 188- 000159 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix M Pharmacokinetics in Male and Female Rats Following Oral Gavage with 0.1 mg/kg PFOA (extended time course) * ' ? i - 189- 000190 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/mL): 0.024 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 0.1 Male Animal 659277 659278 659279 659280 Mean SD Animal Weight (b) 267.0 274.0 280.0 269.0 272.5 5.8 Dose Volume (mL) 1.10 1.10 1.10 1.10 1.10 0.00 DuPont-7473 Actual Dose Rate (mg/kg) 0.099 0.097 0.095 0.099 0.097 0.002 Female Animal 659283 659285 659286 659288 Mean SD Animal Weight (g) 184.0 209.0 209.0 202.0 201.0 11.8 Dose Volume (mL) 0.74 0.84 0.84 0.81 0.81 0.05 Actual Dose Rate (mg/kg) 0.097 0.097 0.097 0.097 0.097 0.000 - 190- 000131 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/xg/mL) in serial plasma in male rats following oral gavage with 0.1 mg/kg PFOA (extended time course) Time (h) 0 1 2 4 8 12 16 24 48 72 96 120 144 168 192 240 288 336 384 432 480 528 576 624 672 720 768 816 864 912 960 1008 1056 1104 1152 1200 1248 1296 1344 1392 1440 1488 1536 1584 1632 1680 1728 1776 1824 1872 1920 1968 2016 659277 <LOQ 0.557 1.692 0.641 0.569 0.418 0.539 0.502 0.395 0.360 0.276 0.358 0.277 0.324 0.310 0.260 0.211 0.235 0.155 0.196 0.126 0.126 0.145 0.128 0.112 0.075 0.071 0.062 0.064 0.050 0.040 0.037 0.036 0.030 0.031 0.029 0.023 0.037 0.028 0.030 0.023 0.019 0.015 0.021 0.006 0.014 <LOQ <LOQ 0.006 0.001 <LOQ <LOQ <LOQ Animal 659278 659279 0.001 0.836 0.912 0.726 0.759 0.824 0.748 0.770 0.642 0.356 0.516 0.448 0.429 0.442 0.314 0.304 0.322 0.251 0.230 0.215 0.169 0.106 0.116 0.146 0.143 0.083 0.086 0.063 0.067 0.068 0.070 0.049 0.038 0.040 0.037 0.036 0.039 0.017 0.047 0.042 0.038 0.030 0.028 0.027 0.020 0.026 0.011 0.011 0.013 0.008 0.008 0.006 0.005 <LOQ 0.619 0.634 0.635 0.488 0.502 0.722 0.633 0.462 0.274 0.317 0.305 0.267 0.273 0.247 0.193 0.134 0.119 0.104 0.079 0.073 0.036 0.031 0.053 0.045 0.028 0.021 0.018 0.015 0.010 0.007 0.004 0.003 0.001 0.003 <LOQ <LOQ 0.008 0.006 0.003 0.008 0.003 0.004 0.003 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 659280 <LOQ 0.844 1.004 0.640 0.764 0.927 0.771 0.668 0.566 0.495 0.413 0.490 0.380 0.485 0.304 0.321 0.272 0.260 0.272 0.200 0.155 0.148 0.123 0.138 0.157 0.099 0.083 0.069 0.061 0.049 0.041 0.041 0.034 0.028 0.026 0.026 0.028 0.042 0.042 0.037 0.031 0.027 0.026 0.019 0.012 0.012 0.012 0.008 0.014 0.009 0.004 0.006 0.004 Mean 0.001 0.714 1.060 0.660 0.645 0.668 0.695 0.643 0.516 0.371 0.381 0.400 0.338 0.381 0.294 0.269 0.235 0.216 0.191 0.172 0.131 0.104 0.104 0.116 0.114 0.071 0.065 0.053 0.052 0.044 0.039 0.033 0.028 0.025 0.024 0.030 0.030 0.026 0.031 0.028 0.025 0.020 0.018 0.017 0.013 0.017 0.012 0.009 0.011 0.006 0.006 0.006 0.005 SD NA 0.148 0.449 0.044 0.139 0.246 0.106 0.111 0.110 0.092 0.107 0.084 0.079 0.099 0,031 0.057 0.081 0.066 0.075 0.063 0.042 0.048 0.050 0.043 0.050 0.031 0.030 0.023 0.025 0.024 0.026 0.020 0.017 0.017 0.015 0.005 0.008 0.016 0.018 0.017 0.013 0.012 0.011 0.010 0.007 0.007 0.001 0.003 0.005 0.004 0.003 0.000 0.001 - 191 - 0001S2 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration {fig equiv/g) in serial plasma in female rats following oral gavage with 0.1 mg/kg PFOA (extended time course) Time(h) 0 1 2 4 8 12 16 24 36 48 72 96 120 144 168 192 240 312 659283 <LOQ 0.427 0.344 0.349 0.101 NS 0.028 0.005 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ Animal 659285 659286 <LOQ 0.587 0.503 0.408 0.134 0.066 0.021 0.004 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 0.575 0.570 0.436 0.121 0.050 0.011 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ 659288 <LOQ 0.478 0.479 0.427 0.139 0.065 0.021 <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ <LOQ Mean NA 0.517 0.474 0.405 0.124 0.060 0.020 0.005 NA NA NA NA NA NA NA NA NA NA SD NA 0.077 0.095 0.039 0.017 0.009 0.007 0.001 NA NA NA NA NA NA NA NA NA NA ft * P. -192- 000193 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Pharmacokinetic parameters in male and female rats following oral gavage with 0.1 mg/kg PFOA (extended time course) Male Animal 659277 659278 659279 659280 Mean SD Lambda z AUCxnf AUCjni/D Cip Tma* (hr) (ME/mL) (1/hr) Tw (hr) (hr*/ie/mL) (hr/xg/mL/mg/kg) (mL/kg-hr) 