Document 2j5p7m3513RgNBxKG0GvdD9d5

Perfluorooctanesulfoinate: PK During Pregnancy in Recovery + Analytical Protocol I i I I Li FINAL REPORT PROTOCOL 418-015 ORAL (GAVAGE) PHARMACOKINETIC RECOVERY IN RATS STUDY OF PFOS SPONSOR'S STUDY NUMBER: T-6295.14 FINAL REPORT DATE: 23 JULY 1999 PROTOCOL 418-015 ORAL (GAVAGE) PHARMACOKINETIC RECOVERY IN RATS STUDY OF PFOS SPONSOFRS STUDY NUMBER: T-6295.14 TABLE OF CONTENTS SUBJECT PAGE 1. SUMMARY AND CONCLUSION 1-1 A. Methods 1-1 B. Results 1-3 C. Conclusion 1-4 li. DESCRIPTION OF TEST PROCEDURES 11-1 A. Conduct ofStudy 11-1 A.l. Sponsor 11-1 A.2. TestingFacility 11-1 A.3. Study Number 11-1 A.4. Sponsoes Study Number 11-1 A.5. Purpose ofthe Study 11-1 A.6. Study Design 11-1 SUBJECT A.7. RegulatoryCompliance A.8. OwnershipoftheStudy A.9. Study Monitor A.10. AlternateStudy Monitor A.11. Study Director A.12. TechnicalPerformance A.13. Report Preparation A.14. ReportReview A.15. Date ProtocolSigned A.16. Dates ofTechnicalPerformance A.17. Records Maintained B. Test ArticlIenformation B.1. Description B.2. LoVBatch Number B.3. Date Received and Storage Conditions B.4. SpecialHandlingInstructions B.5. AnalysisofActivity C. VehicleInformation C.l. Description C.2. Lot Numbers C-3. Dates Received,Source and StorageConditions C.4. SpecialHandlingInstructions ii PAGE 11-1 11-2 11-2 11-2 11-2 11-2 11-2 11-2 11-2 11-3 11-3 11-3 11-3 11-3 11-4 11-4 11-4 11-4 11-4 11-4 11-4 11-4 .SUBJECT C.5. AnalysisofPurity D. Test ArticlePreparationand StorageConditions D.l. Sample Information D.2. AnalyticaRlesults E. Test System E.l. Species E.2. Strain E.3. Supplier(Source) E.4. Sex E.5. RationaleforTest System E.6. Test System Data E.7. Breeder Male Rat Data E.8. Method of Randomization E.9. System of Identification F. Husbandry F.l. Research FacilitRyegistration F.2. Study Rooms F.3. Housing F.4. Lighting F.5. Sanitization F.6. Feed F.7. Feed Analysis PAGE 11-5 11-5 11-5 11-5 11-5 11-5 11-6 11-6 11-6 11-6 11-6 11-6 11-6 11-7 11-7 11-7 11-7 11-7 11-8 11-8 11-8 11-8 SUBJECT F.8. Water F.9. Water Analysis F.10. Bedding F.11. Bedding Analysis G. Methods G.l. Dosage Administration G.2. RationaleforDosage Selection G.3. Route ofAdministration G.4. RationaleforRoute ofAdministration G.5. Frequency of Administration G.6. Length of Study G.7. Method ofStudy Performance G.8. PharrnacokineticSample Collection G.9. Gross Necropsy G. 10. StatisticAanlalyses 111. RESULTS A. MortalityC,linicaalnd Necropsy Observations A.l. Mortality A.2. ClinicaOlbservations A.3. Necropsy Observations B. Body Weights and Body Weight Changes B.1. Precohabitation iv PAGE 11-8 11-8 11-9 11-9 11-9 11-9 11-9 11-10 11-10 11-10 11-10 11-11 11-12 11-13 11-14 111-1 111-1 111-1 111-1 111-1 111-1 111-1 SUBJECT B.2. Gestation PAGE 111-2 B.3. Lactation 111-2 C. Absolut(eg/daya)nd Relati(vge/kg/daFye)edConsumptioVnalues 111-2 C.1. Precohabitation 111-2 C.2. Gestation 111-2 C.3. Lactation 111-3 D. NaturaDlelivearnydLiftOebrservations 111-3 E. ClinicOablservatiofnrsomBirtthoDay21 Postpartuamnd NecropsOybservations 111-3 REFERENCES 111-4 APPENDIXA -REPORT FIGURE Figur1e. BodyWeight-s Fo GeneratioFnemaleRats A-1 APPENDIX B -REPORT TABLES Table1. ClinicOablservatio-nSsummary-Fo Generation FemaleRats B-1 Table2. NecropsOybservatio-nSsummary-Fo Generation FemaleRats B-4 Table3. BodyWeights-Precohabitat-iSounmmary-Fo Generation FemaleRats B-5 Table4. BodyWeightChanges-Precohabfta-tSiuomnmary- Fo GeneratioFnemaleRats B-6 Table5. MatemalBodyWeights-Gestati-oSnummary- Fo GeneratioFnemaleRats B-7 Table6. MatemalBodyWeightChanges-Gestatio-nSummary- Fo GeneratioFnemaleRats B-9 v SUBJECT Table 7. Matemal Body Weights - Lactation- Summary Fo Generation Female Rats PAGE B-10 Table 8. Matemal Body Weight Changes - Lactation- Summary - Fo Generation Female Rats B-1 2 Table 9. Absolute Feed Consumption Values (g/day)Precohabitation-Summary - Fo Generation Female Rats B-13 Table 10. RelativeFeed Consumption Values (g/kg/day-) Precohabitation-Summary - Fo GenerationFemale Rats B-14 Table 11. Matemal AbsoluteFeed Consumption Values (g/day)Gestation- Summary - Fo GenerationFemale Rats B-1 5 Table 12. Matemal RelativeFeed Consumption Values (g/kg/day)- Gestation- Summary - Fo GenerationFemale Rats B-16 Table 13. Matemal AbsoluteFeed Consumption Values (g/day)- Lactation- Summary - Fo Generation Female Rats B-1 7 Table 14. Matemal RelativeFeed Consumption Values (g/kg/day)Lactation- Summary - Fo Generation Female Rats B-18 Table 15. NaturalDeliveryObservations-Summary - Fo Generation Female Rats B-1 9 Table 16. LitteOrbservations(NaturallDyeliveredPups) - Summary Fl Generation Lifters B-20 Table 17. ClinicaOlbservationsfrom Birthto Day 21 Postpartum Summary - Fl GenerationPups B-23 Table 18. Necropsy Observations-Summary - Fl GenerationPups B-24 Table 19. ClinicaOlbservations- IndividuaDlata - Fo Generation Female Rats B-25 Table 20. Necropsy Observations- IndividuaDlata - Fo Generation Female Rats B-28 vi SUBJECT Table21. Body Weights - Precohabitatio-nIndividuaDlata Fo GenerationFemale Rats PAGE B-30 Table 22. Matemal BoctyWeights - Presumed Gestation- Individual Data - Fo GerieratioFnemale Rats B-36 Table 23. Matemal Body Weights -Lactation-IndividuaDlata - Fo GenerationFemale Rats B-39 Table24. Feed Consumption Values - Precohabftatio-nIndividual Data - Fo GenerationFemale Rats B-42 Table 25. Matemal Feed Consumption Values- Presumed Gestation- IndividuaDlata - Fo GenerationFemale Rats B-44 Table 26. Matemal Feed Consumption Values - Lactation- Individual Data - Fo GenerationFemale Rats B-47 Table27. NaturalDeliveryI,mplantatioSnites,and Pup Viability and Sex - IndividuaDlata - Fo GenerationFemale Rats/ Fl GenerationLitters B-49 Table 28. Pup Body Weight LifteArverages from Birthto Day 21 Postpartum- IndividuaDlata - Fl GenerationLifters B-51 Table 29. Pup Body Weights from Birthto Day 21 Postpartum- IndividuaDlata - Fl GenerationPups B-53 Table 30. Pup VitalStatusand Sex from Birthto Day 21 Postpartum- IndividuaDlata - Fl GenerationPups B-65 Table 31. ClinicaOlbservationsfrom Birthto Day 21 Postpartum- IndividuaDlata - Fl GenerationPups B-67 Table 32. Necropsy Observations- IndividuaDlata - Fl Generation Pups B-68 APPENDIX C - PROTOCOL AND AMENDMENT C-1 to C-34 APPENDIX D - DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY D-1 vii SUBJECT APPENDIX E - TEMPERATURE REPORTS AND RELATIVE HUMIDITY APPENDIX F - STATEMENT OF THE STUDY DIRECTOR APPENDIX G - QUALITY ASSURANCE STATEMENT UNIT FINAL REPORT PAGE E-1 to E-9 F-1 G-1 toG-6 viii 418-015:PAGE 1-1 TITLE: ORAL (GAVAGE) PHARMACOKINETIC PFOS IN RATS RECOVERY STUDY OF ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-015 SPONSOR'S STUDY NUMBER: T-6295.14 1. SUMMARY AND CONCLUSION A. Methods' Twenty-fourpresumed pregnant CH:CD@BR VAF/Plus@ (Sprague-Dawley)rats were assigned tothreedosage groups (Groups Ithrough 111)e,ightratsper dosage group. The ratswere administeredthe testarticleP,FOS (FC-95),or the vehicle,0.5% Tween@ 80 inreverseosmosis membrane processed deionizedwater(R.O.deionizedwater),orall(yviagavage),once dailybeginning 43 days priorto cohabitationunticlonfirmedevidenced of mating. Dosages of 0 (Vehicle)0,.1 and 1.6mg/kg/day were administeredata dosage volume of 5 mukg. A.I. Fo Generation Rats: The female ratswere observed forviabiliattyleasttwiceeach day ofthe study. The ratswere examined forclinicaolbservationsof effectsofthe testarticle, abortions,premature deliverieasnd deaths beforeand approximatelyone hour afterdosage. Rats were observed forclinicaolbservationsand general appearance once dailyduringallotherperiodsofstudy. Body weightswere recordeddailyduringthe dosage and postdosage periodsand atsacrificeF.eed consumption valueswere recordedweekly priortocohabitationd,ailyduring gestationand on days 1,4, 7, 10 and 14 oflactatio(nDLs 1,4,7, 10 and 14). The female ratswere evaluatedfordurationofgestationl,iftesrizesand pup viabiliattybirth.Pups thateitherappeared stillbomor thatdied before initial examinationofthe litterfsorviabiliwteyre examined forvitalstatusat birth. Matemal behaviorofthe dams was evaluateddailywhen the pups were examined duringthe 21-day postpartum period.Observed maternalbehavior was recordedon DLs 1,4, 7, 10, 14 and 21. a. Detaileddescriptionosfallprocedures used inthe conduct ofthisstudy are providedinthe appropriatesectionsofthisreportand inAPPENDIX C (PROTOCOL AND AMENDMENT). 418-015:PAGE 1-? Urineand fecalsample were collectedfrom female ratsforthe following intervalso:ne day priorto initiatioofncohabitatiotno the followingmorning,days 6 to 7, 14 to 15 and 20 to21 of presumed gestation(DGs 6 to 7, 14 to 15 and 20 to21),and DLs 21 to22. Followingeach 24-hourcollectioinntervals,amples were shipped tothe Sponsor foranalysis.Blood samples were collectedfrom each ofthe matemal ratson the day cohabitatiownas initiate(dpriotro cohabitation)D,Gs 7, 15 and 21, and DLs 14 and 22. Serum samples were shipped tothe Sponsor foranalysis. Allsurvivingratsassigned tothe studywere sacrificeodn DL 22 followingthe finalcollectioinntervaflorurineand fecalsamples and blood sample collection and a gross necropsy ofthe thoracica,bdominal and pelvicviscerawas performed.Tissueswithgross lesionswere retained.The number and distributiofn implantatiosniteswas recorded.A liversectionfrom each dam was collectedand shipped tothe Sponsor foranalysis. A.2. Fl Generation Lifters: Day 1 of lactatio(npostpartum)was definedas the day ofbirthand was alsothe firsdtay on which allpups ina liftewrere individualwleyighed. The lifterwsere observed forviabiliattyleasttwiceeach day duringthe postpartum period. Lifterwsere observed forclinicaolbservationsand generalappearance once dailyduringthe postpartum period.The pups ineach liftewrere counted once daily.Body weightswere recordedon DLs 1 (birth4), 7, 14 and 21. Pups found dead were examined forgross lesionsand forthe cause ofdeath. For allpups found dead on DLs 2 to 4, alllungswere preserved. On DL 4, lifterwsere culledtofivemale pups and fivefemale pups per lifter, where possible.The lungs and the liverswere collectedfrom the firstten pups culleddeterminedto be atDL 4 from each dosage group (irrespectiovfelifter). The lungs and the liverswere individualrleytained.Remaining culledpups were sacrificeadnd discardedwithoutevaluation. Allremainingpups on studywere sacrificeadnd examined forgross lesionson DL 21. Gross lesionswere retained.The livefrrom each pup was collected, pooled (perliftearn)d shipped to the Sponsor foranalysis.Blood samples were collectedand pooled (perlifterS)e.rum samples were shipped tothe Sponsor foranalysis. B. Results 418-015:PAGE 1-3 No deaths or premature deliveriewsere attributetdo PFOS treatment. One vehiclecontrolgroup ratwas injuredand died afterorbitaslinusbleedingon DG 15. Allotherratssurvivedto scheduled sacrificeA.lladverse clinical observationsdudng the precohabitationg,estationand lactatiopneriodswere r,onsidereudnrelatedto thetestarticleA. red substance was found inthe thoraciccavityofthe ratthatdied. Allotherratsappeared normal atnecropsy. Rats administeredthe 1.6mg/kg/day dosage ofthe testarticlhead reduced body weightgains or body weight lossesineach week ofthe precohabitatiopneriod. Dams inthe 1.6 mg/kg/day dosage group had reduced body weight gains during thefirswteek ofthe gestationperiod(DGs 0 to 7). Body weightgainswere then increasedas compared tothe controlgroup value on DGs 7 to 10, 13 to 15 and 15 to 18. Reflectingthisrebound effectb,ody weight gainswere increasedfor the entiregestationperiod(DGs 0 to 20) inthe 1.6mg/kg/day dosage group,as compared tothe controlgroup value. Body weight gains fordams administered the 1.6mg/kg/day dosage ofthe testarticldeuringthe precohabitatiopneriod generallycontinuedto be increasedduringthe lactatiopneriod. Absolute(g/day)and relativ(eg/kg/dayf)eed consumptionvalueswere reduced inthe 1.6mg/kg/day dosage group ineach week ofthe precohabitation period.Dams inthe 1.6mg/kg/day dosage group had reduced absoluteand relativfeeed consumption valuesduringthe firswteek ofthe gestationperiod (DGs 0 to7). Feed consumption valueswere then generallycomparable to controlgroup values forthe remainderofthe gestationperiod.Absoluteand relativfeeed consumption values inthisdosage group were slightlryeduced,as compared tocontrolgroup values,forthe entiregestationperiod(DGs 0 to20). Absoluteand relativfeeed consumption valuesduringthe lactatiopnedod were reduced inthe groups administered0.1 or 1.6 mg/kg/day ofthetestarticldeuring the precohabitatiopneriod,however the reductionswere not strictdloysagedependent. Administratioonfthe testarticlaetdosages as highas 1.6mg/kg/day didnot adverselyaffectany parameter evaluatedat naturaldeliveryor dudng the 22-day lactatiopneriod.No clinicaolr necropsy observationsinthe Fl generationpups were attributablteo dosages ofthe testarticlaes high as 1.6mg/kg/day. 418-015:PAGE 1-4 C. Conclusion The purpose ofthe study,as statedinthe protocol,was to evaluatethe pharmaco kineticosf PFOS inFo generationpregnantfemale ratsfollowing PFOS treatmentduringthe pre-cohabitatiopneriod.This reportincludesstudy data from the in-lifpeortionwhich was conducted atArgus Research laboratoriesI,nc.The pharmacokineticsamples thatwere collecteduringthe in-lifpeortionof the study were sent tothe Sponsor foranalysisand willbe reportedseparately.Itisthe responsibiliotfythe Sponsor, 3M Corporate Toxicology,to combine the in-lifreesultswiththe analyticarlesultsintoa final pharmacokineticreport. MildredS. ChristianP,h.D.,Fellow,ATS ExecutiveDirectorof Research 23 -rv)9y Date Alan M. Hoberman, Ph.D.,DABT Directorof Research Date exond G. York@Ph@..,DABT Date Associate Directc e@ irchand Study Director 418-015:PAGE 11-1 li. DESCRIPTION OF TEST PROCEDURES A. Conduct of Stud A.l. Sponso: 3M Corporate Toxicoloigy,3M Center,Building220-2E-02, St.Paul, Minnesota 55144-1000 A.2. TestingFacilit Argus Research LaboratoriesI,nc.,905 Sheehy Ddve, BuildingA, Horsham, Pennsylvania 19044-1297 A.3. Study Number: 418-015 A.4. Sponsoes Study Number: T-6295.14 A.5. Purpose of the Study: The purpose ofthisstudy was toevaluatethe pharmacokineticsof PFOS in Fo generationand Fl generationratsdudng gestationand lactatiofnollowing cessationofPFOS treatmentof Cd:CD@)BR VAF/Plus@ female ratsat confirmed mating. A.6. Study Design: The requirementsofthe U.S. Food and Drug Administration(FDA)(')were used as the basisofstudy design. A.7. Regulato[y Comr)liance: The studywas conducted incompliance withGood LaboratoryPractice(GLP) regulationosfthe U.S. Food and Drug Administratio(nFDA)(2), the Japanese Ministryof Healthand Welfare (MHW)(1)and the European Economic Community (EEC)(4)T.here were no deviationsfrom the GLP regulationtshataffectedthe qualityor integritoyfthe study. QualityAssurance Unitfindingsderivedfrom the inspectionsduringthe conduct ofthisstudyare documented and have been providedtothe Study Directorand the TestingFacilitMyanagement. 418-015:PAGE 11-2 A.8. OwnershiR of the Stud The Sponsor owns the study. Allraw data, analyses, reportsand preserved tissuesare the propertyof the Sponsor. A.g. Study Monitor: Marvin T. Case, D.V.M., Ph.D. A.10. Alternate Study Monito Andrew M. Seacat, Ph.D. A.11. Study Director: Raymond G. York, Ph.D.,DABT (AssociateDirectorof Research) A.12. Technical Performance: John F. Barnett,B.S. (Directorof Laboratory Operations) Paul E. Ciott(iTeam Leader) Todd J.KillinoB,.S. (LaboratoryTechnician) A.13. Rer)ortPreg)aration: Raymond G. York,Ph.D.,DABT Jo Ann Frazee, M.S. (StudyCoordinator) Denise P. Gasiorowski (Data Management Specialist) Karen G. Parker,A.A. (ReportAdministrator) A.14. Report Review: MildredS. Christian,Ph.D., Fellow,ATS (ExecutiveDirectorof Research) Alan M. Hoberman, Ph.D, DABT (Directorof Research) A.15. Date Protocol Signed: 16 November 1998 418-015:PAGE 11-3 A.16. Dates of Technical Performance: Rat ArrivalDate Dosage Period [43days priorto the first day of cohabitationu'ntilconfirmed mating(DG 0)] CohabitationPeriod DG b 0 DeliveryPeriod(DLc 1) Sacrificoef Fo generationratsnot selected forcontinued evaluation DL 4 Culling(pups not selectedforcontinued observation) DG 25 Sacrifice(dam withno confirmeddate ofmating) DL 21 Scheduled Sacrific(eFl generationpups) DL 22 Scheduled Sacrific(eFo generationdams) 17 NOV 98 23 NOV 98 -08 JAN 99 04 JAN 99 PM - 09 JAN 99 AM 05 JAN 99 - 08 JAN 99 26 JAN 99 - 30 JAN 99 15 JAN 99 29 JAN 99 -02 FEB 99 02 FEB 99 15 FEB 99 - 19 FEB 99 16 FEB 99 - 20 FEB 99 A.17. Records Maintained: The originarleport,raw data and reservesamples ofthe testarticlaend vehicle components are retainedinthe archivesof Argus Research Laboratories,Inc. Any preservedtissuesare retainedinthe archivesofthe TestingFacilitfyorone year afterthe mailingofthefinalreport,afterwhich time the Sponsor willdecide theirfinaldispositionA.llunused prepared formulationswere discardedatthe TestingFacilityA.llremainingbulktestarticlweas returnedto the Study Monitor upon completionof allwork withthe testarticle. B. Test ArticleInformation: B.1. Description: PFOS (FC-95)-off-whitpeowder B.2. Lot/Batch Number: 217 (Expiratiodnate: May 2000) a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. b. DG isused as an abbreviatiofnorday of(presumed) gestation. C. DL isused as an abbreviationforday of lactatioonrday postpartum. 418-015:PAGE 11-4 B.3. Date Received and Storage Conditions: The testarticlweas receivedon 21 October 1998, and storedatroom temperature. B.4. Special Handling Instructions: Standard safetyprecautions(use of protectivcelothingg,loves,dust-mist respirators,afetygogglesor safetyglassesand a face-shieldw)ere taken when handlingthe bulk'tesatrticlaend prepared suspensions. B.5. Analysis of Activi : Informationregardingthe identityc,omposition,strengthand purityof thetest articlieson filweiththe Sponsor. C. Vehicle Information: C.l. DescriRtion: 0.5% Tween@ 80 inReverse Osmosis Membrane Processed DeionizedWater (R.O.DeionizedWater). Tween@ 80 -a clearoryellowviscousliquid C.2. Lot Numbers: Tween@ 80 - M29477 and M03HO5 C.3. Dates Received, Source and Storaae Conditions: Tween@ 80 was receivedfrom J.T.Baker, PhillipsburgN,ew Jersey,on 17 September 1998 (lotM29477) and 3 December 1998 (lotM03HO5), and storedat room temperature.The R.O. deionizedwater isavailablefrom a continuoussource atthe TestingFacilitaynd ismaintainedatroom temperature. C.4. Special Handling Instructions: Standard safetyprecautions(use of protectivcelothingg,loves,dust-mist respirators,afetygoggles or safetyglassesand a face-shieldw)ere taken when handlingthe vehicle. 418-015:PAGE 11-5 C.S. AnalysisofPuri Neitherthe Sponsor nor the Study Directorwas aware ofany potential contaminantslikeltyobe presentinthe vehiclethatwould interferweiththe resultsof thisstudy. D. Test ArticlePreRaration and Storaue Conditions: Suspensions of PFOS (PC-95) were prepared dailyatconcentrationsof0,0.02, and 0.32 mg/mL. Prepared formulationswere storedatroom temperature. D.l. Sample Information: Sample Type Concentration(all levels) Size 2 mL8 Date Retained 22 NOV 98 07 JAN 99 Storage Conditions Frozen Shipped To Sponsor Date Shipped 23 NOV 98 12 JAN 99 Bulk TestArticle Reserve 1g VehicleComponents Reserve Tween@ 80 Lot M29477 5 mL Lot M03HO5 5 mL R.O. Water 5 mL 19 NOV 98 Room temperature Testing Facility Archives 19 NOV 98 10 DEC 98 19 NOV 98 Room temperature 1 Testing Facility Archives 1 27 JAN 99 27 JAN 99 27 JAN 99 1 27 JAN 99 a. Duplicatesamples were takenfrom the firsatnd lastpreparation.One sample of each setwas shippedfor analysist:heremainingsamples were retainedas backups. Backup samples were storedfrozen(-70*Cor below)and willbe discardedat theTesbng Facilituypon the requestoftheSponsor. D.2. AnalyficalResults: Informationon the stabiliatnyd homogeneity ofthe preparedformulationsand of the stabilitoyfthe bulktestarticlaere on filweiththe Sponsor. Data verifyintghe stabilitoyfthe testarticlienthe vehiclefor48 hours under the conditionsof administratioanre on filewiththe Sponsor. Records were maintainedto document how the testarticlfeormulationswere prepared. Resultsofthe concentrationanalyseswere notavailableatthe time ofthe writing of thisreport. E. Test System: E.l. Species: Rat 418-015:PAGE 11-6 E.2. Strain: CH:CD@BR VAF/Plus@ (Sprague-Dawiey) E.3. SuRplier(Source): Chades- RiverLaboratories,Inc.,Raleigh,North Carolina E.4. Sex: Female (Note: Male ratswere used onlyforthe purposes ofbreedingand are notconsideredpartofthe Test System.) E.5. RationaleforTest System: The CH:CD@)BR VAF/Plus@ (Sprague-Dawley) ratwas selectedas the Test System because: 1)thisstrainof ratwas used inthe reproductiveand developmental toxicitsytudies;2) historicadlata and experienceexistatthe TestingFacilit/")a;nd 3)the testarticliespharmacologicallayctiveinthe species and strain. E.6. Test System Data: Number ofRats Approximate Date of Birth Approximate Age atArrival Weight (g)on the Day AfterArrival Weight (g)at Study Assignment 37 14 SEP 98 65 days 181-222 192-231 E.7. Breeder Male Rat Data: Number of Rats Approximate Date of Birth Approximate Age at Arrival Weight (g)on the Day AfterArrival Weight (g)atCohabitation 112 13 JAN 98 78 days 300-356 515-893 E.8. Method of Randomization: Upon arrivalr,atswere assigned to individuahlousing on the basisofcomputergenerated random units.Afteracclimation3,6 virginfemale ratswere placed intogroups on the basisof physicalappearance and body weightsrecorded duringacclimation.Female ratswere assigned tothreedosage groups (Groups Ithrough111)1,2 ratsper dosage group,using a computer-generated (weight-orderedr)andomizationprocedure. Afterthese 36 ratswere cohabitated, 418-015:PAGE 11-7 eightmated female rats(thosewithconfirmedevidenceof mating)were selected forthe study from each dosage group. On DL 4, a tableof random unitswas used to cullthe liftertsofivemale pups and fivefemale pups per liftewrh,ere possible. E.9. System of Identification: E.9.a.Fo Generation Rats: Male ratswere given unique permanent identificatinounmbers upon assignment tothe TestingFacility'bsreedermale ratpopulation.Female ratswere assigned temporary numbers atreceiptand given unique permanent identification numbers beforecohabitation.Each ratwas individualliydentifiewditha Monel@ self-piercienagr tag (Gey Band and Tag Co., Inc.,No. MSPT 2010 1). E.9.b.Fl Generation Pups: Pups were not individualliydentifiedduringlactationa;llparameterswere evaluatedinterms ofthe lifter. F. Husbandry: F.l. Research FacililRyegistration: USDA RegistratioNno. 23-R-099 under theAnimal Welfare Act,7 U.S.C. 2131 etseq. F.2. Study Rooms: The study rooms were maintainedunder conditionsof positiveairflowrelativteo a hallwayand independentlysuppliedwitha minimum often changes per hour of 100% freshairthathad been passed through99.97% HEPA filtersR.oom temperatureand humiditywere monitoredconstantlythroughoutthe study. Room temperaturewas targetedat 64OF to 790F (180C to260C); relative humiditywas targetedat30% to70%. See APPENDIX E (TEMPERATURE AND RELATIVE HUMIDITY REPORTS). F.3. Housing: Allcage sizesand housing conditionswere incompliance withthe Guide forthe Care and Use of LaboratoryAnimals(').Fo generationratswere individually housed instainlessteel,wire-boftomedcages, except duringthe cohabitation and postpartum periods.During cohabitatione,ach pairofratswas housed in the male rat'scage. Beginning no laterthan DG 20, Fo generationfemale rats 418-015:PAGE 11-8 were individualhloyused innestingboxes, except duringcollectioinntervalfsor udne and fecalsamples. Duringthese collectioinntervalst,he female ratswere housed individualilnymetabolism cages. Each dam and deliveredliftewrere housed ina common nestingbox duringthe postpartum period. F.4. Lighting: An automatically-controlflleudorescentlightcyclewas maintainedat 12-hours light:12-hourdsark,witheach dark pedod beginningat 1900 hours EST. F.5. Sanitization: Cage pan linerswere changed approximatelythreetimes each week. Cages were changed approximatelyevery otherweek. F.6. Feed: Ratswere givenad libituamccesstoCertifieRdodent Diet@#5002 (PMI NutritioInnternationaSlt,.Louis,Missouri)inindividuafleeders. F.7. Feed Analysis: Analyses were routinelpyerformed by the feed supplier.No contaminants at levelsexceeding the maximum concentrationor deviationsfrom expected nutritionraelquirementswere detectedby these analyses. Copies ofthe results ofthe feed analysesare availableinthe raw data. Neitherthe Sponsor nor the Study Directorwas aware ofany potential contaminantslikeltyo have been presentinthe feed thatwould have interfered withthe resultsofthisstudy. F.8. Water: Localwater thathad been processed by passage througha reverseosmosis membrane (R.O.water)was availabletothe ratsad libitumfrom an automatic wateringaccess system and/or individuawlater bottles.Chlorinewas added to the processed water as a bactedostat. F.9. Water Analysis: The processed water isanalyzed twiceannuallyforpossiblechemical contamination(LancasterLaboratoriesL,ancaster,Pennsylvania)and monthly forpossiblebacteriaclontamination(AnalyticaLlaboratodes,Inc.,Chalfont, Pennsylvania).Copies ofthe resultsofthe water analysesare availableinthe raw data. 418-015:PAGE 11-9 Neitherthe Sponsor nor the Study Directornor the Sponsor was aware ofany potentiaclontaminants likeltyo have been presentinthe water thatwould have interferewdiththe resultsofthisstudy. F.10. Bedding: Bed-o'cobs@ was used as the nestingmaterial(The Andersons' Industrial ProductsGroups, Maumee, Ohio). F.11. Beddina Analysis: Bedding was changed as oftenas necessary to keep the animalsdryand clean. Analyses forpossiblecontaminationwere conducted annuallyand documented inthe raw data. Neitherthe Study Directornor the Sponsor was aware ofany agent presentin the bedding thatwas known to interferweiththe resultsofthisstudy. G. Methods: G.l. Dosage Administration: Dosage Group 1 Dosage Concentration (mg/kg/dayr (mg/mL) 0 (Vehicle) 0 11 0.1 0.02 1 1 111 1 1.6 1 0.32 Dosage Volume (mukg) 5 Number of Female Rats 8 5 8 1 1 5 181 Assigned Rat Numbers 13726 - 13728,13731, 13734 -13737 13739 - 13741, 13744 - 13746, 13748- 13749 13751 - 13756, 13758 - 13859 a. The testarticlweas considered 100% pure forthe purpose ofdosage calculations. G.2. RationaleforDosage Selection: Dosages were selectedon the basisof a previousstudyconducted withthe test articl(eArgus Research LaboratoriesI,nc.,Protocol418-008). Inthisstudythe Fo generationmatemal and paternalno-observable-effect-le(vNeOlEL) of PFOS was 0.1 mg/kg/day (0.4mg/kg/day and higherdosages caused reductionsin body weight gain and reduced feed consumption values). The Fo generationreproductiveNOEL was greaterthan 3.2 mg/kg/day;no effectson mating,fertiliotryestrouscyclingoccurred.The NOEL forviabiliatnyd growth inthe Fl generationoffsprinwgas 0.4 mg/kg/day (1.6mg/kg/day and higherdosages caused preimplantatiolnossand reductionsinliftesrize,pup viabilitgyr,owth and survival). 