Document x12wd5792n7790pLvGJanjopg

FINAL REPORT PROTOCOL 418-013 ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.12 FINAL REPORT DATE: 24 JUNE 1999 PROTOCOL 418-013 ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.12 TABLE OF CONTENTS SUBJECT 1. SUMMARY AND CONCLUSION A. Methods B. Results C. Conclusion I[. DESCRIPTION OF TEST PROCEDURES A. Conduct of Study A.l. Sponsor A.2. TestingFacility A.3. Study Number A.4. Sponsoes Study Number A.5. Purpose ofthe Study A.6. Study Design PAGE 1-1 1-1 1-2 1-4 11-1 11-1 11-1 11-1 11-1 11-1 11-1 11-1 SUBJECT A.7. RegulatoryCompliance A.8. Ownership of the Study A.9. Study Monitor A. 10. AlternateStudy Monitor A. 11. Study Director A. 12. Technical Performance A. 13. Report Preparation A. 14. Report Review A. 15. Date ProtocolSigned A. 16. Dates of Technical Performance A. 17. Records Maintained B. Test ArticleInformation B. 1. Description B.2. Lot/Batch Number B.3. Date Received and Storage Conditions B.4. Special Handling Instructions B.5. Analysisof Purity C. Vehicle Information C. 1. Description C.2. Lot Numbers C.3. Date Received and Storage Conditions C.4. Special Handling Instructions 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 11-3 11-3 11-4 11-4 11-4 11-4 11-4 11-4 SUBJECT C.5. AnalysisofPurity D. Test ArticlPereparationand Storage Conditions D. 1. Sample Information D.2. AnalyticalResults E. Test System E. 1. 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.1. Research FacilitRyegistration F.2. Study Rooms F.3. Housing F.4. Lighting F.5. Sanitization F.6. Feed F.7. Feed Analysis PAGE 11-4 11-4 11-5 11-5 11-5 11-5 11-5 11-5 11-5 11-6 11-6 11-6 11-6 11-7 11-7 11-7 11-7 11-7 11-7 11-8 11-8 11-8 SUBJECT PAGE F.8. Water 11-8 F.9. WaterAnalysis 11-8 G. Methods 11-9 G.1. DosageAdministration 11-9 G.2. RationalfeorDosageSelection 11-9 G.3. RouteofAdministration 11-9 G.4. RationalfeorRouteofAdministration 11-9 G.5. FrequencyofAdministration 11-9 G.6. LengthofStudy 11-9 G.7. MethodofStudyPerformance 11-10 G.8. GrossNecropsy 11-11 G.9. StatistiAcnaallyses 11-13 Ill. RESULTS 111-1 A. MortalitPyr,ematureDeliverCyl,inicaanld NecropsyObservations 111-1 A.1. Mortaliatnyd PrematureDelivery 111-1 A.2. ClinicOablservations 111-1 A.3. NecropsyObservations 111-1 B. Body WeightsU,terinWeeightsand Body WeightChanges 111-1 B.1. Precohabitation lii-i B.2. Gestation 111-2 C. Absolut(eg/daya)nd Relativ(eg/kg/daFye)ed Consumption Values 111-2 C.1. Precohabitation iv 111-2 SUBJECT C.2. Gestation D. Caesarean-Sectioning,Liftearnd FetalGross External Observations D. 1. Day 15 of Gestation D.2. Day 21 ofGestation REFERENCES APPENDIX A -REPORT FIGURES Figure 1. Body Weights -Precohabitation Figure 2. Maternal Body Weights - Rats Caesarea n-Sectioned on Day 15 of Gestation Figure3. MaternalBody Weights - Rats Caesarean-Sectioned on Day 21 of Gestation APPENDIX B - REPORT TABLES Table 1. Clinicaalnd Necropsy ObservationsSummary Table 2. Body Weights - Precohabitation- Summary Table 3. Body Weight Changes -Precohabitation-Summary Table 4. MaternalBody Weights and Gravid UterineWeights Gestation- Summary Table 5. Matemal Body Weight Changes -Gestation- Summary Table 6. AbsoluteFeed Consumption Values (g/day)Precohabitation- Summary Table 7. RelativeFeed Consumption Values (g/kg/day)Precohabitation- Summary Table 8. MaternalAbsoluteFeed Consumption Values (g/day)Gestation- Summary v PAGE 111-2 111-3 111-3 111-3 111-4 A-1 A-2 A-3 B-1 B-4 B-5 B-6 B-10 B-12 B-13 B-14 SUBJECT Table 9. Matemal RelativeFeed Consumption Values (g/kg/day)Gestation- Summary PAGE B-16 Table 10. Caesarea n-Sectioning Observations -Summary B-1 8 Table 11. LifteOrbservations(Caesarean-DeliveredEmbryos or Fetuses)- Summary B-21 Table 12. ClinicaOlbservations- IndividuaDlata B-22 Table 13. Necropsy Observations- IndividuaDlata B-29 Table 14. Body Weights -Precohabitation- IndividuaDlata B-34 Table 15. Matemal Body Weights - Presumed Gestation- IndividuaDlata B-44 Table 16. Feed Consumption Values - Precohabitation- IndividuaDlata B-49 Table 17. Matemal Feed Consumption Values - Presumed Gestation- IndividuaDlata B-54 Table 18. Caesarean-SectioningObservations- IndividuaDlata B-59 Table 19. LifteOrbservations(Caesarean-DeliveredEmbryos or Fetuses)- IndividuaDlata B-63 Table 20. Embryonal VitalStatusor FetalSex, VitalStatusand Body Weight - IndividuaDlata B-67 Table 21. PlacentalWeights - IndividuaDlata B-73 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 APPENDIX E - TEMPERATURE REPORTS AND RELATIVE HUMIDITY E-1 to E-3 APPENDIX F - HISTORICAL CONTROL DATA F-1 to F-13 vi SUBJECT APPENDIX G - STATEMENT OF THE STUDY DIRECTOR APPENDIX H - QUALITY ASSURANCE STATEMENT UNIT FINAL REPORT PAGE G-1 H-1 to H-5 vii 418-013:PAGE 1-1 TITLE: ORAL (GAVAGE) PHARMACOKINETIC RATS STUDY OF PFOS IN ARGUS RESEARCH LABORATORIES, INC. PROTOCOL NUMBER: 418-013 SPONSOR'S STUDY NUMBER: T-6295.12 1. SUMMARY AND CONCLUSION A. Methodsa Eighty virgin female Crl:CDOBR VAF/PlusO (Sprague-Dawley) rats were assigned tofivedosage groups (Groups IthroughV), 16 ratsper dosage group. The ratswere administeredthe testarticleP,FOS (FC-95),or the vehicle, 0.5% Tween@ 80 inReverse Osmosis Membrane Processed DeionizedWater (R.O.DeionizedWater),orall(yviagavage),once dailybeginning42 days priorto cohabitatiotnhrougheitherday 14 or 20 ofpresumed gestation(DG 14 or DG 20). Dosages were 0 (Vehicle)0,.1,0.4,1.6and 3.2 mg/kg/day. The dosage volume was 5 mukg. The ratswere observed forviabilitaytleasttwiceeach day ofthe study and for clinicasligns ofeffectsofthe testarticlbeeforeand approximatelyone hour after dosage and on the day sacrificedB.ody weightswere recorded dailyduringthe dosage periodand atsacrificeF.eed consumption values were recorded weekly tocohabitationand dailyduringpresumed gestation. Urine and fecalsamples were collectedone day priorto initiatioofncohabitation tothe followingmorning and DGs 6 to 7, 14 to 15 and 20 to 21. Samples were frozenand shipped tothe Sponsor foranalysis.Blood samples were collected on the day cohabitatiownas initiateadnd on DGs 7, 15 and 21. Blood was centrifugedand the serum was frozenand shipped tothe Sponsor foranalysis. Rats were sacrificeodn DG 15 or 21, as describedbelow, and a gross necropsy of thethoracica,bdominal and pelvicviscerawas performed. A liversectionand the milk-secretingglands from the axillaryt,horacic,abdominal and inguinal a. Detaileddescriptionosfallprocedures used inthe conduct ofthisstudy are providedinthe appropriatesectionsofthisreportand inAPPENDIX C (PROTOCOL AND AMENDMENT). 418-013:PAG'-:71-2 regionsofeach dam were collectedf,rozenand shipped tothe Sponsor for analysis. Eightrandomly selectedfemale rats,witha confirmeddate ofmating,per dosage group were Caesarean-sectionedon DG 15. The graviduteruswas excisedand weighed. Rats were examined fornumber and distributionf corporalutea,implantatiosnites,viableand nonviableembryos. A sample ofthe amnioticfluidofeach viableembryo was collectedand pooled by litterE.ach viableembryo was removed from the uteruswiththe attachedplacentaand pooled by lifterT.hese samples were frozenshipped to the Sponsor foranalysis. Allremainingfemale ratsineach dosage group were Caesarean-sectioned on DG 21 orestimated DG 21. The graviduteruswas excisedand weighed. Individuapllacentalweightswere recorded. Rats were examined fornumber and distributiofncorpora lutea,implantationsites,liveand dead fetusesand early and lateresorptions.Each fetuswas removed from the uterus,weighed and examined forsex and gross externalalterationsS.amples of the amnioticfluid were collectedwhen possible,and pooled by lifterP.lacentaewere collected and pooled by lifterB.lood samples were collectedfrom each fetus,pooled by liftearnd centrifuged.Allsamples were frozenand shipped tothe Sponsor for analysis. Lung and liversamples were collectedfrom threefetusesper liftefrrom two to fivelifterpser dosage group. Sectionsfrom the halfofthe lungsand one lobeof the livewrere shipped to PathologyAssociates InternationaDlu,rham, NorthCarolina,forpossiblefutureevaluation.The otherhalfofthe lungs and the otherlateralobe ofthe liverwere frozenand shipped tothe Sponsor for analysis. The liversofthe remainingfetuses,plusany remainingliverfrom the three selectedfetuses,were collectedp,ooled by liftearnd frozen.The remaining carcass(and head) ofeach fetuswas frozenand shipped tothe Sponsor for analysis. B. Results No deaths or premature deliveriewsere attributetdo PFOS treatment. Three rats(one each inthe 0.1,0.4 and 1.6mg/kg/day dosage groups)were sacrificed because the eye was injuredduringorbitaslinusbleeding.One ratinthe vehicle controlgroup did not have a confirmedmating date and deliveredon day 66 of the study. Two ratsinthe 3.2 mg/kg/day dosage group deliveredon DG 21. All otherratssurvivedto scheduled sacrifice. 418-013:PAGE 1-3 Alladverse clinicaolbservationsduringthe precohabitationand gestationperiods were consideredunrelatedtothe testarticlaend no gross lesionswere revealed by necropsy. Body weight gains forthe entireprecohabitationperiod (DSs 1 to42) were 88.8%, 80.8%, 66.3% and 17.4% ofthe controlgroup value inthe 0.1,0.4,1.6 and 3.2 mg/kg/day dosage groups,respectivelyB.ody weightswere 98.0%, 96.3%, 93.6% and 85.3% ofthe controlgroup value inthese fourrespective dosage groups on DS 42. As the resultof random selectionforassignment to Caesarean-sectioningon eitherDG 15 or DG 21, groups assigned toCaesarean-sectioningon DG 15 weighed lesson DG 0 than groups assigned to Caesarean-sectioningon DG 21. Reflectingthe random selectionand differencesinaverage body weights ofthe two subsets,as wellas the relativelsymallgroup sizes,body weight gains differedinthe two subsets. Inboth subsets,body weightgainswere reduced on DGs 0 to 7 ingroups administered0.4 mg/kg/day and higherdosages ofPFOS, afterwhich body weight changes did notdemonstrate a cleardosage-dependent pattern. Absolute(g/day)and relativ(eg/kg/dayf)eed consumption valueswere reduced inthe 0.4,1.6and 3.2 mg/kg/day dosage groups ineach week ofthe precohabitatiopneriod.Reflectintghese effectsofthe testarticlea,bsoluteand relativfeeed consumption values were reduced forthe entireprecohabitation period(DSs 1 to 42) inthe 0.4,1.6and 3.2 mg/kg/day dosage groups. Groups administered0.4 mg/kg/day and higherdosages ofthe testarticlhead reduced absoluteand relativfeeed consumption values duringthe firswteek of the gestationperiod.Absolutefeed consumption values continuedto be reduced forthe remainder ofthe dosage periodinthe 0.4 and 1.6 mg/kg/day dosage groups assigned to DG 15 Caesarea n-sectionnig and inthe 3.2 mg/kg/day dosage group forratsassigned toeitherDG 15 or DG 21 Caesarea n-sectioning. No Caesarean-sectioningor lifteprarameters were affectedby dosages ofthe testarticlaes high as 3.2 mg/kg/day administeredto ratsCaesarea n-sectioned on DG 15. The liftearverages forimplantationsl,iftesrizesand livefetuseswere reduced in the 3.2 mg/kg/day dosage group Caesarean-sectionedon DG 21. Values were below the ranges observed historicalaltythe TestingFacilityF.etalbody weights were alsoreduced inthe 3.2 mg/kg/day dosage group. No otherCaesareansectioningor lifteprarameters were affectedby dosages ofthe testarticlaes 418-013:PAGE 1-4 highas 3.2 mg/kg/day administeredto ratsCaesarean-sectionedon DG 21. No fetalgross externalalterationwsere observed. C. Conclusion The purpose ofthisstudy,as statedinthe protocolw,as to evaluatethe pharmacokineticsof PFOS inF6 generationdams and Fl generationfetuses followingPFOS treatmenttofemale ratsduringprematingand gestation.This reportsincludesstudy data from the in-Iffpeortionwhich was conducted atArgus Research Laboratories,Inc.The PK samples thatwere collectedduringthe inlifpeortionofthe study were senttothe Sponsor foranalysisand willbe reported separately.Itisthe responsibiliotfythe Sponsor, 3M CorporateToxicology,to combine the in-lifreesultswiththe analyticarlesultsintoa finalpharmacokinetic report.The resultsforthisstudymimic those reportedinthe Combined Oral (Gavage) FertilitDye,velopmental and PednataVPostnatalReproductionToxicity Study of PFOS inRats (Argus Research LaboratoriesI,nc.,Protocol418-008), reported10 June 