2.0 1.692 0.0024 294.8 197.67 1996.62 2.0 0.912 0.0022 319.5 253.70 2615.48 16.0 0.722 0.0036 193.7 126.28 1329.28 2.0 1.004 0.0023 300.5 247.87 2503.71 0.50 0.38 0.75 0.40 5.5 1.08 0.0026 277.10 206.38 7.0 0.42 0.0007 56.62 59.03 2111.28 586.77 0.51 0.17 Female Animal 659283 659285 659286 659288 Mean SD Cmax Lambda z AUCjnf AUCinf/D Cip Tma, (hr) (fifi/mL) (1/hr) Tm (hr) (hr*Mfi/mL) (hru/mL/me/k) (mL/kg*hr) 1.0 0.427 0.20 3.40 2.87 1.0 0.587 0.13 5.21 3.58 1.0 0.575 0.30 2.31 3.48 2.0 0.479* 0.25 2.82 3.42 29.57 36.89 35.87 35.24 33.8 27.1 27.9 28.4 1.25 0.52 0.22 3.44 3.34 0.50 0.08 0.07 1.26 0.32 34.39 3.29 29.30 3.06 - 193 - 000194 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix N Tissue Distribution at and Tma*/? in Male and Female Rats Following Oral Gavage with 1 mg/kg 14C-PFOA - 194- 0001S5 Perfluorooclanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/g): 0.220 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 1.0 DuPont-7473 Male Animal 401M 402M 403M 404M Mean Standard Deviation Tiru/2 405M 406M 407M 408M Mean Standard Deviation Animal Weight (r) Dose Administered Actual Dose Rate __________ E)__________ _______________________ 201.2 198.2 206.2 203.2 202.2 3.4 0.80 0.79 0.78 0.81 0.80 0.01 0.877 0.876 0.831 0.875 0.865 0.022 R a d io a c tiv ity Received (aCi) 23.684 23.301 23.006 23.861 23.463 0.384 206.4 202.1 203.3 200.9 203.2 2.4 0.84 0.80 0.81 0.79 0.81 0.02 0.889 0.874 0.880 0.869 0.878 0.009 24.628 23.713 24.008 23.418 23.942 0.517 Female Animal Tim 409F 410F 41 IF 412F Mean Standard Deviation Tjm* 413F 414F 415F 416F Mean Standard Deviation Animal Weight (R) 193.1 193.2 194.2 189.2 192.4 2.2 192.1 >80.6 195.5 189.6 189.5 6.4 Dose Administered (R) 0.80 0.80 0.78 0.77 0.79 0.02 0.78 0.73 0.78 0.77 0.76 0.02 Actual Dose Rate (mg/kg) 0.872 0.882 0.878 0.889 0.880 0.007 0.894 0.893 0.871 0.890 0.887 0.011 Radioactivity Received (a Ci) 22.593 22.858 22.888 22.563 22.725 0.171 23.035 21.649 22.858 22.652 22.548 0.620 -195* 0001S6 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity at Tmax in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male Timepoint 401M 402M 403M 404M Mean SD prostate skin' whole blood* brain fat* heart lungs spleen liver kidney G.I. tract G.l. contents thyroid thymus testes adrenals muscle' bone' 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m Total 0.070 16.470 21.816 0.064 1.913 0.487 0.717 0.079 25.170 1.718 1.857 1.564 0.010 0.079 0.761 0.015 11.612 2.973 87.375 0.034 11.648 22.028 0.058 2.260 0.345 0.656 0.086 15.970 2.073 2.502 2.371 0.004 0.080 0.765 0.017 11.994 3.218 76.109 0.115 15.385 23.151 0.098 2.946 0.603 0.953 0.101 17.973 1.550 3.931 2.827 0.013 0.097 0.843 0.025 12.951 4.046 87.608 0.114 15.585 21.596 0.063 2.005 0.369 0.632 0.078 27.718 2.455 3.431 1.568 0.004 0.085 0.650 0.018 11.543 2.853 90.767 0.083 14.772 22.148 0.071 2.281 0.451 0.740 0.086 21.708 1.949 2.930 2.083 0.008 0.085 0.755 0.019 12.025 3.273 85.465 0.039 2.135 0.692 0.018 0.467 0.119 0.147 0.011 5.627 0.402 0.929 0.625 0.005 0.008 0.079 0.004 0.648 0.538 6.426 ' Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood==7.4%, fat=7%, muscle=40.4%. bone=7.3% of body weight. Female Timepoint 409F 41OF 411F 412F Mean SD skin' whole blood' brain fat heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle* bone* 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1hl5m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m Total 0.495 7.509 0.048 0.163 0.199 0.592 0.077 6.724 3.706 4.482 21.336 0.009 0.068 0.046 0.021 0.209 0.225 0.091 46.000 0.631 6.355 0.038 0.161 0.309 0.562 0.095 8.932 4.256 4.365 3.325 0.010 0.066 0.075 0.014 0.186 0.372 0.125 29.877 0.268 4.067 0.033 0.107 0.134 0.285 0.040 6.233 3.154 23.647 29.125 0.013 0.038 0.034 0.012 0.094 0.213 0.086 67.583 0.341 5.028 0.027 0.105 0.151 0.376 0.041 6.349 2.037 10.301 34.039 0.007 0.037 0.034 0.009 0.189 0.161 0.100 59.332 0.434 5.740 0.037 0.134 0.198 0.454 0.063 7.060 3.288 10.699 21.956 0.010 0.052 0.047 0.014 0.170 0.243 0.101 50.698 0.162 1.507 0.009 0.032 0.079 0.148 0.027 1.266 0.948 9.066 13.477 0.003 0.017 0.019 0.005 0.051 0.091 0.017 16.485 ' Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood:=7.4%, fat==7%, muscle=40.4%, bone=7.3% of body