418-015:PAGE 11-10 The Fl generationmatemal and patemal NOEL of PFOS was 0.1 mg/kg/day (0.4mg/kg/day dosage caused reductionsinbody weightgain and reduced feed consumption values).The Fl generationrepmductiveNOEL was greaterthan a dosage of0.4 mg/kg/day;no effectson mating or fertiliotcycurred.The NOEL forviabiliatnyd growth inthe F2 generationoffsprinwgas 0.1 mg/kg/day (0.4mg/kg/day dosage caused stillbirtahnsd reductionsinliftesrize,pup viabilitgyr,owth and survival). G.3. Route ofAdministration: Oral(gavage) G.4. RationaleforRoute of Administration: The oral(gavage)mute was selectedforuse because: 1)thiswas therouteof administratioinnthe developmental and reproductivetoxicologystudies;and 2) it isone ofthe possibleroutesof human exposure. G.5. Freguency of Administration: G.5.a. Fo Generation Female Rats: Appropriatedosagesa ofthe testarticloer vehiclewere administeredorally(via gavage) once dailyto female ratsbeginning43b days priortocohabitationuntil DG 0 (confirmedevidenced of mating,such as observationof spermatozoa ina smear ofthe vaginalcontentsor a copulatorypluginsitu).Female ratswere not given the testarticloer vehicleon DG 0. Dosages were adjusteddailyon the basisofthe individuablody weights recordedbeforeintubationT.he ratswere intubatedonce dailyatapproximatelythe same time each day. G.5.b.Fl Generation Pups: Fl generationpups were not directlgyiven the testarticleb,ut may have been possiblyexposed tothe testarticldeuringmatemal gestation(inuteroexposure) or viamatemal milkduringthe lactatiopneriod. G.6. Length of Study: Approximately14 weeks a. See APPENDIX D, item 2. b. See APPENDIX D, item 1. 418-015:PAGE 11-11 G.7. Method of Study Performance: G.7.a. Fo Generation Rats: Afteracclimationand 43 days of dosage administration3,6 healthyvirginfemale ratswere placed intocohabitationwith36 breedermale rats(one male ratper female ratinthe male rat'scage). The cohabitatiopneriodconsistedofa maximum offivedays. Mating was evaluateddailyduringthe cohabitation period.Female ratswithspermatozoa observed ina smear ofthe vaginal contentsor a copulatorypluginsituwere consideredto be at DG 0 and returned to individuahlousing. Twenty-fourmated female rats,eightper dosage group, were selectedforthe study,as previouslydiscussed. The female ratswere observed forviabilitaytleasttwiceeach day ofthe study and forgeneralappearance at leastonce duringacclimation.The ratswere also examined forclinicaolbservationsof effectsofthe testarticlea,bortions, premature deliverieasnd deaths beforeand approximatelyone hour after dosage. Rats were observed forclinicaolbservationsand generalappearance once dailyduringallotherperiodsofstudy. Body weightswere recordedonce duringacclimation.Body weightswere recordeddailyduringthe dosage and postdosage periodsand atsacrificeF.eed consumption valueswere recordedonce duringacclimationw,eekly priorto cohabitationd,ailyduringgestationand on DLs 1,4, 7, 10 and 14. Feed consumption was nottabulatedafterDL 14,when itwas expected thatpups would begin toconsume matemal feed. The female ratswere evaluatedfordurationofgestation(DG 0 to the day the firsptup was observed),liftesrizes(definedas allpups delivered)l,iveliftesrize (livebom pups only)and pup viabiliattybirth.Pups thateitherappeared stillbom or thatdied beforeinitiaelxaminationofthe lifterfsorviabiliwteyre examined for vitalstatusat birth.The lungswere removed and immersed inwater. Pups with lungsthatsank were consideredstillbomp;ups withlungsthatfloatedwere consideredlivebomand to have died shortlyafterbirth.Matemal behaviorofthe dams was evaluateddailywhen the pups were examined duringthe 21-day postpartum period.Observed matemal behaviorwas recordedon DLs 1, 4,7, 10, 14 and 21. Deviationsfrom expected matemal behaviorwere recorded, ifand when present,on allotherdays duringthe postpartum period. G.7.b. Fl Generation Rats: Day 1 oflactatio(npostpartum)was definedas the day of birthand was alsothe firsdtay on which allpups ina liftewrere individualwleyighed (pup body weights were recordedafterallpups ina liftewrere deliveredand groomed by the dam). 418-015:PAGE 11-12 The litterwsere observed forviabilitaytleasttwiceeach day duringthe postpartum period.Dead pups observed atthese timeswere removed from the nestingbox. Lifterwsere observed forclinicaolbservationsand general appearance once dailyduringthe postpartum period.The pups ineach lifter were counted once daily.Body weightswere recordedon DLs 1 (birth)4, 7, 14 and 21. G.8. Pharmacokinetic SamRie Collection: G.8.a.Fo Generation Rats: Urineand fecalsample were collectedfrom female ratsforthe following intervalso:ne day priorto initiatioofncohabitatiotnothe followingmorning, DGs 6 to 7, 14 to 15 and 20 to21, and DLs 21 to22. Followingeach 24-hour collectioinntervals,amples were collectedintocentrifugteubes,placed on dry iceand storedfrozen(-700Cor below) and shipped,frozenon dry ice,tothe Sponsor foranalysis. Blood samples were collectedfrom each ofthe matemal ratsfollowingremoval from metabolism caging(priortotestarticloer vehicleadministratioonn) each of thefollowingdays: on the day cohabitationisinitiate(dpriotrocohabitation), DGs 7,15 and 21, and DLs 14 and 22. On alldays ofcollectioenxcept DL 22, blood samples (approximately1 mL each) were collectedfrom the orbitaslinus.On DL 22, bloodsamples (approximately 4 mL each)were collectedviathe inferiovrena cava. Blood was collectedand transferreidntoserum separatortubes. The samples were spun ina refrigerated centrifuge.The serum was transferredintopolypropylenetubes labeledwiththe study number, ratidentificatidoant,e of collections,tudyday and collection timepoint.Allsamples were immediatelyfrozenon dry iceand maintained frozen(-700Cor below),and shipped,frozenon dry ice,to the Sponsor for analysis. G.8.b.Fl Generation Lifters: On DL 21, blood samples were collectedfrom allremaining pups on study. Blood samples were collectedviathe inferiovrena cava from each pup, pooled (perliftearn)d transferredintoserum separatortubes. The samples were spun ina refrigeratecdentrifugeT.he serum was transferreidntopolypropylenetubes labeledwiththe study number, ratidentificatidoant,e ofcollections,tudy day and collectiotnimepoint.Allsamples were immediatelyfrozenon dry iceand maintainedfrozen(-700Cor below)and shipped tothe Sponsor foranalysis. 418-015:PAGE 11-13 G.9. Gross Necrops G.9.a. Fo Generation Rats: Female rats,withor withoutconfirmed evidence of mating,thatwere cohabited but not assignedto the study,were sacrificebdy carbon dioxideasphyxiation and discardedwithoutfurtherevaluation. Allsurvivingratsassigned tothe studywere sacrificebdy carbon dioxide asphyxiationon DL 22 followingthe finalcollectioinntervaflorurineand fecal samples and blood sample collectionA. gross necropsy ofthe thoracic, abdominal and pelvicviscerawas performed. Tissueswithgross lesionswere preservedinneutralbuffered10% formalinforpossiblefutureevaluation. Representativephotographs of matemal lesionsare availableinthe raw data. The number and distributiofnimplantationsiteswas recorded.A liversection (rightlateralobe)from each dam was collectedf,rozenand stored(-700Cor below)untilshipment tothe Sponsor foranalysis.Allothermatemal tissues were discarded. The ratthatdid notdelivera liftewras sacrificeodn DG 25 and examined for gross lesions.Uteriwere stainedwith 10% ammonium sulfideto confirmthe absence of implantatiosnites('T)h.e ratthatwas found dead was examined for the cause ofdeath on the day ofthe death. The ratwas examined forgross lesions.A liversection(rightlateralobe)was collectedand stored,as described above. Pregnancy statuswas recorded;fetuseswere examined tothe extent possible. G.9.b. Fl Generation PuRs: Allpups culledon DL 4 were sacrificevdiadecapitation.The lungs and the livers were collectedfrom the firstten pups culleddeterminedto be atDL 4 from each dosage group (irrespectivoefliftearn)d preservedforpossiblefuture histopathologiceavlaluation.The lungswere individualrleytainedinBouin's solutiona,nd the liverswere individuallryetainedinneutralbuffered10% formalinforpossiblefutureevaluation.Remaining culledpups were sacrificed and discardedwithoutevaluation. Pups found dead were examined forgross lesionsand forthe cause ofdeath. For allpups found dead on DLs 2 to 4, alllungswere preservedinBouin's solutionforpossiblefutureevaluation. Allremainingpups on studywere sacrificeodn DL 21 by carbon dioxide asphyxiationand examined forgross lesions.Gross lesionswere preservedin neutralbuffered10% formalin.The livefrrom each pup was collectedp,ooled 418-015:PAGE 11-14 (perlifterf)r,ozenand stored(-700Cor below)and shippedtothe Sponsor for analysis. G.10. StatisticaAlnalyses: Averages and percentages were calculated.Liftevralueswere used where appropriate. Ill. RESULTS 418-015:PAGE 111-1 A. MortalifyC,linicaland Necropsy Observations (Summaries -Tables I and 2: IndividualData -Tables 19 and 20) A.I. Mortalitv No deaths or premature deliveriewsere attributetdo PFOS treatment.One vehiclecontrolgroup ratwas injuredand died afterorbitaslinusbleedingon DG 'l5. Allotherratssurvivedtoscheduled sacrifice. A.2. ClinicaOlbservations Alladverse clinicaolbservationsduringthe precohabitationg,estationand lactatiopneriodswere consideredunrelatedto thetestarticlbeecause the incidenceswere notdosage-dependent and/orthe observationoccurredinonly one ratina group. These observationsincludeddentalproblems (missing, broken and/or misalignedincisors)c,hromodacryorrhea,abrasionon the forepaw,localizedalopeciaon the head or limbs,chromorhinorrhea, exophthalmos,hemorrhagic area on the eye, urine-staineadbdominal fur, missingeye, cornealopacity,swollenarea on chestor around eye, red,dded perioraslubstance,tom ear and softor liquifdeces. A red perinasalsubstance, blue paws and gasping were agonal signsforthe ratthatdied. A.3. Necropsy Observations A red substance was found inthe thoraciccavityofthe ratthatdied. Allother ratsappeared normal atnecropsy. B. Body Weights and Body Weight Changes (Fi-ciu1r:eSummaries Tables 3 throu-qh8: IndividualData -Tables 21 through 2;1 B.I. Precohabitation Rats administered1.6mg/kg/day dosage ofthe testarticlhead reduced body weightgains or body weight lossesineach week ofthe precohabitatiopneriod. Reflectintghese effectsofthe testarticleb,ody weight gains were 46.1% ofthe controlgroup valueforthe entireprecohabitatiopneriod(DSs 1 to 43) inthe 1.6 mg/kg/day dosage group. Precohabitationbody weightsand body weight gains were unaffectedby the 0.1 mg/kg/day dosage ofthe testarticle. 418-015:PAGE 111-2 B.2. Gestation Dams administered 1.6 mg/kg/day dosage of the testarticleduring the precohabitatiopneriod(treatmentended on DG 0) had reduced body weight gainsduringthe firswteek ofthe gestationperiod(DGs 0 to7). Body weight gains were then increasedas compared tothe controlgroup value on DGs 7 to 10, 13 to 15 and 15 to 18. Reflectintghisrebound effectb,ody weight gains were increasedforthe entiregestationperiod(DGs 0 to 20) inthe 1.6 mg/kg/day dosage group, as compared to the controlgroup value. Gestationbody weightsand body weight gains were unaffectedby the 0.1 mg/kg/day dosage ofthe testarticle. B.3. Lactation Body weight gainsfordams administeredthe 1.6 mg/kg/day dosage ofthe test articldeuringthe precohabitatiopneriodgenerallycontinuedto be increased duringthe lactatiopneriod. Lactationbody weightsand body weight gains were unaffectedby the 0.1 mg/kg/day dosage ofthe testarticle. C. Absolute (a/day)and Relative(g/kg/day)Feed Consumption Values (Summaries -Tables 9 through 14: IndividualData -Tables 24 throu-qh26) C.l. Precohabitation Absolute(g/day)and relativ(eg/kg/dayf)eed consumption valueswere reduced inthe 1.6mg/kg/day dosage group ineach week ofthe precohabitation period.Reflectingthese effectsofthe testarticlef,eed consumption valueswere reduced forthe entireprecohabitatiopneriod(DSs 1 to43) inthe 1.6mg/kg/day dosage group. Precohabitatiofneed consumption values were unaffectedby the 0.1 mg/kg/day dosage ofthe testarticle. C.2. Gestation Dams administered1.6 mg/kg/day dosage of thetestarticldeuringthe precohabitatiopneriod(treatmentended on DG 0) had reduced absoluteand relativfeeed consumption valuesduringthe firswteek ofthe gestationperiod (DGs 0 to 7). Feed consumption valueswere then generallycomparable to controlgroup values forthe remainder ofthe gestationperiod.Absolute and 418-015:PAGE 111-3 relativfeeed consumption values inthisdosage group were slightlryeduced,as compared to controlgroup values,forthe entiregestationperiod(DGs 0 to 20). Gestationfeed consumption values were unaffectedby the0.1 mg/kg/day dosage ofthe testarticle. C.3. Lactation Absolute and relativfeeed consumption values duringthe lactatiopneriodwere reduced inthe groups administered0.1or 1.6 mg/kg/day ofthe testarticldeuring the precohabitatiopneriod,however the reductionswere not strictdloysagedependent. D. Natural Delivery and LifterObservations (Summaries -Tables 15 and 16: IndividualData -Tables 27 through 30) Naturaldeliveryobservationswere based on 7,8 and 7 pregnant ratsineach of the threerespectivedosage groups. Administratioonfthe testarticlaetdosages as highas 1.6mg/kg/day didnot adverselyaffectany parameter evaluatedat naturaldeliveryor duringthe 22-day lactatiopneriod(durationofgestationa,verages forimplantationasnd livelifter sizes,numbers ofdams withno livebom pups or allpups dyingduringlactation, gestationv,iabiliatnyd lactatioinndices,survivingpups per liftelri,ftesrizeat weighing,pup weight per liftearnd pup sex ratios). E. ClinicalObservations from Birthto Day 21 Postpartum and Necropsy Observations (Summaries -Tables 17 and 18: IndividualData -Tables 31 and 32) No clinicaolr necropsy observationsinthe Fl generationpups were attributable to maternaldosages ofthe testarticlaes high as 1.6 mg/kg/day because: 1)the incidenceswere not dosage-dependent; and/or2)the observationoccurredin onlyone ortwo pups. The onlyadverse clinicaolbservationwas a blacktipof tailinone 0.1 mg/kg/day dosage group pup. No milkinstomach occurredin0, 1 and 2 pups thatwere found dead inthe threerespectivedosage groups. All pups appeared normal at necropsy on DL 4 or 21. 418-015:PAGE111-4 REFERENCES 1. U.S. Food and Drug Administratio(n1994). InternationCaolnference on Harmonisation;Guidelineon detectionof toxicittyoreproductionfor medicinalproducts.FederalRegister,September 22, 1994, Vol.59, No. 183. 2. U.S. Food and Drug Administration.Good LaboratoryPractice Regulations;FinalRule. 21 CFR Part58. 3. Japanese Ministryof Health and Welfare (1997). Good Laboratory Practice Standard forSafety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. 4, European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendation on compliance withprinciplesof good laboratorypractice. OfficialJournal of the European Communities: Legislation3.2 (No. L 315; 28 October):1-17. 5. Christian,M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental ProtectionAgency, Washington, D.C. NationalTechnicalInformationService,U.S. Department ofCommerce, SpringfieldV,A 22161. 6. ChristianM,.S. (1984). Reproductivetoxicitaynd teratologyevaluationsof naltrexone(Proceedingsof NaltrexoneSymposium, New York Academy of Sciences,November 7, 1983),J.Clin.Psychiat4.5(9):7-10. 7. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity(Teratology) Control Data inthe Chailes River Crl.,CD@)BR Rat. Charles River LaboratoriesI,nc.,Wilmington,MA 01887-0630. (Database providedby Argus Research LaboratoriesI,nc.) 8. Institutoef Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski,E. (1964). Fcirbemethodezum makroskopischen Nachweis von lmplantationssteliaemn Uterus der Rafte. Arch.Pathol.Exp. Pharmakol. 247:367. APPENDIX A REPORT FIGURE BODY WEIGHTS Fo GENERATION FEMALE RATS Figure 1 425 400 375 350 325 300 275 250 n ip IP IP c C) Irv 225 200 15 29 43b 8 22 36 0 2 1 3 6 6 10 12 14 16 18 20 5 7 9 11 13 15 17 19 21 DAY OF STUDY DAY OF GESTATION a. As a resultofan errorincalculationw,hich resultedin an errorin testarticlpereparationG,roup 11ratswere dosed with0.125 ffgfkg/dayand Group IIIratswere dosed with 2 rng/kg/da on days I through4 ofstudy. b. Lostvaluerecordedbeylorecohabitation. v 0 (VEHICLE) MG/KG/DAY 0 @6 1 3 5 7 9 11 1 DAY OF LACTATIO 0.1 MG/KG/DAY a APPENDIX B REPORT TABLES PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE I (PAGE 1): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- PRECORABITATION (DAY 1 OF STUDY TO THE DAY OF COHABITATION): MAXIMUM POSSIBLE INCIDENCE 516/ 12 516/ 12 516/ 12 MORTALITY 0 0 0 INCISORS: TOTAL MISALIGNED MISSING/BROKEN 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 16/ 1 16/ 1 S/ 1 CHROMODACRYORRHEA 0/ 0 0/ 0 10/ 1 RIGHT FOREPAW: ABRASION 0/ 0 0/ 0 5/ 1 LOCALIZED ALOPECLA: HEAD 0/ 0 2/ 1 0/ 0 --------------------------------------------------------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE = (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. NIN = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-629 TABLE I (PAGE 2): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b ---------------------------------------------------------------------------------------------------------------- PRESUMED GESTATION: c MAXIMUM POSSIBLE INCIDENCE 190/ 12 192/ 11 201/ 12 ACCIDENTAL DEATH id 0 0 LOCALIZED ALOPECIA: LIMBS 0/ 0 S/ 2 ll/ 2 CHROMODACRYORRHEA 5/ 1 12/ 3 S/ 1 CHROMORHINORRHEA 1/ 1 9/ 2 3/ 1 INCISORS: MISALIGNED 0/ 0 0/ 0 5/ 1 EXOPHTHALMOS 5/ 1 4/ 1 0/ 0 EYES: HEMORRHAGIC AREA 5/ 1 4/ 1 0/ 0 URINE-STAINED ABDOMINAL FUR 0/ 0 2/ 1 0/ 0 RIGHT EYE: REMOVED 0/ 0 1/ 1 0/ 0 GASPING I/ Id 0/ 0 0/ 0 PAWS! BLUE 1/ Id 0/ 0 0/ 0 RED PERINASAL SUBSTANCE 1/ Id 0/ 0 0/ 0 ---------------------------------------------------------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE = IDAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. NIN = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Restricted to rats with a confirmed mating date. d. Dam 13731 had an accidental death on day 15 of gestation. Il ra PROTOCOL 418-015: ORAI. (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 1 (PAGE 3): CLINICAL OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS ------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 111 DOSAGE IMG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b ------------------------------------------------------------------------------------------------------------- LACTATION: MAXIMUM POSSIBLE INCIDENCE MORTALITY CHROMODACRYORRHEA CORNEAL OPACITY 154/ 7 0 0/ 0 0/ 0 176/ 8 0 10/ 2 0/ 0 154/ 7 0 1/ 1 7/ 1 CHROMORHINORRHEA 0/ 0 20/ 4 0/ 0 RIGHT EYE: MISSING 0/ 0 22/ 1 0/ 0 CHEST OR AROUND RIGHT EYE: SWOLLEN RED DRIED PERIORAL SUBSTANCE RIGHT EAR: TORN 1/ 1 0/ 0 3/ 1 7/ 1 1/ 1 0/ 0 0/ 0 0/ 0 0/ 0 SOFT OR LIQUID FECES 3/ 1 0/ 0 0/ 0 ------------------------------------------------------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE - (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. NIN = TOTAL NUMBER OF OBSERVATIONS/NUMBER OF RATS WITH OBSERVATION. a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 2 (PACE 1): NECROPSY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b -------------------------------------------------------------------------------------------------------------- RATS EXAMINED c N ad Bd ad ACCIDENTAL DEATH N le 0 0 APPEARED NORMAL N 7 8 8 THORACIC CAVITY: RED SUBSTANCE N le 0 0 -------------------------------------------------------------------------------------------------------------- a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Refer to the individual clinical observations table (Table 19) for external observations confirmed at nec d. Excludes values for rats that were not selected for continuation on study. e. Dam 13731 had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 3 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 111 DOSAGE (MG/KG/DAY)a 0 (VEHICLE) o.ib 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 BODY WEIGHT (G) DAY 1 MEAN+S.D 225.0 + 10.2 227.2 + 8.9 226.3 + 12.3 DAY a MEAN+S.D 235.4 + 10.8 236.6 + 10.S 233.9 + 13.0 DAY 15 MEAN+S.D 246.2 + 10.9 253.1 + 12.0 242.3 + 14.4 DAY 22 MEAN+S.D 252.2 + 13.4 257.5 + 14.4 245.2 + 15.7 DAY 29 MEAN+S.D 259.6 + 12.7 262.0 + 1S.5 246.9 + 14.5 DAY 36 MEAN+S.D 267.6 + 15.8 269.6 + 15.8 250.3 + 18.6 DAY 43c MEAN+S.D 274.5 + 17.9 276.4 + 18.6 249.2 + 21.5 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Last value recorded before cohabitation. PROTOCOL 418-015: ORAL IGAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-62 TABLE 4 (PAGE 1): BODY WEIGHT CHANGES - PRECOHABITATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 111 DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 BODY WEIGHT CHANGE (G) DAYS 1 8 MEAN+S.D. +10.4 + 4.9 +9.4 + 3.3 +7.6 + 6.7 DAYS 8 15 MEAN+S.D. +10.8 + 5.6 +16.5 + 6.8 +8.4 + 4.4 DAYS 15 22 MEAN+S.D. +6.1 + 6.6 +4.4 + 6.5 +2.9 + 3.8 DAYS 22 29 MEA.N+S.D. +7.3 + 6.9 +4.5 + 6.8 +1.7 + 3.0 DAYS 29 36 MEAN+S.D. +8.0 + 6.3 +7.6 + 6.8 +3.4 + S.8 DAYS 36 43c MEAN+S.D. +6.9 + 6.6 +6.8 + 6.9 -1.2 + 5.3 DAYS 1 43c MEAN+S.D. +49.5 + 14.5 +49.2 + 14.0 +22.8 + 16.3 --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. C. Last value recorded before cohabitation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 5 (PAGE 1).- MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 111 DOSAGE (MG/KG/DAY)a 0 (VEHICLE) o.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N ac ac ac PREGNANT N 8 8 7 MATERNAL BODY WEIGHT (G) DAY 0 MEAN+S.D. 279.6 + 20.2 281.1 + 20.9 261.0 + 19.7 DAY 1 MEAN+S.D. 284.1 + 18.0 290.8 + 18.6 268.7 + 19.9 DAY 2 MEAN+S.D. 289.S + 16.7 297.2 + 17.5 272.7 + 21.0 DAY 3 MEAN+S.D. 296.5 + 19.4 300.2 + 19.0 273.6 + 20.2 DAY 4 MEAN+S.D. 299.9 + 19.7 301.1 + 19.2 275.1 + 19.4 DAY 5 MEAN+S.D. 302.6 + 18.9 303.5 + 17.6 277.0 + 16.7 DAY 6 MEA.N+S.D. 307.2 + 21.3 307.6 + 16.6 279.7 + 15.0 DAY 7 MEAN+S.D. 305.9 + 18.0 309.5 + 15.5 277.1 + 15.0 DAY 8 MEAN+S.D. 305.1 + 21.1 306.9 + 17.7 279.4 + 16.2 DAY 9 MEAN+S.D. 310.2 + 20.3 311.5 + 17.3 287.0 + 16.1 DAY 10 MEAN+S.D. 314.1 + 19.2 318.1 + 19.3 291.6 + 17.5 DAY 11 MEAN+S.D. 322.0 + 24.2 323.8 + 18.9 298.8 + 21.1 DAY 12 MEAN+S.D. 328.1 + 20.8 330.6 + 16.3 301.7 + 15.9 DAY 13 MEAN+S.D. 330.8 + 22.9 331.5 + 18.6 307.6 + 21.7 ------------------------------------------------------------------------------------------------------------- DAY . DAY OF GESTATION a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group Il r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. C. Excludes values for rate that were not selected for continuation on study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 5 (PAGE 2): MATERNAL BODY WEIGHTS - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b -------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N sc oc ac PREGNANT N 8 8 7 MATERNAL BODY WEIGHT (G) DAY 14 MEAN+S.D. 334.5 + 22.6 338.5 + 17.9 311.7 + 19.2 DAY 15 DAY 16 DAY 17 DAY 18 DAY 19 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 342.2 t 341.1 1 355.6 ( 367.1 ( 383.7 1 + 24.1 7]d + 23.5 7]d + 25.8 7]d + 27.2 7]d + 29.5 7]d 347.1 + 20.9 348.5 + 18.6 361.2 + 17.2 376.9 + 21.2 390.1 + 20.5 321.8 + 22.0 328.6 + 20.8 341.6 + 19.3 359.3 + 19.6 369.4 + 18.0 DAY 20 MEAN+S.D. 398.0 + 31.0 [ 7)d 399.4 + 21.7 390.0 + 25.1 DAY 21 MEAN+S.D. 412.4 + 34.5 413.4 + 31.2 402.3 + 20.6 ( 7)d -------------------------------------------------------------------------------------------------------------- DAY = DAY OF GESTATION ( ) = NUMBER OF VALUES AVERAGED a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group 11 with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. C. Excludes values for rats that were not selected for continuation on study. d. Excludes values for dam 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL IGAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 6 (PAGE 1): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) o.