1999. z t -Tvrv f MildredS. ChristianP,h.D.,Fellow,ATS Date Executive DirectorofResearch Alan M. Hoberman, Ph.D.,DABT Directorof Research )4@ ond G. York,P(i@, DABT As,c@;o*-c'- Director of F@psearch and Study Director 7-V Date Date ii. DESCRIPTION OF TEST PROCEDURES A. Conduct of Slqdy: 418-013:PAGE 11-1 A.I. SRonso: 3M CorporateToxicology,3M Center,Building220-2E-02, St.Paul, Minnesota 55144-1000 A.2. Testing Facilily: Argus Research Laboratories,Inc.,905 Sheehy Drive,BuildingA, Horsham, Pennsylvania 19044-1297 A.3. Study Number: 418-013 A.4. St)onsor'sStudy Number: T-6295.12 A.5. PurRose of the Study: The purpose ofthisstudy was toevaluatethe pharmacokineticsofPFOS in Fo generationdams and Fl generationfetusesfollowingPFOS treatmentof Crl:CDODBR VAF/PlusO female ratsduringpremating and gestation. A.6. Study Design: The requirementsof the U.S. Food and Drug Administration(FDA)(')were used as the basisofstudy design. A.7. Regulato[y Compliance: The study was conducted incompliance withGood LaboratoryPractice(GLP) regulationosfthe U.S. Food and Drug Administratio(nFDA)(')t,he Japanese Ministroyf Healthand Welfare (MHW)(3)and the European Economic Community (EEC)(")T.here were no deviationsfrom the GLP regulationtshataffectedthe qualityor integritoyf the study. QualityAssurance Unitfindingsderivedfrom the inspectionsduringthe conduct of thisstudy are documented and have been providedtothe Study Directorand the TestingFacilitMyanagement. A.8. Ownershit) of the Study: 418-013:PAGE 11-2 The Sponsor owns the study. Allraw data, analyses, reportsand preserved tissuesare the propertyofthe Sponsor. A.g. Study Monitor: Marvin T. Case, D.V.M.,Ph.D. A.10. AlternateStudy Monitor: 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 LaboratoryOperations) MargaretM. Martin(Research Associate) CorrieL.Pabst (QualityControlAssociate) A.13. Report Preparation: Raymond G. York,Ph.D.,DABT Jo Ann Frazee,M.S. (StudyCoordinator) Susan K. Bradshaw, B.S.(Data Management Specialist) Karen G. Parker,A.A. (ReportAdministrator) A.14. Report Review: MildredS. ChristianP,h.D.,Fellow,ATS (ExecutiveDirectorof Research) AllanM. Hoberman, Ph.D, DABT (Directoorf Research) A.15. Date ProtocolSigned: 9 November 1998 418-013:PAG7- 11-3 A.16. Dates of Technical Performance: Rat ArrivalDate Dosage Period(42 days priorto cohabitation untilDGa 14 or 20) CohabitationPeriod Caesarea n-SectioningPeriod (DG 15) Caesarean-SectioningPeriod(DG 21) 10 NOV 98 16 NOV 98 - 21 JAN 99 27 DEC 98 PM - 01 JAN 99 AM 12 JAN 99 - 13 JAN 99 18 JAN 99 - 22 JAN 99 A.17. Records Maintained: The originarleport,raw data and reservesamples ofthe bulktestarticlaend vehiclecomponents are retainedinthe archivesofArgus Research Laboratories, Inc.Any preservedtissuesare retainedinthe archivesofthe TestingFacilitfyor one year afterthe mailingofthe draftfinalreport,afterwhich time the Sponsor willdecidetheirfinaldispositionA.llunused prepared formulationswere discardedatthe TestingFacilityU.nused bulktestarticlweillbe returnedtothe Study Monitorupon completionofallwork withthe testarticle. B. Test ArticleInformation: B.I. Description: PFOS (FC-95)- an off-whitpeowder B.2. Lot/BatchNumber: 217 (Expiratiodnate: May 2000) B.3. Date Received and Storage Conditions: The testarticlweas receivedon 21 October 1998, and storedat room temperature. B.4. SpecialHandling Instructions: Standard safetyprecautions(use ofprotectiveclothingg,loves,dust-mist respirators,afetygoggles or safetyglassesand a face-shieldw)ere taken when handlingthe bulktestarticlaend prepared suspensions. a. DG isused as an abbreviatiofnorday of(presumed) gestation. 418-013:PAGE 11-4 B.5. Analysis of Puft: Informationregardingthe identityc,omposition,strengthand purityofthe test articlieson filewiththe Sponsor. C. Vehicle Information: C.I. Description: 0.5% Tween(D 80 inReverse Osmosis Membrane Processed Deionized Water (R.O.DeionizedWater). C.2. Lot Numbers: Tween@ 80 - M29477 and M03HO5 C.3. Date Received and Storage Conditions: The Tween@ 80 was receivedfrom J.T.Baker,PhillipsburNge,w Jersey,on 17 September 1998 (LotNumber M29477) and 3 December 1998 (LotNumber M03HO5), and storedatroom temperature.The R.O. deionizedwater is availablefrom a continuoussource atthe TestingFacilitaynd ismaintained at room temperature. C.4. Special Handling Instructions: Standard safetyprecautions(use of protectiveclothingg,loves,dust-mist respirators,afetygoggles or safetyglassesand a face-shieldw)ere taken when handlingthe vehiclecomponents and prepared vehicle. C.5. Analysis of Puft: Neitherthe Sponsor northe Study Directorwas aware ofany potential contaminants likeltyo be presentinthe vehiclethatwould interferweiththe resultsof thisstudy. D. Test ArticlePreparation and Storaae Conditions: Suspensions of PFOS were prepared dailyat concentrationsof 0, 0.02,0.08, 0.32 and 0.64 mg/mL. Prepared formulationswere storedat room temperature. D.I. Sample Information: 418-013:PAGE li-5 Sample Type Concentration (allevels) Bulk Test Article Reserve VehicieComponents Size 2 mL2 1g Date Retained 15 NOV 98 20 JAN 99 12 NOV 98 Storage/Shipping Conditions Frozen Shipped To Sponsor TestingFacility Room temperatu@e Archives Date Shipped 16 NOV 98 20 JAN 99 25 NOV 98 Reserve TweenO 80 5 mL R.O. deionized water 5 mL 12 NOV 98b 10 DEC 98c 12 NOV 98 Room temperature Room temperature TestingFacility Archives Testing Facility 25 NOV 98 27 JAN 99 25 NOV 98 a. Duplicate samples were taken from the first and last preparation Archives on the day prepared. I One sample I of each set was shipped for analysis. The remaining samples were retained at the Testing Facility as backups. Backup samples the Sponsor. are stored frozen (-70*C or below) and will be discarded at the request of b. Lot Number M29477. c. Lot Number M03HO5. D.2. Anal tical Results: Data verifying the stability of the test article in the vehicle for 48 hours under the conditionsofadministratioannd the stabilitoyfthe bulktestarticlaere on filewith the Sponsor. Homogeneity ofprepared formulationsison filweiththe Sponsor. Resultsofthe concentrationanalysiswere notavailableatthe time ofthe writing of thisreport. E. Test System: E.I. Species: Rat E.2. Strain: Crl:CDOBR VAF/PlusO (Sprague-Dawley) E.3. Supplier(Source): Charles RiverLaboratories,Inc.,Raleigh,North Carolina E.4. Sex: Female (Note: Male ratswere used onlyforthe purposes of breedingand are notconsideredpartofthe Test System.) 418-013:PAGE 11-6 E.5. Rationale for Test System: The Crl:CDOBR VAF/PlusID (Sprague-Dawley) ratwas selectedas the Test System because: 1)thisstrainof ratwas used inthe reproductiveand developmentaltoxicitsytudies;2) historicadlata and experienceexistatthe TestingFacility(5a-n7d);3)the testarticliespharmacologicallyactiveinthe species and strain. E.6. Test System Data: Number of Rats Approximate Date of Birth Approximate Age atArrival Weight (g)on the Day AfterArrival Weight (g)at Study Assignment 122 07 SEP 98 65 days 193-223 208-224 E.7. Breeder Male Rat Data: Number of Rats Approximate Date of Birth Approximate Age atArrival Weight (g)on the Day AfterArrival Weight (g)at Cohabitation 112 13 JAN 98 78 days 300-356 558-871 E.8. Method of Randomization: Upon arrivalr,atswere assigned to individuahlousing on the basisofcomputergenerated random units.Afteracclimationv,irginfemale ratswere selectedfor studyon the basisof physicalappearance and body weight recordedduring acclimation.Female ratswere assigned tofivedosage groups (Groups Ithrough V), 16 ratsper dosage group,usinga compute r-generated(weight-ordered) randomizationprocedure.Withineach dosage group, consecutiveorderwas used to assignfemale ratstocohabitationwith breeder male rats,one male rat per female rat.A tableof random unitswas used to selecteightfemale ratswith confirmedmating dates per dosage group forCaesarean-sectionexaminations on DG 15. The remainingfemale ratsineach dosage group were Caesareansectionedon DG 21a. a. See APPENDIX D (DEVIATIONS FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY), item 1. E.9. System of Identification: 418-013:PAGE 11-7 Male ratswere given unique permanent identificatinounmbers upon assignment to the TestingFacilityb'rseeder male ratpopulation.Female ratswere assigned temporary numbers atreceiptand given unique permanent identification numbers when assigned tothe study. Each ratwas individualliydentifiewditha Monel@ self-piercienagr tag (Gey Band and Tag Co.,Inc.,No. MSPT 20101) inscribedwiththe rat'sdesignated unique permanent number. F. Husbandry: F.I. Research FacililyRegistration: USDA RegistrationNo. 23-R-099 under the Animal Welfare Act, 7 U.S.C. 2131 et seq. F.2. Study Rooms: The study rooms were maintained under conditions of positiveairflowrelativeto a hallwayand independentlysuppliedwitha minimum often changes per hour of 100% freshairthathad been passed through 99.97% HEPA filtersR.oom temperatureand humiditywere monitored constantlythroughoutthe study. Room temperaturewas targetedat640F to 790F (180C to 260C); relative humiditywas targetedat 30% to 70%. See APPENDIX E (TEMPERATURE AND RELATIVE HUMIDITY REPORTS.) F.3. Housinci: Allcage sizesand housing conditionswere incompliancewiththe Guide forthe Care and Use ofLaboratoryAnimals(8).Female ratswere individuallhyoused in stainlessteel,wire-boftomedcages except duringthe cohabitationperiodand duringcollectioinntervalfsorurineand fecalsamples. Duringcohabitatione,ach pairof ratswas housed inthe male rat'scage. Duringcollectioinntervalfsor urineand fecalsamples the female ratswere individuallhyoused inmetabolism cages. No nestingmaterialswere suppliedbecause the female ratswere sacrificebdefore parturitiownas expected. F.4. Lighting: An automatically-controlflleudorescentlightcyclewas maintained at 12-hours light.-12-houdrasrk,witheach dark period beginningat 1900 hours EST. F.5. Sanitization: 418-013:PAGE ii-8 Cage pan linerswere changed approximatelythreetimeseach week. Cages were changed approximatelyevery otherweek. F.6. Feed: Ratswere givenad libituamccesstoCertifieRdodent DietS#5002 (PMI NutritioInnternationaSlt,.Louis,Missouri)inindividuafleeders. F.7. Feed Analysis: Analyses were routinelpyerformed by the feed supplier.No contaminantsat levelsexceeding the maximum concentrationlimitfsorcertifiefdeed or deviationsfrom expected nutritionarlequirementswere detectedby these analyses. Copies ofthe resultsofthe feed analyses are availableinthe raw data. Neitherthe Study Directornor the Sponsor was aware of any agent presentin the feed thatwas known to interferweiththe resultsofthisstudy. F.8. Water: Localwater thathad been processed by passage through a reverseosmosis membrane (R.O.water)was availabletothe ratsad libitumfrom individuawlater bottlesand/orfrom an automaticwateringaccess system. Chlorinewas added tothe processed water as a bacteriostat. F.9. Water Analysis: The processed water isanalyzed twiceannuallyforpossiblechemical contamination(LancasterLaboratoriesL,ancaster,Pennsylvania)and monthly forpossiblebacteriaclontamination(AnalyticaLlaboratoriesI,nc.,Chalfont, Pennsylvania).Copies ofthe resultsofthe water analyses are availableinthe raw data. Neitherthe Study Directornor the Sponsor was aware ofany agent presentin thewater thatwas known to interferweiththe resultsofthisstudy. G. Methods: 418-013:PAGE 11-9 G.I. Dosage Administration: Dosage Group 1 11 111 IV v Dosage ConcentrationDosage Volume (mg/kg/day)a (mg/mL) (mukg) 0 (Vehicle) 0 5 0.1 0.02 5 0.4 0.08 5 1.6 0.32 5 3.2 0.64 5 Number of Female Rats 16 16 16 16 16 AssignedRat Numbers 19501 - 19516 19517 - 19532 19533 - 19548 19549 - 19564 19565 - 19580 a. The testarticlweas considered100% pureforthepurposeofdosage calculations. G.2. Rationale for Dosage Selection: Dosages were selected on the basis of a previous study conducted with the test articl(eArgusResearch LaboratoriesI,nc.,Protocol418-008). G.3. Route of Administration: Oral (gavage) G.4. RationaleforRoute of Administration: The oral(gavage)routewas selectedforuse because: 1)thiswas the routeof administratioinnthe developmental and reproductivteoxicologystudies;and 2) it isone ofthe possibleroutesof human exposure. G.5. Freguency of Administration: Appropriatedosages