weight. - 196- OO137 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity at Tmax/2 in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male Timepoint 405M 406M 407M 408M Mean SD prostate skin* whole blood* brain fat* heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle* bone* 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171b 171 h 171 h 171 h Total 0.030 5.843 7.624 0.021 0.438 0.174 0.322 0.045 37.037 0.841 0.649 0.203 0.003 0.036 0.301 0.008 4.460 1.161 59.196 0.028 6.460 9.975 0.020 0.753 0.210 0.404 0.053 30.266 1.329 1.332 0.244 0.002 0.075 0.375 0.010 6.089 1.229 58.854 0.032 5.939 8.108 0.025 0.535 0.220 0.323 0.039 34.029 1.256 1.105 0.249 0.009 0.037 0.378 0.011 4.681 1.068 58.044 0.030 6.002 7.219 0.020 0.645 0.175 0.313 ,0.041 29.176 1.134 0.835 0.260 0.003 0.056 0.368 0.009 4.704 1.020 52.010 0.030 6.061 8.232 0.022 0.593 0.195 0.341 0.045 32.627 1.140 0.980 0.239 0.004 0.051 0.356 0.010 4.984 1.120 57.026 0.002 0.274 1.218 0.002 0.136 0.024 0.043 0.006 3.601 0.215 0.300 0.025 0.003 0.018 0.037 0.001 0.745 0.094 3.379 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fal=7%, muscle=40.4%, bone=7.3% of body weight. Female skin* whole blood* brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle* bone* Timepoint_______ 413F__________ 414F 4 h 0.270 0.396 4 h 3.406 6.470 4 h 0.047 0.040 4 h 0.070 0.141 4 h 0.252 0.316 4 h 0.512 0.667 4 h 0.050 0.060 4b 4.571 7.984 4 h 2.047 3.147 4h 2.950 13.010 4h 1.158 5.193 4 h 0.006 0.047 4 h 0.079 0.027 4 h 0.057 0.045 4b 0.022 0.020 4h 0.322 0.289 4 h 0.262 0.290 4 b 0.116 0.226 Total 16.197 38.368 415F 0.456 4.995 0.059 0.186 0.261 0.519 0.063 7.582 2.192 2.781 2.731 0.005 0.075 0.049 0.018 0.126 0.741 0.115 22.954 416F 0.488 2.882 0.043 0.259 0.183 0.484 0.058 7.131 3.688 15.108 6.480 0.006 0.049 0.042 0.013 0.296 0.204 0.155 37.569 Mean 0.403 4.438 0.047 0.164 0.253 0.546 0.058 6.817 2.769 8.462 3.891 0.016 0.058 0.048 0.018 0.258 0.374 0.153 28.772 SD 0.096 1.625 0.008 0.079 0.055 0.082 0.006 1.537 0.784 6.519 2.395 0.021 0.024 0.006 0.004 0.089 0.247 0.052 10.976 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. - 197- OOOlS Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (jig equiv/g) at with 1 mg/kg 14C-PFOA in tissues in male and female rats following oral gavage Male Timepoint prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes adrenals muscle bone 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 b 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 401M 0.473 0.760 2.586 0.058 0.240 0.978 1.074 0.319 4.808 1.512 0.397 0.191 0.419 0.325 0.578 0.482 0.252 0.357 Female Timepoint skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle bone 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 409F 0.336 2.040 0.042 0.149 0.484 0.879 0.246 1.964 3.585 0.750 7.648 0.256 0.217 0.438 0.319 0.744 0.134 0.160 402M 0.306 0.537 2.607 0.061 0.283 0.854 1.128 0.325 4.814 1.739 0.435 0.223 0.279 0.275 0.558 0.466 0.260 0.386 410F 0.441 2.355 0.038 0.228 0.635 0.855 0.267 2.294 3.879 0.754 1.575 0.370 0.225 0.652 0.287 0.384 0.161 0.245 403M 0.572 0.673 2.601 0.083 0.350 1.200 1.379 0.422 3.589 1.464 0.734 0.250 0.489 0.279 0.560 0.725 0.266 0.461 404M 0.464 0.718 2.553 0.062 0.251 0.862 1.048 0.289 5.298 2.126 0.547 0.209 0.410 0.230 0.465 0.452 0.250 0.342 Mean 0.454 0.672 2.587 0.066 0.281 0.973 1.157 0.339 4.627 1.710 0.528 0.218 0.399 0.277 0.540 0.531 0.257 0.386 SD 0.110 0.097 0.024 0.012 0.050 0.161 0.151 0.058 0.729 0.302 0.151 0.025 0.088 0.039 0.051 0.130 0.008 0.053 411F 0.188 1.242 0.028 0.081 0.314 0.521 0.152 1.788 2.863 5.365 15.001 0.540 0.143 0.314 0.172 0.240 0.091 0.162 412F 0.228 1.231 0.026 0.109 0.326 0.551 0.138 1.707 1.967 1.869 12.457 0.376 0.114 0.283 0.173 0.271 0.091 0.118 Mean 0.298 1.717 0.033 0.142 0.440 0.701 0.201 1.938 3.073 2.184 9.170 0.385 0.175 0.422 0.238 0.410 0.119 0.171 SD 0.114 0.569 0.008 0.064 0.151 0.192 0.065 0.260 0.852 2.185 5.910 0.117 0.055 0.167 0.076 0.231 0.035 0.053 - 198- OOOS9 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (jig equiv/g) at T ^ xn . in tissues in male and female rats following oral gavage with 1 mg/kg 14C-PFOA Male Timepoint prostate skin whole blood brain fat heart lungs spleen liver kidney G.L tract