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N oc sc ec PREGNANT N 8 8 7 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 - 7 MF.AN+S.D. +26.2 + 7.0 +28.4 + 9.4 +16.1 + 7.0 DAYS 7 - 10 MEAN+S.D. +8.2 + 6.1 +8.6 + 9.0 +14.4 + 7.8 DAYS 10 - 13 DAYS 13 - 15 MEAN+S.D. MEAN+S.D. +16.6 + 6.5 +11.5 + 6.9 +13.4 + 8.8 +15.6 + 12.1 +16.0 + 7.9 +14.3 + 8.6 DAYS 15 - IS MEAN+S.D. +28.7 + 7.4 +29.8 + 14.1 +37.4 + 5.3 DAYS 18 - 20 MEAN+S.D. ( 7)d +30.8 + 6.1 +22.5 + 8.7 +30.7 + 6.9 DAYS 0 - 20 MEAN+S.D. ( 7)d +119.6 + 16.3 +118.2 + 18.4 +129.0 + 16.8 1 7]d --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION I I - NUMBER OF VALUES AVERAGED a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group 11 r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Excludes values for rats that were not selected for continuation on study. d. Excludes values for dam 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (PONSOR'S STUDY NUMBER: T-62 TABLE 7 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE IMG/KG/DAY)a 0 (VEHICLE) o.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N Sc 8c BC PREGNANT N 8 8 7 DELIVERED LITTERS N 7d 8 7 MATERNAL BODY WEIGHT (G) DAY 1 DAY 2 DAY 3 MEAN+S.D. MEAN+S.D. MEAN+S.D. 299.6 + 20.6 302.6 + 25.8 307.7 + 23.5 311.1 + 19.7 302.4 + 23.3 308.1 + 28.4 293.6 + 23.8 289.0 + 20.8 292.0 + 15.9 DAY 4 MEAN+S.D. 305.3 + 22.9 307.8 + 31.7 291.4 + 19.6 DAY 5 MEAN+S.D. 308.7 + 23.5 312.8 + 18.6 297.1 + 16.7 DAY 6 MEAN+S.D. 310.4 + 23.1 314.1 + 16.5 299.7 + 15.4 DAY 7 MEAN+S.D. 314.4 + 17.9 319.2 + 12.5 304.4 + 17.6 DAY 8 MEAN+S.D. 321.0 + 21.2 328.1 + 14.4 309.8 + 14.3 DAY 9 MEAN+S.D. 324.3 + 20.3 333.8 + 12.7 316.0 + 13.3 DAY 10 MEAN+S.D. 328.8 + 19.6 341.4 + 13.1 321.1 + 14.6 DAY 11 MEAN+S.D. 333.4 + 21.4 343.6 + 13.1 329.1 + 11.8 DAY 12 MEAN+S.D. 341.8 + 21.2 346.4 + 11.5 333.6 + 12.6 --------------------------------------------------------------------------------------------------------------- DAY m DAY OF LACTATION a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group 11 r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Excludes values for rats that were not selected for continuation on study. d. Excludes values for dam 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-01S: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 7 (PAGE 2): MATERNAL BODY WEIGHTS - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b -------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N sc Bc Bc PREGNANT N 8 8 7 DELIVERED LITTERS N 7d 8 7 MATERNAL BODY WEIGHT (G) DAY 13 MEAN+S.D. 342.4 + 23.5 342.2 + 17.0 339.8 + 23.5 DAY 14 MEAN+S.D. 331.4 + 16.9 344.1 + 20.7 337.8 + 16.2 DAY 15 MEAN+S.D. 342.6 + 19.2 348.9 + 19.1 335.8 + 17.8 DAY 16 MEAN+S.D. 341.8 + 19.4 353.0 + 15.5 341.4 + 19.9 DAY 17 MEAN+S.D. 338.8 + 21.2 3S6.8 + 14.0 334.7 + 23.1 DAY 18 MEAN+S.D. 348.8 + 19.2 357.1 + 16.8 343.4 + 21.7 DAY 19 MEAN+S.D. 346.0 + 11.1 355.5 + 13.7 337.7 + 19.6 DAY 20 MEAN+S.D. 344.8 + 14.5 351.8 + 19.4 346.0 + 17.4 DAY 21 MEAN+S.D. 339.1 + 14.0 348.8 + 20.5 339.1 + 20.9 DAY 22 MEAN+S.D. 331.7 + 16.7 338.5 + 22.5 337.8 + 23.3 -------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION a. Dosage occurred on day I of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group 11 with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. C. Excludes values for rats that were not selected for continuation on study. d. Excludes values for dam 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T- TABLE 8 (PACE 1): MATERNAL BODY WEIGHT CHANGES - LACTATION - SUMMARY - Fo GENERATION FEMALE RATS ------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b ------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N Bc ac ac PREGNANT N 8 8 7 DELIVERED LITTERS N 7d 8 7 MATERNAL BODY WEIGHT CHANGE (G) DAYS 1 4 MEAN+S.D. +5.7 + 6.7 -3.4 + 30.1 -2.1 + 11.4 DAYS 4 7 MEAN+S.D. +9.1 + 6.3 +11.S + 2S.5 +13.0 + 8.8 DAYS 7 10 MEAN+S.D. +14.4 + 4.2 +22.1 + 4.7 +16.7 + 5.4 DAYS 10 14 MEAN+S.D. +2.6 + 12.7 +2.8 + 13.0 +16.7 + 5.5 DAYS 14 22 MEAN+S.D. +0.3 + 14.5 -5.6 + 24.3 +0.0 + 11.5 DAYS 1 22 MEAN+S.D. +32.1 + 10.0 +27.4 + 22.2 +44.3 + 21.7 ------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group Il with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Excludes values for rats that were not selected for continuation on study. d. Excludes values for dam 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 9 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PRECOHABITATION - SUMMARY - Fo GENERATION FEM -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b -------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 FEED CONSUMPTION (G) DAYS 1 8 MEAN+S.D. 19.6 + 1.2 18.9 + 1.2 18.3 + 1.7 ( II)c I lolc DAYS 8 15 MEAN+S.D. 18.8 + 1.5 20.2 + 1.9 17.8 + 1.4 ( lolc DAYS 15 22 MEAN+S.D. 19.6 + 1.8 19.2 + 1.4 17.5 + 1.3 DAYS 22 29 MEAN+S.D. 1 lolc 18.4 + 2.2 [ 10]c 18.1 + 2.1 [ 11-ic 15.8 + 1.5 ( lolc I lolc DAYS 29 36 MEAN+S.D. 19.1 + 2.0 19.2 + 1.6 16.6 + 1.6 1 lolc I 9]c t 1-11C DAYS 36 - 43d MEAN+S.D. 20.4 + 1.9 20.3 + 2.1 17.5 + 2.8 1 illc ( lllc DAYS 1 - 43d MEAN+S.D. 19.4 + 1.5 19.6 + 1.8 17.4 + 1.6 ( lljc ( illc -------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF STUDY t I = NUMBER OF VALUES AVERAGED a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group Il with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Excludes values that were associated with spillage. d. Last value recorded before cohabitation. PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 10 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PRECOHABITATION - SUMMARY - Po GENERATION ------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b -------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 FEED CONSUMPTION (G/KG/DAY) DAYS 1 8 MEAN+S.D. 85.0 + 5.2 82.3 + 3.8 79.3 + 4.9 ( Illc ( lolc DAYS 8 15 MEAN+S.D. 77.4 + 5.4 82.2 + 5.6 74.8 + 4.5 ( lolc DAYS 15 22 MEAN+S.D. 78.8 + 6.7 76.1 + 4.2 71.7 + 3.3 DAYS 22 29 MEAN+S.D. i IO)c 71.7 + 7.8 ( lolc 69.1 + 5.3 ( lilc 64.4 + 5.2 1 lolc ( IO)c DAYS 29 36 MEAN+S.D. 72.2 + 5.9 72.4 + 4.5 66.8 + 4.6 i 10]c I 9]c I illc DAYS 36 43d MEAN+S.D. 74.5 + 4.5 73.6 + 5.8 70.0 + 9.2 ( illc ( illc DAYS 1 43d MEAN+S.D. 76.7 + 4.9 76.6 + 4.7 71.7 + 4.7 ( illc I illc -------------------------------------------------------------------------------------------------------------- DAYS m DAYS OF STUDY I I = NUMBER OF VALUES AVERAGED a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group 11 with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Excludes values that were associated with spillage. d. Last value recorded before cohabitation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 11 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY - Fo GENERATION I --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 111 DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N oc Bc ac PREGNANT N 8 a 7 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 - I DAYS 7 - 10 DAYS 10 - 13 DAYS 13 - 15 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 25.2 1 23.8 + 1.2 7]d + 2.4 26.5 + 2.6 24.9 + 3.0 24.2 + 2.0 24.1 + 2.3 26.4 + 1.4 25.5 + 2.3 21.1 + 2.4 22.1 + 3.0 22.5 + 2.8 26.1 + 2.7 DAYS 15 - 18 MEAN+S.D. 26.0 + 1.9 27.5 + 2.8 26.2 + 1.6 ( 7)e DAYS 18 - 20 MEAN+S.D. 25.4 + 3.3 27.1 + 2.4 23.8 + 3.7 1 7]e DAYS 0 - 20 MEAN+S.D. 2S.2 + 1.3 2S.4 + 0.8 23.0 + 2.0 1 7]e --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION ( ) = NUMBER OF VALUES AVERAGED a. Dosage occurred on day I of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group Il with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Excludes values for rats that were not selected for continuation on study. d. Excludes values that were associated with spillage. e. Excludes values for rat 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 12 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY - Fo GENERATIO -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- PATS TESTED N 12 12 12 INCLUDED IN ANALYSES N oc 8c ac PREGNANT N 8 8 7 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 - 7 DAYS 7 - 10 MEAN+S.D. MEAN+S.D. 84.5 [ 76.9 + 6.6 7]d + 5.3 81.1 + 7.6 77.3 + 6.2 77.9 + 11.9 78.1 + 12.3 DAYS 10 - 13 MPAN+S.D. 81.9 + 5.5 81.0 + 6.5 75.0 + 9.0 DAYS 13 - 15 MEAN+S.D. 74.6 + 10.5 75.2 + 6.1 83.4 + 8.8 DAYS 15 - 18 MEAN+S.D. 74.1 + 4.5 77.0 + 10.3 77.8 + 5.9 ( 7]e DAYS 18 - 20 HEAN+S.D. 66.3 + 5.9 70.1 + 8.9 63.7 + 7.1 t 7]e DAYS 0 - 20 MEAN+S.D. 78.6 + 4.3 77.8 + 5.0 76.1 + 7.8 1 7]e --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION I I a NUMBER OF VALUES AVERAGED a. Dosage occurred on day I of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group Il r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Excludes values for rats that were not selected for continuation on study. d. Excludes values that were associated with spillage. e. Excludes values for rat 13731, which had an accidental death on day 15 of gestation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 13 (PAGE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - LACTATION - SUMMARY - Po GENERATION F --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANALYSES N ac Bc sc PREGNANT N 8 8 7 DELIVERED LITTERS N 7d 8 7 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 1 - 4 MEAN+S.D. 31.3 + 5.1 30.1 + 18.9 23.2 + 3.5 DAYS 4 - 7 DAYS 7 - 10 MEAN+S.D. MEAN+S.D. 47.7 1 54.4 + 10.7 6)e + 3.1 42.1 + 12.4 ( 7]e 54.8 + 8.6 40.1 + 5.6 51.0 + 6.3 DAYS 10 - 14f MEAN+S.D. 68.1 + 2.9 61.4 + 5.2 59.5 + 3.4 ( 6)e f 7]e DAYS 1 - 14f MEAN+S.D. 52.9 + 2.8 41.7 + 8.3 44.7 + 3.9 ( 6]e ( 7]e --------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF LACTATION ( I = NUMBER OF VALUES AVERAGED a. Dosage occurred on day I of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Excludes values for rats that were not selected for continuation on study. d. Excludes values for rat 13731, which had an accidental death on day 15 of gestation. e. Excludes values that were associated with spillage. f. Because it is presumed that the pups begin to consume maternal feed after day 14 of lactation, maternal fe were not tabulated on days 14 to 22 of lactation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 14 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - LACTATION - SUMMARY - Fo GENERATION ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii ill DOSAGE (MG/KG/DAY)a 0 (VEHICIE) O.lb 1.6b ---------------------------------------------------------------------------------------------------------------- RATS TESTED N 12 12 12 INCLUDED IN ANAIJYSES N ac ec ec PREGNANT N a 8 7 DELIVERED LITTERS N 7d a 7 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 1 4 MEAN+S.D. 103.6 + 20.3 98.7 + 65.9 80.1 + 14.9 DAYS 4 7 DAYS 7 10 MEAN+S.D. MEAN*S.D. 152.0 + 33.1 ( 6]e 169.3 + 12.9 134.8 I 166.2 + 44.5 7]e + 28.4 135.2 + 23.8 163.2 + 21.1 DAYS 10 14f MEAN+S.D. 202.0 + 17.6 178.4 + 16.5 179.3 + 13.1 i 6]e I 7)e DAYS 1 14f MEAN+S.D. 165.3 + 16.6 144.5 + 28.9 144.1 + 16.6 1 6]e ( 7)e ---------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF LACTATION t I = NUMBER OF VALUES AVERAGED a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II ra with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 Of Btudy, C. Excludes values for rats that were not selected for continuation on study. d. Excludes values for rat 13731, which had an accidental death on day 15 of gestation. e. Excludes values that were associated with spillage. f. Because it is presumed that the pups begin to consume maternal feed after day 14 of lactation, maternal fee were not tabulated on days 14 to 22 of lactation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 15 (PAGE 1): NATURAL DELIVERY OBSERVATIONS - SUMMARY - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- RATS ASSIGNED TO NATURAL DELIVERY N a 8 8 PREGNANT N 8 8 7 DELIVERED LITTERS DURATION OF GESTATION N d MEAN+S.D. IMPLANTATION SITES PER DELIVERED LITTER N MEAN+S.D. DAMS WITH STILLBORN PUPS N(t) DAMS WITH NO LIVEBORN PUPS N GESTATION INDEX f t NIN 7c 22.6 + 0.5 ( Sle 106 15.1 + 0.9 1( 14.3) 0 100.0 7/ 7 a 23.0 + 0.0 6]e 119 14.9 + 2.0 1( 12.S) 0 100.0 8/ 8 7 22.2 + 0.4 6)e 108 15.4 + 2.2 0( 0.0) 0 100.0 7/ 7 DAMS WITH ALL PUPS DYING DAYS 1-4 POSTPARTUM N 0 0 0 DAMS WITH ALL PUPS DYING DAYS 5-21 POSTPARTUM N 0 0 0 --------------------------------------------------------------------------------------------------------------- ( I = NUMBER OF VALUES AVERAGED a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Excludes values for dam 13731, which had an accidental death on day 15 of gestation. d. Calculated as the time (in days) elapsed between confirmed mating (arbitrarily defined as day 0) and the t the first pup was delivered. e. Excludes values for rats that did not have delivery observations recorded. f. Number of rate with live offspring/number of pregnant rats. PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 16 (PAGE 1): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I ii III MATERNAL DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 7 7 PUPS DELIVERED (TOTAL) N MEAN+S.D. 97 13.8 + 1.6 103 12.9 + 3.0 97 13.8 + 1.7 LIVEBORN MEAN+S.D. N(i) 13.7 + 1.4 96( 99.0) 12.8 + 3.0 102( 99.0) 13.8 + 1.7 97(100.0) STILLBORN MEAN+S.D. N(i) 0.1 + 0.4 1( 1.0) 0.1 + 0.4 1( 1.0) 0.0 + 0.0 0( 0.0) CULLED N 23 26 24 PUPS FOUND DEAD OR PRESUMED CANNIBALIZED DAY 1 DAYS 2- 4 DAYS 5- 7 DAYS B-14 DAYS 15-21 VIABILITY INDEX c NIN(II) MIN(%) NIN(T) NIN(L) NIN(T) t NIN 0/ 96( 3/ 96( 0/ 70( 0/ 70( 0/ 70( 0.0) 3.1) 0.0) 0.0) 0.0) 96.9 93/ 96 0/102( 1/102( 0/ 75( 0/ 75( 0/ 75( 0.0) 1.0) 0.0) 0.0) 0.0) 99.0 101/102 1/ 97( 2/ 96( 0/ 70( 0/ 70( 0/ 70( 1.0) 2.1) 0.0) 0.0) 0.0) 96.9 94/ 97 LACTATION INDEX d t 100.0 100.0 100.0 NIN 70/ 70 75/ 75 70/ 70 --------------------------------------------------------------------------------------------------------------- DAY(S) = DAY(S) POSTPARTUM a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Number of live pups on day 4 (preculling) postpartum/number of liveborn pups on day 1 postpartum. d. Number of live pups on day 21 (weaning) postpartum/number of live pups on day 4 (postculling) postpartum. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 16 (PAGE 2): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I ii III MATERNAL DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b --------------------------------------------------------------------------------------------------------------- DELIVERED LITTERS WITH ONE OR MORE LIVEBORM PUPS N 7 a 7 SURVIVING PUPS/LITTER c DAY I d MEAN+S.D. 13.7 + 1.4 12.8 + 3.0 13.8 + 1.7 DAY 4 PRECULLING MEAN+S.D. 13.3 + 1.0 12.6 + 3.0 13.4 + 1.8 DAY 4 POSTCULLING MEAN+S.D. 10.0 + 0.0 9.4 + 0.9 10.0 + 0.0 DAY 7 MEAN+S.D. 10.0 + 0.0 9.4 + 0.9 10.0 + 0.0 DAY 14 HEAN+S.D. 10.0 + 0.0 9.4 + 0.9 10.0 + 0.0 DAY 21 MEAN+S.D. 10.0 + 0.0 9.4 + 0.9 10.0 + 0.0 PERCENT MALE PUPS PER NUMBER OF PUPS SEXED DAY I d MEAW+S.D. 44.9 + 17.9 46.6 + 11.2 50.9 + 9.7 DAY 4 PRECULLING MEAN+S.D. 46.2 + 17.9 47.3 + 12.1 50.4 + 10.9 DAY 4 POSTCULLING MEAN+S.D. 48.6 + 12.1 46.5 + 5.2 48.6 + 3.8 DAY 7 MEAN+S.D. 48.6 + 12.1 46.5 + 5.2 48.6 + 3.8 DAY 14 MEAN+S.D. 48.6 + 12.1 46.5 + 5.2 48.6 + 3.6 DAY 21 MEAN+S.D. 48.6 + 12.1 46.5 + 5.2 48.6 + 3.8 --------------------------------------------------------------------------------------------------------------- DAY = DAY POSTPARTUM a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group 11 r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. c. Average number of live pups per litter, including litters with no surviving pups. d. Includes pups born alive, found dead day 1 postpartum. PROTOCOL 418-015: ORAL (GAVAGE) PHARRACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-629 TABLE 16 (PAGE 3): LITTER OBSERVATIONS (NATURALLY DELIVERED PUPS) - SUMMARY - Fl GENERATION LITTERS -----------------------------------------------------------------I ------------------------------------------- MATERNAL DOSAGE GROUP I ii III MATERNAL DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b ---------------------------------------------------------------------------------------------------------------- DELIVERED LITTERS WITH ONE OR MORE LIVEBORN PUPS N 7 8 7 LIVE LITTER SIZE AT WEIGHING DAY I MEAN+S.D. DAY 4 PRECULLING MEM+S.D. DAY 4 POSTCULLING MEAN+S.D. DAY 7 MEAN+S.D. DAY 14 MEAN+S.D. DAY 21 MEAN+S.D. PUP WEIGHT/LITTER (GRAMS) 13.7 + 1.4 13.3 + 1.0 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 12.8 + 3.0 12.6 + 3.0 9.4 + 0.9 9.4 + 0.9 9.4 + 0.9 9.4 + 0.9 13.6 + 1.7 13.4 + 1.8 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 10.0 + 0.0 DAY 1 DAY 4 PRECULLING DAY 4 POSTCULLING DAY 7 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 6.4 + 0.4 8.9 + 0.9 9.0 + 0.9 14.1 + 1.3 6.5 + 0.5 8.5 + 1.4 0.5 + 1.4 13.2 + 1.8 6.2 + 0.6 8.2 + 0.8 8.3 + 0.8 12.8 + 1.3 DAY 14 MEAN+S.D. 29.8 + 1.6 29.4 + 2.4 26.6 + 2.6 DAY 21 MEAN+S.D. 47.1 + 3.6 47.5 + 2.9 42.4 + 3.9 ---------------------------------------------------------------------------------------------------------------- DAY c DAY POSTPARTUM a. Dosage occurred on day 1 of study until day 0 of presumed gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II ra with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 17 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 21 POSTPARTUM - SUMMARY - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I ii III MATERNAL DOSAGE (MG/KG/DAY)a 0 (VEHICLE) O.lb 1.6b LITTERS EXAMINED (N) 7 0 7 --------------------------------------------------------------------------------------------------------------- PERSISTENT CLINICAL OBSERVATIONS: c TIP OF TAIL, BLACK NIN 0/ 0 15/ 1 0/ 0 --------------------------------------------------------------------------------------------------------------- NIN - TOTAL FREOUENCY (DAYS x PUPS)/LITTERSWITH OBSERVATIONS a. Dosage occurred on day I of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days 1 through 4 of study. C. Tabulation restricted to adverse observations; all other pups appeared normal. II r PROTOCOL 418-01S: OPAL (GA"GE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 18 (PAGE 1): NECROPSY OBSERVATIONS - SUMMARY - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I ii III MATERNAL DOSAGE (MG/KG/DAY)a 0 (VEHICLE) o.ib 1.6b LITTERS EXAMINED (N) 7 8 7 --------------------------------------------------------------------------------------------------------------- TOTAL PUPS STILLBORN OR FOUND DEAD c N 2 2 2 STILLBORN N 1 1 0 FOUND DEAD N 1 1 2 NO MILK IN STOMACH e N(t) 0( 0.0) 1(ioo.o) 2(100.0) --------------------------------------------------------------------------------------------------------------- PUPS SACRIFICED AND NECROPSIED ON DAY 4 AND/OR 21 POSTPARTUM d LITTERS EXAMINED PUPS EVALUATED N 7 8 7 N 80 85 so APPEARED NOR14AL LITTER INCIDENCE N(t) 7(100.0) B(IOO.O) 7(100.0) PUP INCIDENCE N(l) BO(loo-0) 85(loo.0) 80(100.0) --------------------------------------------------------------------------------------------------------------- a. Dosage occurred on day 1 of study until day 0 of gestation. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II r with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Restricted to pups in which complete necropsies were performed. Complete necropsies were not performed on autolysis or cannibalization precluded evaluation. d. Refer to the individual pup clinical observations table (Table 31) for external clinical observations conf e. Analysis restricted to pups found dead and necropsied. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 19 (PAGE I)- CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 13726 NO ADVERSE FINDINGS 13727 NO ADVERSE FINDINGS 13728 13729 13730 DL( 22 DS( 44- 45) DS( 44- 45) DS( 44- 45) DG( 0- 4) DG( 0- 4) DG( 0- 4) DG( 4 DG( 4 CHEST: SWOLLEN (RIGHT SIDE, 2.0 CM X 1.5 CM; LEFT SIDE, 2.0 CM X 1.0 CM)a CHROMODACRYORRHEA EXOPHTHALMOS RIGHT EYE: HEMORRHAGIC AREA CHROMODACRYORRHEA EXOPHTHAL140S RIGHT EYE: HEMORRHAGIC AREA EXCLUDED FROM STUDY EXCLUDED FROM STUDY 13731 DG( 15 RED PERINASAL SUBSTANCE a DG( 15 FORE AND HINDPAWS: BLUE DG( 15 GASPING DG( 15 ACCIDENTAL DEATH 13732 DG( 4 EXCLUDED FROM STUDY 13733 DG( 4 EXCLUDED FROM STUDY 13734 NO ADVERSE FINDINGS 13735 NO ADVERSE FINDINGS 13736 DG( 14 CHROMORRINORRHEA DL( 16- 18) SOFT OR LIQUID FECES DL( 20- 22) RIGHT EAR: TORN a 13737 NO ADVERSE FINDINGS ------------------------------------------------------------------------------------------------------------- DS a DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL = DAY OF LACTATION a. observation confirmed at necropsy. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 19 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 LOW DOSAGE 0.1 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION -------------------------------------------------------------------------------------------------------------- 13738 13739 DS( DG( DG( 45 ) 0- 1) 4 URINE-STAINED ABDOMINAL FUR URINE-STAINED ABDOMINAL FUR EXCLUDED FROM STUDY NO ADVERSE FINDINGS 13740 DG( 11- 12) CHROMORHINORRHEA 13741 DL( 2- 11) CHROMORHINORRHEA NO ADVERSE FINDINGS 13742 DG( 4- 6) LOCALIZED ALOPECIA: LIMBS 13743 DG( 6) DS( I- 2) EXCLUDED PROM STUDY LOCALIZED ALOPECIA: HEAD DS( 3 ALOPECIA NO LONGER APPARENT DS( 50 EXCLUDED FROM STUDY 13744 DL( 16 CHROMORHINORRHEA 13745 DL( 16 DG( 16- 19) DG( 16- 19) RED DRIED PERIORAL SUBSTANCE EXOPHTHAL.MOS RIGHT EYE: HEMORRHAGIC AREA DG( 16- 20) DG( 20 ) DL( I- 22) CHROMODACRYORRHEA RIGHT EYE: REMOVED RIGHT BYE: MISSING b DL( I- 7) DL( I- 7) CHROMODACRYORRHEA AROUND RIGHT EYE: SWOLLEN DL( a) SWELLING RESOLVED 13746 DL( 18- 20) DL( 18- 22) CHROMODACRYORRHEA CHROMORHINORRHEA b 13747 DG( 3- 4) DG( 4) LOCALIZED ALOPECIA: EXCLUDED FROM STUDY LIMBS 13748 DG( 9- 13) CHROMODACRYORRHEA DG( 10- 16) CHROMORHINORRHEA DL( 4- 7) CHROMORHINORRHEA 13749 DG( 4- 5) CHROMODACRYORRHEA -------------------------------------------------------------------------------------------------------------- DS - DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL - DAY OF LACTATION a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days I through 4 of study. b. observation confirmed at necropsy. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 19 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION -------------------------------------------------------------------------------------------------------------- 13750 DS( 28- 32) INCISORS: MISSING/BROKEN DS( 33 ) INCISORS: GROWN IN DS( 28- 45) INCISORS: MISALIGNED DS( 34- 45) CHROMODACRYORRHEA DG( 0- 4) CHROMODACRYORRHEA DG( 0- 4) INCISORST MISALIGNED DG( 4 EXCLUDED FROM STUDY 137Sl NO ADVERSE FINDINGS 13752 DL( 16- 22) RIGHT EYE: CORNEAL OPACITY b 13753 NO ADVERSE FINDINGS 13754 NO ADVERSE FINDINGS 13755 NO ADVERSE FINDINGS 13756 DG( 9- 16) LOCALIZED ALOPECIA: LIMBS DG( 17 ALOPECIA NO LONGER APPARENT DL( 17 CHROMODACRYORRHEA 13757 DS( 36- 40) FOREPAW: RIGHT, ABRASION (0.2 CM X 0.3 CM) DS( 41 ) ABRASION HEALED DG( 4- 6) LOCALIZED ALOPECIA: LIMBS DG( 6 EXCLUDED FROM STUDY 13758 NO ADVERSE FINDINGS 13759 NO ADVERSE FINDINGS 13760 13761 DG( 6 DG( I- 3) EXCLUDED FROM STUDY CHROMORHINORRHEA DG( 3) EXCLUDED FROM STUDY -------------------------------------------------------------------------------------------------------------- DS - DAY OF STUDY DG = DAY OF PRESUMED GESTATION DL - DAY OF LACTATION a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days 1 through 4 of study. b. Observation confirmed at necropsy. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 20 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Po GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 13726 DL 22 p 45 ALL TISSUES APPEARED NORMAL. 13727 DL 22 p 43 ALL TISSUES APPEARED NORMAL. 13728 DL 22 p 43 CERVICAL MAMMARY GLAND: LARGE. ALL OTHER TISSUES APPEARED NORMAL. 13731 DG 15 p 44 ACCIDENTAL DEATH ON DAY 15 OF GESTAT THORACIC CAVITY: RED SUBSTANCE. ALL OTHER TISSUES APPEARED NORMAL. UTERINE CONTENTS: 17 IMPLANTATION S AND ONE EARLY RESORPTION).B 13734 DL 22 p 45 ALL TISSUES APPEARED NORMAL. 13735 DL 22 p 43 ALL TISSUES APPEARED NORMAL. 13736 DL 22 p 46 ALL TISSUES APPEARED NORMAL. 13737 DL 22 p 46 ALL TISSUES APPEARED NORMAL. O.Ic 13739 DL 22 p 44 ALL TISSUES APPEARED NORMAL. 13740 DL 22 p 43 ALL TISSUES APPEARED NORMAL, 13741 DL 22 p 45 ALL TISSUES APPEARED NORMAL. 13744 DL 22 p 46 ALL TISSUES APPEARED NORMAL. 13745 DL 22 p 43 ALL TISSUES APPEARED NORMAL. 13746 DL 22 p 46 ALL TISSUES APPEARED NORMAL. 13748 DL 22 p 43 ALL TISSUES APPEARED NORMAL. 13749 DL 22 p 43 ALL TISSUES APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT DG = DAY OF GESTATION DL = DAY OF LACTATION a. Refer to the individual clinical observations table (Table 19) for external observations confirmed at necr b. Viability of fetuses unable to be determined because of maternal death. C. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 20 IPAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- III 1.6b 13751 DL 22 p 44 ALL TISSUES 13752 DL 22 p 45 ALL TISSUES 13753 DL 22 p 44 ALL TISSUES 13754 DL 22 p 46 ALL TISSUES 13755 DG 25 NP 46 ALL TISSUES 13756 DL 22 p 45 ALL TISSUES 13758 DL 22 p 46 ALL TISSUES 13759 DL 22 p 43 ALL TISSUES --------------------------------------------------------------------------------------------------------------- APPEARED APPEARED APPEARED APPEARED APPEARED APPEARED APPEARED APPEARED NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. NORMAL. P = PREGNANT NP = NOT PREGNANT DG - DAY OF GESTATION DL - DAY OF LACTATION a. Refer to the individual clinical observations table (Table 19) for external observations confirmed at nec b. As a result of an error in calculation, which resulted 2 mg/kg/day on days 1 through 4 of study. in an error in test article preparation, rate were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 21 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14 ---------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- 13726 203. 