ofthe testarticloer vehiclewere administeredorally(via gavage) once dailytofemale ratsbeginning42 days priorto cohabitation througheitherDG 14 or DG 20. The dosage volume was 5 mukg, adjusteddaily on the basisofthe individuablody weights recordedbefore intubation.The rats were intubatedonce dailyatapproximatelythe same time each day. G.6. Length of Study: Approximately11 weeks G.7. Method of Study Performance: 418-013:PAGE 11-10 The female ratswere observed forviabilitayt leasttwice each day ofthe study and forgeneralappearance at leastonce duringacclimation.The ratswere also examined forclinicaolbservationsofeffectsof the testarticlea,bortions, premature deliverieasnd deaths before and approximatelyone hour after dosage and once priorto sacrifice. Body weights were recordedat leastonce duringacclimationd,ailyduringthe dosage periodand atsacrificeF.eed consumption valueswere recorded at least once duringthe acclimationperiod,weekly to cohabitationand dailyduring presumed gestation. After42 days oftestarticlaedministration8,0 healthyvirginfemale ratswere placed intocohabitatiownith80 breeder male rats(one male ratper female ratin the male rat'scage)a.Mating was evaluateddailyduringthe cohabitationperiod and confirmedby the observationof spermatozoa ina smear ofthe vaginal contentsand/or a copulatoryplug insitu.Female ratsconsideredto be at DG 0 were returnedto individuahlousing. Urine and fecalsamples were collectedfrom female ratsatthe following intervalso:ne day priorto initiatioofncohabitatiotno the followingmorning and from each ofthe mated female ratson DGs 6 to 7, 14 to 15 and 20 to 21. Followingeach 24-hour collectioinntervals,amples were collectedintocentrifuge tubes,placedon dry iceand maintainedfrozen(-700Cor below) untilshipment (on dry ice)to the Sponsor foranalysis. Blood samples were collectedfrom each ofthe female ratsfollowingremoval from metabolism caging(priortotestarticloer vehicleadministrationo)n the day cohabitationwas initiate(dpriotro cohabitationa)nd from each ofthe mated female ratson DGs 7, 15 and 21. On alldays of collectioenxcept DGs 15 and 21 (dams attheirterminalcollectioinntervalb)l,oodsamples (approximately1 mL each) were collectedfrom the orbitaslinus.On DG 21, blood samples (approximately4 mL each) were collectedviathe inferiovrena cava. When blood collectioonn DG 15 was a finalbleedforthe respectivedam (i.e.t,he dam was Caesarean-sectionedthatday),the sample was approximately4 mL and was collectedviathe inferiovrena cava. Blood was transferredintoserum separatortubes and spun ina refrigeratecdentrifuge.The serum was transferredintopolypropylenetubes labeledwiththe study number, rat identificatidoant,e ofcollections,tudy day and collectiotnimepoint.Allsamples were immediatelyfrozenon dry iceand stored(-70"Cor below) untilshipment (on dry ice)to the Sponsor foranalysis. a. See APPENDIX D, item 1. G.8. Gross Necropsva: 418-013:PAGE11-11 Ratswere sacrificbeydcarbondioxidaesphyxiatioonnDG 15 or21,as describedbelow. A gross necropsy ofthe thoracica,bdominal and pelvicviscera was performed. A livesrection(righltaterallobe)and the milk-secretingglands from the axillaryt,horacica,bdominal and inguinalregions(lefstideonly)of each dam (exceptr-atswithoutconfirmeddates of mating)were collectedf,rozenand stored(-70*Cor below) untislhipment (on dry ice)tothe Sponsor foranalysis. To confirmthe pregnancy status,uterifrom ratsthatappeared nonpregnant were stainedwith10% ammonium sulfide('T)i.ssueswithgross lesionswere preserved inneutralbuffered10% formalinforpossiblefutureevaluation;all othermaternaltissueswere discarded.Representativephotographs of maternal lesionsare availableinthe raw data. G.8.a.DG 15 Caesarean-Sectioning Eightrandomly selectedfemale rats,witha confirmeddate of mating,per dosage group were Caesarean-sectionedon DG 15. The graviduteruswas excisedand weighed. Rats were examined fornumber and distributionf corporalutea,implantatiosnites,viableand nonviableembryos. A viable embryo was definedas ovalor crescentshaped, pink,firmand enclosed inan amnioticsac fillewdithclearfluid.A nonviableembryo isamorphous, small,pale pinktotan ordeep red to black,softand enclosed inan amnioticsac fillewdith clear,cloudy or opaque fluid. A sample ofthe amnioticfluidof each viableembryo was collectedand pooled per liftefrr,ozenand stored(-700Cor below) untilshipment,on dry ice,to the Sponsor foranalysis.Each viableembryo was removed from the uteruswiththe attachedplacenta,pooled per liftefrozenand stored(-700Cor below) until shipment,on dry ice,tothe Sponsor foranalysis. G.8.b.DG 21 Caesarean-Sectionin-q Allremainingfemale ratsineach dosage group were Caesarean-sectioned on DG 21 (ratswithconfirmeddatesof mating)or estimatedDG 21 (ratswithout confirmeddates ofmating).The graviduteruswas excisedand weighed. Individuapllacentalweightswere recorded. Rats were examined fornumber and distributiofncorpora lutea,implantationsites,liveand dead fetusesand early and lateresorptions.An earlyresorptionwas definedas one inwhich organogenesiswas notgrosslyevident.A lateresorptionwas definedas one in a. A tableofrandom unitswas used to selectone controlgroup ratfrom which alltissuesexamined atnecropsy were retained,inorderto provide controltissuesforany possiblehistopathologicaelvaluationsofgross lesions. 418-013:PAGE 11-12 which the occurrenceof organogenesis was grosslyevident.A livefetuswas definedas a term fetusthatresponded tostimuli.Nonresponding term fetuses are consideredto be dead (therewere no dead fetuses).Dead fetusesand late resorptionsare differentiatbeyd the degree ofautolysispresent;marked to extreme autolysisindicatedthatthe fetuswas a lateresorption. Each fetuswas removed from the uterus,placed inan individuaclontainerand identifiewditha tag notingthe study number, liftenrumber, and uterine distributioEna.ch fetuswas subsequentlyweighed and examined forsex and gross externalalterationsL.ivefetuseswere sacrificevdiadecapitation. Samples ofthe amnioticfluid(when possible)and the placentaof each fetus were collectedi,ndividuallpyooled per liftefrr,ozenand stored(-700C or below) untilshipment tothe Sponsor foranalysis.Blood samples were collectedfrom each fetusviadecapitationp,ooled per liftearnd transferredintoserum separator tubes. The samples were spun ina refrigeratecdentrifugeT.he serum was transferredintopolypropylenetubes labeledwiththe study number, lifter identificatidoant,e of collections,tudy day and collectiotnimepoint.Allsamples were immediatelyfrozenon dry iceand stored(-700Cor below) untilshipment (on dry ice)tothe Sponsor foranalysis. Lung and liversamples were collectedfrom threefetusesper liftefrom two to fivelifterpser dosage group. One halfof the lungsand one lateralobe of the liverwere individuallryetainedinneutralbuffered10% formalininscintillation vials.Priortofixations,everal(minimum oftwo)sections(approximately1 mm thick)ofthishalfofthe lungsand thislobe ofthe liverwere removed using a scalpeland retainedinMcDowell-Trump's Fixative(storedunder refrigeratiion scintillatviioanls)T.he lung and liversectionsinMcDowell-Trump's Fixativeand the lungand liversamples inneutralbuffered10% formalinwere shipped (on cold packs),forpossiblefutureevaluationby electronmicroscopy or light microscopy,respectivelyt,o PathologyAssociatesInternationaDlu,rham, North Carolina.The otherhalfof the lungsand the otherlateralobe ofthe liverwere flashfrozeninliquidnitrogenand storedinheat-sealablepouches on dry ice. These flashfrozenlung and liversamples were maintainedfrozen(-700Cor below) untislhipment (on dry ice)to the Sponsor foranalysis. The liverosfthe remainingfetuses,plusany remainingliverfrom the three selectedfetuses,were collectedp,ooled by liftearnd frozen.The remaining carcass(and head) ofeach fetuswas frozen.These samples were maintained frozen(-700Cor below) untislhipment (on dry ice)to the Sponsor foranalysis. G.8.c. Moribund -Sacrificoer Premature Deliverv Rats thatwere sacrificebdecause of moribund conditionor premature delivery were examined forthe cause on the day the observationwas made. The rats 418-013:PAGE 11-13 were examined forgross lesions.A livesrection(righltaterallobe)and the milksecretingglands from the axillaryt,horacic,abdominal and inguinalregionsof each dam (lefstideonly)were collectedf,rozenand stored(-700C or below) until shipment (on dry ice)tothe Sponsor foranalysis.Pregnancy statusand uterine contentswere recorded. Deliveredpups were examined tothe extentpossible, usingthe same methods describedforterm fetuses.Uteriofapparently nonpregnant ratswere stainedwith 10% ammonium sulfidetoconfirmthe absence of implantatiosnites('). G.9. StatisticaAlnalyses: Averages and percentageswere calculated.Liftevralueswere used where appropriate. 111. RESULTS 418-013:PAGE III-", A. MortalityP.remature Delivery,Clinicaland Necro sy Observations (Summa[y -Table 1:IndividualData -Tables 12 and 13) A.l. Mortalilya d Premature Delivery No deathsorprematuredeliveriewsere attributetdoPFOS treatment.Three rats(one each inthe 0.1,0.4and 1.6mg/kg/day dosage groups)were sacrificed because the eye was injuredduringorbitaslinusbleeding.One ratinthe vehicle controlgroup didnot have a confirmedmating date and deliveredon day 66 of the study.Two ratsinthe 3.2 mg/kg/day dosage group deliveredon DG 21. All otherratssurvivedto scheduled sacrifice. A.2. ClinicaOlbservations Alladverse clinicaolbservationsduringthe precohabitatioannd gestationperiods were consideredunrelatedto the testarticlbeecause: 1)the incidenceswere not dosage-dependent;and/or2) the observationoccurred inonlyone ortwo rats.These observationsincludedmissing orswollenforepaw digits,cabs on forepaw,swollenforepaw,localizedalopeciaon the limbs,head, back and/or underside,chromorhinorrhea,chromodacryorrhea, swollenears,dentalproblems (missing,broken and/ormisalignedincisors)s,wollensnout,corneal hemorrhage, cornealopacity,axillarmyass, exophthalmos, enophthalmos, lenticulaorpacity,lacrimationu,ngroomed coat,traumatizedcornea,cloudy, whitefilmon eye, missingrighteye, largeeye, dark red eye, and lidsunable to close. A.3. Necr:)psyObservations No gross lesionswere revealedby necropsy. B. Body Weights, UterineWeights and BoAy-Weiqht Changes (FiluresI through-3- Summaries -Tables 2 throuah 5: IndividualData - Tables 14 and ISJ B.1. Precohabitation Body weightgainsforthe entireprecohabitatiopneriod(DSs 1 to 42) were 88.8%, 80.8%, 66.3% and 17.4% ofthe controlgroup value inthe 0.1,0.4,1.6 and 3.2 mg/kg/day dosage groups,respectivelyB.ody weightswere 98.0%, 96.3%, 93.6% and 85.3 % ofthe controlgroup value inthese fourrespective dosage groups on DS 42. B.2. Gestation 418-013:PAGE lli-2 As the resultofrandom selectionforassignment to Caesarean-sectioningon eitherDG 15 or DG 21, groups assigned to Caesarean-sectioningon DG 15 weighed lesson DG 0 than groups assigned to Caesarea n-sectioningon DG 21. Reflectingthe random selectionand differencesinaverage body weights ofthe two subsets,as wellas the relativelsymallgroup sizes,body weight gains differedinthetwo subsets. Inboth subsets,body weightgainswere reduced on DGs 0 to7 ingroups administered0.4 mg/kg/day and higherdosages of PFOS, afterwhich body weight changes didnot demonstrate a cleardosage-dependent pattern. Gestationbody weightsand body weight gains were unaffectedby the 0.1 mg/kg/day dosage ofthe testarticleG.ravid uterineweightswere unaffected by dosages ofthe testarticlaes high as 3.2 mg/kg/day. C. Absolute (g/day) and Relative (g/kci/day)Feed ConsumDtion Values (Summaries - Tables 6 through 9: Individual Data - Tables 16 and 171 C.I. Precohabitation Absolute (g/day)and relative(g/kg/day)feed consumption values were reduced inthe 0.4,1.6and 3.2 mg/kg/day dosage groups ineach week ofthe precohabitatiopneriod.Reflectingthese effectsofthe testarticlea,bsolutefeed consumption valueswere 94.8%, 92.2% and 83.8% of the controlgroup value, and relativfeeed consumption valueswere 95.1%, 94.3% and 90.7% ofthe controlgroup valueforthe entireprecohabitatiopneriod(DSs 1 to42) inthe 0.4, 1.6and 3.2 mg/kg/day dosage groups. Absoluteand relativfeeed consumption values duringthe precohabitationperiod were unaffectedby the 0.1 mg/kg/day dosage ofthe testarticle. C.2. Gestation Groups administered0.4 mg/kg/day and higherdosages ofthe testarticlhead reduced absoluteand relativfeeed consumption values duringthe firswteek of the gestationperiod(no valueswere availableforevaluationforthe 1.6 mg/kg/day dosage group assigned to Caesarean-sectioningon DG 15). Absolute feed consumption values continuedto be reduced forthe remainder of the dosage periodinthe 0.4 and 1.6mg/kg/day dosage groups assigned to DG 15 Caesarea n-sectioning and inthe 3.2 mg/kg/day dosage group forrats assigned to eitherDG 15 or DG 21 Caesarean-sectioning.Relativefeed consumption valuestended tobe increasedover the controlgroup values forthe 1.6and 3.2mg/kg/day dosage groups afterDG 10. 