G.l. contents thyroid thymus testes adrenals muscle bone 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 405M 0.132 0.273 0.916 0.021 0.056 0.312 0.418 0.127 5.070 0.606 0.129 0.024 0.216 0.130 0.196 0.144 0.098 0.141 Fem ale Timepoint skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle bone 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 413F 0.491 1.530 0.041 0.141 0.512 0.803 0.211 1.379 1.991 0.609 0.577 0.238 0.321 0.422 0.296 0.663 0.130 0.172 406M 0.126 0.297 1.178 0.020 0.094 0.363 0.445 0.135 3.828 0.955 0.147 0.028 0.198 0.189 0.255 0.187 0.132 0.147 414F 0.461 1.990 0.037 0.187 0.632 1.000 0.269 2.538 3.144 2.522 2.074 4.430 0.089 0.558 0.375 0.494 0.181 0.207 407M 0.150 0.275 0.964 0.022 0.067 0.362 0.417 0.118 4.161 0.827 0.148 0.024 0.578 0.115 0.253 0.180 0.102 0.129 415F 0.337 1.626 0.049 0.149 0.507 0.810 0.215 2.020 2.587 0.529 1.024 0.244 0.295 0.447 0.323 0.286 0.306 0.183 408M 0.204 0.274 0.847 0.020 0.080 0.318 0.383 0.121 3.663 0.703 0.134 0.024 0.199 0.130 0.220 0.189 0.101 0.121 416F 0.371 1.866 0.037 0.150 0.482 0.799 0.213 2.319 3.868 2.610 2.880 0.244 0.171 0.434 0.240 0.817 0.131 0.192 Mean 0.153 0.280 0.976 0.021 0.074 0.339 0.416 0.125 4.180 0.773 0.140 0.025 0.298 0.141 0.231 0.175 0.108 0.135 Mean 0.415 1.753 0.041 0.157 0.533 0.853 0.227 2.064 2.897 1.567 1.639 1.289 0.219 0.465 0.308 0.565 0.187 0.188 SD 0.035 0.011 0.143 0.001 0.017 0.027 0.025 0.008 0.628 0.151 0.010 0.002 0.187 0.033 0.029 0.021 0.016 0.012 SD 0.073 0.212 0.005 0.020 0.067 0.098 0.028 0.503 0.800 1.154 1.039 2.094 0.109 0.063 0.056 0.228 0.083 0.015 -199- 000200 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix O Tissue Distribution at Tma* and Tniax/2 in Male and Female Rats Following Oral Gavage with 5 mg/kg 14C-PFOA - 200- 000201 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/g): 1.171 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 5.0 DuPont-7473 Male Animal Tin* 417M 418M 419M 420M Mean Standard Deviation Tpw/2 421M 422M 423M 424M Mean Standard Deviation Animal Weight Dose Administered Actual Dose Rate __________ (g)___________________ E)_______ ___________<ngfltg)_________ 233.2 220.1 232.4 230.8 229.1 6.1 0.94 0.88 0.93 0.95 0.92 0.03 4.727 4.667 4.667 4.811 4.718 0.068 216.2 234.5 223.4 226.4 225.1 7.6 0.87 0.94 0.89 0.92 0.90 0.03 4.708 4.690 4.688 4.739 4.706 0.024 Radioactivity Received (mC) 28.944 26.976 28.483 29.160 28.391 0.985 26.730 28.883 27.499 28.175 27.822 0.922 Female Animal T|ttn 425F 426F 427F 428F Animal Weight Dose Administered _______________ (g) __________ (g)__________ 187.7 190.6 194.5 200.1 0.75 0.76 0.78 0.79 Mean Standard Deviation 193.2 5.4 0.77 0.02 T max/2 429F 430F 431F 432F 204.9 184.7 184.8 194.8 0.83 0.74 0.75 0.77 Mean Standard Deviation 192.3 9.6 0.78 0.04 Actual Dose Rate (mg/kg) 4.687 4.677 4.722 4.642 4.682 0.033 4.751 4.718 4.760 4.654 4.721 0.048 Radioactivity Received ____________ ifii_____________ 23.100 23.408 24.115 24.392 23.754 0.601 25.563 22.885 23.100 23.808 23.838 1.214 i .H - 201- 000202 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity at Tma* in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male Timepoint 417M 418M 419M 420M Mean SD prostate skin* whole blood* brain fat* heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle* bone* 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m Total 0.020 14.877 23.248 0.053 3.046 0.400 0.753 0.101 22.078 1.833 1.984 2.699 0.019 0.101 0.638 0.020 13.827 3.445 89.142 0.049 14.875 27.499 0.061 2.919 0.459 0.846 0.117 16.355 1.912 2.644 1.316 0.008 0.080 0.899 0.017 12.851 3.243 86.150 0.123 16.766 23.612 0.068 2.773 0.485 0.740 0.089 18.794 2.372 1.941 3.031 0.009 0.086 0.759 0.025 14.188 3.257 89.118 0.090 15.742 25.317 0.020 2.521 0.426 0.034 0.077 17.773 2.563 3.463 3.480 0.007 0.073 0.475 0.024 13.394 2.242 87.721 0.071 15.565 24.919 0.051 2.815 0.443 0.593 0.096 18.750 2.170 2.508 2.632 0.011 0.085 0.693 0.022 13.565 3.047 88.033 0.045 0.899 1.942 0.021 0.225 0.037 0.376 0.017 2.434 0.354 0.713 0.934 0.006 0.012 0.180 0.004 0.576 0.544 1.420 * Percent recovery scaled to whole animal assuming the following: skin=I9%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. Female Timepoint 425F 426F 427F 428F Mean