198. 206. 210. 213. 210. 215. 216. 214. 212. 218. 221. 224. 223 13727 242. 248. 249. 251. 246. 244. 251. 252. 245. 247. 252. 2SS. 252. 253 13720 229. 231. 226. 235. 238. 242. 238. 244. 243. 250. 244. 244. 252. 260 13729 225. 219. 225. 227. 229. 228. 235. 238. 237. 237. 243, 245. 246. 247 13730 219. 214. 219. 224. 221. 220. 226. 228. 226. 225. 230. 232. 233. 230 13731 229. 237. 239. 239. 236. 241. 243. 244. 238. 241. 245. 251. 257. 257 13732 218. 216. 222. 225. 228. 226. 221. 232. 228. 230. 228. 228. 236. 237 13733 228. 237. 236. 236. 233. 242. 246. 240. 244. 247. 250. 246. 247. 257 13734 222. 218. 218. 227. 232. 233. 234. 230. 235. 239. 236. 233. 235. 244 13735 227. 229. 227. 220. 228. 230. 232. 224. 226. 232, 234. 229. 234. 239 13736 219. 220. 216. 224. 222. 218. 221. 228. 222. 223. 224. 231. 232. 236 13737 239. 248. 250. 253, 258. 247. 247. 249. 247. 242. 239. 245. 2S2. 252 ---------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 2S 26 27 28 29 30 ---------------------------------------------------------------------------------------------------------------- 13726 229. 226. 232. 235. 235. 234. 239. 241. 237. 237. 243. 246. 244. 244 13727 258. 258. 254. 260. 262. 257. 253. 249. 253. 258. 259. 260. 253. 256 13728 252. 2S4. 266. 268. 266. 261. 271. 270. 273. 272. 266. 268. 271. 272 13729 258. 256. 253. 254. 259. 259. 262. 262. 263. 265. 267. 269. 268. 269 13730 241. 233. 239. 245. 242. 241. 242. 248. 250. 246. 255. 2S6. 258. 256 13731 270. 268. 262. 262. 269. 273. 270. 265. 258. 266. 272. 273. 268. 267 13732 236. 244. 243. 246. 241. 241. 247. 248. 250. 246. 248. 2SO. 255. 256 13733 258. 256. 265. 269. 263. 267. 263. 263. 263. 262. 263. 273. 273. 272 13734 244. 247. 251. 255. 255. 2Sl. 252. 252. 258. 259. 266, 268. 260. 261 13735 239. 241. 241. 234. 240. 243. 240. 233. 242. 244. 244. 238, 244. 247 13736 236. 234. 226. 232. 236. 234. 225. 232. 239. 237. 234. 236. 241. 238 13737 256. 256. 256. 260. 268. 266. 261. 270. 268. 269. 269. 277. 280. 280 ---------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 21 (PAGE 2): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- 13726 248. 244. 248. 256. 255. 252. 257. 260. 258. 251. 251. 13727 260. 259. 268. 269. 267. 266. 262. 271. 280. 278. 274. 13728 276. 280. 278. 284. 290. 268. 269. 287. 288. 290. 289. 13729 277. 273. 279. 282. 284. 261. 284. 287. 290. 284. 287. 13730 261. 258. 267. 268. 263. 260. 267. 271. 272. 270. 272. 13731 280. 276. 276. 282. 291. 293. 284. 285. 289. 290. 288. 13732 255. 257. 260. 255. 255. 261. 262. 260. 261. 259. 270. 13733 271. 275. 271. 273. 280. 282, 276. 277. 264. 278. 294. 13734 269. 268. 268. 266. 272. 277. 274. 269. 271. 275. 278. 13735 251. 254. 250. 247. 250. 253. 255. 248. 256. 258. 254. 13736 242. 241. 234. 239. 243. 243. 238. 242. 248. 245. 241. 13737 285. 284. 285. 290. 292. 295. 292. 298. 298. 296. 296. ---------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 21 (PAGE 3): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a --------------------------------------------------------------------------------------------------------------- 13738 13739 208. 209. 208. 205. 214. 216. 216. 215. 213. 216. 219. 218. 221. 22 242. 240. 235. 239. 249. 253. 252. 2Sl. 249. 256. 260. 259. 255. 26 13740 13741 13742 13743 13744 13745 233. 241. 235. 234. 236. 242. 240. 238. 238. 241. 244. 243. 248. 2S 232. 228. 231. 240. 239. 239. 240. 247. 243. 240. 246. 252. 249. 24 218. 219. 219. 222. 229. 230. 224. 227. 236. 242. 242. 241. 246. 25 233. 23S. 242. 23S. 241. 249. 253. 248. 2S4. 258. 259. 255. 266. 27 225. 228. 226. 228. 234. 237. 241. 237. 239. 245. 247. 246. 246. 25 228. 233. 227. 226. 235. 241. 242. 237, 242, 244. 246. 244. 249. 25 13746 221. 221. 219. 226. 229. 234. 232. 226. 234. 239. 242. 238. 236. 24 13747 235. 231. 235. 242. 239. 236. 240. 24S. 247. 242. 247. 251. 251. 24 13748 224. 223. 220. 226. 228. 232. 227. 232. 234. 240. 241. 238. 239. 24 13749 227. 219. 228. 237. 234. 229. 226. 236. 244. 242. 242. 237. 247. 25 --------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 3 --------------------------------------------------------------------------------------------------------------- 13738 229. 228. 22S. 228. 236. 234. 233. 228. 233. 237. 237. 23S. 232. 23 13739 259. 268. 268. 266. 262. 272. 269. 268. 263. 271. 268. 272. 267. 27 13740 254. 255. 257. 253. 255. 261. 260. 2S5. 258. 265. 263. 264. 264. 26 13741 259. 256. 259. 26S. 259. 258. 260, 266. 265. 261. 272. 272. 271. 26 13742 247. 250. 252. 256. 253. 250. 255. 2S9. 258. 255. 255. 259. 264. 26 13743 284. 275. 280. 278. 286. 290. 285. 280. 286. 290. 291. 283. 292. 29 13744 261. 259. 263. 261. 263. 26S. 264. 261. 267. 270. 273. 268. 276. 27 13745 254. 255. 256. 2Sl. 257. 263. 256. 255. 260. 264. 263. 261. 266. 27 13746 246. 242. 247. 252. 252. 244. 243. 248. 251. 249. 243. 240. 245. 25 13747 254. 250. 251. 257. 258. 253. 256, 258. 259. 256. 259. 264. 260. 25 13748 255. 255. 252. 248. 250. 252. 246. 241. 244. 248. 244. 242. 247. 24 13749 245. 254. 256. 257. 250. 248. 254. 2S9. 256. 256. 258. 260. 260. 26 --------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NLTMBER: T-62 TABLE 21 (PAGE 4): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAYb -------------------------------------------------------------------------------------------------------------- 13738 241. 240. 249. 252. 247. 251. 246. 251. 252. 251. 254. 13739 274. 286. 290. 283. 284. 291. 294. 288. 289. 302. 298. 13740 271. 279. 277. 273. 276. 281. 276. 276. 280. 282. 269. 13741 276. 276. 284. 281. 281. 281. 287. 291. 292. 287. 295. 13742 264. 268. 268. 265. 266. 267. 267. 265. 274. 274. 269. 13743 299. 302. 301. 296. 302. 308. 311. 306. 312. 316. 314. 13744 274. 283. 286. 284. 279. 280. 285. 293. 297. 290. 286. 13745 266. 270. 275. 269. 272. 275. 273. 266. 275. 271. 271. 13746 246. 252. 251. 249. 254. 251. 252. 259. 263. 25S. 2S7. 13747 264. 260. 265. 272. 268. 268. 268. 272. 273. 269. 278. 13748 244. 247. 246. 242. 248. 252. 247. 242. 251. 252. 256. 13749 270. 266. 267. 269. 274. 272. 268. 273. 279. 276. 270. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 21 (PAGE 5): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 6 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAYa --------------------------------------------------------------------------------------------------------------- 13750 213. 206. 214. 217. 215. 212. 218. 216. 214. 212. 217. 223. 218. 21 13751 256. 260. 266. 263. 256. 259. 260. 261. 257. 259. 262. 268. 262. 27 13752 225. 223. 227. 237. 231. 233. 239. 238. 237. 236. 244. 245. 245. 24 13753 227. 233. 225. 239. 240. 237. 233. 237. 233. 238. 235. 247. 250. 24 13754 219. 224. 217. 224. 227. 224. 222. 226. 227. 227. 228. 229. 232. 23 13755 236. 23S. 231. 238. 240. 239. 235. 240. 236. 241. 235. 238. 244. 24 13756 218. 227. 228. 232. 230. 230. 236. 240. 238. 242. 239. 246. 248. 25 13757 233. 241. 235. 235. 239, 248. 246. 247. 243. 246. 250. 254, 255. 25 13758 219. 219. 218. 221. 226. 229. 223. 226. 223. 228. 227. 233. 239. 23 13759 235. 238. 232. 232. 239. 240. 240. 231. 223. 233. 233. 236. 236. 23 13760 212. 213. 214. 211. 220. 215. 215. 214. 215. 216. 215. 220. 224. 22 13761 223. 227. 220. 226. 228. 228. 230. 231. 231. 231. 232. 236. 238. 24 --------------------------------------------------------------------------------------------------------------- DAY 17 is 19 20 21 22 23 24 25 26 27 28 29 3 --------------------------------------------------------------------------------------------------------------- 13750 13751 221. 214. 222. 226. 223. 218. 223. 220. 216. 214. 221. 215. 221. 21 268. 277. 277. 278. 271. 274. 277. 274. 261. 267. 275. 277. 271. 27 137S2 250. 247. 252. 256. 253. 250. 252. 253. 252. 249. 255. 255. 256. 25 13753 250. 246. 245. 248. 252. 254. 248. 251. 253. 254. 254. 257. 2S9. 26 13754 237. 234. 228. 231. 235. 232. 232. 236. 235. 232. 233. 237. 234. 23 13755 248. 244. 240. 242. 244. 244. 240. 240. 239. 241. 239. 240. 24S. 24 13756 13757 254. 257. 250. 260. 252. 259. 253. 260. 247. 266. 253. 26S. 254. 257, 254. 256. 2Sl. 263. 252. 267. 257. 269. 253. 265. 253. 2S 265. 2 13758 13759 239. 243. 236. 239. 238. 244. 238. 241. 241. 242. 243. 245. 232. 238. 232. 239. 234. 238. 234. 237. 228. 237. 234. 241. 241. 23 244. 2 13760 229. 222. 225. 225. 232. 226. 226. 232. 231. 229. 230. 232. 231. 2 13761 240. 238. 237. 242. 246. 239. 234. 240. 241. 241. 237. 243. 243. 24 -------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 21 (PAGE 6): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42 43a -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAYb -------------------------------------------------------------------------------------------------------------- 13750 217. 219. 226. 223. 222. 219. 218. 224. 223. 223. 212. 13751 269. 279. 282. 286. 278. 288. 291. 289. 285. 284. 287. 13752 259. 257. 262. 265. 267. 265. 267. 267. 266. 262. 269. 13753 262. 264. 264. 259. 2SG. 261. 264. 265. 260. 263. 263. 13754 236. 235. 229. 232. 238. 233. 229. 233. 236. 234. 231. 13755 240. 239. 241. 246. 244. 246. 242. 244. 243. 242. 239. 13756 254. 255. 2S3. 253. 2S9. 260. 255. 254. 256. 262. 2S6. 13757 269. 266. 275. 277. 276. 272. 269. 280. 286. 282. 276. 13758 238. 235. 238. 238. 240. 238. 237. 239. 243. 240. 232. 13759 245. 243. 245. 244. 246. 246. 239. 244. 238. 242. 240. 13760 239. 239. 239. 237. 240. 236. 234. 236. 242. 242. 244. 13761 243. 243. 242. 244. 246. 243. 239. 246. 247. 247. 241. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recored before cohabitation. b. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 22 (PAGE 1): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 ---------------------------------------------------------------------------------------------------------------- 13726 P 259. 265. 270. 275. 282. 283. 282. 281. 278. 283. 293. 13727 P 281. 292. 297. 296. 303. 304. 320. 305. 316. 316. 325. 13728 P 29S. 289. 293. 308. 309. 314. 32S. 323. 320. 320. 329. 13729 a 281. 297. 302. 302. 303. 13730 a 281. 281. 285. 288. 296. 13731 P 288. 291. 302. 306. 309. 323. 322. 310. 322. 328. 330. 13732 a 278. 282. 282. 288. 294. 13733 a 298. 309. 311. 310. 311. 13734 P 279. 283. 292. 300. 302. 303. 305. 316. 313. 312. 320. 13735 P 259. 262. 270. 272. 274. 277. 281. 285. 284. 289. 286. 13736 P 260. 273. 275. 284. 284. 288. 288. 295. 279. 293. 296. 13737 P 316. 318. 317. 331. 336. 329. 335. 332. 329. 341. 334. ---------------------------------------------------------------------------------------------------------------- DAY 13 14 15 16 17 18 19 20 21 22 23 ---------------------------------------------------------------------------------------------------------------- 13726 P 13727 P 13728 P 13729 a 13730 a 13731 P 13732 a 306. 335. 342. 358. 308. 348. 353. 357. 312. 354. 364, 369. 315. 348. 368. 326. 366. 384. 337. 379. 399. 352. 397. 427. 360. 413. 441. ACCIDENTAL DEATH ON DAY IS OF GESTATION 370. 421. 464. 372. 13733 a 13734 P 334. 332. 344. 347. 363. 367. 385. 399. 406. 13735 P 298. 309. 314. 313. 324. 333. 346. 362. 373. 360. 13736 P 314. 310. 318. 327. 340. 355. 371. 385. 409. 13737 P 359. 359. 363. 370. 386. 400. 408. 426. 444. ----------------------------------------------------------------------------------------------------------- P - PREGNANT NP = NOT PREGNANT DAY - DAY OF PRESUMED GESTATION (VALUES EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Rat was not selected for continuation on study; values excluded from group averages. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 22 (PAGE 2): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS -------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 1 -------------------------------------------------------------------------------------------------------------- 13738 b 261. 265. 269. 272. 282. 13739 P 294. 308. 315. 313. 309. 316. 314. 305. 319. 322. 33 13740 P 262. 280. 297. 298. 298. 300. 313. 317. 310. 310. 31 13741 P 310. 314. 319. 325. 333. 331. 323. 331. 326. 327. 34 13742 b 282. 284. 291. 296. 295. 291. 300. 13743 b 13744 P 302. 311. 311. 318. 318. 320. 321. 326. 326. 332. 33 13745 P 280. 281. 288. 294. 292. 294. 307. 308. 297. 304. 31 13746 P 252. 268. 275. 279. 278. 282. 276. 292. 281. 287. 29 13747 b 293. 295. 296. 295. 305. 13748 P 254. 267. 273. 270. 277. 283. 289. 286. 284. 288. 29 13749 P 275. 289. 300. 305. 304. 302. 318. 311. 312. 322. 32 -------------------------------------------------------------------------------------------------------------- DAY 13 14 is 16 17 is 19 20 21 22 2 -------------------------------------------------------------------------------------------------------------- 13738 b 13739 P 347. 348. 364. 367. 379. 401. 415. 436. 455. 13740 P 322. 334. 341. 337. 347. 356. 368. 374. 385. 13741 P 349. 352. 364. 364. 377. 397. 415. 407. 437. 13742 b 13743 b 13744 P 358. 363. 366. 374. 385. 402. 410. 42S. 448. 13745 P 322. 339. 350. 336. 347. 358. 369. 384. 392. 13746 P 318. 314. 308. 332. 346. 362. 379. 305. 405. 13747 b 13748 P 303. 312. 326. 324. 344. 355. 374. 387. 369. 387. 13749 P 333. 346. 358. 354. 365. 384. 391. 397. 416. -------------------------------------------------------------------------------------------------------------- PDAY ALL a. b. PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) = DAY OF PRESUMED GESTATION WEIGHTS WERE RECORDED IN GRAMS (C). As a result of an error in calculation, which resulted in an error in test 0.125 mg/kg/day on days 1 through 4 of study. Rat was not selected for continuation on study; values excluded from group article preparation, averages. rats were PROTOCOL 418-015: OPAL (GAVAGE) PHARf4ACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 22 (PAGE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 a 9 10 --------------------------------------------------------------------------------------------------------------- 13?SO b 232. 221. 229. 237. 241. 13751 P 295. 302. 31S. 314. 312. 309. 310. 305. 312. 321. 326 13752 P 277. 287. 278. 279. 282. 278. 284. 289. 281. 289. 290 13753 P 263. 274. 276. 278. 280. 287. 284. 270. 281. 289. 292 13754 P 246. 254. 256. 259. 259. 263. 267. 264. 264. 275. 271 13755 NP 252. 255. 258. 263. 264. 263. 266. 269. 265. 273. 266 13756 P 261. 262. 271. 271. 276. 274. 274. 278. 282. 282. 298 13757 b 284. 296. 302. 302. 305. 305. 321. 13758 P 243. 254. 254. 256. 256. 263. 269. 265. 267. 275. 281 13759 P 242. 248. 2S9. 2S8. 261. 265. 270. 269. 269. 278. 283 13760 b 236. 243. 250. 250. 2S2. 258. 261. 13761 b 252. 257. 255. 266. --------------------------------------------------------------------------------------------------------------- DAY 13 14 15 16 17 18 19 20 21 22 23 --------------------------------------------------------------------------------------------------------------- 13750 b 13751 P 13752 P 13753 P 13754 P 13755 NP 13756 P 13757 b 352. 349. 361. 370. 380. 398. 39S. 436. 432. 437. 297. 304. 324. 326. 348. 359. 371. 384. 411. 318. 321. 334. 335. 345. 367. 382. 405. 413. 290. 288. 294. 302. 321. 336. 337. 355. 366. 279. 275. 276. 274. 276. 282. 278. 284. 287. 278. 289 306. 308. 319. 322. 336. 351. 364. 380. 394. 13758 P 13759 P 13760 b 294. 296. 302. 310. 301. 320. 321. 324. 330. 331. 348. 356. 364. 373. 381. 389. 394. 406. 13761 b --------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY e DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days 1 through 4 of study. b. Rat was not selected for continuation on study; values excluded from group averages. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 23 (PAGE 1): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 a 9 10 11 --------------------------------------------------------------------------------------------------------------- 13726 13727 13728 13731 270. 270. 278. 280. 282. 284. 293. 299. 298. 303. 310 309. 325. 332. 325. 331. 338. 327. 335. 339. 336. 346 324. 316. 329. 327. 333. 333. 337. 348. 348. 352. 358 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 310. 313. 317. 320. 326. 323. 324. 332. 331. 341. 326 13735 276. 265. 279. 275. 280. 286. 295. 293. 298. 308. 312 13736 292. 299. 294. 289. 291. 290. 300. 306. 317. 316. 321 13737 316. 328. 325. 321. 318. 319. 325. 334. 339. 346. 361 --------------------------------------------------------------------------------------------------------------- DAY 14 15 16 17 18 19 20 21 22 --------------------------------------------------------------------------------------------------------------- 13726 323. 321. 32S. 336. 332. 332. 337. 328. 312. 13727 352. 360. 360. 358. 359. 357. 356. 342. 327. 13728 344. 367. 362. 356. 375. 358. 352. 354. 353. 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 325. 337. 338. 343. 353. 350. 337. 327. 330. 13735 312. 328. 327. 319. 329. 341. 339. 328. 313. 13736 314. 325. 318. 303. 328. 332. 325. 333. 334. 13737 350. 360, 363. 357. 366. 352. 368. 362. 353. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 23 (PAGE 2): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 a 9 10 11 --------------------------------------------------------------------------------------------------------------- 13739 316. 302. 317. 324. 325. 329. 323, 335. 337. 345. 350 13740 310. 297. 303. 296. 307. 307. 313. 317. 324. 338. 337 13741 320. 315. 333. 333. 323. 324. 317. 336. 342. 346. 343 13744 327. 339, 340. 342. 336. 322. 343. 350. 358. 367. 364 13745 313. 279. 253. 242. 282. 299. 312. 312. 325. 330. 338 13746 284. 272. 293. 302. 298. 295. 307. 319. 325. 333. 348 23748 281. 288. 296. 297. 301. 298. 308. 314. 320. 325. 319 13749 338. 327. 330. 326. 330. 339. 331. 342. 339. 347. 350 --------------------------------------------------------------------------------------------------------------- DAY 14 is 16 17 le 19 20 21 22 --------------------------------------------------------------------------------------------------------------- 13739 352. 356. 352. 347. 342. 355. 350. 362. 362. 13740 340. 334. 341. 343. 343. 338. 333. 335. 306. 13741 366. 359. 359. 366. 382. 364. 352. 340. 338. 13744 367. 375. 379. 378. 371. 378. 389. 386. 374. 13745 309. 357. 368. 363. 360. 365. 357. 362. 348.b 13746 324. 316. 331. 340. 339. 340. 327. 320. 316. 13748 336. 334. 342. 348. 346. 346. 342. 338. 331. 13749 359. 360. 352. 369. 374. 358. 364. 347. 333. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN CRJKMS (G). a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. b. Dead body weight. PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 23 (PAGE 3): MATERNAL BODY WEIGHTS - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE PATS -------------------------------------------------------------------------------------------------------------- RAT DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 1 -------------------------------------------------------------------------------------------------------------- 13751 13752 13753 13754 13755 333. 321. 323. 331. 333. 331. 341. 341. 342. 353. 35 278. 289. 298. 293. 292. 298. 312. 307. 315. 319. 33 299. 300. 290. 293. 300. 305. 300. 309. 320. 321. 32 264. 253. 272. 271. 281. 285. 297. 303. 319. 314. 32 NOT PREGNANT 13756 311. 296. 292. 290. 292. 299. 296. 309. 305. 316. 32 13758 296. 264. 283. 288. 291. 292. 289. 302. 309. 309. 32 13759 274. 280. 286. 274. 291. 288. 294. 298. 302. 316. 32 -------------------------------------------------------------------------------------------------------------- DAY 14 15 16 17 18 19 20 21 22 -------------------------------------------------------------------------------------------------------------- ..1,1751 13752 13753 13754 13755 13756 370. 371. 333. 333. 340. 333. 322. 324. NOT PREGNANT 340. 328. 380. 342. 321. 340. 344. 372'@.. 340. 333. 338. 295. 389. 343. 339. 334. 323. 370. 334. 343. 304. 341. 376. 325. 344. 333. 342. 367. 319. 342. 332. 356. 38 6. 330. 326. 336. 337. 13758 322. 317. 320. 323. 329. 330. 340. 308. 311. 13759 338. 345. 343. 342. 347. 342. 362. 350. 339. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days I through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 24 (PAGE 2): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Po GENERATION FEMALE RATS --------------------------------------------------------------------------------------------------------------- DAYS 1- 0 8- 15 15- 22 22- 29 29- 36 36- 43a --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 13726 140. b 138. 131. 137. 137. 13727 141. 129. 128. 106. 130. 137. 13728 135. 149. 139. 143. 141. ls9. 13729 131. 143. 141. 136. 142. 142. 13730 140. 135. 146. 147. 154. 155. 13731 144. 138. 144. 120. 130. 146. 23732 127. 114. 120. 117. lie. 124. 13733 144. 138. b b b 141. 13734 134. 129. 147. 136. 129. 144. 13735 b b b b b b 13736 122. 119. 114. 107. 108. 120. 13737 151. 126. iss. 143. 150. 163. --------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY c --------------------------------------------------------------------------------------------------------------- 13738 116. 114. 124. 114. 128. 128. 13739 b 147. 140. 122. 144. 144. 13740 138. 13S. 138. 126. 146. 141. 13741 137. 130. 139. 132. 142. 153. 13742 131. iso. 137. 137. 142. 146. 13743 b 164. b iss. b 163. 13744 139. 143. 146. 130. 143. 139. 13745 139. 144. 133. 126. 126. 135. 13746 128. 133. 121. 99. 116. 120. 13747 142. 153. b b b 168. 13748 13S. iso. 148. b b b 13749 119. 131. 121. 124. 122. 126. --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINSTIC RECOVERY STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 24 (PAGE 2): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RATS ------------------------------------------------------------------------------------I -------------------------- DAYS I- 8 8- 15 IS- 22 22- 29 29- 36 36- 43a -------------------------------------------------------------------------------------------------------------- RAT 0 DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY b -------------------------------------------------------------------------------------------------------------- 13750 114. 110. ill. 98. 100. 89. 13751 148. 134. 135. Ill. 124. 133. 13752 234. 126. 124. 114. 117. 123. 13753 131. 128. 123. 122. 131. 171. 13754 123. 119. 121. 108. 125. 116. 13755 121. 113. 115. 105. 113. 114. 137S6 129. 128. 121. ios. 107. 115. 13757 131. 133. 133. 124. 135. 134. 13758 124. 130. 133. 105. 100. 117. 13759 149. 141. c 134. c 137. 13760 109. 113. 108. 99. ill. 112. 13761 122. 122. 123. 107. ill. 110. -------------------------------------------------------------------------------------------------------------- DAYS ALL a. b. c. m DAYS OF STUDY WEIGHTS WERE RECORDED IN GRAMS (G). Last value recorded before cohabitation. As a result of an error in calculation, which resulted 2 mg/kg/day on days I through 4 of study. Spilled feed precluded the calculation of this value. in an error in test article preparation, rats were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 25 (PACE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION -------------------------------------------------------------------------------------------------------------- RAT 0 DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - I I - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 -------------------------------------------------------------------------------------------------------------- 13726 P 13727 P 25. 24. 28. 22. 23. 22. 41. 19. 22. 23. 26. 24. 21. 27. 25. 25. 24. 21. 24. 25. 13728 P 18. 41. 14. 28. 28. 27. 23. 24. 25. 28. 13729 a b 24. 25. 18. 13730 a 24. 25. 28. 25. 13731 P 16. 28. 26. 27. 25. 27. 13. 29. 26. 27. 13732 a 25. 22. 24. 20. 13733 a 28. 27. 25. 21. 13734 P 23. 27. 27. 22. 27. 28. 30. 27. 28. 24. 23735 P 24. 25. 28. 23. 29. 24. b 24. 22. 20. 13736 P 28. 23. 23. 23. 25. 24. 24. 16. 20. 27. 13737 P 24. 25. 26. 29. 24. 27. 28. 21. 25. 24. ------------------------------------------------------------------------------------------------------------ DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 --------------------------------------------------------------------------------------------------------------- 13726 P 13727 P 13728 P 13729 a 13730 a 13731 P 13732 a 22. 33. 22. 23. 23. 24. 20. b 14. Is. 22. 19. 29. 27. 30. 26. 23. 20. 25. 26. 29. 29. 34. 29. 24. 27. 29. ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13733 a 13734 P 27. 27. 25. 27. 31. 31. 21. 26. 13735 P 20. 32. 25. 29. 25. 24. 24. 29. 10. 13736 P 24. 20. 25. 26. 23. 26. 20. 25. 13737 P 24. 29. 24. 32. 26. 22. 25. 2S. -------------------------------------------------------------------------------------------------------------- P= DAYS ALL a. b. PREGNANT = DAYS WEIGHTS Rat was Spilled NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) OF PRESUMED GESTATION WERE RECORDED IN GRAMS (G). not selected for continuation on study; values excluded feed precluded the calculation of this value. from group averages. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 25 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION FE ---------------------------------------------------------------------------------------------------------------- rtAT # DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a ---------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - I I - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 a - 9 9 - 10 10 --------------------------------------------------------------------------------------------------------------- 13738 b 24, 21. 22. 21. 13739 P 13740 P 13741 P 13742 b 22. 28. 26. 23. 20, 25. 19. 2S. 24. 27, 2 24. 24. 27. 25. 26. 26. 23. 25. 25. 2 26. 27. 30. 26. 24. 24. 27. 20. 21. 23. 3 24. 24. 28. 27. 27. 24. 13743 b 13744 P 13745 P 25. 23. 22. 22. 26. 24. 27. 26. 21. 32. 2 19. 23. 27. 23. 25. 23. 24. 21. 27. 26. 2 13746 P 13747 b 24. 22. 15. 16. 20. 22. 23. 20. 19. 25. 1 29. 24. 28. 23. 13748 P 25. 23. 28. 39. 29. 23. 22. 21. 24. 20. 2 13749 P 27. 22. 29. 26. 26. 24. 14. 22. 31. 28. 2 --------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - 15 IS - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 --------------------------------------------------------------------------------------------------------------- 13738 b 13739 P 13740 P 13741 P 13742 b 24. 34. 23. 27. 28. 27. 27. 39. 23. 25. 24. 26. 28. 26. 27. 20. 23. 26. 26. 27. 28. 28. is. 28. 13743 b 13744 P 27. 22. 25. 29. 27. 22. 29. 27. 1374S P 25. 30. 19. 24. 26. 27. 34. 25. 13746 P 27. 17. 28. 34. 35. 30. 25. 28. 13747 b 13748 P 16. 34. 26. 32. 31. 30. 30. 10. 14. 13749 P 22. 33. 21. 33. 33. 24. 30. 29. --------------------------------------------------------------------------------------------------------------- P= DAYS ALL a. b. PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) - DAYS OF PRESUMED GESTATION WEIGHTS WERE RECORDED IN GRAMS (G). As a result of an error in calculation, which resulted in an error in test 0.125 mg/kg/day on days 1 through 4 of study. Rat was not selected for continuation on study; values excluded from group article preparation, averages. rats were PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 25 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA - Fo GENERATION -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 -------------------------------------------------------------------------------------------------------------- 13750 b S. is. 21. 12. 13751 P 20. 29. 22. 23. 16. 21. 18. 20. 26. 23. 13752 P 28. 15. 16. 16. 16. 17. 23. 11. 20. 15. 13753 P 23. 21. 23. 23. Is. 18. 16. 19. 23. 23. 13754 P 22. 22. le. 20. 20. 20. 24. 20. 20. 24. 13755 NP 22. 21. Is. 22. 22. 20. 29. 25. 23. 26. 13756 P 17. 20. Is. 16. 16. 16. 26. is. 20. 23. 13757 b 26. 22. 25. 26. 26. 26. 13758 P 23. 22. Is. 21. 22. 22. 45. 28. 22. 30. 13759 P 20. 25. 22. 24. 25. 23. 28. 23. 23. 27. 13760 b is. 18. 19. 20. 19. 17. 13761 b 20. 19. 20. -------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 21 - 22 22 - 23 23 -------------------------------------------------------------------------------------------------------------- 13750 b 13751 P 25. 29. 33. 25. 25. 23. 35. 24, 23. 13752 P 19. 32. 25. 25. 26. 22. 22. 30. 13753 P 24. 32. 23. 29. 24. 27. 26. 13754 P 22. 23. 24. 27. 29. 14. 23. 27. 137S5 NP 21. 21. 22. 25, 25. 15. 38. 27. 17. 20. 13756 P 20. 25. 24. 24. 24. 21. 21. 28. 13757 b 13758 P 27. 30. 28. 26. 24. 23. 23. 33. 13759 P 24. 34. 28. 28. 30. 28. 26. 32. 13760 b 13761 b -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS = DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. As a result of an error in calculation, which resulted in an error in test 2 mg/kg/day on days 1 through 4 of study. b. Rat was not selected for continuation on study; values excluded from group article preparation, averages. rats were PROTOCOL 418-015: ORAL IGAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 26 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Po GENERATION FEMALE RA -------------------------------------------------------------------------------------------------------------- DAYS 1 - 4 4 - 7 7 - 10 10 - 14a -------------------------------------------------------------------------------------------------------------- RAT 0 DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 13726 85. 144. 171. 281. 13727 106. 128. 152. 252. 13728 91. 136. 173. 282. 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 93. 136. 170. 268. 1373S 119. c 161. 280. 13736 71. 110. ISO. c 13737 92. 204. 165. 271. -------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY b -------------------------------------------------------------------------------------------------------------- 13739 49. 108. 146. 223. 13740 74. 130. 149. 233. 13741 99. 127. 169. 268. 13744 97. 109. 175. 265. 13745 20, ill. 166. 250. 13746 121. 206. 219. c 13748 206. c 159. 262. 13749 57. 93. 133. 218. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. It is presumed that the pups begin to consume the maternal feed after day 14 of lactation. b. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. C. Spilled feed precluded the calculation of this value. PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PPOS IN PATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 26 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - LACTATION - INDIVIDUAL DATA - Fo GENERATION FEMALE RA -------------------------------------------------------------------------------------------------------------DAYS I - 4 4 - 7 7 - 10 10 - 14a -------------------------------------------------------------------------------------------------------------- RAT 0 DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY b -------------------------------------------------------------------------------------------------------------- 13751 65. 110. 157. 234. 13752 72. 119. 140. 232. 13753 69. 115. 161. 244. 13754 70. 151. 181. 240. 13755 NOT PREGNANT 13756 52. 99. 121. 228. 13758 72. 115. 150. 223. 13759 87, 133. 161. 264. -------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF LACTATION ALL WEIGHTS WERE RECORDED IN GRAMS (C). a. It is presumed that the pups begin to consume the maternal feed after day 14 of lactation. b. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days I through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 27 (PAGE 1): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA Fo GENERATION FEMALE RATS/Fl GENERATION LITTERS -------------------------------------------------------------------------------------------------------------- DURATION OF LITTER DELIVERED NUMBER OF LIVE PUPS AT RAT/ LITTER GESTATION (DAYS) LIVE BORN STILLBORN TOTAL BORN COMPLETION OF DAY POSTPARTUM I 4a 4b 7 14 21 NUMBER N N N N mp mp mF mF mF mp -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 13726 23 13727 23 13 0 13 14 0 14 49 77 49 77 46 55 46 55 46 55 46 55 13728 23 is 1 16 7a 76 55 55 55 55 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 23 13 0 13 76 76 55 55 55 55 13735 23 12 0 12 39 39 37 37 37 37 13736 c 13 0 13 10 3 10 3 73 73 73 73 13737 c 16 0 16 5 11 5 10 55 55 55 55 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY d -------------------------------------------------------------------------------------------------------------- 23739 23 16 0 16 97 97 55 55 55 55 13740 23 9 0 9 45 45 45 45 4s 45 13741 23 16 0 16 79 79 55 55 55 55 13744 c 14 0 14 4 10 4 10 46 46 46 46 13745 c 12 0 12 75 74 44 44 44 44 13746 23 13 1 14 85 85 55 55 55 55 13748 23 14 0 14 68 68 55 55 55 55 13749 23 8 0 8 35 35 35 35 35 35 -------------------------------------------------------------------------------------------------------------- M = MALE F = FEMALE a. Number of live pups preculling. b. Number of live pups postculling. c. Delivery observations were not recorded, values excluded from group averages. d. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days I through 4 of study. PROTOCOL 418-015; ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 27 (PAGE 2): NATURAL DELIVERY, IMPLANTATION SITES, AND PUP VIABILITY AND SEX - INDIVIDUAL DATA Po GENERATION FEMALE RATS/Fl GENERATION LITTERS ---------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a ---------------------------------------------------------------------------------------------------------------- DURATION OF LITTER DELIVERED NUMBER OF LIVE PUPS AT RAT/ GESTATION LIVE STILL- TOTAL COMPLETION OF DAY POSTPARTUM LITTER (DAYS) BORN BORN BORN 1 4b 4c 7 14 21 NUMBER N N N N mF mp mF mp mF MF ---------------------------------------------------------------------------------------------------------------- 13751 13752 13753 13754 13755 23 22 22 d NOT PREGNANT 13(l) 0 13 15 0 15 14 0 14 11 0 11 75 69 77 47 75 59 77 47 5s 55 55 46 55 55 55 46 55 s5 55 46 55 55 55 46 13756 22 13 0 13 76 66 55 55 55 55 137SO 22 is 0 15 96 96 55 55 5s 55 13759 22 16 0 16 10 6 10 6 55 55 55 55 ---------------------------------------------------------------------------------------------------------------- M MALE F - FEMALE ( - NUMBER OF PUPS DYING PRIOR TO WEIGHING ON DAY 1 POSTPARTUM. a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were d 0.125 mg/kg/day on days I through 4 of study. b. Number of live pupa preculling. C. Number of live pups postculling. d. Delivery observations were not recorded, values excluded from group averages. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 28 (PAGE I).- PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GEN ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER DAY I DAY 4 DAY 7 DAY 14 DAY 21 NUMBER m p T m F T m F T m F T m F T ---------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- 13726 7.0 6.3 6.5 9.8 9.2 9.4 15.4 14.7 15.0 31.2 30.4 30.8 50.7 48.3 49.3 13727 6.2 6.0 6.1 9.4 9.0 9.2 15.2 14.1 14.7 30.3 28.7 29.5 48.6 48.0 48.3 13728 7.0 6.3 6.6 10.2 9.6 9.9 15.8 14.9 15.3 33.3 31.7 32.5 55.0 51.8 53.4 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 6.3 5.7 6.0 9.0 8.3 8.7 15.3 13.9 14.6 29.6 28.2 28.9 44.3 41.1 42.7 13735 6.4 5.8 . 6.0 9.0 8.4 8.5 14.0 12.2 12.8 29.7 27.2 28.0 46.5 43.2 44.2 13736 7.3 6.8 7.2 9.6 9.1 9.4 14.9 14.3 14.7 30.8 29.0 30.3 46.4 45.6 46.2 13737 6.5 6.1 6.2 7.4 6.9 7.0 11.9 ll.S 11.7 28.7 27.9 28.3 45.6 45.1 45.4 ---------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a ---------------------------------------------------------------------------------------------------------------- 13739 6.4 6.2 6.3 7.8 7.5 7.7 11.6 11.6 11.6 25.9 26.2 26.0 42.8 42.2 42.5 13740 7.3 6.6 6.9 10.6 9.6 10.0 15.6 13.8 14.6 32.4 29.9 31.0 51.8 47.2 49.3 13741 6.4 6.0 6.2 9.4 9.2 9.3 16.0 15.3 25.7 32.6 31.6 32.1 49.9 47.7 48.8 13744 6.8 6.4 6.5 7.2 6.9 7.0 11.5 11.1 11.3 31.6 31.0 31.2 46.0 44.0 44.8 13745 5.7 5.1 5.4 6.5 6.3 6.4 11.3 10.8 11.0 28.8 27.8 28.3 50.2 48.6 49.4 13746 6.9 6.5 6.7 9.0 8.3 0.8 14.2 13.5 13.8 26.9 25.3 26.1 47.3 46.6 47.0 13748 7.0 6.5 6.7 8.7 8.4 8.5 13.1 13.0 13.0 30.0 28.5 29.2 48.2 45.2 46.7 13749 7.3 6.9 7.1 11.0 10.4 10.6 15.7 14.7. 15.0 32.6 30.9 31.5 54.1 49.9 51.5 ---------------------------------------------------------------------------------------------------------------- N - MALE ALL WEIGHTS F = FEMALE WERE RECORDED T - TOTAL (SUM OF PUP WEIGHTS/NUMBER OF LIVE PUPS) DAY = DAY POSTPARTUM IN GRAMS (C). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were d 0.125 mg/kg/day on days I through 4 of study. PROTOCOL 418-015: OPAL (GAVAGE) PRARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 28 (PAGE 2): PUP BODY WEIGHT LITTER AVERAGES FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl G -------------------------------------------------------------------------------------------------------------- RAT/ LITTER DAY 1 DAY 4 DAY 7 DAY 14 DAY 21 NUMBER m F T m F T m F T m F T m F T -------------------------------------------------------------------------------------------------------------- 14ATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- 13751 7.3 6.6 7.0 10.0 8.9 9.5 15.0 13.4 14.2 30.9 28.0 29.4 45.3 48.5 46.9 13752 6.1 5.9 6.0 8.7 8.0 8.3 13.5 12.3 12.9 26.7 25.8 26.2 42.4 40.0 41.2 13753 6.5 6.4 6.5 8.6 8.7 8.7 13.1 13.6 13.3 28.6 28.7 28.6 45.4 45.2 45.3 13754 7.1 6.8 6.9 8.6 8.6 8.6 14.2 13.9 14.0 29.3 28.9 29.1 47.9 46.0 46.8 13755 NOT PREGNANT 13756 6.1 5.7 5.9 7.9 7.3 7.6 10.8 10.3 10.5 23.7 20.8 22.3 38.5 35.3 36.9 13758 5.5 5.6 5.6 7.1 7.2 7.1 11.6 11.8 11.7 24.3 24.9 24.6 38.9 39.0 39.0 13759 6.0 5.6 5.8 8.0 7.5 7.8 12.9 12.4 12.7 26.4 25.3 25.8 42.0 40.2 41.1 -------------------------------------------------------------------------------------------------------------- N - 14ALS F = FEMALE T - TOTAL (SUN OF PUP WEIGHTS/NUMBER OF LIVE ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE a. As a result of an error in calculation, which resulted in an error 2 mg/kg/day on days I through 4 of study. PUPS) DAY - DAY POSTPARTUM ONLY WEIGHTS OF LIVE PUPS. in test article preparation, rats were PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 1): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 B 9 10 11 12 13 14 is 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY POSTPA ------------------------------------------------------------------------------------------------------------ 13726 7.6 6.9 6.6 7.0 6.2 6.1 6.4 6.3 6.0 5.8 6.4 7.2 6.1 13727 6.3 6.4 6.1 6.1 6.4 6.3 6.2 5.8 6.1 5.9 6.1 5.7 6.2 6.0 13728 7.1 6.9 6.9 7.2 7.4 6.4 7.2 6.3 4.S 7.1 6.4 6.1 6.0 6.4 7.0 FS 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 6.4 6.3 5.5 6.8 6.2 6.4 6.8 6.0 5.7 5.6 6.0 5.3 5.5 13735 6.3 7.0 5.9 6.3 6.2 6.0 5.6 6.0 5.6 S.7 5.0 5.9 13736 7.7 7.6 6.6 7.6 7.6 6.9 7.7 7.4 7.2 7.1 7.0 6.9 6.4 13737 6.8 6.3 6.2 6.8 6.3 6.4 5.7 6.0 6.1 6.1 6.7 6.1 6.0 5.9 5.8 5.9 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP Il LOW DOSAGE 0.1 MG/KG/DAY a POSTPA -------------------------------------------------------------------------------------------------------------- 13739 6.3 6.8 5.7 5.9 6.7 6.2 6.8 6.3 6.5 6.1 6.2 6.4 6.3 6.4 6.1 6.0 13740 7.2 7.8 6.8 7.5 6.8 6.6 6.6 6.9 6.2 13741 6.1 6.1 6.7 6.7 6.3 6.5 6.2 5.6 6.2 6.2 S.7 6.3 6.0 5.7 6.4 6.3 13744 7.0 7.1 6.8 6.5 6.0 6.2 6.8 6.4 6.3 6.0 6.2 6.8 6.5 6.6 13745 5.8 5.8 5.8 5.7 5.8 5.7 S.1 4.0 S.3 5.4 5.4 5.5 13746 6.6 6.8 7.2 7.4 6.9 6.8 7.5 5.9 ms 6.6 6.0 6.2 6.7 6.9 13748 7.3 7.2 7.6 6.6 6.8 6.6 6.6 7.0 6.9 6.6 6.2 5.8 6.0 6.6 13749 7.3 7.5 7.1 6.6 6.9 6.8 7.0 7.3 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days I through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 2): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 1 8 9 10 11 12 13 14 is 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a POSTPA -------------------------------------------------------------------------------------------------------------- 13751 7.0 7.9 7.2 7.0 7.6 7.4 7.2 6.3 6.6 6.4 6.6 7.0 FD 1 13752 4.1 6.5 6.2 6.8 6.4 6.6 6.0 6.0 5.2 5.9 6.0 6.1 6.4 5.5 5.8 13753 6.9 6.8 7.0 6.2 6.8 5.5 6.5 6.8 6.6 6.3 6.2 6.2 7.1 6.0 13754 7.0 7.4 7.0 7.1 6.8 6.6 7.0 7.1 6.7 6.6 7.1 13755 NOT PREGNANT 137S6 6.1 6.4 5.8 5.9 6.0 6.2 6.2 S.7 6.3 5.7 5.4 5.5 5.7 13758 5.4 5.7 5.4 5.6 5.5 5.6 5.5 5.3 MA 5.5 S.4 S.3 6.0 5.7 6.0 13759 5.7 6.1 6.2 6.2 6.2 4.9 6.5 5.9 6.1 6.1 5.2 5.9 4.6 5.8 5.9 6.0 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITRER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER SECOND LETTER M-MALE, F-FEKALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. As a result of an error in calculation, which resulted in an error in test article preparation, 2 mg/kg/day on days I through 4 of study. rate were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 3): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP 0 1 2 3 4 5 6 7 a 9 10 11 12 13 14 is 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY POSTPARTUM D -------------------------------------------------------------------------------------------------------------- 13726 9.7 9.2 10.5 9.8 8.9 9.9 9.2 9.2 9.0 9.5 9.2 9.4 9.0 13727 9.5 9.4 9.1 9.4 9.2 9.7 9.4 8.5 9.4 8.8 9.2 9.1 9.2 8.9 13728 10.3 10.6 9.9 10.0 10.4 9.9 10.4 10.3 9.8 9.3 9.0 9.0 10.0 FD 3 FM 2 FS 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 9.0 9.2 9.5 8.8 9.6 9.3 7.8 8.9 7.8 8.8 8.6 7.1 8.4 13735 9.4 9.0 8.5 8.7 8.8 7.9 8.3 8.7 8.7 B.S 7.2 8.4 13736 9.1 8.8 9.6 9.8 10.1 8.8 10.0 9.1 10.2 10.1 9.6 8.4 9.3 13737 7.6 7.6 7.2 6.9 7.7 7.1 6.9 6.9 7.5 7.1 6.8 7.0 5.7 6.7 7.0 FM 4 -------------------------------------------------------------------------------------------------------------- 14ATERNAL DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a POSTPARTUM D -------------------------------------------------------------------------------------------------------------- 13739 8.1 8.4 8.5 7.9 7.7 7.2 7.8 7.5 7.4 7.9 7.7 7.3 7.9 7.0 7.4 7.3 13740 10.7 10.8 10.2 10.9 9.S 9.4 9.7 9.5 9.8 13741 9.7 9.1 9.9 8.8 9.0 9.2 10.2 8.6 9.3 9.8 9.8 8.3 9.8 9.5 8.2 9.4 13744 6.1 7.5 7.5 7.5 7.7 7.5 6.0 7.0 6.7 6.8 7.4 6.7 5.9 7.0 13745 6.6 5.8 6.5 6.8 6.6 6.8 6.6 6.3 6.2 6.4 6.3 FD 2 13746 9.3 9.0 8.6 9.2 9.6 8.8 9.7 8.2 ms 8.4 9.0 9.1 7.4 7.6 13748 9.3 9.7 8.8 7.8 6.5 7.9 8.3 8.1 7.9 9.0 8.9 9.3 7.1 8.6 13749 10.9 11.2 10.8 10.6 10.8 10.2 10.2 10.4 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE FIRST LETTER SECOND LETTER NUMBER FOLLOWING a. As a result RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), *SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 Of study. PROTOCOL 418-015! ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 4): PUP BODY WEIGHTS FRBM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY POSTPARTUM D -------------------------------------------------------------------------------------------------------------- 13751 10.1 10.1 10.0 9.6 10.0 10.5 9.8 9.0 9.2 8.1 9.8 8.2 FD 1 13752 9.2 7.6 8.5 8.9 9.1 MD 4 7.1 8.1 8.1 8.6 8.1 7.0 8.3 8.8 8.3 13753 7.7 8.8 9.2 9.4 8.8 8.4 8.3 8.2 9.2 8.9 8.3 8.4 8.4 9.6 13754 8.4 8.9 8.5 8.8 9.6 8.1 9.2 8.7 8.8 7.7 8.0 13755 NOT PREGNANT 13756 8.2 7.7 7.8 8.0 8.0 7.8 MM 3 6.6 6.9 7.3 7.0 7.4 8.5 13758 7.1 6.6 7.1 7.7 7.1 7.4 7.3 6.9 6.5 7.5 6.9 7.2 6.7 7.7 6.9 13759 8.0 8.9 8.3 6.6 8.3 7.4 8.2 8.8 8.0 8.0 5.9 8.0 8.2 7.3 7.6 8.1 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER SECOND LETTER NUMBER FOLLOWING a. As a result 2 mg/kg/day N-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. of an error in calculation, which resulted in an error in test article preparation, on days I through 4 of study. rats were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629 TABLE 29 (PAGE 5): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS ---------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 ---------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY POSTPARTUM DAY ---------------------------------------------------------------------------------------------------------------- 13726 9.7 9.2 10.5 9.8 FC 4 9.9 9.2 9.2 FC 4 9.5 9.2 FC 4 9.0 13727 9.5 MC 4 9.1 9.4 9.2 9.7 MC 4 FC 4 9.4 8.8 9.2 9.1 FC 4 8.9 13728 10.3 10.6 9.9 MC 4 MC 4 9.9 10.4 10.3 FC 4 9.3 9.0 9.0 10.0 PD 3 FM 2 FS 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 9.0 9.2 MC 4 8.8 9.6 9.3 MC 4 8.9 FC 4 8.8 8.6 7.1 8.4 13735 9.4 9.0 8.5 8.7 FC 4 7.9 8.3 8.7 8.7 8.5 7.2 FC 4 13736 MC 4 MC 4 9.6 MC 4 10.1 8.8 10.0 9.1 10.2 10.1 9.6 8.4 9.3 13737 7.6 7.6 7.2 6.9 7.7 7.1 6.9 FC 4 7.5 7.1 FC 4 7.0 PC 4 PC 4 PC 4 FM 4 --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 11 LOW DOSAGE 0.1 MG/KG/DAY a POSTPARTUM DA --------------------------------------------------------------------------------------------------------------- 13739 MC 4 MC 4 MC 4 7.9 MC 4 7.2 7.8 7.5 7.4 7.9 7.7 7.3 FC 4 7.0 7.4 FC 4 13740 10.7 10.8 10.2 10.9 9.5 9.4 9.7 9.5 9.8 13741 MC 4 9.1 9.9 8.8 MC 4 9.2 10.2 8.6 9.3 FC 4 FC 4 8.3 9.8 FC 4 FC 4 9.4 13744 6.1 7.5 7.5 7.5 7.7 7.5 6.0 7.0 FC 4 6.8 7.4 FC 4 FC 4 FC 4 13745 MC 4 MC 4 6.5 MC 4 6.6 6.8 6.6 6.3 6.2 6.4 6.3 FD 2 13746 MC 4 MC 4 8.6 9.2 9.6 8.8 MC 4 8.2 ms 8.4 9.0 9.1 7.4 7.6 13748 9.3 9.7 8.8 7.8 8.5 MC 4 FC 4 8.1 FC 4 9.0 PC 4 9.3 7.1 8.6 13749 10.9 11.2 10.8 10.6 10.8 10.2 10.2 10.4 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE FIRST LETTER SECOND LETTER NUMBER FOLLOWING a. As a result RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. M-MALE, F-FEMALE, U-SEX UNDETERMINABLE A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. of an error in calculation, which resulted in an error in test article preparation, rats X-OTH were 2 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 6): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY POSTPARTUM D ------------------------------------------------------------------------------------------------------------ 13751 13752 13753 13754 13755 MC 4 10.1 MC 4 9.6 10.0 10.5 9.8 9.0 9.2 8.1 9.8 8.2 FD 1 9.2 7.6 6.5 8.9 9.1 MD 4 FC 4 PC 4 8.1 PC 4 FC 4 7.0 8.3 8.8 8.3 7.7 8.8 MC 4 9.4 MC 4 8.4 8.3 FC 4 9.2 8.9 6.3 FC 4 8.4 9.6 8.4 8.9 8.5 8.8 9.6 8.1 9.2 8.7 8.8 7.7 PC 4 NOT PREGNANT 13756 8.2 MC 4 7.8 8.0 8.0 7.8 MM 3 6.6 6.9 7.3 7.0 7.4 PC 4 13758 MC 4 MC 4 7.1 MC 4 7.1 7.4 7.3 MC 4 6.5 7.5 6.9 7.2 PC 4 7.7 6.9 13759 MC 4 0.9 8.3 MC 4 MC 4 MC 4 8.2 8.8 8.0 MC 4 FC 4 8.0 8.2 7.3 7.6 8.1 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER FIRST LM ER M-NALE, F-FENALE, U-SEX UNDETERMINABLE WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. SECOND NUMBER LETTER FOLLOWING A-ALIVE, S-STILLBORN, U-UNCERTAIN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), 'SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. X-OT a. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days I through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 7)@ PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 -------------------------------------------------------------------------------------------------------------- PAT/ LITTER MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY POSTPA -------------------------------------------------------------------------------------------------------------- 13726 15.3 15.1 16.6 14.4 FC 4 14.7 15.1 14.3 FC 4 14.5 15.0 FC 4 14.6 13727 15.6 MC 4 14.7 15.4 14.9 15.5 MC 4 FC 4 14.6 14.7 13.7 13.4 FC 4 14.3 13728 15.1 15.6 15.9 MC 4 MC 4 16.0 16.2 15.6 FC 4 14.8 15.6 14.2 14.4 FD 3 FM 2 FS 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 15.6 15.6 MC 4 14.5 15.1 15.6 MC 4 12.6 FC 4 14.4 14.3 13.6 14.7 13735 14.2 13.2 14.5 11.7 FC 4 10.7 13.9 11.2 12.0 13.3 12.8 FC 4 13736 MC 4 MC 4 15.6 MC 4 15.9 14.6 13.3 15.1 15.9 13.8 14.8 14.5 13.5 13737 12.9 13.0 11.1 10.6 12.0 12.6 11.7 FC 4 11.3 10.5 FC 4 11.4 FC 4 FC 4 PC 4 FM 4 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP 11 0.1 MG/KG/DAY a POSTPARTUM DAY 7 -------------------------------------------------------------------------------------------------------------- 13739 MC 4 MC 4 MC 4 11.4 MC 4 12.3 12.2 11.4 10.8 10.9 12.4 10.8 FC 4 12.5 11.3 FC 4 13740 16.1 14.7 15.2 16.2 14.4 13.6 13.1 13.8 13.9 13741 MC 4 15.4 15.9 16.9 MC 4 16.4 15.5 14.8 15.9 FC 4 FC 4 14.4 14.8 FC 4 FC 4 16.6 13744 12.0 11.9 10.3 11.7 11.1 11.6 12.2 11.2 FC 4 10.6 10.0 FC 4 FC 4 FC 4 13745 MC 4 MC 4 11.2 MC 4 11.8 11.0 11.2 10.8 11.1 10.6 10.7 FD 2 13746 MC 4 MC 4 14.1 14.3 14.2 15.3 MC 4 13.2 ms 11.9 12.4 14.4 14.9 13.8 13748 12.7 14.0 12.8 10.9 15.1 MC 4 FC 4 13.1 FC 4 13.6 FC 4 10.2 14.5 13.5 13749 15.5 15.4 16.1 14.2 14.4 15.0 14.4 15.3 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE FIRST LETTER SECOND LETTER NUMBER FOLLOWING a. As a result RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), X-OTHER "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL IGAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 29 (PAGE 8): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 s 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a POSTPAR ---------------------------------------------------------------------------------------------------------------- 13751 MC 4 15.7 MC 4 1S.2 IS.2 14.2 14.8 13.4 13.3 13.7 12.2 14.4 FD 1 13752 14.0 14.0 12.1 14.2 13.3 MD 4 FC 4 FC 4 10.4 FC 4 FC 4 12.9 13.1 12.8 12.4 13753 13.1 13.2 MC 4 14.1 MC 4 13.2 11.8 FC 4 14.4 12.S 13.9 FC 4 12.9 14.2 13754 13.5 14.4 14.7 14.0 14.4 14.1 13.7 12.3 15.0 13.8 FC 4 13755 NOT PREGNANT 13756 13.0 MC 4 12.3 8.6 11.6 8.4 MM 3 10.8 8.7 11.0 11.0 9.8 FC 4 13758 MC 4 MC 4 11.5 MC 4 11.5 11.8 11.4 MC 4 12.0 12.6 11.6 12.1 FC 4 11.3 11.2 13759 MC 4 12.5 12.4 MC 4 MC 4 MC 4 12.6 13.2 14.0 MC 4 FC 4 12.9 11.5 12.7 12.2 12.7 --------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. As a result of an error in calculation, which resulted in an error in test article preparation, 2 mg/kg/day on days I through 4 of study. rats were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 9): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 is 16 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY POSTPA -------------------------------------------------------------------------------------------------------------- 13726 31.2 29.8 31.1 32.9 FC 4 30.4 30.4 29.5 FC 4 30.6 31.1 FC 4 30.6 13727 29.8 MC 4 30.5 29.0 31.7 30.5 MC 4 FC 4 28.9 27.5 29.3 28.6 FC 4 29.0 13728 32.8 33.6 33.0 MC 4 MC 4 33.9 33.0 32.8 FC 4 30.8 30.0 32.9 31.8 FD 3 FM 2 FS 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 30.7 30.3 MC 4 29.8 28.1 29.2 MC 4 28.6 FC 4 31.3 28.3 24.1 28.9 13735 30.8 30.0 28.4 28.3 FC 4 27.7 24.5 30.1 27.7 26.3 26.1 FC 4 13736 MC 4 MC 4 32.2 MC 4 32.1 29.1 29.8 30.2 31.6 31.0 29.1 28.4 29.6 13737 30.1 29.7 26.3 28.2 29.0 27.9 27.S FC 4 27.7 30.0 FC 4 26.4 FC 4 FC 4 FC 4 FM 4 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a POSTPAR -------------------------------------------------------------------------------------------------------------- 13739 MC 4 MC 4 MC 4 25.0 MC 4 26.9 25.4 26.9 25.5 25.9 27.0 27.0 FC 4 24.9 26.0 FC 4 13740 31.7 31.3 