418-013:PAGE 111-3 Absoluteand relativfeeed consumption valuesduringgestationwere unaffected by the 0.1 mg/kg/day dosage ofthe testarticle. D. Caesarean-Sectioning, Lifterand FetalGross ExternalObservations (Summaries -Tables 10 and 11: IndividualData - Tables 18 through Ul D.l. Day 15 of Gestation Pregnancy occurredin6 (75.0%),7 (87.5%),8 (100.0%),6 (85.7%) and 8 (100.0%) ofthe ratsineach dosage group. No Caesarean-sectioningor lifteprarameters were affectedby dosages ofthe testarticlaes highas 3.2 mg/kg/day administeredto ratsCaesarean-sectioned on DG 15. The liftearverages forcorporalutea,implantationsv,iableand nonviableembryos and percentnonviableembryos per liftewrere comparable among the fivedosage groups. No dam had a liftecronsistingofonly nonviable embryos. D.2. Day 21 of Gestation Pregnancy occurred in7 (100.0%), 7 (100.0%), 5 (83.3%),2 (50.0%) and 6 (100.0%) ratswitha confirmedmating date ineach dosage group. One ratin each ofthe 0.1 and 0.4 mg/kg/day dosage groups was moribund sacrificeodn DG 0 and the pregnancy statuscould not be determined and two ratsinthe 3.2 mg/kg/day dosage group deliveredbefore Caesarean-sectioningon DG 21, as previouslydescribed.As a resultC,aesarean-sectioningobservationswere based on 7,7, 5,2 and 4 pregnant ratswithone or more livefetusesinGroups I throughV, respectively. The liftearverages forimplantationsl,iftesrizesand livefetuseswere reduced in the 3.2 mg/kg/day dosage group. Values were below the ranges observed historicalaltythe TestingFacilityaF.etalbody weightswere also reduced inthe 3.2 mg/kg/day dosage group. No otherCaesarean-sectioningor lifter parameters were affectedby dosages ofthe testarticlaes highas 3.2 mg/kg/day administeredto ratsCaesarean-sectionedon DG 21. The liftearverages for corporalutea,earlyand lateresorptionsp,lacentalweights,percentlivemale fetusesand percentresorbedconceptuses were comparable among the five dosage groups. No dam had a littecronsistingofonly resorbedconceptuses, and therewere no dead fetuses.No fetalalterationwsere identifieadtgross externalexamination. a. See APPENDIX F (HISTORICAL CONTROL DATA). REFERENCES 418-013:PAGE 111-4 1. U.S. Food and Drug Administratio(n1994).InternationaClonference on Harmonisation;Guidelineon detectionoftoxicittyo reproductionfor medicinalproducts.FederalRegister,September 22, 1994, Vol.59, No. 183. 2. U.S. Food and Drug Administration.Good LaboratoryPractice Regulations;FinalRule. 21 CFR Part 58. 3. Japanese Ministryof Health and Welfare (1997). Good Laboratory Practice Standard for Safety 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 Protection Agency, Washington, D.C. NationalTechnicalInformationService,U.S. Department of Commerce, SpringfieldV,A 22161. 6. ChristianM,.S. (1984).Reproductivetoxicitaynd teratologyevaluationsof naltrexone(Proceedingsof NaltrexoneSymposium, New York Academy ofSciences,November 7, 1983),J.Clin.Psychiat.45(9):7-10. 7. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the ChaHes River Cri.-CDOBR Rat. Charles River LaboratoriesI,nc.W,ilmington,MA 01887-0630. (Data base providedby Argus Research LaboratoriesI,nc.) 8. Instituteof Laboratory Animal Resources (1996). Guide for the Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 9. Salewski,E. (1964).Fcjrbemethodezum makroskopischen Nachweis von lmplantationsstellaemn Uterusder Rafte.Arch. Pathol.Exp. Pharmakol. 247:367. APPENDIX A REPORT FIGURES BODY WEIGHTS PRECOHABITATION Figure 1 300 290 280 270 260 A 250 240 230 220 15 22 29 DAY OF STUDY a. Last valuerecordedbeforecohabitation 36 4@a MATERNAL BODY WEIGHTS RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTAI Figure2 360 340 320 -F I 300 Ak 280 260 240 220 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 DAY OF GESTATION MATERNAL BODY WEIGHTS RATS CAESAR EAN-SECTION ED ON DAY 21 OF GESTA Figure3 450 400 350 300 A 250 j v Ak A A A AAA AA A IF, A A 200 1 2 3 4 5 6 7 8 1'0 I'l 1'2 13 14 15 16 17 18 19 20 21 DAY OF GESTATION APPENDIX B REPORT TABLES PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE I (PACE 1): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY (See footnotes on the last page of this table.) ----------------- DOSAGE GROUP ------------------------------------------------------------------------------------------- DOSAGE (MG/KG/DAY) 1 11 111 IV --------------------------------------------------------0--(-V-E-H-IC-L-E-)-------0-.-1 -----------0-.-4-----------1-.-6----------- MORTALITY DELIVERED AND SACRIFICED MORIBUND SACRIFICED 1 1 la 0 0 Id PRECOHABITATION (DAY 1 OF STUDY TO THE DAY F COHABITATION): 1 1 0 0 le if MAXIMUM POSSIBLE INCIDENCE FOREPAW: THIRD DIGIT MISSING 672/ 16 672/ 16 672/ 16 672/ 16 LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE FOREPAW: THIRD DIGIT SWOLLEN FOREPAWS: SCAB(S) 0/ 0 40/ 1 40/ 1 0/ 0 0/ 0 0/ 0 43/ 2 43/ 2 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 26/ 2 14/ 1 12/ 1 0/ 0 CHROMORHINORRHEA 0/ 0 0/ 0 0/ 0 14/ 1 CHROMODACRYORRHEA 0/ 0 0/ 0 1/ 1 S/ 4 EARS: SWOLLEN 0/ 0 S/ 1 0/ 0 0/ 0 FOREPAWS: SWOLLEN 0/ 0 0/ 0 0/ 0 3/ 1 INCISORS: TOTAL 0/ 0 0/ 0 0/ 0 2/ 1 SNOUT: MISSING/BROKEN MISALIGNED SWOLLEN 0/ 0 0/ 0 0/ 0 21/ 1 12/ 1 9/ 1 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 0/ 0 1/ 1 0/ 0 0/ 0 -M-A-X-IM-U-M---P-OS-S-I-BL-E---I-N-C-ID-E-N-C-E ---=--(-DA-Y-S--x--R-A-TS-)-/-NU-M-B-E-R ---O-F--R-AT-S---EX-A-M-I-NE-D---P-E-R--G-RO-U-P-.---------------------------------------- NIN = TOTAL NUMBER OF OBSERVATIONS/NLIMBER OF RATS WITH OBSERVATION. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S TABLE 1 (PAGE 2): CLINICAL AND NECROPSY OBSERVATIONS - SUMMARY (See footnotes on the last page of this table.) STUDY NUMBER: T-6295.12) -D-O-SA-G-E ---GR-O-U-P ---------------------------------------------------------------------------------------------------- DOSAGE (MG/KG/DAY) I ii III 0 (VEHICLE) 0.1 IV -P-R-E-SU-M-E-D ---GE-S-TA-T-I-ON-:----g-----------------------------------------------------------------0-.-4-----------1-.-6----------- MAXIMUM POSSIBLE INCIDENCE LOCALIZED ALOPECIA: TOTAL LIMBS UNDERSIDE HEAD BACK FOREPAW: THIRD DIGIT MISSING CORNEAL HEMORRHAGE 282/ 15 3/ 1 3/ 1 0/ 0 0/ 0 0/ 0 0/ 0 283/ 16 37/ 3 23/ 2d 0/ o 14/ 1 0/ 0 0/ 0 261/ 15 3/ 1 0/ 0 3/ 1 0/ 0 0/ 0 0/ 0 216/ 12 45/ 3 37/ 2 16/ 1 0/ 0 7/ 1 0/ 0 CORNEAL OPACITY 0/ 0 0/ 0 0/ 0 6/ 1 RIGHT AXILLA: MASS 0/ 0 27/ 3d 20/ 2e 0/ 0 EXOPHTHALMOS G/ 0 o/ o 0/ o 0/ 0 CHROMODACRYORRHEA FOREPAWS: SCAB(S) ENOPHTHALMOS 0/ 0 11/ 1 0/ 0 4/ 2d 1/ Id 0/ 0 4/ 2e 1/ le 0/ 0 1/ 1 0/ 0 1/ 1 LENTICULAR OPACITY LACRIMATION 0/ 0 16/ la 0/ 0 o/ o a/ 1 3/ 1 0/ 0 0/ 0 RIGHT EYE: LARGE RIGHT EYE: DARK RED ------------------------------------------------------ 0/ 0 0/ 0 0/ 0 0/ 0 1/ Id 1/ ld 2/ 1 I/ le I/ le 0/ 0 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--O-B-SE-R-V-A-TI-O-N-.------------------------------------------------ PROTOCOL TABLE I 418-013: OPAL (GAVAGE) PHARMACOV (PAGE 3): CLINICAL AND NECROPSY INETIC STUDY OBSERVATIONS OF PFOS IN RATS - SUMMARY (SPONSOR'S STUDY NUMBER! T-6295.12) ---------------------------------------------- DOSAGE GROUP ---------------------------------------------------------- DOSAGE (MG/KG/DAY) I ii III IV ------------------------------------------------ 0 (VEHICLE) 0.1 0.4 1.6 PRESUMED GESTATIO,: g ------------------------------------------------------------- MAXIMUM POSSIBLE INCIDENCE RIGHT EYE: LIDS UNABLE TO CLOSE CHROMORHINORRHEA 282/ 15 0/ 0 283/ 16 1/ ld 261/ 15 I/ le 216/ 12 0/ 0 SNOUT: SWOLLEN 7/ 2 0/ 0 1/ 1 0/ 0 UNGROOMED COAT 0/ 0 3/ 2d I/ le 0/ 0 TRAUMATIZED CORNEA 0/ 0 0/ 0 1/ 1 0/ 0 RIGHT EYE: CLOUDY, WHITE FILM RIGHT EYE: MISSING 0/ 0 0/ 0 33/ 3 11/ 1 0/ 0 0/ 0 0/ 0 0/ 0 INCISORS: MISSING/BROKEN 0/ 0 11/ 1 0/ 0 0/ o 0/ 0 7/ 1 0/ 0 0/ 0 PERSISTENT ADVERSE CLINICAL OBSERVATIONS WERE CONFIRMED AT ---------------------------------------------------------------N-EC-R-O-PS-Y-,---N-O--A-D-DI-T-I-ON-A-L---G-R-OS-S---L-E-SI-O-N-S ---WE-R-E--I-l-------- MNAIXNIMUM POSSIBLE INCIDENCE - (DAYS x RATS)/NUMBER OF RATS EXAMINED PER GROUP. a. = TOTAL NUMBER OF OBSERVATIONS/NUMBER Dam 19503 did not have OF RATS WITH OBSERVATION. b. Dam 19571 delivered a confirmed mating date and delivered on day 21 of gestation. on day 66 of study. C. Dam 19579 delivered on day 21 of gestation. d. Rat 19520 was moribund e. Rat 19541 was moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. f. Rat 19557 was moribund sacrificed on day 0 of presumed gestation; pregnancy status could not be determined. g. sacrificed on day 43 of study. Restricted to rats with a confirmed mating date. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS TABLE 2 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - SUMMARY (SPONSOR'S STUDY NUMBER: T-6295.12) -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 --------------------------------------------I ------------------------------------------------------------------ RATS TESTED N 16 16 16 16 BODY WEIGHT (G) DAY 1 HEAN+S.D 227.3 + 7.2 227.4 + 4.3 226.9 + 6.2 226.9 + 6.9 DAY 8 MEAN+S.D 243.4 + 10.2 244.6 + 6.8 242.0 + 9.4 241.7 + 10.1 DAY 15 MEAN+S.D 2S2.9 + 11.6 252.1 + 8.6 249.0 + 11.7 249.8 + 11.2 DAY 22 MEAN+S.D 266.3 + 12.8 263.9 + 10.5 261.1 + 15.1 258.7 + 12.3 DAY 29 MEAN+S.D 270.9 + 16.0 266.9 + 11.5 264.0 + 17.1 258.4 + 14.4 DAY 36 MEAN+S.D 273.3 + 18.6 272.2 + 12.7 270.3 + 17.9 263.0 + 1S.0 DAY 42a MEAN+S.D 278.9 + 18.2 273.2 + 15.4 268.6 + 23.4 261.1 + 15.9 ------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY a. Last value recorded before cohabitation. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PPOS IN RATS TABLE 3 (PAGE 1): BODY WEIGHT CHANGES - PRECOHABITATION - SUMMARY (SPONSOR'S STUDY NUMBER: T-6295.12) -D-O-SA-G-E---GR-O-UP---------------------------------------------------------------------------------------------------- DOSAGE (MG/KG/DAY) I ii III IV 0 (VEHICLE) 0.1 0.4 -R-A-TS---T-ES-T-E-D -------------------------------------------------------------------------------------1-.6--------------- N 16 16 16 BODY WEIGHT CHANGE (G) 16 DAYS 1 8 MEAN+S.D. +16.1 + 5.6 +17.2 + 5.1 +15.1 + 5.1 +14.8 + 5.7 DAYS 8 15 MEAN+S.D. +9.5 + 5.6 +7.5 + 4.7 +7.0 + 4.4 DAYS 15 22 MEAN+S.D. +13.4 + 5.8 +8.1 + 4.7 DAYS 22 29 MEAN+S.D. +4,6 + 6.6 +11.8 + 4.8 +12.1 5.2 +8.9 + 4.2 +3.0 + 5.7 +2.9 + 6.1 -0.2 + 4.8 DAYS 29 36 MEAN+S.D. +2.4 + 7.1 +5.3 + 5.4 +6.3 + 6.4 DAYS 36 42a MEANS.D. +4.6 4.4 DAYS I 42a +5.6 + 5.2 +0.9 + 6.1 -1.7 23.6 -1.9 + 6.0 MEJkN+S.b. +51.6 + 16.9 +45.8 1 14.6 -------------------------------------------------------------------------+-4-1-.-7 --+--2-2.