SD skin* whole blood* brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle* bone* 1 h 15 m 1hl5m 1 h 15 m 1 h 15m 1 h 15 m 1 h 15 m 1 h 15 m 1hl5m 1 h 15 m 1 h 15 m 1h 15m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1h 15 m 1 h 15 m I h 15 m 0.786 9.284 0.086 0.154 0.425 0.863 0.093 11.414 3.676 3.863 0.722 0.011 0.103 0.079 0.022 0.336 0.335 0.146 0.444 6.015 0.042 0.196 0.308 0.683 0.076 9.858 3.825 9.744 4.915 0.006 0.087 0.056 0.022 0.318 0.300 0.191 0.665 10.381 0.077 0.148 0.431 0.923 0.122 14.190 4.289 6.376 1.086 0.009 0.081 0.082 0.029 0.315 0.404 0.114 0.600 6.675 0.060 0.383 0.388 0.838 0.113 9.296 5.380 8.584 4.862 0.007 0.147 0.066 0.031 0.331 0.375 0.246 0.624 8.089 0.066 0.220 0.388 0.827 0.101 11.190 4.293 7.142 2.896 0.008 0.105 0.071 0.026 0.325 0.354 0.174 0.142 2.080 0.019 0.111 0.057 0.102 0.021 2.192 0.771 2.594 2.305 0.002 0.030 0.012 0.005 0.010 0.046 0.057 Total 32.398 37.086 39.722 38.382 36.897 3.187 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood==7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. - 202- j 000203 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity at Tmax/2 in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male Timepoint 421M 422M 423M 424M Mean SD prostate skin* whole blood* brain fat* heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus testes adrenals muscle* bone* 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 0.045 7.317 10.510 0.012 0.670 0.245 0.075 0.062 27.111 1.064 0.790 0.255 0.005 0.052 0.400 0.011 5.754 1.180 0.071 7.072 10.810 0.025 1.138 0.235 0.339 0.053 24.563 1.222 1.238 0.240 0.002 0.049 0.445 0.008 6.018 1.287 0.077 6.761 11.936 0.030 0.839 0.296 0.522 0.057 24.990 1.215 0.997 0.304 0.007 0.054 0.310 0.008 6.822 1.535 0.035 7.780 11.305 0.024 1.016 0.232 0.441 0.069 24.260 1.346 1.181 0.282 0.003 0.056 0.332 0.009 7.123 1.496 0.057 7.233 11.140 0.023 0.916 0.252 0.344 0.060 25.231 1.212 1.052 0.270 0.004 0.053 0.372 0.009 6.429 1.375 0.020 0.430 0.624 0.008 0.205 0.030 0.194 0.007 1.289 0.115 0.202 0.028 0.002 0.003 0.062 0.001 0.648 0.169 Total 55.558 54.815 56.760 56.990 56.031 1.025 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood==7.4%, fal=7%, muscle=40.4%. bone=7.3% of body weight. Female Timepoint 429F 430F 431F 432F Mean SD skin* whole blood* brain fat heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle* bone* 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h Total 0.336 6.530 0.045 0.040 0.275 0.529 0.055 7.010 2.439 4.052 0.352 0.003 0.053 0.043 0.013 0.229 0.231 0.101 22.336 0.326 4.650 0.054 0.131 0.250 0.638 0.072 7.008 1.970 5.558 2.739 0.007 0.074 0.051 0.020 0.199 0.314 0.212 24.273 0.138 3.722 0.031 0.072 0.161 0.345 0.045 6.163 2.621 21.515 14.420 0.008 0.050 0.023 0.010 0.214 0.160 0.053 49.751 0.426 6.740 0.051 0.196 0.259 0.767 0.068 8.522 3.710 1.893 4.894 0.004 0.094 0.046 0.016 0.272 0.284 0.203 28.445 0.307 5.411 0.045 0.110 0.236 0.570 0.060 7.176 2.685 8.255 5.601 0.006 0.068 0.041 0.015 0.229 0.247 0.142 31.201 0.121 1.466 0.010 0.069 0.051 0.179 0.012 0.982 0.736 8.967 6.165 0.002 0.021 0.012 0.004 0.031 0.068 0.078 12.626 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood==7.4%, fat==7%, muscle=40.4%, bone=7.3% of body weight. H f -203- 000204 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/ig equiv/g) at Tnm in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male Timepoint prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes adrenals muscle bone 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 417M_______418M 0.291 3.701 14.849 0.282 2.056 4.850 6.579 1.847 24.514 8.938 2.449 1.698 4.105 2.274 2.567 2.736 1.618 2.230 1.792 3.654 17.344 0.316 1.947 5.682 6.683 2.036 20.607 9.924 2.586 1.047 2.362 1.890 2.943 3.289 1.485 2.073 419M 3.041 4.118 14.892 0.346 1.849 5.550 6.610 1.973 21.231 11.480 2.425 1.902 2.387 1.576 2.742 3.332 1.639 2.082 420M 2.926 3.986 16.461 0.297 1.733 5.461 8.110 2.199 27.471 13.730 3.695 2.352 3.315 1.948 2.764 4.441 1.595 1.478 Mean 2.012 3.865 15.886 0.310 1.896 5.386 6.996 2.014 23.456 11.018 2.789 1.750 3.042 1.922 2.754 3.449 1.584 1.966 SD 1.278 0.224 1.227 0.028 0.138 0.369 0.744 0.146 3.178 2.089 0.609 0.542 0.836 0.286 0.154 0.714 0.069 0.333 F em ale