33.5 33.3 29.8 30.8 30.2 28.8 29.7 13741 MC 4 32.9 33.4 33.8 MC 4 31.2 31.6 31.4 33.9 FC 4 FC 4 30.3 33.0 FC 4 FC 4 29.6 13744 31.9 31.3 28.7 34.5 32.3 27.5 31.5 33.2 FC 4 28.9 32.6 FC 4 FC 4 FC 4 13745 MC 4 MC 4 28.7 MC 4 29.2 28.4 28.7 28.5 28.1 26.5 28.2 FD 2 13746 MC 4 MC 4 27.4 26.5 26.6 27.5 MC 4 26.6 ms 24.6 26.9 24.6 26.5 24.1 13748 30.7 26.9 32.4 29.6 30.3 MC 4 FC 4 30.3 FC 4 29.4 FC 4 29.9 28.1 24.7 13749 33.1 32.5 32.2 29.8 30.0 31.9 32.8 29.9 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), NUMBER FOLLOWING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. As a result of an error in calculation, which resulted in an error in test article preparation, 0.12S mg/kg/day on days 1 through 4 of study. rats were PROTOCOL 418-015: ORAL IGAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 29 (PAGE 10): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 1 --------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a POSTPAT --------------------------------------------------------------------------------------------------------------- 13751 MC 4 32.0 MC 4 30.9 31.2 30.6 29.6 28.4 27.6 30.0 26.3 27.8 FD 1 13752 28.6 26.4 27.4 25.7 25.3 MD 4 FC 4 FC 4 26.2 FC 4 FC 4 27.0 26.2 23.0 26.5 13753 29.8 29.7 MC 4 29.0 MC 4 27.8 26.7 FC 4 29.4 27.8 29.0 FC 4 29.6 27.6 13754 29.9 30.2 28.2 28.8 29.9 30.2 29.0 28.7 27.2 28.6 FC 4 1375S NOT PREGNANT 13756 18.5 MC 4 24.7 27.3 28.5 19.6 MM 3 19.5 26.2 21.4 17.7 19.3 FC 4 13758 MC 4 MC 4 26.0 MC 4 25.6 23.8 22.3 MC 4 24.0 26.2 23.7 26.6 FC 4 24.9 23.3 13759 MC 4 26.5 27.9 MC 4 MC 4 MC 4 26.5 25.7 25.4 MC 4 FC 4 23.4 2S.1 25.2 26.9 25.9 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE FIRST LETTER SECOND LETTER NUMBER FOLLOWING a. As a result 2 mg/kg/day RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE of an error in calculation, which resulted in an on days 1 through 4 of study. INCLUDE ONLY WEIGHTS OF LIVE PUPS. (PRESUMED CANNIBALIZED), EVENT OCCURRED. error in test article preparation, rats were PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 29 (PAGE 11): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 1 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER N MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY POSTPA --------------------------------------------------------------------------------------------------------------- 13726 49.7 49.6 52.8 50.6 FC 4 48.7 48.0 48.1 FC 4 47.5 48.4 FC 4 49.3 13727 48.2 MC 4 48.4 48.9 52.3 45.1 MC 4 FC 4 47.7 48.2 47.9 47.6 FC 4 48.4 13728 54.4 55.6 54.1 MC 4 MC 4 55.0 55.9 52.2 FC 4 52.4 53.4 50.5 50.4 FD 3 FM 2 FS 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 45.8 46.3 MC 4 41.8 42.4 45.4 MC 4 42.1 FC 4 41.0 43.3 40.4 38.6 13735 48.2 45.3 46.1 44.3 FC 4 42.9 38.3 41.1 43.9 48.7 43.6 FC 4 13736 MC 4 MC 4 47.8 MC 4 48.3 45.9 45.6 48.7 45.2 43.4 45.7 45.1 46.0 13737 46.7 48.5 44.4 41.2 47.4 44.9 42.4 FC 4 45.5 49.0 FC 4 43.6 FC 4 FC 4 FC 4 FM 4 -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP Il LOW DOSAGE 0.1 MG/KG/DAY a POSTPAR -------------------------------------------------------------------------------------------------------------- 13739 MC 4 MC 4 MC 4 43.6 MC 4 46.1 40.5 42.2 41.5 39.2 41.8 44.1 FC 4 44.0 41.8 PC 4 13740 48.6 53.7 54.2 50.9 47.5 44.8 49.7 47.6 46.4 13741 MC 4 51.4 49.0 49.2 MC 4 51.0 49.0 46.7 49.S FC 4 FC 4 Sl.9 45.1 FC 4 FC 4 45.5 13744 45.0 46.6 45.9 46.4 41.9 42.6 44.2 47.1 FC 4 46.1 41.8 FC 4 FC 4 FC 4 13745 MC 4 MC 4 49.7 MC 4 52.2 49.9 49.0 48.3 47.9 49.2 48.8 FD 2 13746 MC 4 MC 4 50.9 47.7 43.3 47.6 MC 4 47.1 ms 42.4 46.4 50.4 50.4 43.3 13748 48.5 47.6 44.8 49.7 50.4 MC 4 FC 4 40.8 FC 4 45.1 FC 4 47.2 46.7 46.2 13749 53.8 55.4 53.2 47.3 51.0 52.5 50.9 47.7 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. FIRST LETTER M-MALE, F-FEMALE SECOND LETTER A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), NUMBER FOLLOWING *SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. As a result of an error in calculation, which resulted in an error in test article preparation, 0.125 mg/kg/day on days 1 through 4 of study. rats were PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS ISPONSOR'S STUDY NUMBER: T-6 TABLE 29 (PAGE 12): PUP BODY WEIGHTS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a POSTPA -------------------------------------------------------------------------------------------------------------- 13751 MC 4 46.7 MC 4 48.4 43.4 44.0 44.1 51.6 48.3 46.3 50.9 45.4 FD 1 13752 41.3 46.4 42.5 40.5 41.5 MD 4 FC 4 FC 4 40.4 FC 4 FC 4 35.4 40.4 40.8 42.9 13753 46.5 45.3 MC 4 45.6 MC 4 47.8 41.9 FC 4 47.5 46.5 43.4 FC 4 43.4 45.2 13754 46.8 50.4 46.4 48.1 46.8 44.2 44.9 47.5 46.1 46.7 FC 4 13755 NOT PREGNANT 13756 39.7 MC 4 42.4 33.8 33.4 43.0 MM 3 33.9 41.6 30.7 34.1 36.4 FC 4 13758 MC 4 MC 4 39.6 MC 4 38.7 39.0 38.4 MC 4 39.0 36.9 39.0 40.7 FC 4 41.3 37.3 13759 MC 4 41.9 44.1 MC 4 MC 4 MC 4 39.0 40.8 43.2 MC 4 FC 4 41.8 39.3 36.7 41.2 42.2 -------------------------------------------------------------------------------------------------------------- ALL WEIGHTS WERE FIRST LETTER SECOND LETTER NUMBER FOLLOWING a. As a result 2 mg/kg/day RECORDED IN GRAMS (G). MEAN LITTER WEIGHTS INCLUDE ONLY WEIGHTS OF LIVE PUPS. H-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), *SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. of an error in calculation, which resulted in an error in test article preparation, on days 1 through 4 of study. rats were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 30 (PAGE 1): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATIO --------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 17 18 19 20 --------------------------------------------------------------------------------------------------------------- PAT/ LITTER MATERNAL DOSAGE GROUP I VEHICLE CONTROL 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 13726 M A N A M A M A FC 4 F A F A F A FC 4 F A F A F@ 4 F A 13727 M A MC 4 M A M A M A M A MC 4 FC 4 P A F A F A F A FC 4 P A 13728 M A M A N A MC 4 MC 4 M A M A F A FC 4 F A F A F A F A FD 3 FM 2 F S 13731 ACCIDENTAL DEATH ON DAY 15 OF GESTATION 13734 M A M A MC 4 M A M A M A MC 4 F A FC 4 F A F A F A F A 13735 M A M A M A P A FC 4 F A F A F A F A F A F A FC 4 13736 MC 4 MC 4 M A MC 4 M A M A M A M A M A M A F A F A F A 13737 M A M A M A M A M A F A F A FC 4 F A F A FC 4 F A FC 4 FC 4 FC 4 FM 4 --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP II LOW DOSAGE 0.1 MG/KG/DAY a --------------------------------------------------------------------------------------------------------------- 13739 MC 4 MC 4 MC 4 N A MC 4 M A M A M A M A F A F A F A FC 4 F A F A FC 4 13740 M A M A M A M A FA F A F A F A F A 13741 MC 4 M A M A M A MC 4 M A M A F A F A FC 4 FC 4 F A F A FC 4 FC 4 F A 13744 N A M A M A M A F A F A F A F A FC 4 F A F A FC 4 FC 4 FC 4 13745 MC 4 MC 4 M A MC 4 M A M A M A F A F A F A F A FD 2 13746 MC 4 MC 4 M A M A M A M A MC 4 M A M S F A F A F A F A F A 13748 M A M A M A M A M A MC 4 FC 4 F A FC 4 F A FC 4 F A F A F A 13749 M A M A M A FA F A F A FA F A --------------------------------------------------------------------------------------------------------------- FIRST LETTER SECOND LETTER M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), NUMBER FOLLOWING "SECOND LETTER- INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. a. As a result of an error in calculation, which resulted in an error in test article 0.125 mg/kg/day on days 1 through 4 of study. preparation, rats were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 30 (PAGE 2): PUP VITAL STATUS AND SEX FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATI -------------------------------------------------------------------------------------------------------------- PUP # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 is 19 20 -------------------------------------------------------------------------------------------------------------- RAT/ LITTER MATERNAL DOSAGE GROUP III HIGH DOSAGE 1.6 MG/KG/DAY a -------------------------------------------------------------------------------------------------------------- 137Sl MC 4 M A MC 4 M A M A M A M A F A F A F A F A P A FD 1 13752 M A M A M A M A M A MD 4 FC 4 PC 4 F A FC 4 FC 4 F A F A F A F A 13753 M A M A MC 4 M A MC 4 M A M A FC 4 F A F A F A FC 4 F A F A 13754 M A M A M A M A F A F A F A F A F A F A FC 4 13755 NOT PREGNANT 137S6 M A MC 4 M A M A M A M A MM 3 F A F A F A F A F A FC 4 13758 MC 4 MC 4 M A MC 4 M A M A M A MC 4 M A F A P A F A FC 4 F A F A 13759 MC 4 M A M A MC 4 MC 4 MC 4 M A M A M A MC 4 FC 4 F A F A F A F A F A -------------------------------------------------------------------------------------------------------------- FIRST LETTER SECOND LETTER NUMBER POLL#OWING a. As a result 2 mg/kg/day M-MALE, F-FEMALE A-ALIVE, S-STILLBORN, D-DIED, C-CULLED, M-MISSING (PRESUMED CANNIBALIZED), "SECOND LETTER" INDICATES THE DAY POSTPARTUM THE EVENT OCCURRED. of an error in calculation, which resulted in an error in test article preparation, on days I through 4 of study. rats were PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 31 (PAGE 1): CLINICAL OBSERVATIONS FROM BIRTH TO DAY 21 POSTPARTUM - INDIVIDUAL DATA - Fl GENERATION -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP LITTER MATERNAL DOSAGE (MG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATION a -------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) - - ii O.lb 13746 7-21 1/10 PUPS: TIP OF TAIL, III 1.6b - - -------------------------------------------------------------------------------------------------------------- a. Tabulation restricted to adverse observations; all other pups appeared normal. b. As a result of an error in calculation, which resulted in an error in test article preparation, Group II with 0.125 mg/kg/day and Group III rats were dosed with 2 mg/kg/day on days I through 4 of study. c. Observation confirmed at necropsy. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 32 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP LITTER MATERNAL DOSAGE (MG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATIONS a -------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 13726 13727 4, 21 4, 21 ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. 13728 1 3 4, 21 1 PUP: STILLBORN. ALL TISSUES 1 PUP: FOUND DEAD. ALL TISSUE ALL PUPS APPEARED NORMAL. 13734 4, 21 ALL PUPS APPEARED NORMAL. 13735 4, 21 ALL PUPS APPEARED NORMAL. 13736 4, 21 ALL PUPS APPEARED NORMAL. 13737 4, 21 ALL PUPS APPEARED NORMAL. -------------------------------------------------------------------------------------------------------------- a. complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation individual pup clinical observations table (Table 31) for external observations confirmed at necropsy. PROTOCOL 418-015: OPAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6 TABLE 32 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS -------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP LITTER MATERNAL DOSAGE (MG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATIONS a -------------------------------------------------------------------------------------------------------------- ii O.lb 13739 13740 13741 4, 21 21 4, 21 ALL PUPS APPEARED ALL PUPS APPEARED ALL PUPS APPEARED NORMAL. NORMAL. NORMAL. 13744 4, 21 ALL PUPS APPEARED NORMAL. 13745 2 4, 21 1 PUP: FOUND DEAD. NO MILK IN TISSUES APPEARED NORMAL ALL PUPS APPEARED NORMAL. 13746 1 4, 21 1 PUP: STILLBORN. ALL TISSUES ALL PUPS APPEARED NORMAL. 13748 4, 21 ALL PUPS APPEARED NORMAL. 13749 21 ALL PUPS APPEARED NORMAL. -------------------------------------------------------------------------------------------------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation individual pup clinical observations table (Table 31) for external observations confirmed at necropsy. b. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 0.125 mg/kg/day on days 1 through 4 of study. PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-62 TABLE 32 (PAGE 3); NECROPSY OBSERVATIONS - INDIVIDUAL DATA - Fl GENERATION PUPS --------------------------------------------------------------------------------------------------------------- MATERNAL DOSAGE GROUP LITTER 14ATERNAL DOSAGE IMG/KG/DAY) NUMBER DAY(S) POSTPARTUM OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- III 1.6b 13751 1 1 PUP: FOUND DEAD. NO MILK IN 4, 21 TISSUES APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 137S2 4 21 1 PUP: FOUND DEAD. NO MILK IN TISSUES APPEARED NORMAL. ALL OTHER PUPS APPEARED NORMAL. ALL PUPS APPEARED NORMAL. 13753 4, 21 ALL PUPS APPEARED NORMAL. 13754 4, 21 ALL PUPS APPEARED NORMAL. 13756 4, 21 ALL PUPS APPEARED NORMAL. 13758 4, 21 ALL PUPS APPEARED NORMAL, 13759 4, 21 ALL PUPS APPEARED NORMAL. -----------------------------------------------------------------------------------------:--------------------- a. Complete necropsies were not performed on pups in which autolysis or cannibalization precluded evaluation. individual pup clinical observations table (Table 31) for external observations confirmed at necrOPBY. b. As a result of an error in calculation, which resulted in an error in test article preparation, rats were 2 mg/kg/day on days I through 4 of study. APPENDIX C . PROTOCOL AND AMENDMENT 418-015:PAGE C-1 t=IC A 905 Argus Research Sheehy Laboratories,Inc. Drive, BuildingA Homham, PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587 PROTOCOL 418-015 SPONSOR'S STUDY NUMBER: T-6295.14 STUDY TITLE: Oral(Gavage) PhaffnacokinetiRcecovery Study ofPFOS in Rats PURPOSE: The purpose ofthisstudy istoevaluatethe pharmacokineticsofPFOS inFo generationand Fl generationratsduringgestationand lactatiofnollowing cessationof PFOS treatmentof CH:CDOBR VAF/PlusQD female ratsat confirmedmating. TESTING FACILITY: Argus Research LaboratoriesI,nc. 905 Sheehy Drive,BuildingA Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Teleftx: (215)443-8587 STUDY DIRECTOR: Raymond G. York,Ph.D.,DABT AssociateDirectorof Research SPONSOR: 3M CorporateToxicology 3M Center,Building220-2E-02 St.Paul,Minnesota 55144-1000 STUDY MONITOR: MarvinT. Case, D.V.M.,Ph.D. Telephone: (651)733-5180 Telefax: (651)733-1773 ALTERNATE STUDY MONITOR: Andrew M. Seacat,Ph.D. Telephone: (651)575-3161 Telefax: (651)733-1773 418-015:PAGE C-2 Pmtocol 41MI 5 Page 2 REGULATORY CITATIONS: U.S. Food and Drug Administration(1994).InternationaClonference on Harmonisation; Guidelineon detectionof toxicittyo reproductionformedicinalproducts. Federal Register,September 22, 1994, Vol. 59, No. 183. U.S. Food and Drug Administration.Good Laboratory PracticeRegulations;FinalRule. 21 CFR Part 58. Japanese Ministryof Health and Wetfare (1997). Good Laboratory PracticeStandard forSafety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendation on compliance withprinciplesof good laboratorypractice.OfficiaJlournal of the European Communities: Legislation.32 (No. L 315; 28 October): 1-17. REGULATORY COMPLIANCE: This study willbe conducted incompliance withthe Good Laboratory Practice(GLP) regulationscitedabove. Allchanges or revisionsof thisprotocolshallbe documented, signed by the Study Directorand the Sponsor, dated and maintained with the protocol. The QualityAssurance Unit(QAU) willauditthe protocol,the raw data and the report, and willinspectcriticaplhases of the study in accordance with the Standard Operating Procedures of Argus Research Laboratories,Inc. The finalreportwillincludea statement signed by the Study Directorthatthe report accuratelyreflecttshe raw data obtained during the performance of the study and that allapplicableGLP regulationswere followed inthe conduct of the study. Should significandteviationsfrom GLP regulationsoccur, each willbe described indetail, togetherwith how the deviationmight affectthe qualityor integritoyf the study. SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 to the protocol. 4lMl5: PAGE C-3 PMtoCol 41 B-015 Page 3 TEST ARTICLE AND VEHICL Identification: Test Article: Name: Physical Description: Lot/BatchNumber SpecificGravity: Purity: ExpirationDate: PFOS (Synonym: FC-95). Light-mlored powder. 217. -0.6. 98.9%. May 2000. Informationon the identityc,omposiflon,strengthand purityof the testarticleison file withthe Sponsor. Vehicle: 0.5% TweenO 80 in Reverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplierand lotidentificatiofnTweengD 80 to be documented in the raw data. Neitherthe Sponsor nor the Study Directoris aware of any potentialcontaminants likely to be present inthe vehiclethatwould interferewith the resultsof thisstudy. Therefore, no analyses otherthan those mentioned inthisprotocolwillbe conducted. Safely Precautions: Gloves, mask, appropriateeye protectionand a uniform/labcoat are to be wom during formulationpreparationand administration.The MaterialSafety Data Sheet (MSDS) is attached to the protocol(ATTACHMENT 2). Storage: Bulk Test Article: Vehicle Components: Prepared Vehicle: Prepared Formulations: Room Room Room Room temperature. temperature. temperature. temperature. Alltestarticleshipments to the Testing Facilitsyhould be addressed to the attentionof JulianGulbinski,Manager of Formulations,at the previouslycitedaddress and telephone number. Shipments should includeinformationconcerning storage conditionsand shipping cartons should be labeledappropriately.The recipientshould be notffiedin advance of shipment. 418-015:PAGE C-4 PMtoCol 41 " 15 Page 4 FORMULATION: Freguency of Preparabon: Formulations (suspensions) willbe prepared daily at the Testing Facility. Data verifying the stabilitoyf the test articleinthe vehicle for48 hours under the conditions of administrationare on filewith the Sponsor. Detailedpreparationproceduresare attachedtothisprotocol(ATTACHMENT 3). Ad8ustment forPuri : The test articlewillbe considered 100% pure forthe purpose of dosage calculations. Testing FacililyReserve SamRIO : The Sponsor willreserve a sample (1 g) of each lotof the bulk testarticleused during the course of thisstudy. The Testing Facilitywillreserve a sample (5 mL) of each lotof the vehicle components used during the course of thisstudy. Samples willbe stored under the previously cited conditions. ANALYSES: Samples additionalto those described below may be taken ifdeemed the course of the study. necessary during Bulk Test Article SamRiin No analyses of the bulk test articlewillbe conducted during the course of thisstudy. Information on the stabilitoyf the bulk test articleison filewith the Sponsor. Analyses of Prepared Formulation : Homogeneity and stabilitoyf prepared formulations ison filewith the Sponsor. However, records willbe maintained to document how the test articleformulations were prepared. Concentration of Test ArticleFormulations Concentration of the prepared formulations willbe verifiedduring the course of this study. Duplicate samples (2 mL each) willbe taken from the firstand lastpreparation on the day prepared. One sample of each set willbe shipped for analysis;the remaining samples willbe retained at the Testing Facilityas backup samples. Backup samples willbe stored frozen (-70*C or below) and discarded at the Testing Facility upon request of the Sponsor. 4lMl5:PAGE C-5 PMtoCol 41 MI 5 Page 5 Shig)RingInstructions: Samples to be analyzed willbe shipped (frozenon dry ice)to: KrisJ. Hansen, Ph.D. 3M Environmental Technology and 935 Bush Avenue Building2-3E-09 St.Paul,Minnesota 55133-3331 Telephone: (612)778-6018 Telefax: (612)778-6176 Safety Services The recipientwillbe notifiedin advance of sample shipment. DISPOSITION: Prepared formulationswillbe discarded at the Testing FacilityA.llremaining bulk test articlweillbe returnedto the Study Monitor at the previouslycitedaddress upon completion of allwork with the testarticle. TEST SYSTEM: SRecies/Strain and Reason for Selection: The Crl:CEMBR VAF/PlusS (Sprague-Dawley) ratwas selectedas the Test System because: 1)thisstrainof ratwas used inthe reproductiveand developmental toxicity studies;2) historicadlata and experience existatthe Testing Facilv'-3;)and 3) the test articleispharmacologicallyactivein the species and strain. Number: Initiaplopulationacclimated: Populationselected forstudy: Body Weight and Age: 36 virginfemale rats. 24 mated female rats(8 per dosage group). Female ratswillbe ordered to have body weights of 200 g to 225 g each atreceipt,at which time they willbe expected to be at least60 days of age. Actual body weights will be recorded the day afterreceiptand willbe documented in the raw data. The weight range willbe includedin the finalreport. Sex: Female ratswillbe given the testarticle.Male ratsof the same source and strainwill be used only as breeders and are not considered part of the Test System. 418-015:PAGE C-6 Protocol418-015 Page 6 Source: Charles RiverLaboratoriesI,nc. The ratswillbe shipped infifterecdartons by airfreightand/or truckfrom Charles River Laboratories,Inc.,to the Testing Facility. Identification: Fo Generation: Rats are permanently identifieudsing MoneND self-piercinegar tags (Gey Band and Tag Co., Inc.,No. MSPT 20101). Male ratsare given unique permanent identification numbers upon assignment to the Testing Facility'bsreeder male ratpopulation. Female ratsare assigned temporary numbers at receiptand given unique permanent identificationnumbers when assigned to the study. Fl Generation: Pups willnot be individualliydentifiedduring lactationa;llparameters willbe evaluated in terms of the litter. ANIMAL HUSBANDRY: Allcage sizes and housing conditionsare incompliance with the Guide forthe Care and Use of Laboratory Animale'). Housing: Fo Generation Rats/Fl Generation Litters: Fo generation ratswillbe individuallhyoused in stainlesssteel,wire-bottomed cages, except during the cohabitationand postpartum periods. During cohabitation,each pair of ratswillbe housed inthe male rat'scage. Beginning no laterthan day 20 of presumed gestation,Fo generation female ratswillbe individuallhyoused in nesting boxes, except during collectionintervalsforurineand fecalsamples. During these collectioinntervalst,he female ratswillbe housed individualliynmetabolism cages. Each dam and deliveredliftewrillbe housed ina common nesting box during the postpartum period. 418-015:PAGE C-7 Protocol418-015 Page 7 Nestina Material: Nesting material (bed-o'cobsID) willbe provided. Bedding willbe changed as oftenas necessary to keep the animals dry and clean. Analyses forpossiblecontamination are conducted annually and documented inthe raw data. Room Air,Temperature and Humidity: The animal room isindependently supplied with at leastten changes per hour of 100% freshairthathas been passed through 99.97% HEPA filter(sAiroClean4D room). Room temperature willbe maintained at64"F (180C) to 79OF (260C) and monitored constantly.Room humiditywillalso be monitored constantlyand maintained at 30% to 70%. Liam: An automaticallycontrolled12-hour light:12-houdrark fluorescentlightcyclewillbe maintained. Each dark period willbegin at 1900 hours EST. Diet: Rats willbe given CertifiedRodent Diett #5002 (PMI NutritioInnternationala)vailable ad libitumfrom individualfeeders. Water: Water willbe availablead libitumfrom individualboftlesattached to the cages or from an automatic watering access system. Allwater willbe from a localsource and passed through a reverseosmosis membrane before use. Chlorinewillbe added to the processed water as a bacteriostatp;rocessed water isexpected to containno more than 1.2 ppm chlorineat the time of analysis.Water isanalyzed monthly forpossible bacterialcontamination and twice annually forpossible chemical contamination. Contaminants: Neitherthe Sponsor nor the Study Directorisaware of any potentialcontaminants likely to be present inthe certifiediet,the drinkingwater or the nestingmaterialat levelsthat would interferweith the resultsof thisstudy. Therefore, no analyses otherthan those routinelyperformed by the feed supplieror those mentioned inthisprotocolwillbe conducted. 41 Ml 5:PAGE C-8 Protocol418-015 Page 8 RANDOMIZATION AND COHABITATION: Upon arrival,male and female rats willbe assigned to individual housing on the basis of computer-generated random units.Afteracclimation,36 virginfemale ratswillbe selectedforstudy on the basis of physicalappearance and body weights recorded duringacclimation.The female ratswillbe assigned to dosage groups (12 female rats per dosage group) based on computer-generated (weight-ordered)randomization procedures and treatedwith eitherthe vehicleor the testarticlfeor42 days priorto cohabitation. Within each dosage group, consecutive order willbe used to assign female ratsto cohabitationwith breeder male rats,one male ratper female rat.The cohabitation period willconsistof a maximum of fivedays. Female ratswith spermatozoa observed ina smear of the vaginalcontents and/or a copulatoryplug observed in situwillbe considered to be at day 0 of presumed gestationand assigned to individualhousing. Eightmated female rats(thosewithconfirmed evidence of mating) willbe assigned to each dosage group. Any remaining female ratswith or withoutconfirmed evidence of mating thatwere assigned to eitherof the treatedgroups or the controlgroup priorto cohabitationwillbe sacrificedand discarded withoutfurtherevaluationat the discretion of the Study Directorand the Study Monitor. Day 1 of lactatio(npostpartum)isdefined as the day of birthand isalsothe firsdtay on which allpups in a liftearre individuallwyeighed (pup body weights willbe recorded afterallpups in a littearre deliveredand groomed by the dam). On day 4 postpartum, lifterwsillbe culledto fivemale pups and fivefemale pups per liftewrh,ere possible.Pups not selectedforcontinued evaluationwillbe sacrificedvia decapitationt;he lungs (saved inBouin's solution)and the liver(saved inneutral buffered10% formalin)willbe collectedfrom the firsten culledpups from each dosage group (irrespectivoef liftedre)termined to be atday 4 postpartum and preserved for possiblefuturehistopathologicaelvaluation.Remaining culledpups willbe sacrificed viadecapitationand discarded without necropsy evaluation. ADMINISTRATION: Route and Reason for Choice: The oral(gavage) routewas selected foruse because: 1) thiswas the route of administrationinthe developmental and reproductivetoxicologystudies;and 2) itisone of the possible routes of human exposure. 