Timepoint skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle bone 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 425F 2.776 13.308 0.402 1.236 5.117 7.439 1.792 16.406 20.184 4.674 1.357 2.414 1.602 3.712 3.122 4.717 1.053 1.560 426F 2.232 11.813 0.237 0.902 3.446 5.688 1.327 16.200 22.608 8.584 7.769 2.034 1.614 3.083 1.905 3.327 0.835 1.044 427F 2.717 15.618 0.371 1.428 5.073 7.518 1.902 21.643 30.506 8.056 1.240 2.467 1.666 3.990 3.065 4.713 1.109 1.253 428F 3.387 14.816 0.309 1.513 4.370 7.324 1.876 13.487 30.869 10.441 9.079 2.641 1.876 4.047 3.150 7.061 1.269 1.584 Mean 2.778 13.889 0.330 1.270 4.501 6.992 1.725 16.934 26.042 7.938 4.861 2.389 1.689 3.708 2.810 4.955 1.067 1.360 SD 0.474 1.683 0.073 0.271 0.783 0.873 0.269 3.409 5.457 2.405 4.149 0.256 0.127 0.442 0.605 1.549 0.179 0.259 -204- r- > 0002C5 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (/tg equiv/g) at Tmax/2 in tissues in male and female rats following oral gavage with 5 mg/kg 14C-PFOA Male Timepoint 421M prostate skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus testes adrenals muscle bone 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 1.051 1.813 6.687 0.132 0.450 2.285 2.743 0.808 17.572 3.740 0.844 0.139 1.142 0.853 1.192 ' 1.157 0.671 0.761 Female Timepoint skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle bone 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 429F 2.479 9.103 0.246 0.936 3.213 4.703 1.144 11.674 15.742 4.921 0.637 2.023 0.986 2.540 2.122 4.998 0.686 1.145 422M 1.227 1.746 6.852 0.180 0.763 2.447 2.679 0.887 17.541 4.463 0.873 0.168 1.514 1.064 1.548 1.253 0.699 0.827 430F 2.921 9.556 0.318 0.793 3.285 5.630 1.337 11.509 12.594 4.924 11.051 2.323 1.492 3.635 2.405 4.197 0.939 1.223 423M 1.003 1.668 7.561 0.164 0.562 2.742 3.841 0.945 20.646 4.515 1.100 0.197 1.745 1.394 1.063 0.847 0.792 0.986 431F 1.458 7.136 0.133 0.639 2.030 3.077 0.911 9.767 14.933 27.291 30.066 2.174 0.983 1.986 1.609 3.318 0.461 0.547 424M 0.846 1.941 7.240 0.144 0.688 2.294 3.142 0.831 17.207 5.119 1.035 0.195 2.206 1.258 1.172 1.202 0.836 0.971 432F 3.270 10.466 0.269 1.107 3.026 5.867 1.403 12.229 19.679 2.482 7.052 1.439 1.475 3.359 2.076 5.799 0.982 1.364 Mean 1.032 1.792 7.085 0.155 0.616 2.442 3.101 0.868 18.242 4.459 0.963 0.175 1.652 1.142 1.244 1.115 0.749 0.886 Mean 2.532 9.065 0.241 0.868 2.889 4.819 1.199 11.295 15.737 9.904 12.202 1.990 1.234 2.880 2.053 4.578 0.767 1.070 SD 0.157 0.116 0.393 0.021 0.138 0.213 0.534 0.061 1.611 0.564 0.124 0.027 0.445 0.236 0.211 0.183 0.077 0.110 SD 0.786 1.406 0.078 0.200 0.583 1.265 0.221 1.064 2.948 11.648 12.659 0.387 0.288 0.756 0.330 1.065 0.242 0.360 -205 - 000200 Periluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Appendix P Tissue Distribution at Tni2Xand Tma/2 in Male and Female Rats Following Oral Gavage with 25 mg/kg 14C-PFOA OV/-. -206- 000207 Perfluorooctanoic Acid: Toxicokinetics in the Rat Dosing information for male and female rats Dose Concentration (mg/g) : 6.166 Nominal Dose Volume (mL/kg): 4.0 Nominal Dose Rate (mg/kg): 25.0 DuPont-7473 Male Animal Tpu 433M 434M 435M 436M Animal Weight ______________ (E> 226.3 211.6 214.1 212.9 Mean Standard Deviation 216.2 6.8 Tmu/2 437M 438M 439M 440M 216.5 215.3 229.3 212.1 Mean Standard Deviation 218.3 7.6 Dose Administered Actual Dose Rate (r) _______ Cmg^e)________ Radioactivity Received (Ci) 0.92 25.096 0.86 25.090 0.87 24.970 0.86 24.937 0.88 25.023 0.03 0.082 24.972 23.345 23.508 23.345 23.792 0.790 0.87 24.864 0.88 25.060 0.93 25.036 0.85 24.828 0.88 24.947 0.03 0.118 23.670 23.724 25.243 23.155 23.948 0.900 Female Animal Tom 441F 442F 443F 444F Mean Standard Deviation Xnwx/2 445F 446F 447F 448F Mean Standard Deviation Animal Weight __________ ()________ Dose Administered Actual Dose Rate Radioactivity Received (8)__________ _______ (mg/kg)________ ___________ (Mi)____________ 189.4 194.7 188.6 188.5 190.3 3.0 0.78 0.78 0.76 0.76 0.77 0.01 25.297 24.735 24.685 24.894 24.902 0.278 21.067 21.176 20.471 20.634 20.837 0.338 187.2 194.1 194.9 196.4 193.2 4.1 0.76 0.78 0.79 0.79 0.78 0.02 24.902 24.875 25.026 24.803 24.901 0.093 20.498 21.230 21.447 21.420 21.149 0.444 ? ?* -207- 000208 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity at T^x in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Tiznepoint 433M 434M 435M 436M Mean SD prostate skin* whole blood* brain fat* heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus testes adrenals muscle* bone* 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m Total 0.093 12.804 23.459 0.072 2.748 0.431 0.994 0.120 16.443 2.178 2.107 2.927 0.011 0.129 0.559 0.024 13.506 3.363 81.968 0.064 15.036 24.118 0.057 2.178 0.501 0.802 0.113 17.822 2.654 3.559 4.029 0.007 0.126 0.696 0.031 13.583 3.462 88.838 0.058 14.136 21.382 0.063 1.904 0.509 0.762 0.085 18.576 1.982 2.885 6.087 0.007 0.142 0.520 0.023 12.496 2.908 84.525 0.053 13.367 22.659 0.059 1.782 0.402 0.893 0.104 17.272 2.359 2.583 3.702 0.009 0.084 0.716 0.025 11.836 2.513 80.418 0.067 13.836 22.905 0.063 2.153 0.461 0.863 0.106 17.528 2.293 2.784 4.186 0.009 0.120 0.623 0.026 12.855 3.062 83.937 0.018 0.969 1.177 0.007 0.430 0.053 0.103 0.015 0.900 0.286 0.608 1.349 0.002 0.025 0.098 0.004 0.841 0.438 3.680 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood==7.4%, fat=7%, muscle=40.4%, bone=7.3% of body weight. Female Timepoint 441F 442F 443F 444F Mean SD skin* whole blood* brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle* bone* 1h 15m 1h 15m 1hl5m 1hl5m 1h 15m 1h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m Total 0.222 8.178 0.060 0.110 0.342 0.692 0.091 9.798 6.648 5.309 2.845 0.008 0.104 0.084 0.029 0.438 0.209 0.093 35.260 0.386 4.221 0.049 0.168 0.304 0.580 0.086 9.253 4.828 8.941 3.229 0.005 0.070 0.056 0.028 0.410 0.347 0.149 33.110 0.303 9.428 0.068 0.097 0.273 0.722 0.086 10.023 5.302 8.036 3.666 0.008 0.060 0.076 0.030 0.597 0.363 0.126 39.264 0.608 6.803 0.055 0.212 0.348 0.718 0.101 13.076 6.689 5.405 0.225 0.013 0.130 0.067 0.038 0.319 0.513 0.259 35.579 0.380 7.158 0.058 0.147 0.317 0.678 0.091 10.538 5.867 6.923 2.491 0.009 0.091 0.071 0.031 0.441 0.358 0.157 35.803 0.166 2.232 0.008 0.053 0.035 0.067 0.007 1.723 0.946 1.846 1.548 0.003 0.032 0.012 0.005 0.116 0.124 0.072 2.554 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood:=7.4%, fat==7%, muscle=40.4%, bone=7.3% of body weight. -208- 000203 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Percent recovery of radioactivity at Tmax/2 in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Timepoint 437M 438M 439M 440M Mean SD prostate skin* whole blood* brain fat* heart lungs spleen liver kidney G.I. tract G.l. contents thyroid thymus testes adrenals muscle* bone" 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h Total 0.034 5.384 8.988 0.020 0.781 0.207 0.367 0.046 24.179 1.135 0.915 0.287 0.006 0.055 0.252 0.013 4.773 0.943 48.385 0.014 5.100 7.418 0.020 0.607 0.141 0.317 0.036 20.536 1.038 1.018 0.163 0.005 0.051 0.247 0.009 4.184 1.005 41.909 0.040 5.635 8.500 0.019 0.606 0.170 0.298 0.045 19.067 0.996 0.631 0.273 0.004 0.039 0.207 0.007 3.961 0.769 41.267 0.022 5.557 6.709 0.015 0.518 0.139 0.230 0.041 16.799 0.843 0.667 0.116 0.003 0.035 0.189 0.005 4.095 0.905 36.888 0.028 5.419 7.904 0.019 0.628 0.164 0.303 0.042 20.145 1.003 0.808 0.210 0.005 0.045 0.224 0.009 4.253 0.906 42.112 0.012 0.237 1.032 0.002 0.110 0.032 0.057 0.005 3.098 0.122 0.189 0.084 0.001 0.010 0.031 0.003 0.358 0.100 4.740 * Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.4%, bone=73 % of body weight. Female Timepoint_______ 445F__________446F__________ 447F__________448F_________ Mean__________ SD skin* whole blood* brain fat heart lungs spleen liver kidney G.l. tract G.l. contents thyroid thymus ovaries adrenals uterus muscle" bone* 4h 4b 4h 4h 4h 4h 4h 4h 4h 4h 4h 4b 4h 4h 4h 4h 4h 4h Total 0.238 7.097 0.055 0.091 0.282 0.803 0.072 8.299 4.990 4.880 2.590 0.009 0.080 0.055 0.022 0.435 0.338 0.104 30.440 0.505 4.332 0.056 0.209 0.376 1.020 0.071 9.076 4.942 4.357 0.535 0.008 0.104 0.084 0.021 0.195 0.406 0.305 26.602 0.302 7.408 0.083 0.091 0.283 0.753 0.068 10.334 4.903 2.914 0.967 0.004 0.060 0.074 0.020 0.289 0.352 0.125 29.030 0.614 6.790 0.039 0.199 0.208 0.524 0.067 8.611 4.162 2.036 0.391 0.007 0.062 0.068 0.020 0.297 0.363 . 