418-015:PAGE C-9 PMt=l 41 " 15 Page 9 Method and Freauency: Fo GenerationFemale Rats: Female ratswillbe given the testarticleonce dailybeginning 42 days priorto cohabitationuntilday 0 of presumed gestation(confirmedevidence of mating, such as observationof spermatozoa in a smear of the vaginalcontents or a copulatoryplug in situ).Female ratswillnot be given the testarticleor the vehicleon day 0 of presumed gestation.Dosages willbe adjusted dailyforbody weight changes and given at approximately the same time each day. Fl Generation Pur)s: Fl generation pups willnot be directlgyiven the testarticleb,ut may be possibly exposed to the testarticleduring matemal gestation(inutero exposure) or via matemal milkduring the lactationperiod. Rationale for Dosace Selection: Dosages were selected on the basis of a previous study conducted with the testarticle (Argus Research Laboratories,Inc.,Protocol418-008). Dosage Levels,Concentrations and Volumes: Number of Dosage Female Groug) Rats Dosage a 0 (Vehicle) a I o.i a1 1.6 concentration (mWmL) Dosage Volume (mL/kg) 0 5 0.02 5 0.32 5 !@2usBatchNumber 5418-015-A(Day.Month.Year) B41"1 "(Day.Month.Year) E"1&015-C(Day.Month.Year) The test article will be considered ioo% pure for Ow punxme of dosage calculations. TESTS, ANALYSES AND MEASUREMENTS - Fo GENERATION: Viab lity: AllPeriods: At leasttwicedaily. ClinicalObservations and/or General APRearance: AcclimationPeriod: At leastonce. 418-015: PAGE C-11@'@ PMtocol 418-015 Page 10 Dosage Period: Twice daily.Priortoadministratioannd once approximatelyone hour postdosage. AllOther Pedods: Once daily. Matemal Behavior: Days 1, 4, 7, 10, 14 and 21 postpartum. Observed abnormal behavior willbe recorded daily. Clinicalobservationsmay be recorded more frequentlythan citedabove, ifdeemed appropriateby the Study Directorand/or Study Monitor. Body Weights: Acclimation Period: At leastonce. Dosage Period: Daily. AllOther Periods: Daily. Sacrifice: Terminal weight. Feed ConsumRfion Values (recordedand tabulated): Acclimation Period: At leastonce. Dosage Period: Weekly to cohabitation. AllOther Periods: Dailyduring presumed gestationand days 1,4, 7, 10 and 14 postpartum. Feed consumption willnot be tabulatedafterday 14 postpartum, when itis expected thatpups willbegin to consume matemal feed. Feed consumption values may be recorded more frequentlyifitisnecessary to replenishthe feed. These intervalswillnot be tabulated. Mating Performance: Mating willbe evaluated dailyduring the cohabitationperiod and confirmed by observationof spermatozoa in a smear of the vaginalcontents and/or a copulatoryplug observed insitu. 418-015: PAGE C-1 1 Protocol41 " 15 Page 11 Natural Delivery: Female ratswillbe evaluated for: Duration of Gestation (day 0 of presumed gestationto the day the firstpup is observed). LitterSize (definedas allpups delivered). Live LifteSrize (livebom pups only). Pup Viabilitayt Birth. Pharmacokinetic SamRIO Collecton: Urine and Fecal Samples: Female ratswillbe housed individualliynmetabolism cages forcollectionof urineand fecalsamples forthe followingintervals:one day priorto initiationf cohabitationto the followingmorning and days 6 to 7, 14 to 15 and 20 to 21 of presumed gestation,as well as days 21 to 22 postpartum. Followingeach 24-hour collectionintervals,amples will be collectedintocentrifugetubes,placed on dry iceand storedfrozen (-700C or below) untilshipment foranalysis. In the event that a dam begins to deliverbefore completion of urineand fecalsample collectioonn day 20 to day 21 of presumed gestation,the dam willbe removed from the metabolism cage and placed ina nestingbox with sufficienbtedding. Blood Samr)les: Blood samples willbe collectedfrom each of the matemal ratsfollowingremoval from metabolism caging (priorto administrationo)n each of the followingdays: on the day cohabitationisinitiate(dpriorto cohabitation)a,nd on days 7, 15 and 21 of presumed gestation,as well as on days 14 and 22 postpartum. The time of blood collectiownill be recorded inthe raw data. 418-015:PAGE C-12 ProtoCol41 "l 5 Page 12 On alldays ofcollectioenxceptday 22 postpartum,bloodsamples (approximatelIy mL each)willbe collectefdromthe orbitaslinus.Ifnecessary,whole bloodmay be collectefdrom an aftematesite;ifso,the aftematesitewillbe documented inthe raw data. On day 22 postpartum, blood samples (approximately4 mL each) willbe collectedviathe inferiovrena cava. Blood willbe collectedand transferredintoserum separatortubes. The samples willbe spun ina refrigeratecdentrifuge.The serum will be transferredintopolypropylene tubes labeledwith the study number, animal identificatiodna,te of collections,tudy day and collectiontimepoint.Allsamples willbe immediately frozenon dry iceand maintained frozen (-70*C or below) untilshipment to the Sponsor foranalysis. Shippina Instructions: Allsamples willbe maintained frozen(-700C or below) untilshipment foranalysis. Samples willbe shipped on frozen on dry icevia overnightmail. A packing listwillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitor willbe notifiedin advance of sample shipment. METHOD OF SACRIFICE - Fo GENERATION RATS: Rats willbe sacrificedby carbon dioxideasphyxiation. NECROPSY - Fo GENERATION RATS: Gross lesionswillbe retainedinneutralbuffered10% formalinforpossiblefuture evaluation(a tableof random unitswillbe used to selectone controlgroup ratfrom which alltissuesexamined at necropsy willbe retained,in order to provide control tissuesforany possiblehistopathologicaelvaluationsof gross lesions).Unless specificallcyitedbelow, allothertissueswillbe discarded. Rats Not Selected for Continued Evaluation: Any remaining female ratswith or withoutconfirmed evidence of mating thatwere assigned to eitherof the treatedgroups or the controlgroup priorto cohabitationwillbe sacrificedand discarded withoutfurtherevaluationat the discretionof the Study Directorand the Study Monitor. Rats Not Delivering a Lifter: Rats thatdo not delivera littewrillbe sacrificedon day 25 of presumed gestationand examined forgross lesions.Uteriwillbe stainedwith 10% ammonium the absence of implantationsites(s). sulfideto confirm 41"15:PAGE C-13 PMtoool 418-015 Page 13 Scheduled SacrWice: On day 22 postpartum, following the finalcollection intervalfor urine and f6cal samples and blood sample collectionf,emale ratswillbe sacrificeda,nd a gross necropsy of the thoracic,abdominal and,pelvicviscerawillbe performed. The number and distribution ofimplantationsiteswillbe recorded. A liversection(rightlateralobe)from each dam willbe collectedf,rozen and stored(-70*C or below) untilshipment to the Sponsor for analysis. Dams with No Surviving Pups: Dams with no survivingpups willbe sacrificedafterthe lastpup isfound dead, missing or presumed cannibalized. Priorto sacrificea,blood sample (approximately4 mL) willbe collectedfrom the matemal ratviathe inferiovrena cava and transferredintoserum separatortubes. The sample willbe spun ina refrigeratecdentrifuge.The serum willbe transferredintoa polypropylene tube labeledwith the study number, animal identificatiodna,te of collections,tudy day and collectiontimepoint.The sample willbe immediately frozen on dry iceand maintained frozen(-700C or below) untilshipment tothe Sponsor for analysis. A gross necropsy of the thoracic,abdominal and pelvicviscerawillbe performed. A liversection(rightlateralobe)from each dam willbe collected,frozen and stored (-700C or below) untilshipment tothe Sponsor foranalysis.Postpartum data forthese dams willbe excluded from summary tables. Rats Found Dead or Moribund: Rats thatdie or are sacrificedbecause of moribund condition,abortionor premature deliverywillbe examined forthe cause of death or moribund conditionon the day the observationismade. The ratswillbe examined forgross lesions.A liversection(right lateralobe)from each dam willbe collectedf,rozen and stored(-70*C or below) until shipment to the Sponsor foranalysis.Pregnancy statusand uterinecontents of female ratswillbe recorded. Aborted fetuses and/or deliveredpups willbe examined to the extent possible.Uteriof apparently nonpregnant ratswillbe stainedwith 10% ammonium sulfideto confirmthe absence of implantationsites(s). Shii)t)ingInstructions: Allsamples willbe maintained frozen (-70"C or below) untilshipment foranalysis. Samples willbe shipped frozen on dry icevia overnightmail. A packing liswtillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitor willbe notifiedin advance of sample shipment. 418-015:PAGE C-14 PMtoCol 418-015 Page 14 TESTS, ANALYSES AND MEASUREMENTS - Fi GENERATIO Viabilily: Postpartum Period: Lifterswillbe observed fordead pups at leasttwice daily.The pups in each littewrillbe counted once daily. ClinicalObservations and/or General Ag)g)earance: Postpartum Period: Once daily. Clinicalobservationsmay be recorded more frequentlythan citedabove, ifdeemed appropriateby the Study Directorand/or the Study Monitor. Body Weights: Postpartum Period: Days I (birth)4,, 7, 14 and 21 postpartum. Sacrifice: Terminal weight. METHOD OF SACRIFICE - Fl GENERATION RATS: As previouslycitedforFo generation rats. NECROPSY - Fl GENERATION RAT : Gross lesionswillbe retainedin neutralbuffered 10% formalinforpossiblefuture evaluation(a tableof random unitswillbe used to selectone controlgroup ratof each sex from which alltissuesexamined at necropsy willbe retained,in order to provide controltissuesforany possiblehistopathologicaelvaluationsofgross lesions).Unless specificallcyitedbelow, allothertissueswillbe discarded. Caps and labeledtubes willbe weighed (combined weight,tothe nearest .001 gram) before and afterretentionof pooled pup samples. These weights willbe documented in the raw data,and copies of these weights willbe includedwith the packing lisptriorto shipment. 41 Ml 5:PAGE C-1 ProtoCol418-015 Page 15 PuRs Found Dead on Day I PosIRartum: Pups thatdiebeforeexaminationofthe liftefrorpup viabiliwtiyllbe evaluatedforvital statusat birth.The lungs willbe removed and immersed inwater. Pups with lungs that sink willbe identifieads stillbom;pups with lungs thatfloatwillbe identifieads livebom, and tohave diedshortlyafterbirth.Pups withgrosslesionswillbe preservedinBouin's solutionforpossiblefutureevaluation.Alllungs willbe preserved in Bouin's solutionfor possiblefutureevaluation. PuRs Found Dead or Moribund on Days 2 to 21 Pos!Rartum: Pups found dead or sacrificedbecause of moribunditywillbe examined forgross lesionsand forthe cause of death or the moribund condition.Pups with gross lesions found on days 2 to 4 postpartum willbe preserved in Bouin'ssolutionforpossiblefuture evaluation;gross lesionsof pups found on days 5 to 21 postpartum willbe preserved in neutralbuffered10% formalin. For allpups found dead on days 2 to 4 postpartum, alllungs willbe preserved in Bouin'ssolutionforpossiblefutureevaluation.For allpups found dead on days 5 to 21 postpartum, alllungs willbe preserved in neutralbuffered 10% formalinforpossible futureevaluation. Pug)s Not Selected for Continued Observafion - Day 4 Pos!Rartum: Allpups culledon day 4 postpartum willbe sacrificedvia decapitation.Alllungs and liverswillbe collectedfrom the firstten pups culled(irrespectivoef littefrr)om each dosage group; the lungs willbe individuallryetainedinBouin's solution,and the livers willbe individuallryetainedin neutralbuffered 10% formalinforpossiblefuture evaluation.Remaining culledpups willbe sacrificedand discarded withoutevaluation. Scheduled Sacrifice: On day 21 postpartum, blood samples willbe collectedfrom allremaining pups on study. Blood samples willbe collectedviathe inferiovrena cava from each pup, pooled (per littearn)d transferredintoserum separator tubes. The samples willbe spun ina refrigeratecdentrifuge.The serum willbe transferredintopolypropylene tubes labeled with the study number, animal identificatiodna,te of collections,tudy day and collection timepoint.Allsamples willbe immediately frozen on dry iceand maintained frozen (-700C or below) untilshipment tothe Sponsor foranalysis. The pups willbe examined forgross lesions.The liverfrom each pup willbe collected, pooled (perlitterf)r,ozen and stored(-700C or below) untilshipment to the Sponsor for analysis. 4lMl5:PAGE C-16 PMtoool 418-015 Page 16 Shipping Instructions: Allsamples willbe maintainedfrozen(-700C or below) untilshipment foranalysis. Samples willbe shippedfrozenon dry iceviaovernightmail.A packingliswtillbe includedwith the samples and sent to KrisJ. Hansen, Ph.D., at the previouslycited address. Both the recipientand the Study Monitor willbe notifiedin advance of sample shipment. STATISTICAL EVALUATION: Averages and percentages willbe calculated.Lfttervalues willbe used where appropriate.Additionalprocedures and/or analyses may be performed, ifdeemed appropriate. DATA ACQUISITION, VERIFICATION AND STORAG Data willbe hand- and/or computer-recorded. Records willbe reviewed by the Study Directorand/or appropriatemanagement personnel within21 days aftergeneration. All originalrecords willbe stored inthe archives of the Testing Facility.Alloriginaldata will be bound and indexed. A copy of allraw data willbe suppliedto the Sponsor upon request. Preserved tissueswillbe stored at the Testing Facilitayt no charge forone year aftermailingof the draftfinalreport,afterwhich time the Sponsor willbe contacted to determine the dispositionof these materials. 418-015:PAGE C-17 PMtoool 41 "15 Page 17 RECORDS TO BE MAINTAINED: Protocol and Amendments. Test ArticleV,ehicle and/or Reagent Receipt,Preparationand Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (ifprescribedby StaffVeterinarian). General Comments. ClinicalObservations and/or General Appearance. Tissue and Sample Collection,Processing and Shipment. Cap and Labeled Tube Weights. Body Weights. Feed Consumption Values. Natural DeliveryObservations. LitterObservations. Gross Necropsy Observations. Organ Weights (ifrequired). Photographs (ifrequired). Study Maintenance (room and environmental records). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. KEY PERSONNEL: Executive Directorof Research: Mildred S. Christian,Ph.D., Fellow,ATS Directorof Research: Alan M. Hoberman, Ph.D., DABT Associate Directorof Research and Study Director:Raymond G. York, Ph.D., DABT Directorof Laboratory Operations: John F. Barnett,B.S. Directorof Study Management: ValerieA. Sharper, M.S. Manager of Animal Operations and Chairperson, InstitutionAanlimal Care and Use Committee: Dena C. Lebo, V.M.D. Directorof Operations and Compliance: Barbara J. Patterson,B.A. Consuftant,VeterinaryPathology: W. Ray Brown, D.V.M., Ph.D., ACVP 41Ml 5:PAGE C-18 Pmtocol 418-015 Page 18 FINAL REPORT: A comprehensive draftfinalreportwillbe prepared on completionofthe study and will be finalizedfollowingconsultationwith the Sponsor. The reportwillincludethe following: Summary and Conclusion. Experimental Design and Method. Evaluationof Test Resufts. Appendices: Figures,Summary and IndividuaTlables Summarizing the Above Data, Protocoland Associated Amendments and Deviations,Study Directoes GLP Compliance Statement, Reports of Supporting Data (ifappropriate)and QAU Statement. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described in thisprotocolhave been reviewed by the Testing Facility's InstitutionAanlimal Care and Use Committee. Allprocedures described inthisprotocol thatinvolvestudy animals willbe conducted in a manner to avoid or minimize discomfort,distressor pain to the animals. The Sponsors signaturebelow documents the factthat informationconcerning the necessityforconducting thisstudy and the factthatthisisnot an unnecessarily duplicativestudy may be obtained from the Sponsor. No alternative(invitro) procedures were availableformeeting the stated purposes of the study. 418-015:PAGEC-19 PMtocol41M 15 Page 19 REFERENCES: 1. Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Protection Agency, Washington, D.C. National Technical Information Service, U.S. Department of Commerce, Springfield,VA 22161. 2. Christian, M.S. (1984). Reproductive toxicityand teratology evaluations of nattrexone (Proceedings of Nattrexone Symposium, New York Academy of Sciences, November 7, 1983), J. Clin.Psychiat. 45(g):7-1 0. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity(Teratology) Control Data in the Chailes River Ctl.*CDWR Rat Charles River Laboratories, Inc.,Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories, Inc.) 4. Instituteof Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski, E. (1964). Farbemethode zum makroskopischen Nachweis von lmplantationsstelien am Uterus der Ratte. Arch. Pathol. Exp. Pharmakol. 247:367. PROTOCOL APPROVAL: FOR THE TESTING FACILITY AlanM. Hoberman,Ph.D.,DABT Directorof Research 418-015:PAGE C-20 Protocol41 Ml 5 Page20 6 - ,vuv-rb Date 4y-lm-pndG. York, PkD), DABT Associate Directorof Research Study Director Date Dena C. Lebo, V.M.D. Chairperson, InstitutionAanlimal Care and Use Committee FOR THE SPONSOR Marvin T. Case, D.V.M., Ph.D. Study Monitor Date 17 Al..,j Date 418-015:PAGE C-21 ATTACHMENT I SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE A'TTACHMENT 1 418-015:PAGE C-22 Protocol418-015 Page I of 2 STUDY SCHEMATIC PHARMACOKINETIC RECOVERY STUDY8 StartOf Dosage Female Rats - Premating Period (42 Days) End of DosaW Scheduied Sacrffice Cohabitation Period (5 days) Presumed Gestation Period Posipartunv Lactabon Period Dosage Period. a. For additionaldetailssee "Tests,Analyses and Measurementsm section of the protocol. b. Fo generation female ratswillreceivetestarticloer vehicleuntilmating isconfirmed (day 0 of presumed gestation). ATRACHMENT 1 418-015:PAGE C-23 Pmtocol 418-015 Page 2 of 2 SCHEDULEO 17 NOV 98 23 NOV 98 04 JAN 99 PM - 09 JAN 99 AM 05 JAN 99 09 JAN 99 26 JAN 99 03 FEB 99 29 JAN 99 06 FEB 99 30 JAN 99 03 FEB 99 15 FEB 99 - 23 FEB 99 16 FEB 99 - 24 FEB 99 29 JUN 99 Animals Arrive-AcclimationBegins. Dosage Period - Female Rats [42 days priorto cohabitationuntilconfirmed mating (day 0 of presumed gestation)]. Cohabitation Period. FirstPossible Day 0 of Presumed Gestation. Last Possible Day 0 of Presumed Gestation. FirstPossible Delivery(Day 21 of presumed gestation). Last Possible Delivery(Day 25 of presumed gestation). FirstPossible Day 4 Postpartum Culling. Last Possible Day 4 Postpartum Culling. FirstPossible Day 25 of Presumed Female Sacrifice. Last Possible Day 25 of Presumed Female Sacrifice. Gestation Gestation Scheduled Sacrifice- Fl Generation Pups (Day 21 postpartum). Scheduled Sacrifice- Fo Generation Dams (Day 22 postpartum). DraftFinalReport. a. The study initiatiodnate isthe day the Study Directorsigns the protocol. 418-015:PAGE C-24 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 418-015:PAGE C-25 14ATERIAL SAFETY DATA $MEET 3M 3H Center at. Paul, Minnesota 551"-1000 1-600-364-3577 or (612) 737-6501 (24 hours) Copyright, logg, Minnesota Mining end Manufacturing Company. All rights reserved. Copying and/or downloading of this information for the purpose of properly Utilizing 3M products is allowed provided that: 1) the information is copied in fu.U with no changes unless prior agreement2is obtained frm 3M, and 2) neither the copy nor the original is resold or otherwise distributed with the intention of earning a profit thereon. DIVISION: 3M CHEMICALS TRADE NAME: FC-95 FLUORAD Brand Fluorochanical Surfactant ID NUMBER/U.P.C.: 96-0207-0103-7 00-SI13S-09054-1 98-0207-0104-S 96-0211-0886-5 00-51135-09362-7 98-0211-3916-1 ZF-0002-10"-l ISSUED: January 29, 1998 SUPERSEDES: November 05, 1997 DOCUMENT: 10-3796-9 00-51135-09056-8 00-51135-02311-2 ----------------------------------------------------------------------------- 1. INGREDIENT C.A.S. NO. PERCENT ----------------------------------------------------------------------------- POTASSIUM PERFLUORDALKYL SULFONATE ...... 2795-39.3 82 as POTASSIUM PERFLUOROALKYL SULFONATE ...... 3871-99-6 3 a POTASSIUM PERFLUOROALKYL SULFONATE ...... 29-420-49-3 3 7 POTASSIUM PERFLUOROALKYL SULFONATE ...... 60270-SS-6 2 6 POTASSIUM PERFLUOROALKYL SULFONATE ...... 3872-2S-1 1 3 ........................................................................... 2. PHYSICAL DATA ----------------------------------------------------------------------------- BOILING POINT: ...............N./A. VAPOR PRESSURE: ..............N./.A VAPOR DENSITY: ...............N./.A EVAPORATION RATE: .............N/.A SOLUBILITY IN WATER: ..........s.light SPECIFIC GRAVITY: .............c.a. 0.6 Water-I (Bulk) PERCENT VOLATILE: .............0.% PH: ........................7..-.B. (0.1% Aqueous) VISCOSITY: ..................N./.D. MELTING POINT: ...............N./.D APPEARANCE AND ODOR: Light colored, free flowing powder. ----------------------------------------------------------------------------Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-015:PAGE C-26 JOS: lr,-gspLuoRAD Brand irluorochosical Surfactant January 29, 1998 PAGE 2 ........................................................................... 3. FIREAND EXPLOSIONHAZARDDATA ...................a ....................................................... FLASH POINT:....a .............No.ne FLAMMABLE LIMITS - LEL: ......:. MIA FLAMMABLE LIMITS - UEL: ....... MIA Au-roiGNrTION TEMPERATURE: ......N/A EXTINGUISHING NOIA: Water, Carbon dioxide, Dry chemical, Foam SPECIAL FIRE FIGHTING PROCEDURES: Wear full protective clothing, including haimt, self-contained, positive pressure or pressure demand breathing apparatus, bunker coat and pants, bands around arms, waist and 1*ga, face mask, and protective covering for exposed areas of the head. UNUSUAL FIRE AND EXPLOSION HAZARDS: See hazardous Decomposition section for products of combustion. ----------------------------------------------------------------------------4. REACTIVITY DATA ----------------------------------------------------------------------------- STABILITY: Stable INCOMPATIBILITY - MATERIALS/CONDITIONS Not applicable. TD AVOID: HAZARDOUS POLYMERIZATIDN: hazardous polymerization will not occur. HAZARDOUS DECOMPOSITIDN PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, hydrogen Fluoride, Toxic Vapors, Gases or Particulates. ----------------------------------------------------------------------------S. ENVIRONMENTAL INFORMATION ----------------------------------------------------------------------------- SPILL RESPONSE: Observe precautions from other sections. Vacuum, use wet sweeping compound or water to avoid dusting. CAUTIONI A vacuum cleaner could be an ignition source. Clean up residue with water. Place in an approved metal container. Seal the container. RECOMMENDED DISPOSAL: Do not release to waterways or sewer. Do not use in products or processes that could result in aquatic concentrations greater than 1/10 of the lowest ECSO or LCSC concentration. Incinerate in an industrial or commercial facility in the presence of a combustible material. Combustion products will include MF. Disposal alternative: Dispose of waste product in a facility permitted to ----------------------------------------------------------------------------- Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-015:PAGE C-27 W.M: M. gs IrLuoRAD Brand friucrochemical Surfac'tant january 29, ION PAGE 3 ----------------------------------------------------------------------------S. ENVIRONMENTAILNFORMATION (continued) ----------------------------------------------------------------------------- accept chemical waste. ENVIRONMENTAL DATA: 96-mr. Aquatic Fish LCSO, fathead Hinnow(Pinsphales prowelas)=38 mg/l, gluagill Sunfish(Lepmis macrochirus)-IM mg/l, Rainbow Trout(Saloo gairdneri)-Il mg/l; 48-Mr. ECSO, Daphnis Magna - So moll; CDD=.004 g/g; BOD20 - Nil. REGULATORY INFORMATION: Volatile organic Compounds: N/A. VOC Less M20 & Exempt Solvents: N/A. Since regulations vary, consult applicable regulations or authorities before disposal. U.S. EPA hazardous Waste Number a None (Not U.S. EPA Hazardous). This product complies with the chemical registration requirements of TSCA, EINECS, CDSL, AICS, MITI and Korea. EPCRA HAZARD CLASS: FIRE MAZARD: No PRESSURE: No REACTIVITY: No ACUTE: Yes CHRONIC: Yes ----------------------------------------------------------------------------6. SUGGESTED FIRST AID ------------------------------------------------------------------------ EYE CONTACT: Immediately flush eyes with large amounts of water for at least 15 minutes. Get immediate medical attention. SKIN CONTACT: Immediately flush skin with large amounts of water. Remove contaminated clothing. If irritation persists, call a physician. Wash contaminated clothing before rouse. INHALATION: If signs/symptoms occur, remove person to fresh air. If signs/symptoms continue, call a physician. IF SWALLOWED: Drink two glasses of water. Call a physician. ----------------------------------------------------------------------------7. PRECAUTIONARY INFORMATION ----------------------------------------------------------------------------- EYE PROTECTION: Avoid eye contact. Wear vented gaggles. ----------------------------------------------------------------------------Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-015:PAGE C-28 peds: re.S IRLUDW arand fluorochosimSaurfactant isnuary 29, 1998 PAGE 4 --------------------------------------------------w -.