0.189 24.647 0.415 6.407 0.058 0.148 0.287 0.775 0.070 9.080 4.749 3.547 1.121 0.007 0.077 0.070 0.021 0.304 0.365 0.181 27.680 0.175 1.406 0.018 0.065 0.069 0.204 0.002 0.895 0.393 1.306 1.010 0.002 0.020 0.012 0.001 0.099 0.029 0.090 2.569 7 Percent recovery scaled to whole animal assuming the following: skin=19%, whole blood=7.4%, fat=7%, muscle=40.49b, bone=7.3Sb of body weight. -209 - 000210 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (n g equiv/g) at Tma, in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Tiroepoint prostate skin whole blood brain fat heart lungs spleen liver kidney G.l. tract G.I. contents thyroid thymus testes adrenals muscle bone 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 10 h 30 m 433M_______ 434M 16.536 16.911 79.555 2.028 9.853 25.768 39.555 10.878 104.969 54.316 13.998 10.379 14.447 10.679 15.564 19.135 8.390 11.560 10.556 19.856 81.775 1.727 7.806 29.032 35.354 9.963 104.531 65.723 19.390 12.655 9.435 11.774 15.111 18.001 8.436 11.901 435M 12.205 18.578 72.151 1.731 6.792 27.495 34.800 9.519 96.916 53.015 17.098 19.618 10.757 7.675 12.606 14.601 7.724 9.946 436M 8.895 17.544 76.359 1.744 6.348 25.962 38.938 10.589 112.946 62.540 15.253 14.793 17.845 9.400 14.049 17.347 7.306 8.586 Mean 12.048 18.222 77.460 1.807 7.700 27.064 37.162 10.237 104.841 58.898 16.435 14.361 13.121 9.882 14.333 17.271 7.964 10.498 SD 3.283 1.288 4.180 0.147 1.560 1.522 2.431 0.612 6.548 6.203 2.346 3.941 3.797 1.762 1.314 1.927 0.546 1.534 Fem ale Timepoint skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle bone 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 1 h 15 m 441F 16.938 72.495 1.458 6.606 22.800 35.975 10.116 91.459 156.202 30.144 41.312 12.153 11.542 16.842 13.847 19.232 5.131 6.791 442F 13.873 68.907 1.264 5.883 19.379 29.665 8.980 78.829 139.072 46.167 55.170 9.705 10.574 17.827 14.588 32.332 5.050 5.857 443F 16.616 71.931 1.703 11.868 20.280 34.466 9.289 97.762 169.866 53.284 29.353 17.651 9.866 20.687 15.937 24.272 5.224 6.928 444F 20.719 79.054 1.476 9.355 22.636 36.153 10.955 102.318 171.518 27.417 9.838 22.662 15.270 21.164 18.536 19.429 6.349 6.609 Mean 17.036 73.097 1.475 8.428 21.274 34.065 9.835 92.592 159.165 39.253 33.918 15.543 11.813 19.130 15.727 23.816 5.439 6.546 SD 2.814 4.272 0.179 2.738 1.709 3.029 0.888 10.198 15.051 12.486 19.210 5.794 2.405 2.121 2.063 6.137 0.611 0.478 - 210- OOQ^J.1 Perfluorooctanoic Acid: Toxicokinetics in the Rat DuPont-7473 Concentration (fig equiv/g) at Tmax^ in tissues in male and female rats following oral gavage with 25 mg/kg 14C-PFOA Male Timepoint prostate skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus testes adrenals muscle bone 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 171 h 437M_______ 438M 4.283 7.046 30.200 0.561 2.774 9.524 11.723 3.221 56.472 20.305 4.008 0.629 7.776 3.371 4.662 4.914 2.938 3.213 5.072 6.726 25.120 0.592 2.175 7.347 11.473 3.031 59.199 21.206 3.406 0.598 12.532 3.920 4.424 4.993 2.596 3.449 439M 3.680 7.425 28.758 0.549 2.166 8.858 10.834 3.101 57.258 18.655 4.150 0.907 5.498 3.579 3.820 4.941 2.455 2.639 440M 3.004 7.261 22.510 0.444 1.838 7.039 8.427 2.677 45.887 16.448 2.619 0.466 5.297 2.815 3.182 3.795 2.516 3.078 Mean 4.010 7.114 26.647 0.536 2.238 8.192 10.614 3.007 54.704 19.153 3.546 0.650 7.776 3.421 4.022 4.661 2.626 3.095 SD 0.880 0.302 3.490 0.064 0.390 1.192 1.505 0.234 5.989 2.090 0.697 0.186 3.364 0.463 0.663 0.578 0.216 0.341 F em ale Timepoint skin whole blood brain fat heart lungs spleen liver kidney G.I. tract G.I. contents thyroid thymus ovaries adrenals uterus muscle bone 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 4h 445F 13.924 61.232 1.488 5.748 18.813 36.513 8.529 69.455 139.298 53.753 31.034 11.702 7.010 17.229 12.446 19.110 4.643 6.286 446F 19.713 60.929 1.437 7.054 22.809 35.611 8.108 65.237 134.958 22.141 8.983 24.562 8.218 21.763 10.894 19.728 6.105 8.955 447F 18.605 65.639 2.184 5.817 16.459 34.476 8.977 78.901 141.409 15.616 6.624 16.474 7.760 15.614 14.362 27.286 5.470 7.238 ISO ON NIV1NOO 448F 14.371 52.779 1.095 6.008 14.601 23.884 7.924 75.252 121.609 9.548 3.804 16.897 6.703 20.279 11.972 20.002 4.971 5.094 Mean 16.653 60.145 1.551 6.157 18.171 32.621 8.385 72.211 134.318 25.265 12.611 17.409 7.423 18.721 12.419 21.531 5.297 6.893 SD 2.934 5.361 0.457 0.608 3.540 5.884 0.469 6.062 8.888 19.676 12.463 5.319 0.691 2.802 1.449 3.854 0.637 1.631 -211 - 00021%