-.-.-.-.-.-...a.-.------ 7. PRErAUrIONARY INFORMATION (continued) ----------------------------------------------------------------------------- SKIN PROTECRION: Avoid skin contact. woor appropriate gloves when handling this material. A pair of gloves made from the following mterial(s) are recommended: butyl rubber. Us* one or nor* of the following personal protectlon items as necessary to prevent skin contact: head covering, coveralls. Protective garments (other than gloves) should be made of either of the following materials: polyethylene/polyvinylidene chloride (Baranex). RECOMMENDED VENTILATION: Use with appropriate local exhaust ventilation. Use in a wellventilated area. Provide sufficient ventilation to maintain emissions below recommended exposure limits. If exhaust ventilation is not adequate, use appropriate respiratory protection. RESPIRATORY PROTECTION: Avoid breathing of dust. Select one of the following MIDSH approved respirators based on airborne concentration of contaminants and in accordance with OSHA regulations: half-sask dust and mist respirator, half-mask supplied air respirator, full-face dust and mist respirator, full-face supplied air respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or smoke when using this product. Wash exposed areas thoroughly with soap and water. Wash hands after handling and before eating. RECOMMENDED STORAGE: Keep container dry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonflammable. OTHER PRECAUTIONARY INFORMATION: No smoking: Smoking while using this product can result in contamination of the tobacco and/or smoke and lead to the formation of the hazardous decomposition products mentioned in section 4 of this MSOS. mmrs HAZARD RATINGS: HEALTH: 2 FLAMMABILITY: 0 REACTIVITY: 0 PERSONAL PROTECTION: X (See precautions, section 7.) EXPOSURE LIMITS INGREDIENT VALUE UNIT TYPE AUTH SKIN* ----------------------------------------------------------------------------- POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MGIM3 TWA 3M y POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUDROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MG/f43- TWA 3M y ----------------------------------------------------------------------------- Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-015:PAGE C-29 MSD,S: Fr,.@FLUDW January 211,1998 Brand fluorocheaica3. Surtactant EXPOSURE LIMITS (continued) PAW 5 VALUE UNIT TYPE AUTH SKINO INGREDIENT ........................................................................... poTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 NG/N3 ThA 3m y - SKIN NOTATION: Listed substances indicated with 'Y' under SKIN refer to -thepotential contribution to the overall exposure by the cutaneous route including mucous membrane and my*, either by airborne or, more particularly, by direct contact with the substance. Vehicles can alter skin absorption. SOURCE OF EXPOSURE LIMIT DATA: - 3m: 3M Recommended Exposure Guidelines ----------------------------------------------------------------------------- S. HEALTH HAZARD DATA ----------------------------------------------------------------------------- EYE CONTACT: Mild Eye Irritation: signs/symptoms can include redness, swelling, pain, and tearing. SKIN CONTACT: Mild Skin Irritation (after prolonged or repeated contact): signs/symptoms can include redness, swelling, and itching. May be absorbed through the skin and persist in the body for an extended time. INHALATION: May be harmful if inhaled. May be absorbed by inhalation and persist in the body for an extended time. Single overexposure, above recommended guidelines, may cause: Irritation (upper respiratory): signs/symptoms can include soreness of the nose and throat, coughing and sneezing. IF SWALLOWED: Ingestion is not a likely route of exposure to this product. Illness may result from a single swallowing of a moderate quantity of this material. May be harmful if swallowed. MUTAGENICITY: Mutagenicity assays indicate the product is not mutagenic. ----------------------------------------------------------------------------Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately 418-015:PAGE C-31-1, MSDS: pe.gs IRLWW isnuary 29, 1998 Brand Fluarachosical Surfactant PAGE 6 ------------------------------------------------------------------------ 8. ME&TH MA2MD [ATA (continued) ----------------------------------------------w .-.-.-........................ FtEPRODUCTIVE/DEVELDPMWAL TOXINS: Not toratogenic in the rat at oral domes below maternally toxic levels. OTHER HEALTH HAZARD INFORMATION: This product is not known to California Proposition SS. contain any substances regulated under A Product Toxicity Summary Sheet is available. ---------------------------------------------------------w ------------------SECTION CHMGE DATES -------------------------------------w--------------------------------------- HEADING SECTION CHANGED SINCE November 05, 1997 ISSUE ----------------------------------------------------------------------------Abbreviations: N/D - Not Determined N/A - Not Applicable CA - Approximately ------------w -------------------------------------------w ------------------- The information in this Material Safety Date Sheet (MSDS) is believed to be correct as of the date issued. 3M MAKES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILrTy OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE OF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whether the 3M product is fit for a particular purpose and suitable for user's method of use or application. Given the variety of factors that can affect the use and application of a 3M product, some of which are uniquely within the user's knowledge and control, it is essential that the user evaluate the 3M product to determine whether it is fit for a particular purpose and suitable for user's method of use or application. 3M provides information in electronic form as a service to its customers. Due to the remote possibility that electronic transfer may have resulted in errors, omissions or alterations in this information, 3M makes no representation* as to its completeness or accuracy. In addition, information obtained from a database may not be as current as the information in the 14SDS available directly from 3f4. 418-015:PAGE C-31 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 418-015:PAGE C-32 ATRACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION Pmtocol 418-015 Version:418-015 (06NOV 98) Page I of2 PROCEDURE Test Article: Vehicle: PFOS 0.5% TweeriG 80 in R.O. Deionized Water A. Purpose: The purpose of thisprocedure isto providea method forthe preparation of dosage suspensions of PFOS and the vehiclefororaladministrationto ratson Argus Study 418-015. B. General Information: 1 . Allsuspension containerswillbe labeledand colorcoded. Each labelwill specifythe protocolnumber, testarticleidentificatioAnr,gus batch number, concentration,dosage level,preparationdate,expirationdate and storage conditions. 2a. Suspensions willbe prepared: X Daily Weekly -,For days of use 2b. Vehicle willbe prepared: Daily X Weekly For days of use 3. Suspensions willbe prepared at a finaldosage volume of 5 mukg. 4. Safety: X Gloves, lab coat,goggles or safetyglasses and faceshield X Dust-Mist Respirator - Half-Face Respirator - Full-FaceRespirator/PosftivePressure Hood Tyvek Suit/Apron 5. Dosage suspensions adjusted forFree base and % Purity. Yes X No (Calculationsbased on 100%) FreeBase _ Purity 6. Sampling requirements: Cited in protocol. 7. Storage: Cited inprotocol. 418-015:PAGE C-33 A77ACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION Pmtocol 418-015 Version:418-015 (06NOV 98) Page 2 of 2 PROCEDURE NOTE: Test articlewillbe prepared as a serialdilutiofnrom the high dosage to the low dosage. Once the finalvolumes are achieved, stirbars are to be added to the containers;mixing should occur during sampling and/or administration. C. Preparation of Vehicle 1 Add the requiredamount of R.O. deionized water to an appropriately labeledcontainer.Heat the water to 500C t 5*C, add the required amount of Tweene 80 and mix untiluniform (See TEST ARTICLE CALCULATIONS). D. Test ArticleSuspension Preparation: 1 . To prepare the 0.32 mg/mL, Group IIIsuspension, add the required amount oftestarticl(eSee TEST ARTICLE CALCULATIONS) intoan appropriatelysized,labeledcontainer. QS ad to the requiredamount with vehicleand heat the mixture to 80 0C 5 0C forapproximately 30 minutes or untilthe TA/S dissolves. 2. Once the testarticlheas dissolved;spin whilethe suspension cools. (Be sure there isa visiblevortex,thiswillachieve the desired emulsion. This may be prepared the day before use.) 3. To prepare the 0.02 mg/mL, Group 11suspension, remove the required amount of stock suspension (Group 111()See TEST ARTICLE CALCULATIONS), QS ad with the vehicleand mix. 4. To prepare the 0 mg/mL, Group Isuspension, add requiredamount of vehicleto an appropriatelysized,labeledcontainer(See TEST ARTICLE CALCULATIONS) and mix. Written by: Approved by: Date: /(,-Wa"9r Clarification: o Qes (See attached clarificatifoonrm.) Initials/Date 418-015P:AGE C-34 PRIMFDICA ArLzuRsesearcLhaboratoriIensc.. 905 Sheehy Drive.BuildingA Horsham. PA 19044 Telephone:(215)443-8710 Telefax:(215)443@-8597 PROTOCOL 418-015 Oral(Gavage) Pharmacokinetc Recovery Study ofPFOS inRats SPONSOR'S STUDY NUMBER: T-6295.14 Amendment 1 - 07 January 1999 1. Analyses ofPrer)aredFormulations,ConcentrationofTest ArticlFeormulations (Page 4 ofthe protocol): [Effectivdeate: 06 January 1999] Duplicatesamples (2mL each) willbe taken from the firsatnd lastpreparationratherthan,the firsatnd lastpreparationon the day prepared. Reason forChange: The preparationoccursthe day beforeitisused fordosage administratioannd, accordingthe toprotocold,ay 0 ofgestationisthe lastday ofdosage administrationS.ince day 0 ofgestationisthe day thatthe ratsare confirmed pregnant,the sample cannot be taken ofthe lastday ofpreparation. Alan M. Hoberman, Ph.D.,DABT Directoorf Research OIL< e??:-O-w99 Date kay;@ond G. York, h. .,DABT Date AssociateDirector1:@:esearch and Study Director Dena C. Lebo,V.M.D. @:ate MarvinT. Case, D.V.M.,Ph.D. Date Chairperson,InstitutioAnnailmal Care and Study Monitor Use Committee APPENDIX D DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY 418-015:PAGE D-1 STANDARD DEVIATIONS FROM THE PROTOCOL AND OPERATING PROCEDURES OF THE TESTING FACILITY Allratswere administeredthe testarticloer vehicleon premating day 43, 4 January 1999. The ratsshould have been placedintocohabitationon premating day 42 afterdosage administrationT.hisdeviationdid not adverselyaffectthe outcome or interpretatiofn the study because no data were lost. 2. From 23 November 1998 to26 November 1998 (days 1 to4 ofthe premating period)t,hefollowingFo generationfemale ratsreceivedthe incorrectlcyalculatedamount oftestarticlienvehicle,resultinginthe rats receiving25% more ofthe testarticltehan required. Dosage Groug 11 111 Dosage (ma/kg/day) Rat Numbers 0.1 13738 -13749 1.6 13750 - 13761 These deviationsdid notadverselyaffectthe outcome or interpretatiofn the study because the dosages were onlyforthe firsftourdays out ofa totalof43 days oftestarticlaedministratiopnriorto mating. Alldeviationsa cumented inthe raw data. nd G. York,P,h@@, D ABT Date AssociateDirectorof e,,e.asr;ch and Study Director TEMPERATURE APPENDIX E AND RELATIVE HUMIDITY REPORTS ARGUS 418-015:PAGE E-1 Temperature and RelativeHumidity Report Location: Room 15 Protocol Number. 418-015 Range of Dates: 17-Nov-1998 14:00 to 27-Nov-1998 08:55 TargetRange: Species:rat TotalNumber of Days: TotalNumber ofHours: TotalNumber of DataPoints: Temperature 64OF to 79*F RelativeHumidity 30% to70% 11 234.72 235 11 234.72 235 Mean (:StD): Maximum: Median: Minimum: Number ofPointsinRange (%): Number ofPointsHigh (%): Number ofPointsLow (%): 67.0 (tC).9) 44.6 4.8) 68.9 53.4 67.2 45.6 63.4 30.3 232 (98.7) 235 (100.0) 0 (0-0) 0 (0.0) 3 (1.3) 0 (0-0) Report Generated:28-Apr-1999at12:33 COMMENTS: REVIEWED BY: DATE: Cumulativeby Location(vO4.01.97) ARGUS 418-015:PAGE E-2 Temperature and RelativeHurrddityReport Location:Room 04 Protocol Number. 418-015 Range of Dates: 27-Nov-1998 08:55 to 30-Nov-1998 09:15 TargetRange: Species:rat TotalNumber of Days: TotalNumber ofHours: TotalNumber of DataPoints: Temperature 64*F to79*F 4 71.99 75 RelativeHumidity 30% to70% 4 71.99 75 Mean (:tSD): 72.4 Maximum: 74.3 Median: 72.3 Minimum: 69.6 Number ofPointsinRange (%): 75 Number ofPointsHigh (%): 0 Number of PointsLow 0 Report Generated:28-Apr-9I99 at12:35 (t0-9) 45.1 68.7 44.1 40.0 (100.0) 75 (0-0) 0 (0-0) 0 4.8) (100.0) (0-0) (0-0) COMMENTS: REVIEVVED BY: ig!(LAak DATE: fzf@f Cumulativeby Location(vO4.01.97) ARGUS 418-015.-PAGE E-3 Temperature and RelativeHumidity Report Location:Room 27 Protocol Number. 418-015 Range of Dates: 30-Nov-1998 09:15 to 09-Dec-1998 14:30 TargetRange: Species:rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber ofData Points: Tomperatum 64'F to79OF 10 221.0 222 RelativeHumidity 30% to70% 10 221.0 222 Mean (SD): 71.3 Maximum: 72.4 Median: 71.3 Minimum: 70.0 Number ofPointsinRange (%): 222 Number ofPointsHigh (%): 0 Number ofPointsLow (%): 0 ReportGenerated:28-Apr-1999 at12:37 (0.5) 59.3 68.8 61.2 37.0 (100.0) 222 (0-0) 0 (0-0) 0 (6.7) (100.0) (0-0) (0-0) COMMENTS: REVIEWED BY: DATE: Cumulativeby Location(vO4.01.97) ARGUS 418-015:PAGE E-4 Temperature and RelativeHumidity Report Location:Room 28-29 Protocol Number. 418-015 Range of Dates: 09-Dec-1998 14:30 to 15-Dec-1998 15:44 TargetRange: Species:rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber ofData Points: Temperature 64*F to79OF 7 145.01 146 RelativeHumidity 30% to70% 7 145.01 146 Mean (:tSD): 70.7 Maximum: 76.5 Median: 70.4 Minimum: 69.1 Number of PointsInRange 146 Number ofPointsHigh (%): 0 Number of PointsLow 0 Report Generated:28-Apr-9I99 at12:39 (t1.1) 49.0 59.4 48.8 39.6 (100.0) 146 (0-0) 0 (0-0) 0 (t3.5) (100.0) (0-0) (0-0) COMMENTS: REVIEWED BY: DATE: !@I/f@f Cumulativeby Location(vO4.01.97) ARGUS 418-015-.PAGE E-5 Temperature and RelativeHumidity Report Location:Room 27 Protocol Number. 418-015 Range of Dates: I5-Dec-I998 15:44 to 21 -Dec-I998 13:45 TargetRange: Species:rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber ofData Points: Temperature 64*F to79*F 7 141.77 143 RelativeHumidity 30% to70% 7 141.77 143 Mean (SD): 70.6 Maximum: 72.5 Median: 70.5 Minimum: 69.0 Number of PointsinRange 143 Number ofPointsHigh (%): 0 Number ofPointsLow (%): 0 Report Generated:28-Apr-1999at12:41 0.8) 59.9 64.8 60.1 49.1 (100.0) 143 (0-0) 0 (0.()) 0 (2.6 (100.0) (0-0) (0-0) COMMENTS: REVIEWED BY: DATE: Cumulativeby Location(vG4.01.97) ARGUS 418-015:PAGEE-6 Temperatureand RelativHeumidityReport LocationR:oom 28-29 ProtocolNumber. 418-015 Range ofDates:21-Dec-I998 13:45to20-Feb-9199 15:00 TargetRange: Species:rat TotalNumber of Days: TotalNumber of Hours: TotalNumber of DataPoints: Temperature RelativeHumidity 64*F to79*F 30% to70% 62 1464.99 1466 62 1464.99 1466 Mean (SD): Maximum: Median: Minimum: Number ofPointsInRange (%): Number ofPointsHigh (%): Number ofPointsLow 69.0 1.1) 49.0 (:k10.6) 71.9 81.6 69.0 48.7 66.9 10.0 1466 (100.0) 1407 0 (0-0) 49 0 (0-0) 10 (96.0) (3.3) (0.7) Report Generated:28-Apr-1999at12:46 COMMENTS: REVIEWED BY: DATE: 5 rlff Cumulativbey Locatio(nvO4.01.97) ARGUS 418-015:PAGE E-7 Temperature DeviationsReport Location:Room 15 Protocol Number: 418-015 Range of Dates: 17-Nov-1998 14:00 to 27-Nov-1998 08:55 Temperature Target Range: Species: rat Date 25-Nov-1998 25-Nov-1998 25-Nov-1998 rime 03:00 06:00 07:00 Temp. 63.8 L 63.7 L 63.4 L 64*F to79*F Date Time Temp. H - Value outof range -High L m Value out ofrange -Low Temp. - Temperature OF Report Generated:28-Apr-1999at12:34 /These deviationdsidnotadverselyaffecttheoutcome orinterpretatoifotnhestudy. The followindgeviation(ism)pactedon theoutcome ofthestudyas described: r StudyDirector TaL, Date @a92 Deviationbsy Location(vO4.01.97) ARGUS 418-015:PAGE E-8 RelativeHumidity DeviationsReport Location:Room 28-29 Protocol Number: 418-015 Range of Dates: 21-Dec-I998 13:46 to 20-Feb-I999 15:00 Humidity Target Range: Species: rat 30% to70% Date 21-Dec-1998 29-Dec-1998 04-Jan-1999 05-Jan-1999 05-Jan-I999 06-Jan-1999 06-Jan-1999 06-Jan-1999 D6-Jan-I999 06-Jan-1999 06-Jan-I999 06-Jan-1999 18-Jan-1999 18-Jan-1999 I8-Jan-I999 18-Jan-1999 18-Jan-1999 Time 18:00 12:00 13:00 22:00 23:00 00:00 01:00 02:00 03:00 04:00 05:00 06:00 11:00 15:00 18:00 20:00 21:00 P. H. 77.2 H 70.3H 29.4L 25.3L 26.8L 19.3L 20.7L 17.0L 14.7L 15.7L 10.0L 10.2L 75.4H 72.4 H 72.0 H 73.4 H 75.4 H Date 21-Jan-1999 22-Jan-I999 22-Jan-I999 22-Jan-I999 22-Jan-I999 22-Jan-1999 23-Jan-1999 23-Jan-I999 23-Jan-1999 23-Jan-1999 23-Jan-1999 23-Jan-1999 24-Jan-1999 24-Jan-1999 24-Jan-I999 24-Jan-1999 24-Jan-1999 Time 18:00 03:00 05:00 07:00 09:00 21:00 06:00 09:00 13:00 18:00 19:00 23:00 00:00 03:00 04:00 05:00 06:00 H Value out ofrange -High L Value out of range -Low R.H.- RelativeHumidity Report Generated:28-Apr-1999 at12:53 R.H. 76.4H 70.9H 71.7H 70.4H 70.4H 70.5H 71.4H 72.3H 73.9H 78.2H 75.1H 77.9H 73.1H 80.7H 72.4H 72.2H 73.4H i,/ These deviationdsidnotadverselyaffectheoutcome orinterpretatoifotnhestudy. The followindgeviation(ism)pactedon theoutcome ofthestudyas described: Study Directo Date: Deviationbsy Location(vO4.01.97) ARGUS 418-015:PAGE E-9 RelativeHumidity DeviationsReport Location:Room 28-29 Protocol Number. 418-015 Range of Dates: 21 -Dec-I998 13:45 to 20-Feb-I999 15:00 Humidity Target Range: Species: rat 30% to 70% Date 24-Jan-1999 24-Jan-1999 24-Jan-I999 24-Jan-I999 24-Jan-I999 24-Jan-1999 24-Jan-I999 24-Jan-1999 24-Jan-I999 24-Jan-I999 24-Jan-I999 26-Jan-I999 26-Jan-I999 26-Jan-1999 26-Jan-1999 26-Jan-1999 26-Jan-1999 rime 07:00 09:00 10:00 11:00 12:C)O 13:00 14:00 15:00 16:00 17:00 18:00 11:00 15:00 16:00 17:00 18:00 20:00 R.H. 70.1 H 70.3 H 75.5 H 75.6 H 75.8 H 78.2 H 81.6 H 75.5 H 80.3 H 78.7 H 75.9 H 70.9 H 74.5 H 75.7 H 76.1 H 73.6 H 70.6 H Date 02-Feb-1999 02-Feb-1999 02-Feb-1999 02-Feb-I999 03-Feb-1999 03-Feb-I999 03-Feb-1999 06-Feb-1999 Time 19:00 20:00 21:00 23:00 01:00 03:00 07:00 12:00 H Value out of range - High L Value out of range -Low R.H. u RelativeHumidity Report Generated: 28-Apr-1999 at 12:54 R.H. 77.7 H 77.7 H 70.7 H 77.7 H 76.7 H 75.4 H 75.2 H 72.9 H These deviationsdid notadverselyaffectthe outcome or interpretationfthe study. The followingdeviation(si)mpacted on the outcome ofthe study as described: Study Directo Date: Deviationsby Location(vD4.01.97) STATEMENT APPENDIX F OF THE STUDY DIRECTOR @riPUMMICA 418-015:PAGE F-1 Argus ResearchLaboratoriesI,nc. 905 Sheehy Drive,BuildingA Horsham, PA 19044 Telephone:(215)443-8710 Teiefax(:215)443-8587 PROTOCOL 418-015: ORAL (GAVAGE) PHARMACOKINETIC RECOVERY STUDY OF PFO$ IN RATS SPONSORS STUDY NUMBER: T-6295.14 STATEMENT OF THE STUDY DIRECTOR This finalreportaccuratelyreflecttshe raw data obtainedduringthe performance of the study. No deviationsfrom the U.S. Food and Drug Administration(FDA) Good Laboratory PracticeRegulations; FinalRuleO,the Japanese Ministryof Health and Welfare (MHW) Good Laboratory Practice Standard for Safety Studies on Druge and the European Economic Community (EEC) Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisiorilrecommendation on compliance with principles of good laboratorypracticecoccurred thataffectedthe qualityor integritoyf the study. Ra@nd G. York (Ph. DABT Associate Director f search and Study Director Date a. U.S. Food and Drug Administration.Good Laboratory Practice Regulations; FinalRule. 21 CFR Part 58. b. Japanese Ministry of Health and Welfare (1988). Good Laboratory Practice Standard for Safaty Studies on Dnigs, MHW Ordinance Number 21, March 26, 1997. C. European Economic Community (1989). Council decision on 28 July 1989 on the acceptance by the European Economic Community of an OECD decisionlrecommendation on compliance with principles of good laboratory practice. Official Journal of the European Communities: Legislation.32(No. L 315; 28 October): 1-17. APPENDIX G QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Pummlc4k 418-015:PAGE G-1 Argus Research Laboratories.Inc. 905 Shechv Drive.BuildingA fiorsham.PA 19044 Telephone:(215)443-87 10 Telefax:(215)443-8587 QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT Study Director:Raymond G. York,Ph.D.,DABT ExecutiveDirectorof Research: MildredS. ChristianP,h.D.,Fellow,ATS Protocol418-015: Oral (Gavage) PharmacokineticRecovery Study of PFOS inRats Sponsor'sStudy Number T-6295.14 The draftprotocolforthisstudywas auditedforadherence to U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations,Japanese Ministroyf Healthand Welfare(MHW); Good LaboratoryPracticeStandard for SafetyStudieson Drugs,and European Economic Community (1989)council decisionon 28 July 1989 on the acceptance by the European Economic Community ofan OECD decision/recommendationon compliancewithprinciples ofgood laboratorypracticeon 18 OCT 98. Criticaplhases ofthisstudywere inspected10 times;studyinformation and raw data were auditedtwice(see tables1 and 2 fordatesand phases/data). The draftfinalreportand the raw data forthisstudywere compared and auditedforaccuracy,foradherence to protocolrequirements,and foradherence to U.S. Food and Drug Administratio(nFDA) Good LaboratoryPractice Regulations,Japanese MinistryofHealthand Welfare(MHW); Good Laboratory PracticeStandard forSafetyStudieson Drugs, and European Economic Community (1989)councildecisionon 28 July1989 on the acceptance by the European Economic Community ofan OECD decision/recommendationon compliancewith principleosf good laboratorypracticebetween 10 MAY 99 and 15 JUN 99, and forrevisionsrequestedby the Sponsor on 20 JUL 99, 22 JUL 99, and forfinalizatioonn 23 JUL 99. 418-015:PAGE G-2 This studywas conducted accordingto U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations,Japanese Ministry of Healthand Weffare(MHW); Good LaboratoryPracticeStandard forSafety Studieson Drugs.And European Economic Community (1989)councildecision on 28 July 1989 on theacceptance by the European Economic Community of an OECD decision/recommendationon compliance withprincipleosf good laboratorypractice. Nancy .Gongliewski QualityAssurance Manager A LIA-el 7 A *@@ A V/-':buomw(/SBk.oS,. 'Date Senior Quali'tuyAssurance Associate and PrincipalAuditor TABLE 1 CRITICAL PHASES INSPECTED 418-015:PAGE G-3 Test ArticleAdministratio-nGavaae Date ofinspection:23 NOV 98 Date resultsreportedtothe Study Directorand Management: 25 NOV 98 Test ArticlePrel?aration Date ofinspection:01 DEC 98 Date resultsreportedtothe Study Directorand Management: 02 DEC 98 Urine/FecalCollection Date of inspection:04 JAN 99 Date resultsreportedtothe Study Directorand Management: 04 JAN 99 Blood Collection Dates ofinspection:04 JAN 99, 17 FEB 99 Dates resultsreportedtothe Study Directorand Management: 04 JAN 99,17 FEB 99 Cohabitation Date of inspection:05 JAN 99 Date resultsreportedto the Study Directorand Management: 07 JAN 99 NaturalDelive!)L/LifWtaetrch Date of inspection:27 JAN 99 Date resultsreportedtothe Study Directorand Management: 02 FEB 99 418-015:PAGE G-4 Culling,Pup ScrificeD:ay 4 Date ofinspection:01 FEB 99 Date resultsreportedto the Study Directorand Management: 03 FEB 99 Necropsy, Fo Dams, El-P-u-2sDates ofinspection:17 FEB 99, 17 FEB 99 Dates resultsreportedto the Study Directorand Management: 22 FEB 99, 22 FEB 99 418-015:PAGE G-5 TABLE 2 RAW DATA AUDIT(S) The followinsgtudyinformatioannd raw datawere audited from 07 APR 99 to 13 APR 99: Protocol. Protocolamendment. Listof personneland computer operatorcodes. Errorcodes and codes forclinicaslignobservations. Animal receiptr,andomizationp,hysicalexaminationand acclimation. In-lifteransactiornecord. Feed consumption. Cohabitation. Naturaldeliveryobservations. Lifteorbservations. Pup body weightsand status. Pup Randomization. Necropsy. Organ weights. Tissuepackinglists. Male breedercolonyrecords. General comments. Study maintenance records. Temperature and relativheumidityreports. Feed, waterand beddinganalyses. Editrequests. Deviations. Data reviewpage. Blood collectiodnataand packinglists. Key fortestingfacilictoymputer backup recordabbreviations. Urine/Fecalcollectiodnataand packinglists. The resultsofthisauditwere reportedto theStudy Directorand Management on 15 APR 99. 418-015:PAGE G-6 The followingstudy informationand raw data were audited on 20 APR 99: Vehiclereceiptp,reparationand use. Test articlreeceiptp,reparationand use. Test articlpeacking lists. Deviations. The resultsofthisauditwere reportedto the Study Directorand Management on 23 APR 99.