-6- +34.2 + 13.2 DAYS - DAYS OF STUDY 7 --------------------------- a. Last value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 1): MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 -------------------------------------------------------------------------------------------------------------- RATS TESTED N 8 8 8 INCLUDED IN ANALYSES N a 8 a 7a PREGNANT N 6 7 a 6 MATERNAL BODY WEIGHT (G) DAY 0 MEAN+S.D. 281.0 + 28.9 276.0 + 10.5 270.4 + 14.4 267.2 + 16.4 DAY 1 MEAN+S.D. 287.0 + 26.0 282.4 + 13.0 275.1 + 20.8 267.2 + 15.1 DAY 2 DAY 3 DAY 4 DAY 5 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 294.3 + 25.1 297.2 + 27.5 302.8 + 28.4 304.2 + 26.2 290.0 + 11.2 289.1 + 11.2 293.6 + 10.5 296.6 + 12.4 282.8 + 19.3 285.0 + 16.9 286.8 + 17.4 289.4 + 16.3 275.3 + 17.0 274.8 + 17.4 275.8 + 15.8 276.8 + 19.0 DAY 6 MEAN+S.D. 308.7 + 27.1 302.0 + 14.2 291.9 + 17.3 278.3 + 16.3 DAY 7 MEAN+S.D. 310.2 + 28.2 302.6 + 12.2 285.5 + 20.6 273.0 + 14.5 DAY 8 MEAN+S.D. 309.2 + 25.0 302.8 + 12.0 295.8 + 21.3 279.0 + 17.7 DAY 9 MEAN+S.D. 315.7 + 23.1 308.0 + 13.7 294.4 + 17.9 280.3 + 16.1 DAY 10 MEAN+S.D. 319.7 + 25.0 313.7 + 17.9 296.9 + 19.0 285.3 + 15.6 DAY 11 MEAN+S.D. 327.3 + 24.S 314.6 + 13.9 302.4 + 18.6 292.2 + 16.8 DAY 12 MEAN+S.D. 330.5 + 23.9 321.7 + 15.0 309.1 + 18.7 297.2 + 18.9 -------------------------------------------------------------------------------------------------------------- DAY = DAY OF GESTATION a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 2): MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY ---- --------------------- --- -- -- --------- RATS CAESAREAN-SECTIONED -------------------------------------------------------------ON DAY 15 OF GESTATION -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) I ii III IV 0 (VEHICLE) 0.1 ------------------------------------------------------------------------------0.-4-----------------1.-6------------- RATS TESTED N 8 INCLUDED IN ANALYSES N 8 a 8 7a PREGNANT N 6 MATERNAL BODY WEIGHT (G) DAY 13 DAY 14 DAY 15 GRAVID UTERINE MEAN+S.D. MEANS.D. MEANS.D. 335.3 + 24.6 339.2 + 25.8 345.2 + 27.8 324.3 + 15.6 328.4 + 15.s 331.1 + 18.6 314.0 + 19.8 317.8 19.2 319.9 + 22.4 298.8 + 17.8 305.5 + 20.8 309.8 + 17.7 WEIGHTS (G) MEAN+S.D. 19.8 + 4.3 17.5 + 5.2 16.3 + 5.6 17.6 + 1.3 DAY 15C b MEAN+S.D. 325.3 + 25.7 313.7 + 15.7 ------------------------------------------------------------------ 303.6 + 19.5 292.2 + 16.9 DAY = DAY OF GESTATION -------------------------------------------- a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. b- ISC - Corrected maternal body weight (day 15 of gestation body weight minus the gravid uterine weight). PROTOCOL 418-013- OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 3): MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) 1 11 111 IV 0 (VEHICLE) 0.1 0.4 1.6 -------------------------------------------------------------------------------------------------------------- RATS TESTED N a a 8 8 INCLUDED IN ANALYSES N 7a 7b 6a,b 4a,c PREGNANT N 7 7 5 2 MATERNAL BODY WEIGHT (G) DAY 0 MEAN+S.D. 281.6 + 14.S 282.3 + 19.1 292.8 + 22.7 273.0 + 1.4 DAY I MEAN+S.D. 290.3 + 15.6 290.4 + 17.9 294.4 + 24.7 277.5 + 0.7 DAY 2 MEAN+S.D. 293.8 + 14.0 298.3 + 16.5 301.2 + 24.8 282.5 + 0.7 DAY 3 MEAN+S.D. 298.4 + 12.6 300.4 + 19.3 302.6 + 23.3 284.5 + 2.1 DAY 4 MEAN+S.D. 300.4 + 13.4 304.0 + 14.2 305.4 + 24.5 283.5 + 0.7 DAY 5 MEAN+S.D. 304.8 + 9.6 307.4 + 16.8 307.8 + 26.1 289.5 + 2.1 DAY 6 MEAN+S.D. 307.6 + 10.7 310.6 + 17.4 310.6 + 26.6 290.0 + 2.8 DAY 7 MEA.N+S.D. 301.0 + 12.8 313.3 + 18.3 307.4 + 25.9 285.5 + 0.7 DAY a MEAH+S.D. 305.8 + 12.8 314.4 + 16.6 309.0 + 26.1 286.0 + 1.4 DAY 9 MEAN+S.D. 311.0 + 10.4 318.7 + 15.4 313.4 + 27.3 292.5 + 2.1 DAY 10 MEAN+S.D. 317.0 + 8.8 323.1 + 16.9 316.6 + 26.3 295.0 + 7.1 DAY 11 MEAN+S.D. 321.7 + 10.1 329.6 + 15.2 320.2 + 23.7 300.S + 7.8 DAY 12 MEAN+S.D. 326.1 + 11.6 330.8 + 16.8 327.2 + 28.2 304.5 + 12.0 --------------------------------------------------------------------------------------------------------------- DAY - DAY OF GESTATION a. Excludes values for rate that did not have a confirmed mating date. b. Excludes values for rate 19S20 and 19541, which were moribund sacrificed on day 0 of presumed gestation; p could not be determined. C. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR,S STUDY NUMBER: T-6295.12) TABLE 4 (PAGE 4): MATERNAL BODY WEIGHTS AND GRAVID UTERINE WEIGHTS - GESTATION - SUMMARY ----------------------RATS CAESAREAN-SECTIONED --O-N--D-AY---21---OF--G-E-S-TA-T-I-ON------------------------------------------------------------------- -D-O-SA-G-E ---GR-O-UP----------------------------------------------------------------------------------------------------- DOSAGE (MG/KG/DAY) I ii 0 (VEHICLE) III IV ------------------------------------------------------------0-.1-----------------0.-4-----------------1.-6-------------- RATS TESTED N 8 8 INCLUDED IN ANALYSES N 7a 7b PREGNANT 6a,b 4a,c N7 7 MATERNAL BODY WEIGHT (G) DAY 13 DAY 14 DAY IS DAY 16 DAY 17 DAY 18 DAY 19 DAY 20 DAY 21 GRAVID UTERINE MEAN+S.D. MEAN+S.D. MEA.N+S.D. MEAN+S.D. MEA.N+S.D. MEAN+S.D. MEA.N+S.D. MEAN+S.D. MEAN+S.D. 329.1 + 10.1 334.0 12.1 336.0 + 16.0 345.7 + 13.8 361.8 16.2 377.4 + 15.5 395.6 + 18.5 410.1 + 20.1 423.0 + 27.4 336.0 + 14.9 340.3 + 17.4 339.7 + 17.5 350.7 + 20.9 366.8 + 22.8 381.4 + 23.2 397.6 + 24.9 413.0 25.2 427.0 + 29.0 331.8 + 28.0 337.0 + 28.7 329.2 + 24.3 349.4 + 29.5 361.0 + 28.1 379.8 + 31.0 391.8 + 29.7 410.2 + 34.7 423..0 + 30.7 312.5 + 4.9 317.0 + 12.7 323.5 + 6.4 339.0 + 15,6 357.5 + 12.0 377.5 + 2o.5 394.S + 24.7 413.5 + 34.6 426.5 + 23.3 WEIGHTS (G) MEAN+S.D. 96.3 + 12.0 92.0 + 27.0 91.3 + 34.0 112.7 + 37.0 DAY 21C d MEAN+S.D. 326.7 22.6 335.0 + 22.4 -D-AY-----------------------------------------------------------------------3-3-1-.-7 --+--26-.-2--------3-1-3.-e---+ --13-.-6------------ = DAY OF GESTATION a. b. Excludes values for rats that did not have a confirmed mating date. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day could not be determined. C. 0 of presumed gestation; pre d- Excludes values for rat 19557, which was moribund sacrificed on day 43 of study 21C - Corrected maternal body weight (day 21 of gestation body weight minus the (day 2 of cohabitation). gravid uterine weight). PROTOCOL 418-013! ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 5 (PACE 1): MATERNAL BODY WEIGHT CHANGES - GESTATION - SUMMARY -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY IS OF GESTATION -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 -------------------------------------------------------------------------------------------------------------- RATS TESTED N 8 8 8 INCLUDED IN ANALYSES N a 8 a 7a PREGNANT N 6 7 a 6 MATERNAL BODY WEIGHT CHANGE (C) DAYS 0 7 DAYS 7 8 DAYS 8 9 DAYS 9 10 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. +29.2 + 7.7 -1.0 + 6.4 +6.5 + 3.9 +4.0 + 3.5 +26.6 + 5.0 +0.3 + 3.9 +5.1 + 2.5 +5.7 + 11.6 +15.1 + 7.6 +10.2 + 7.8 -1.4 + 10.6 +2.5 + 3.2 +5.8 + 9.0 +6.0 + 6.1 +1.3 + 5.5 +S.0 + 2.4 DAYS 7 10 DAYS 10 13 DAYS 13 IS MEAN+S.D. MEAN+S.D. MEAN*S.D. +9.5 + 5.8 +15.7 + 2.1 +9.8 + 3.9 +11.1 + 8.9 +10.6 + 11.0 +6.8 + 12.2 +11.4 + 5.0 +17.1 + 7.6 +5.9 + 9.6 +12.3 + 8.8 +13.5 + 4.3 +11.0 + 4.6 DAYS 0 15 MEAN*S.D. +64.2 + 9.9 +55.1 + 10.2 +49.5 + 12.8 +42.7 + 4.2 DAYS 0 15c b MEAN*S.D. +44.3 + 11.7 +37.7 + 8.0 +33.2 + 11.6 +25.1 + 3.2 -------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF GESTATION a. Excludes values for rat 19561; necropsy and Caesarean-section observation were not recorded. b. JSC = Corrected maternal body weight (day 15 of gestation body weight minus the gravid uterine weight). PROTOCOL 418-013: ORAI- (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S TABLE 5 (PAGE 2): MATERNAL BODY WEIGHT CHANCES - GESTATION - SUMMARY STUDY NUMBER: T-6295.12) --------------- ----------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION -D-OS-A-G-E ---GR-O-UP----------------------------------------------------------------------------------------------------- DOSAGE (MG/KG/DAY) I ii III IV 0 (VEHICLE) 0.1 0.4 -R-A-TS---T-ES-T-E-D -------------------------------------------------------------------------------------1-.-6-------------- N a 8 a INCLUDED IN ANAI.YSES N 7a 7b PREGNANT N 6a,b 4a,c 7 75 MATERNAL BODY WEIGHT CHANGE (G) DAYS 0 7 MEAN+S.D. +19.4 + 9.6 +31.0 + 11.7 +14.6 + 7.6 DAYS 7 8 +12.5 + 0.7 MEAN+S.D. +4.8 10.2 +1.1 + 7.6 +1.6 + 4.0 DAYS 8 9 +0.5 + 2.1 MEAN+S.D. +5.1 + 3.1 +4.3 + 3.4 +4.4 + 8.0 DAYS 9 10 +6.5 + 0.7 MEM+S.D. +6.0 2.9 +4.4 + 4.9 +3.2 + 5.2 +2.5 + 4.9 DAYS 7 10 MEAN+S.D. +16.0 + 11.0 +9.8 + 6.9 +9.2 + 7.2 DAYS 10 13 MEAN+S.D. +12.1 4.0 +12.8 + 3.7 +15.2 + 5,5 +9.5 + 7.8 DAYS 13 - 15 MEAN+S.D. +6.8 + 7.4 +3.7 + 9.6 -2.6 + 15.4 +17.5 + 2.1 DAYS 15 - 18 MEAN+S.D. +41.4 4.5 +41.7 + 14.8 +50.6 + 11.6 +11.0 + 1.4 DAYS 18 - 21 MEAN+S.D. 445.6 + 15.7 +45.6 + 12.2 +43.2 + 12.3 +54.0 + 14.1 DAYS 0 - 21 MEM+S.D. +141.4 + 25.0 +144.7 + 20.3 +130.2 + 23.8 +49.0 + 2.8 +153.5 + 24.7 DAYS 0 - 21C d MEAN+S.D. +45.2 + 18.5 +52.7 + 16.4 -------------------------------------------------------------------------+-3-8-.-9 --+--1-4.-0--------+-4-0-.8---+ --12-.-2------------ DAYS = DAYS OF GESTATION a. b. Excludes values for rats that did not have a confirmed mating date. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day could not be determined. 0 of presumed gestation; pre C. d. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study 21C - Corrected maternal body weight (day 21 of gestation body weight minus the (day 2 of cohabitation). gravid uterine weight). PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 6 (PAGE 1): ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - PRECOHASITATION - SUMMARY -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 ------------------------------------------------------------------------------------------------------------- RATS TESTED N 16 16 16 16 FEED CONSUMPTION (G) DAYS 1 8 DAYS 8 IS DAYS 15 22 DAYS 22 29 DAYS 29 36 MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 19.7 + 1.8 ( 15]a 18.8 + 1.8 ( illa 20.0 + 1.7 1 14]a 18.6 + 1.8 ( 12)a 18.7 + 2.0 19.4 + 1.5 18.8 + 1.6 I 1S)a 19.5 + 1.6 18.9 + 1.9 ( 15]a 18.0 + 1.8 18.8 + 1.8 17.4 + 1.6 I 15]a 18.5 + 2.1 ( l@la 18.1 + 2.3 [ 14)a 18.0 + 1.5 18.9 + 1.5 ( 15]a 18.2 + 1.9 I lila 17.9 + 1,7 ( 13)a 17.2 + 1.7 ( 13)a 17.1 + 1.8 DAYS 36 42b MEAN+S.D. 19.5 + 2.2 18.0 + 1.8 17.9 + 1.7 16.7 + 1.8 DAYS 1 42b MEAN+S.D. I 13)a 19.2 + 1.3 18.8 + 1.4 [ lila 18.2 + 1.8 [ 13-la 17.7 + 1.4 ( 13)a I lila ( lila -------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF STUDY [ I = NUMBER OF VALUES AVERAGED a. Excludes values that were incorrectly recorded, as well as those associated with spillage. b. Last value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 7 (PAGE 1): RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - PRECOHABITATION - SUMMARY --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 11 111 IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 --------------------------------------------------------------------------------------------------------------- RATS TESTED N 16 16 16 16 FEED CONSUMPTION (g) DAYS 1 8 DAYS 8 15 DAYS 15 22 DAYS 22 29 DAYS 29 36 HEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 83.8 + 6.1 ( 15)a 75.8 + 5.4 i II)a 77.0 + S.1 [ 14]a 69.4 + 5.2 ( 12)a 68.8 + 6.2 82.1 + 6.1 75.6 + 6.0 I 15]a 75.5 + 5.8 71.4 + 6.4 t 15]a 67.0 + 5.8 80.0 + 5.7 70.8 + 4.2 ( 15]a 72.8 + 4.8 ( 15]a 68.8 + 4.8 ( 14]a 67.6 + 4.7 80.6 + 5.9 I 15]a 74.3 + 7.6 ( l@la 70.4 + 5.6 ( lila 66.1 + 5.8 i 13]a 65.6 + 6.5 DAYS 36 42b MFAN+S.D. 71.5 + 9.4 65.6 + 5.4 65.9 + 4.8 64.0 + 7.9 t 13]a ( lila I lila DAYS 1 42b MEAN+S.D. 74.2 + 4.9 72.4 + 4.7 70.6 + 4.3 70.0 + 6.0 ( 13]a I 13]a ( lila --------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY I ) = NUMBER OF VALUES AVERAGED a. Excludes values that were incorrectly recorded, as well as those associated with spillage. b. bast value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PVOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 8 (PACE 1): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (G/DAY) - GESTATION - SUMMARY ---- --------------------- --- -- -- --------- RATS CAESAREAN-SECTIONED --------------------------------------------------------------ON DAY 15 OF GESTATION --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) I ii III IV ---------------------------------------0--I-V-E-H-IC-L-E-)------------0-.1-----------------0.-4-----------------1-.6-------------- RATS TESTED N 8 8 INCLUDED IN ANALYSES N 8 8 a 7a PREGNANT N 6 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 7 MEAN+S.D. 24.0 + 2.2 22.5 + 2.0 20.4 + 2.8 b DAYS 7 MEAN+S.D. 22.7 + 1.2 21.4 + 2.7 [ 2)b 21.0 + 2.5 19.5 + 2.8 DAYS a 9 MEAN+S.D. 24.2 + 1.9 24.7 + 3.4 22.1 + 2.4 22.3 + 1.6 DAYS 9 - 10 MEAN+S.D. 25.0 + 2.9 23.0 + 3.5 22.6 3.s 21.7 + 1.4 DAYS 7 - 10 MEAN+S.D. 24.0 + 1.7 23.0 + 3.0 21.9 + 2.3 21.2 + 1.7 DAYS 10 - 13 MEAN*S.D. 24.8 + 2.6 24.3 + 2.2 23.2 + 1.9 23.0 + 2.0 DAYS 13 - 15 MEAN+S.D. 21.9 + 2.1 20.7 + 1.4 18.9 3.0 18.8 + 1.1 DAYS 0 - 15 MEAN+S.D. 23.9 + 2.o I 6]b 23.0 + 1.8 t 7]b 21.0 + 2.2 20.1 + 1.2 t 6)b [ 5]b ---------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION I I - NUMBER OF VALUES AVERAGED a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. b. Excludes values that were not recorded, as well as those associated with spillage. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 8 (PAGE 2): MATERNAL ABSOLUTE FEED CONSUMPTION VALUES (GIDAY) - GESTATION - SUMMARY ---------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION ---------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) I ii III IV ----------------------------------- 0 (VEHICLE) 0.1 0.4 1.6 ---------------------------------------------------------------------------- RATS TESTED N 8 8 a INCLUDED IN ANALYSES N 7a 7b 6a,b 4a,c PREGNANT N7 7 5 MATERNAL FEED CONSUMPTION (G/DAY) DAYS 0 7 DAYS 7 8 MEAN+S.D. MEAN+S.D. 23.6 + 1.4 21.8 + 2.6 23.7 + 2.3 23.0 + 3.1 21.o i 22.2 + 3.3 ild + 2.4 20.3 + i.o 20.5 + 0.7 DAYS 8 9 MEA.N+S.D. 23.6 + 2.4 2S.6 + 1.8 22.8 + 4.5 20.0 + 4.2 DAYS 9 10 MEAN+S.D. 24.3 + 1.5 24.8 + 3.4 22.4 + 4.5 22.5 + 0.7 DAYS 7 10 MEAN+S.D. 23.2 + 1.4 24.4 + 2.3 22.5 + 3.6 21.0 + 1.4 DAYS 10 13 MEAN+S.D. 24.4 + 1.8 25.3 + 2.0 24.5 + 2.8 23.3 + 1.4 DAYS 13 IS DAYS 15 MEAN+S.D. MEAN+S.D. 22.1 + 2.5 2S.3 + 2.1 20.9 1 24.3 + 1.6 6)d + 5,6 19.1 + 2.9 24.9 + 4.7 21.8 + 1.8 26.8 + 0.2 DAYS 18 21 MEAN+S.D. 23.9 + 3.2 25.1 + 2.6 24.9 + 2.4 2S.5 + 0.7 DAYS 0 21 MEAN+S.D. 23.8 + 1.1 24.1 + 1.6 23.0 + 2.6 ---------------------------------------------------------------------------------------------22-.-6-----0-.-1----------- DAYS = DAYS OF GESTATION I ) = NUMBER OF VALUES AVERAGED a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rats 19S20 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pre could not be determined. C. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). d. Excludes values that were not recorded, as well as those associated with spillage. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 9 (PAGE 1): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 -------------------------------------------------------------------------------------------------------------- RATS TESTED N 8 8 8 8 INCLUDED IN ANALYSES N a 8 a 7a PREGNANT N 6 7 8 6 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 7 DAYS 7 8 MEAN+S.D. MEAN+S.D. 80.6 + 8.2 73.6 + 7.3 77.2 + 6.8 70.9 + 9.1 71.6 + 5.2 ( ilb 72.2 + 6.8 b 70.5 + 7.6 DAYS 8 9 MEAN+S.D. 77.6 + 7.0 80.8 + 9.8 75.0 + 5.9 79.9 + 4.5 DAYS 9 10 MEAN+S.D. 78.8 + 8.1 74.0 + 11.1 76.3 + 8.6 76.7 + 4.3 DAYS 7 10 MEAN+S.D. 76.6 + 6.2 75.2 + 9.4 74.7 + 4.2 75.8 + 4.0 DAYS 10 13 MEAN+S.D. 7S.8 + 6.5 76.3 + 6.3 75.7 + 3.3 78.6 + 7.5 DAYS 13 - IS MEAN*S.D. 64.6 + 5.2 63.3 + 4.2 60.2 + 8.6 61.7 + 5.2 6)b 7]b DAYS 0 - 15 MEAN+S.D. 76.5 + 6.2 75.8 + 5.1 71.8 + 3.8 70.9 + 4.7 ( 6)b [ ilb -------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF GESTATION ( ) - NUMBER OF VALUES AVERAGED a. Excludes values for rat 19S61; necropsy and Caesarean-section observations were not recorded. b. Excludes values that were not recorded, as well as those associated with spillage. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 9 (PAGE 2): MATERNAL RELATIVE FEED CONSUMPTION VALUES (G/KG/DAY) - GESTATION - SUMMARY -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) I ii III 0 (VEHICLE) 0.1 IV -----------------------------------------------------------------------------0-.-4----------------1-.6-------------- RATS TESTED N 8 8 a 8 INCLUDED IN ANALYSES N 7a 7b 6a,b 4a,c PREGNANT N 7 7 5 2 MATERNAL FEED CONSUMPTION (G/KG/DAY) DAYS 0 7 DAYS 7 8 DAYS 8 9 DAYS 9 10 DAYS 7 - 10 DAYS 10 - 13 MEANS.D. MEANS.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. MEAN+S.D. 79.6 + 7.2 72.0 + 7.7 76.7 + 10.0 77.4 + 6.1 75.3 + 5.4 75.3 + 5.3 79.0 + 7.5 73.1 + 7.6 80.8 + 5.7 77.3 + 8.2 77.0 + 5.2 76.6 + 4.6 71.6 + 5.0 [ ild 72.2 + 7.9 73.2 + 12.7 70.6 + 9.6 71.9 + 8.7 75.4 + 4.2 71.6 + 3.3 71.8 + 2.6 69.1 14.3 76.6 + 1.2 72.4 + 4.3 76.8 + 2.6 DAYS 13 - 15 DAYS 15 - 18 MEAN+S.D. MEAN+S.D. 66.4 + 6.7 71.4 + 5.2 62.3 [ 67.1 + 4.1 6]d + 14.1 57.4 + 7.6 70.1 + 12.2 68.4 + 3.8 76.9 + 3.5 DAYS 18 - 21 MEAN+S.D. 59.4 + 6.2 62.3 + 7.0 62.1 + 6.7 63.5 + 5.8 DAYS 0 - 21 MEAN+S.D. 72.1 + 4.0 72.0 + 3.2 --------------------------------------------------------------------------6-9-.-1--+---5-.-4--------7-1-.-2 --+---1-.6---------- DAYS = DAYS OF GESTATION I I = NUMBER OF VALUES AVERAGED a. b. Excludes values for rats that did not have a confirmed mating date. Excludes values for rats 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; p could not be determined. C. d. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). Excludes values that were not recorded, as well as those associated with spillage. PROTOCOL 418-013: OPAT, (CAVACE) PHARMACOKINETIC STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 10 (PACE 1): CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 --------------------------------------------------------------------------------------------------------------- RATS TESTED N a 8 8 8 INCLUDED IN ANALYSES N 8 8 a 7a PREGNANT N(t) 6( 75.0) 7( 87.5) B(IOO.O) 6( 85.7) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 15 OF GESTATION N 6 7 6 CORPORA BUTEA MEAN+S.D. 17.3 + 3.4 18.0 + 1.7 17.1 + 3.0 19.8 + 3.2 IMPLANTATIONS MEAN+S.D. 15.8 + 1.9 13.4 + 3.8 13.0 + 3.7 14.2 + 1.3 VIABLE EMBRYOS N MEAN+S.D. 89 14.8 + 2.8 86 12.3 + 3.7 94 11.8 + 4.7 77 12.8 + 1.2 NONVIABLE EMBRYOS N MEAN+S.D. 6 1.0 + 1.3 8 1.1 + 1.3 10 1.2 + 1.4 1.3 + 1.0 DAMS WITH ANY NONVIABLE EMBRYOS N(t) 3( 50.0) 5( 71.4) 4( 50.0) 5( 83.3) DAMS WITH ALL NONVIABLE EMBRYOS N(t) 0( 0.0) 0( 0.0) 0( 0.0) 0( 0.0) DAMS WITH VIABLE EMBRYOS N(t) 6(100.0) 7(100.0) 8(100.0) 6(100.0) t NONVIABLE EMBRYOS/LITTER MEAN+S.D. 6.8 + 8.4 8.6 + 9.3 13.4 + 17.7 9.2 + 6.5 --------------------------------------------------------------------------------------------------------------- a. Excludes values for rat 19561; necropsy and Caesarean-section observations were not recorded. PROTOCOL 418-013: OPAL (GAVA(;E) PHARMACOKINETIC STUDY OF PFOS IN RATS TABLE 10 (PAGE 2): CAESAREAN-SECTIONIN(; OBSERVATIONS - SUMMARY (SPONSOR'S STUDY NUMBER: T-6295.12) --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) I ii III 0 (VEHICLE) 0.1 0.4 IV ------------------------------------------------------------------------------------------------1.-6-------------- PATS TESTED N a 8 INCLUDED IN ANALYSES PREGNANT DELIVERED RATS PREGNANT AND N N(t) N(l) 7a 7(100.0) 0( 0.0) 7b 7(100.0) 0( 0.0) 6a,b 5( 83.3) 0( 0.0) 4a,c 2( 50.0) 0( 0.0) CAESAREAN-SECTIONED ON DAY 21 OF GESTATION CORPORA LUTEA IMPLANTATIONS LITTER SIZES LIVE FETUSES DEAD FETUSES RESORPTIONS EARLY RESORPTIONS LATE RESORPTIONS a. Excludes values for N MEAN+S.D. MEAN+S.D. MEAN+S.D. N MEAN+S.D. N MEAN+S.D. N MEAN+S.D. N MEAN+S.D rats that did 7 18.6 + 4.2 15.1 + 2.1 13.7 1.7 9613.7 + 1.7 0 1.4 + 1.7 10 1.4 + 1.7 0 0.0 + 0.0 --not have a confirmed 7 5 16.1 + 2.3 14.0 + 2.9 12.4 + 4.0 87 12.4 + 4.0 0 21.8 + 14.4 + 13.2 + 66 13.2 + 0 4.8 4.2 5.2 5.2 20.5 + 2.1 17.0 + 7.1 16.0 + 5.6 32 16.0 + 5.6 1.6 + 1.5 1.2 + 1.8 1.0 + 1.4 9 1.3 + 1.1 6 1.2 + 1.8 2 1.0 + 1.4 2 0.3 + 0.8 0 0.0 + 0.0 0.0 0.0 ma-t-i-ng--d-a-t-e-.----------------------------------------- b. cEoxuclldudensot vbaeluedsetefromrinreadt.s 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; pr( C. Excludes values for rat 19557, which was moribund sacrificed on day 43 of study (day 2 of cohabitation). PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 10 (PACE 3)! CAESAREAN-SECTIONING OBSERVATIONS - SUMMARY --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 --------------------------------------------------------------------------------------------------------------- RATS TESTED N a a a 8 INCLUDED IN ANALYSES N 7a 7b 6a,b 4a,c PREGNANT DELIVERED N(l) N(t) 7(100.0) 0( 0.0) 7(100.0) 0( 0-0) 5( 83.3) 0( 0.0) 2( 50.0) 0( 0.0) RATS PREGNANT AND CAESAREAN-SECTIONED ON DAY 21 OF GESTATION N 7 7 5 2 DAMS WITH ANY RESORPTIONS N(t) 5( 71.4) 5( 71.4) 2( 40.0) 1( 50.0) DAMS WITH ALL CONCEPTUSES RESORBED N(l) 0( 0.0) o( o.o) 0( 0.0) 0( 0.0) DAMS WITH VIABLE FETUSES N(t) 7(100.0) 7(100.0) 5(100.0) 2(100.0) PLACENTAE APPEARED NORMAL N(t) 7(100.0) 7(100.0) 5(100.0) 2(100.0) --------------------------------------------------------------------------------------------------------------- a. Excludes values for rats that did not have a confirmed mating date. b. Excludes values for rate 19520 and 19541, which were moribund sacrificed on day 0 of presumed gestation; p could not be determined. c. Excludes values for rat 19SS7. which was moribund sacrificed on day 43 of study (day 2 of cohabitation). PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 11 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - SUMMARY --------------------------------------------------------------------------------------------------------------- RATS CAESAREM-SECTIONED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP I ii III IV DOSAGE (MG/KG/DAY) 0 (VEHICLE) 0.1 0.4 1.6 --------------------------------------------------------------------------------------------------------------- LITTERS WITH ONE OR MORE LIVE FETUSES N 7 7 5 2 IMPLANTATIONS MEAN+S.D. 15.1 + 2.1 14.0 + 2.9 14.4 + 4.2 17.0 + 7.1 LIVE FETUSES N MEAN+S.D. 96 13.7 + 1.7 87 12.4 + 4.0 66 13.2 + 5.2 32 16.0 + 5.6 LIVE MALE FETUSES t LIVE MALE FETUSES/LITTER N MEAN+S.D. 44 46.0 + 11.7 46 52.2 + 14.2 36 59.3 + 19.4 15 45.8 + 5.9 LIVE FETAL BODY WEIGHTS (GRAMS)/LITTER MEAN+S.D. 5.12 + 0.29 5.31 + 0.42 5.06 + 0.09 5.02 + 0.16 14ALE FETUSES MEAN+S.D. 5.24 + 0.29 5.50 + 0.54 5.19 + 0.12 5.12 + 0.20 FEMALE FETUSES MEAN+S.D. 5.02 + 0.33 5.06 + 0.33 4.84 + 0.14 4.95 + 0.16 PLACENTAL WEIGHTS (GRAMS)/LITTER MEAN+S.D. 0.50 + 0.06 0.54 + 0.08 0.48 + 0.06 0.50 + 0.06 14ALE FETUSES MEAN+S.D. 0.50 + 0.06 0.55 + 0.09 0.49 + 0.07 0.52 + 0.09 FEMALE FETUSES MEAN+S.D. 0.49 + 0.05 0.52 + 0.07 0.44 + 0.08 0.47 + 0.04 t RESORBED CONCEPTUSES/LITTER MEAN+S.D. 8.8 + 9.9 13.4 + 14.6 11.0 + 19.2 4.6 + 6.4 NO FETAL ALTERATIONS WERE IDENTIFIED AT GROSS EXTERNAL EXAMINATION ---------------------------------------------------------------------------------------------------------------- I I - NUMBER OF VALUES AVERAGED a. Litter 19568 had no female fetuses. PROTOCOL 418-013: ORAL (GAVAGE) PRARMACOKINETIC STUDY OF VFOS IN RATS (SPONSOR'S STUDY NLTMBER: T-6295.12) TABLE 12 (PAGE 1): CLINICAL OBSERVATIONS - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT 4 DESCRIPTION --------------------------------------------------------------------------------------------------------------- 19501 19502 19503 DS( 43- 66) NO ADVERSE NO ADVERSE LENTICULAR FINDINGS FINDINGS OPACITY 19504 19505 19506 DS( 66 DELIVERED NO ADVERSE NO ADVERSE NO ADVERSE AND SACRIFICED FINDINGS FINDINGS FINDINGS 19507 NO ADVERSE FINDINGS 19508 NO ADVERSE FINDINGS 19509 DG( 17- 21) CHROMORHINORRHEA a 19510 NO ADVERSE FINDINGS 19511 DS( 3- 42) LOCALIZED ALOPECIA: LIMBS DG( 0- 2) LOCALIZED ALOPECIA: LIMBS DG( 3) ALOPECIA NO LONGER APPARENT DG( 0- 15) LENTICULAR OPACITY 19512 19513 19514 19515 19516 DG( 14- 15) DG( 11- 21) CHROMORHINORRHEA a CHROMODACRYORRHEA a NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS ---------------------------------------------------------------------------------------------------------------- DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. observation confirmed at necropsy. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 2): CLINICAL OBSERVATIONS - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 19517 19518 19519 19520 DG( 7- 15) DS( 13- 42) DG( 0- 21) DS( 10- 14) TRAUMATIZED CORNEA a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS LOCALIZED ALOPECIA: LIMBS a CHROMODACRYORRHEA DS( 10- 21) INCISORS: MISSING/BROKEN DS( 22 ) INCISORS: GROWN IN DS( 22- 30) DS( 31 ) INCISORS: INCISORS: MISALIGNED REALIGNED 19521 DS( 30- 42) DS( 42 DG( 0 DG( 0 DG( 0 DG( 0 DG( 0 DG( 0 DG( a DG( 0 DG( 0 DS( 44- 4S) LOCALIZED ALOPECIA: LIMBS SNOUT: SWOLLEN CHROMODACRYORRHEA a EXOPHTHALMOS a SNOUT: SWOLLEN a LOCALIZED ALOPECIA: LIMBS CORNEAL OPACITY RIGHT EYE: LARGE RIGHT EYE: DARK RED RIGHT EYE: LIDS UNABLE TO MORIBUND SACRIFICED EXOPHTHALMOS a CLOSE DG( 0) DG( 4- 5) EXOPHTHALMOS NO LONGER SNOUT: SWOLLEN APPARENT DG( 6) DG( 11- 21) SWELLING RESOLVED CORNEAL OPACITY 19S22 19523 19524 DS( 43- 45) DS( 43- 45) DG( 0) DS( 44- 45) DG( 0) DG( 0- 10) NO ADVERSE FINDINGS NO ADVERSE FINDINGS RIGHT EYE: CLOUDY, WHITE FILM SNOUT: SWOLLEN SWELLING RESOLVED EXOPHTHALMOS EXOPHTHALMOS NO LONGER APPARENT TRAUMATIZED CORNEA DG( 0- 10) DG( 0- 13) RIGHT EYE: CLOUDY, WHITE FILM LOCALIZED ALOPECIA: HEAD DG( 14 ) AWPECIA NO LONGER APPARENT DG( 11- 21) RIGHT EYE: MISSING a --------------------------------------------------------------------------------------------------------------- DS - DAY OF STUDY DG - DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-013: ORAL ((;AVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS TABLE 12 (PAGE 3): CLINICAL OBSERVATIONS - INDIVIDUAL DATA (SPONSOR'S STUDY NUMBER! T-6295.12) -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- RAT N DESCRIPTION -------------------------------------------------------------------------------------------------------------- 19525 NO ADVERSE FINDINGS 19526 NO ADVERSE FINDINGS 19527 NO ADVERSE FINDINGS 19528 DG( 2- 8) INCISORS: MISSING/BROKEN DG( 9 INCISORS: GROWN IN 19529 NO ADVERSE FINDINGS 19530 NO ADVERSE FINDINGS 19531 DG( 0- 2) EXOPHTHALMOS DG( 3) DG( 1- 1S) EXOPHTHALMOS NO LONGER APPARENT CORNEAL OPACITY DG( 3- 15) TRAUMATIZED CORNEA a 19532 NO ADVERSE FINDINGS --------------------------------------------------------------------------------------------------------------- DS - DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-013: ORAL (CA"GE) PHARMACOKINETIC STUDY OF PFOS IN RATS TA13LE 12 (PACE 4): CLINICAL OBSERVATIONS - INDIVIDUAL DATA (SPONSOR'S STUDY NUMBER: T-6295.12) --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 111 0.4 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 19533 DG( 0- 1) L-ACRIMATION DG( 0- 2) LENTICULAR OPACITY 19534 19535 19536 19537 19538 19539 19540 19541 19542 19543 19544 19545 19546 19547 19548 DG( 0- 2) DG( 3) DG( 3- 21) DG( 14- 21) DS( 25 DG( 0 DG( 0 DG( 0 DG( 0 DG( 0 DG( 0 DG( 0 DG( 0 DG( 10- 12) DG( 13 DG( 1 DG( I EXOPHTHABMOS EXOPHTHALMOS NO LONGER APPARENT CORNEAL OPACITY ENOPHTHALMOS a NO ADVERSE FINDINGS CHROMORHINORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMODACRYORRHEA a EXOPHTHALMOS a SNOUT: SWOLLEN a CORNEAL OPACITY RIGHT EYE: LARGE RIGHT EYE: DARK RED RIGHT EYE: LIDS UNABLE TO CLOSE MORIBUND SACRIFICED LOCALIZED ALOPECIA: UNDERSIDE ALOPECIA NO LONGER APPARENT NO ADVERSE FINDINGS NO ADVERSE FINDINGS NO ADVERSE FINDINGS CHROMORHINORRHEA UNGROOMED COAT NO ADVERSE FINDINGS NO ADVERSE FINDINGS ---------------------------------------------------------------------------------------------------------------- DS = DAY OF STUDY DG = DAY OF PRESUMED a. Observation confirmed at necropsy. GESTATION PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PPOS IN RATS TA13LE 12 (PAGE 5): CLINICAL OBSERVATIONS - INDIVIDUAL DATA (SPONSOR'S STUDY NUMBER: T-6295.12) --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 19549 DS( 57- 59) LOCALIZED ALOPECIA: LIMBS DS( 60 ) ALOPECIA NO LONGER APPARENT DS( 62- 63) LOCALIZED ALOPECIA: LIMBS 19550 19551 19552 DS( 64 DS( 44- 50) ALOPECIA NO LONGER NO ADVERSE FINDINGS NO ADVERSE FINDINGS CORNEAL OPACITY APPARENT DS( 50- 68) CORNEAL HEMORRHAGE a 19553 OS(,62- 68) LOCALIZED ALOPECIA: NO ADVERSE FINDINGS LIMBS a 19554 DS( 1 CHROMORHINORRHEA 19555 NO ADVERSE FINDINGS 19556 19557 DS( 29- 43) D(;( 0- 21) DS( 43 LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: CHROMODACRYORRHEA a LIMBS LIMBS a DS( 43 EXOPHTRALmos a DS( 43 SNOUT: SWOLLEN a DS( 43 CORNEAL OPACITY DS( 43 RIGHT EYE: LARGE DS( 43 RIGHT EYE: DARK RED DS( 43 RIGHT EYE: LIDS UNABLE TO CLOSE 19558 DS( 43 DS( 36- 38) MORIBUND SACRIFICED EARS: SWOLLEN 19559 DS( 39 DG( 15 SWELLING RESOLVED EXOPHTHALMOS DG( 16 EXOPHTHALMOS NO LONGER APPARENT 19560 DG( 16- 21) DS( 2) CORNEAL HEMORRHAGE a CHROMORHINORRHEA 19561 19562 DG( 15- 21) DS( 23 LOCALIZED ALOPECIA: NO ADVERSE FINDINGS CHROMORHINORRHEA BACK a 19563 NO ADVERSE FINDINGS ---------------------------------------------------------------------------------------------------------------- DS = DAY OF STUDY DG c DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 12 (PAGE 6): CLINICAL OBSERVATIONS - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PAT # DESCRIPTION --------------------------------------------------------------------------------------------------------------- 19S64 DS4 I CHROMORHINORRHEA DS( 6 CHROMORHINORRHEA DS( 29- 42) DS( 34- 35) DS( 36- 42) DS( 31- 42) LEFT FOREPAW: SCAB (DID NOT EXCEED 0.5 CM X 0.3 CM) RIGHT FOREPAW: SCAB (0.4 CM X 0.3 CM) RIGHT FOREPAW: TWO SCABS (DID NOT EXCEED 0.5 CM X 0.3 CM) LOCALIZED ALOPECIA: UNDERSIDE DS( 36- 37) FOREPAWS: SWOLLEN DS( 38 SWELLING RESOLVED DGI 0 DG( 0 DG( I LEFT FOREPAW: RIGHT FOREPAW: SCABS HEALED SCAB (DID NOT EXCEED 0.5 CM X 0.3 CM) TWO SCABS (DID NOT EXCEED 0.5 CM X 0.3 CM) DG( 0- 15) DG( I- 15) LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: UNDERSIDE a LIMBS a --------------------------------------------------------------------------------------------------------------- DS - DAY OF STUDY DG - DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: TABLE 12 (PAGE 7): CLINICAL OBSERVATIONS - INDIVIDUAL DATA T-6295.12) -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- RAT # DESCRIPTION -------------------------------------------------------------------------------------------------------------- 19565 DG( 0- 15) CORNEAL HEMORRHAGE a DG( I- 2) EXOPHTHALMOS l9s66 DG( 3 DS( 13 EXOPHTHALMOS NO LONGER APPARENT CHROMODACRYORRHEA DG( 0- 15) CORNEAL OPACITY 19567 19568 19569 19570 DG( 13- 15) DS( 9- 11) DS( 12 ) DS( 9- 42) DG( 0- 21) DS( 34- 43) DS( 36- 38) DS( 39 ) LOCALIZED ALOPECIA: UNDERSIDE a NO ADVERSE FINDINGS RIGHT FOREPAW: THIRD DIGIT SWOLLEN SWELLING RESOLVED RIGHT FOREPAW: THIRD DIGIT MISSING RIGHT FOREPAW: THIRD DIGIT MISSING a NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LIMBS RIGHT FOREPAW: SCAB (0.2 CM IN DIAMETER) SCAB HEALED 19571 19572 19573 19574 19575 19S76 19577 19578 19S79 DG( 0- 15) DG( 21 ) DG( 11- 21) DS( 1 DG( 1 DS( S4- 68) DG( 0- 21) LOCALIZED ALOPECIA: LIMBS a DELIVERED AND SACRIFICED LEFT AXILLA: MASS (1.5 CM X 1.0 CM X 1.0 CM)a NO ADVERSE FINDINGS CHROMORHINORRHEA CHROMODACRYORRHEA NO ADVERSE FINDINGS NO ADVERSE FINDINGS LOCALIZED ALOPECIA: LOCALIZED ALOPECIA: BACK a LIMBS a DG( 21 ) DELIVERED AND SACRIFICED 19580 DG( 11- 1S) LOCALIZED ALOPECIA: LIMBS a --------------------------------------------------------------------------------------------------------------- DS = DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Observation confirmed at necropsy. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 13 (PAGE 1): NECROPSY OBSERVATIONS - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a -------------------------------------------------------------------------------------------------------------- I 0 (VEHICLE) 19501 DG 15 p S7 ALL TISSUES APPEARED NORMAL. 19502 DG 15 NP 58 ALL TISSUES APPEARED NORMAL. 19SO3 DS 66 p 66 DELIVERED AND SACRIFICED ON DAY 66 ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 14 IMPLANTATION PUPS).B 19504 DG 21 p 65 ALL TISSUES APPEARED NORMAL. 19505 DG 21 p 64 ALL TISSUES APPEARED NORMAL. 19506 DG 21 p 64 ALL TISSUES APPEARED NORMAL. 19507 DG 21 p 67 ALL TISSUES APPEARED NORMAL. 19508 DG 15 p 58 ALL TISSUES APPEARED NORMAL. 19509 DG 21 p 64 ALL TISSUES APPEARED NORMAL. 19510 DC 15 p 57 ALL TISSUES APPEARED NORMAL. 19sil DG 15 p S7 ALL TISSUES APPEARED NORMAL. 19512 DG 21 p 66 ALL TISSUES APPEARED NORMAL. 19513 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19514 DG 15 NP 57 ALL TISSUES APPEARED NORMAL. 19515 DG 15 p 58 ALL TISSUES APPEARED NORMAL. 19516 DG 21 p 65 ALL TISSUES APPEARED NORMAL. -------------------------------------------------------------------------------------------------------------- P PREGNANT NP = NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at nec b. Appeared normal for their developmental ages at the time maternal death occurred. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 13 (PAGE 2): NECROPSY OBSERVATIONS - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP RAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- ii 0.1 19517 DG 15 NP 58 ALL TISSUES APPEARED NORMAL. 19sle DG 21 p 64 ALL TISSUES APPEARED NORMAL. l9sl9 DG 21 p 63 ALL TISSUES APPEARED NORMAL. 19520 DG 0 43 MORIBUND SACRIFICED ON DAY 0 OF PRES PREGNANCY STATUS COULD NOT BE DETERM ALL TISSUES APPEARED NORMAL. 19521 DG 21 p 66 ALL TISSUES APPEARED NORMAL. 19522 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19523 DG 21 p 67 ALL TISSUES APPEARED NORMAL. 19524 DG 21 p 66 ALL TISSUES APPEARED NORMAL. 19525 DC 15 p 57 ALL TISSUES APPEARED NORMAL. 19526 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19527 DC 15 p sa ALL TISSUES APPEARED NORMAL. 19528 DG 21 p 63 ALL TISSUES APPEARED NORMAL. 19529 DG 21 p 63 ALL TISSUES APPEARED NORMAL. 19S30 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19531 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19532 DG 15 p 57 ALL TISSUES APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- P PREGNANT NP = NOT PREGNANT DS DAY OF STUDY DG - DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necr PROTOCOL, 418-013: ORAI, (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABBE 13 (PACE 3): NECROPSY OBSERVATIONS - INDIVIDUAB DATA --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP PAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- III 0.4 19533 DG 21 p 63 ALI, TISSUES APPEARED NORMAL. 19534 DG 21 p 63 ALL TISSUES APPEARED NORKAL. 19535 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19536 DG 21 p 65 ALL TISSUES APPEARED NORMAL. 19537 DG 15 p S? ALL TISSUES APPEARED NORMAL. 19538 DG 15 p se ALL TISSUES APPEARED NORMAL. 19539 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19540 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19541 DG 0 43 MORIBUND SACRIFICED ON DAY 0 OF PRES PREGNANCY STATUS COULD NOT BE DETERM ALL TISSUES APPEARED NORMAL. 19542 DG 15 p se ALL TISSUES APPEARED NORMAL. 19543 DG 21 p 66 ALL TISSUES APPEARED NORMAL. 19S44 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19545 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19546 DG 21 p 63 ALI, TISSUES APPEARED NORMAL. 19547 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19548 DG 21 NP 63 ALL TISSUES APPEARED NORMAL. --------------------------------------------------------------------------------------------------------------- P PREGNANT NP = NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necr PROTOCOL 418-013- OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-629S.12) TABLE 13 (PACE 4): NECROPSY OBSERVATIONS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------- DOSAGE GROUP DOSAGE (MG/KG/DAY) RAT NUMBER DAY OF NECROPSY PREGNANCY STATUS DOSAGES ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- IV 1.6 19549 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19550 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19551 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19SS2 DS 68 NP 67 ALL TISSUES APPEARED NORMAL. 19553 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19554 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19555 DG 15 NP 57 ALL TISSUES APPEARED NORMAL. 19S56 DG 21 NP 64 ALL TISSUES APPEARED NORMAL. 19557 DS 43 43 MORIBUND SACRIFICED ON DAY 43 OF STU COHABITATION). ALL TISSUES APPEARED NORMAL. 19558 DG IS p 57 19559 DG 21 NP 64 19560 DG 21 p 66 19S61b DG is 57 ALL TISSUES ALL TISSUES ALL TISSUES APPEARED APPEARED APPEARED NORMAL. NORMAL. NORMAL. 19562 DG 21 p 65 ALL TISSUES APPEARED NORMAL. 19563 DC 15 p 57 ALL TISSUES APPEARED NORMAL. 19564 DG 15 p 57 ALL TISSUES APPEARED NORMAL. ---------------------------------------------------------------------------------------------------------------- P PREGNANT NP = NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necro b. Necropsy and Caesarean-section observations were not recorded for rat 19561. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! TABLE 13 (PAGE S): NECROPSY OBSERVATIONS - INDIVIDUAL DATA T-6295.12) --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP PAT DAY OF PREGNANCY DOSAGES DOSAGE (MG/KG/DAY) NUMBER NECROPSY STATUS ADMINISTERED OBSERVATIONS a --------------------------------------------------------------------------------------------------------------- v 3.2 19S65 DG 15 p 57 ALL TISSUES APPEARED NORMAL. 19566 DG 15 p se ALL TISSUES APPEARED NORMAL. 19567 DG 15 p se ALL TISSUES APPEARED NORMAL. 19568 DG 21 p 63 ALL TISSUES APPEARED NORMAL. 19569 19570 DG 21 p DG IS p 66 ALL TISSUES APPEARED NORMAL. 58 ALL TISSUES APPEARED NORMAL. 19S71 DG 21 p 66 DELIVERED ON DAY 21 OF GESTATION. ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 15 IMPLANTATION S DELIVERED PUPS, SIX LIVE FETUSES AND RESORPTIONS).B 19572 19573 19574 19575 19S76 19577 19578 DG 21 p DG 15 p DG 15 p DG 15 p DS 68 NP DG 21 p DS 68 NP 64' ALL TISSUES APPEARED NORMAL. 57 ALL TISSUES APPEARED NORMAL. 57 ALL TISSUES APPEARED NORMAL. 57 ALL TISSUES APPEARED NORMAL. 67 ALL TISSUES APPEARED NORMAL. 63 ALL TISSUES APPEARED NORMAL. 67 ALL TISSUES APPEARED NORMAL. 19579 DG 21 p 65 DELIVERED ON DAY 21 OF GESTATION. ALL TISSUES APPEARED NORMAL. UTERINE CONTENTS: 11 IMPLANTATION S DELIVERED PUPS AND EIGHT LIVE FETUSE 19580 DG 15 p 57 ALL TISSUES APPEARED NORMAL. ---------- ---------------------------------------------------------------------------------------------------- P PREGNANT NP = NOT PREGNANT DS DAY OF STUDY DG = DAY OF PRESUMED GESTATION a. Refer to the individual clinical observations table (Table 12) for external observations confirmed at necr b. Appeared normal for their developmental ages at the time maternal death occurred. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 1): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- 19501 215. 218. 222. 219. 220. 226. 232. 231. 228. 223. 227. 232. 234. 23 19502 221. 231. 234. 240. 230. 240. 243. 243. 240. 244. 248. 247. 249. -24 19503 227. 224. 224. 226. 234. 235. 233. 238. 237. 240. 236. 241. 247. 24 19SO4 231. 233. 235. 239. 242. 240. 251. 252. 252. 254. 256. 259. 264. 26 19505 225. 224. 225. 232. 238. 242. 243. 242. 242. 245. 246. 247. 250. 25 19SO6 240. 236. 240. 243. 248. 253. 251. 254. 255. 255. 253. 260. 263. 26 19507 219. 222. 224. 223. 221. 221. 226. 231. 234. 229. 234. 238. 242. 24 19508 22S. 227. 228. 226. 232. 238. 240. 238. 241. 247. 249. 244. 252. 25 19509 221. 219. 218. 222. 226. 227. 227. 227. 230. 233. 236. 231. 238. 24 19510 236. 234. 234. 238. 243. 238. 243. 250. 250. 252. 245. 253. 258. 25 19sil 229. 229. 230. 236. 242. 243. 241. 251. 256. 251. 248. 258. 262. 25 19512 235. 232. 240. 243. 241. 240. 245. 2Sl. 248. 249. 254. 259. 255. 25 19513 235. 238. 240. 246. 252. 254. 252. 261. 264. 266. 264. 267. 274. 27 19514 234. 228. 238. 240. 244, 240. 250. 252. 257. 247. 249. 256. 259. 26 195is 223. 224. 227. 222. 223. 229. 233. 230. 229. 230. 235. 235. 235. 23 19516 221. 226. 230. 234. 231. 237. 239, 243. 238. 249. 246. 248. 248. 25 --------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 3 --------------------------------------------------------------------------------------------------------------- 19501 237. 237. 236. 239. 244. 244. 238. 244. 246. 246. 245. 247. 2SO. 24 19502 256. 257. 2S4. 260. 266. 264. 263. 262. 268. 273. 273. 268. 268. 27 19503 246. 254. 253. 252. 253. 260. 259. 252. 257. 259. 261. 256. 260. 26 19504 272. 271. 273. 272. 281. 280. 282. 277. 279. 28S. 286. 284. 281. 28 19505 253. 257. 261. 260. 260. 257. 257. 262. 264. 268. 272. 271. 27S. 27 19506 266. 267. 264. 268. 268. 266. 262. 265. 268. 267. 265. 269. 276, 27 19507 250. 251. 251. 251. 259. 261. 262. 255. 257. 263. 263. 264. 260. 26 19508 258. 26S. 264. 260. 267. 272. 268. 267. 270. 273. 271. 268. 274. 27 19509 246. 249. 249. 24S. 253. 257. 256. 251. 256. 257. 2S7. 252. 258. 25 19510 261. 261. 256. 262. 264. 265. 2SS. 262. 264. 258. 259. 262. 261. 25 19511 269. 271. 266. 272. 277. 273. 267. 280. 280. 274. 274. 285. 286. 28 19512 267. 264. 270. 274. 272. 273. 275. 282. 280. 275. 284. 284. 282, 27 19513 264. 288. 284. 294. 300. 299. 292. 297. 304. 303. 305. 310. 311. 30 19514 264. 269. 268. 269. 270. 27S. 274. 278. 276. 273. 282. 287. 284. 27 19515 244. 244. 245. 249. 252. 251. 247. 248. 253. 246. 242. 245. 248. 24 19516 251. 258. 265. 260. 256. 264. 266. 26S. 265. 268. 272. 270. 261. 26 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 2): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT 0 DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42a -------------------------------------------------------------------------------------------------------------- 19501 2SI. 2S2. 249. 2SO. 252. 252. 249. 251. 256. 255, 19502 268. 276. 270. 266. 271. 274. 271. 270. 273. 280. 19503 263. 266. 267. 263. 265. 268. 268. 269. 270. 277. 19504 288. 293. 295. 292. 292. 295. 296. 292. 293. 298. 19505 19506 19507 19508 19509 281. 273. 262. 278. 258. 279. 266. 266. 285. 262. 280. 264. 270. 283. 260. 278. 260. 262. 277. 259. 280. 256. 262. 283. 258. 285. 257. 268. 288. 260. 283. 2S7. 268. 283. 258. 282. 254. 267. 279. 260. 280. 261. 263. 288. 262. 285. 261. 266. 289. 266. 19510 268. 265. 2S7. 259. 254. 260. 260. 263. 261. 260. 19511 290. 287. 282, 291. 292. 284. 284. 292. 296. 292. 19512 285. 281. 284. 286. 286. 284. 287. 292. 288. 288. 19513 317. 314. 308. 317. 318. 317. 312. 322. 325. 324. 19514 288. 283. 286. 288. 287. 286. 290. 298. 293. 290. 19515 250. 248. 245. 246. 2SI. 255. 252. 251. 257. 257. 19516 271. 277. 278. 279. 274. 278. 282. 280. 274. 275. --------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-013! ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 3): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA ----------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP 11 0.1 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14 ---------------------------------------------------------------------------------------------------------------- 19517 221. 219. 225. 228. 230. 227. 233. 237. 235. 232. 238. 244. 244. 240 19518 234. 238. 237. 249. 250. 250. 252. 261. 264. 264. 265. 262. 271. - 273 19519 19520 19521 19522 19523 19524 19525 19s@6 225. 230, 220. 234. 228. 233. 224. 227. 230. 227. 222, 236. 227. 231. 218. 230. 236. 232. 229. 237. 228. 235. 225. 229. 237. 237. 235. 242. 232. 239. 229. 228. 237. 240. 239. 247. 238. 239. 231. 234. 243. 236. 235. 250. 238. 236. 229. 238. 247. 243. 242. 245. 234. 242. 233. 240. 244. 242. 245. 256. 239. 250. 235. 241. 248. 239. 244. 255. 240. 249. 232. 239. 252. 240. 242. 255. 243. 249. 234. 246. 254. 240. 248. 250. 243. 254. 239. 249. 252. 246. 254. 258. 245. 257. 240. 251. 251. 241. 251. 263. 248. 256. 240. 247. 258 240 250 258 251. 256 242. 243. 19527 19528 227. 229. 233. 231. 231. 237. 233. 238. 237. 240. 239. 239. 244. 245. 248. 248. 244. 242. 252. 243. 254. 248. 260. 253. 258. 252. 255 251. 19529 226. 229. 227. 234. 235. 234. 234. 239. 238. 243. 238. 241. 244. 243. 19530 228. 229. 230. 238. 239. 242. 234, 240. 244. 245. 243. 252. 255. 252. 19531 222. 220. 230. 233. 236. 235. 242. 244. 247, 242. 241. 243. 249. 248. 19532 230. 230. 230. 228. 234. 239. 244. 245. 239. 246. 248. 253. 250. 246. ----------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 30 ----------------------------------------------------------------------------------------------------------------- 19517 247. 245. 250. 249. 253. 247. 254. 256. 257. 256. 261. 260. 256. 249. 19518 275. 276. 279. 278. 280. 288. 286. 284. 289. 290. 289. 285. 293. 291. 19519 264. 267. 270. 265. 266. 271. 274. 274. 269. 273. 278. 276, 270. 277. 19520 248. 246. 254. 250. 254. 253. 258. 256. 257. 260. 260. 260. 263. 255. 19521 259. 258. 264. 269. 267. 264. 268. 272. 272. 266. 275. 275. 271. 268. 19522 268. 266. 2S7. 26S. 269. 270. 267. 275. 282. 278. 272. 278. 281. 277. 19523 260. 259. 259. 262. 268. 266. 262. 263. 267. 269. 269. 267. 268. 271. 19524 264. 264. 266. 272. 270. 268. 275. 277. 276. 276. 280. 285. 276. 273. 19525 245. 249. 247. 247. 251. 255. 253. 251. 250. 247. 251. 253. 252. 257. 19526 256. 252. 250. 256. 262. 265. 260. 260. 263. 266. 268. 264. 260. 268. 19527 267. 264. 264. 269, 270. 275. 273. 270. 272. 277. 278. 277. 270. 275. 19528 255. 254. 260. 260. 256. 256. 256. 266. 261. 259. 266. 264. 256. 252. 19529 244. 243. 242. 245. 246. 248. 241. 246. 247. 246. 240. 251. 246. 247. 19530 259. 261. 254. 264. 269. 267. 264. 264. 270. 267. 265. 271. 271. 266. 19531 255. 255. 253. 258. 264, 261. 255. 262. 265. 265. 262. 270. 271. 270. 19532 257. 256. 255. 262. 268. 269. 265. 265. 270. 270. 274. 271. 267. 270. ----------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: OPAL IGA"GE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 4): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA ---------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP Il 0.1 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42a ---------------------------------------------------------------------------------------------------------------- 19517 262. 2S6. 259. 262. 260. 255. 262. 266. 265. 257. 19518 294. 290. 295. 296. 301. 303. 300. 298. 303. 310. 19519 273. 277. 280. 276, 273. 269. 276. 278. 278. 276. 19520 265. 265. 269. 272. 267. 264. 265. 268. 273. 263. 19521 279. 274. 281. 284. 280. 279. 288. 288. 285. 288. 19522 285. 286. 276. 280. 286. 285. 278. 282. 287. 282. 19523 271. 273. 274. 266. 266. 268. 273. 274. 275. 274. 19524 282. 276. 280. 279. 278. 276. 281. 284. 280. 279. 19525 2S3. 254. 254. 251. 257. 252. 254. 255. 252. 255. 19526 262. 262. 268. 270. 269. 268. 271. 270. 272. 272. 19527 278. 276. 282. 286. 286. 284. 279. 283. 285. 293. 19528 257. 253. 260. 261. 258. 256. 262. 266. 259. 257. 19529 251. 247. 242. 247. 249. 250. 244. 254. 253. 251. 19530 274. 270. 267, 271. 274. 272. 267. 270. 277. 269. 19531 273. 269. 266. 277. 283. 276. 273. 280. 282. 276. 19532 272. 277. 281. 278. 271. 271. 282. 278. 280. 269. ---------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 5): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA ---------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP 111 0.4 MG/KC/DAY ---------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 14 ---------------------------------------------------------------------------------------------------------------- 19S33 221. 224. 222. 220. 226. 236. 228. 229. 223. 227. 233. 231. 232. 230 19534 234. 241. 244. 251. 256. 258. 253. 262. 266. 263. 259. 264. 266. -269 19535 221. 220. 226. 223. 226. 230. 227. 229. 230. 230. 233. 230. 230. 232 19536 225. 225. 230. 233. 23S. 230. 230. 242. 242. 243. 240. 243. 246. 246 19537 217. 219. 223. 221. 222. 224. 227. 233. 227. 229. 235. 233. 231. 231 19S38 230. 227. 229. 233. 237. 237. 241. 242. 243. 243. 244. 248. 246. 247 19539 226. 227. 225. 230. 231. 230. 234. 238. 233. 230. 233. 236. 239. 238 19540 230. 229. 230. 234. 236. 236. 233. 241. 243. 243. 241. 246. 250. 250 19541 218. 218. 216. 220, 225. 226. 227. 229. 230. 230. 228. 234. 236. 236 19542 237. 236. 231. 240. 24S. 244. 240. 250. 252. 253. 246. 250. 257. 261 19543 234. 233. 242. 244. 249. 245. 252. 254. 254. 253. 257. 257. 263. 258 19544 231. 231. 232. 235. 241. 245. 246. 248. 245. 254. 253. 258, 257. 253 19545 222. 220. 232. 231. 231. 229. 234. 238. 239. 240. 235. 239. 241. 245 19546 234. 235. 231. 239. 240. 239. 242. 248. 248. 24.8. 250. 2SS. 263. 265 19547 223. 228. 228. 230. 235. 232. 234. 243. 242. 240. 239. 247. 248. 246 19548 228. 231. 23S. 234. 239. 242. 246. 246. 243. 247. 249. 250. 253. 254 ---------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 30 ---------------------------------------------------------------------------------------------------------------- 19533 240. 237. 238. 244. 246. 245. 246. 243. 245. 248. 252. 250. 244. 247 19534 282. 280. 281. 286. 291. 288. 281. 293. 296. 297. 295. 303. 298. 296 19535 239. 238. 242. 243. 244. 248. 244. 247. 245. 247. 251. 251. 254. 255 19536 246. 250. 2S3. 255. 2S2. 255. 259. 255. 255. 258. 261. 260. 256. 262 19537 238. 234. 235. 237. 244. 243. 236. 239. 238. 238. 244. 243. 240. 240 19538 252. 253. 2S2. 255. 254. 252. 253. 255. 255. 256. 256. 256. 254. 254 19539 242. 242. 244. 246. 248. 248. 246. 248. 253. 250. 247. 250. 253. 249 19540 256. 256. 2S3. 264. 263. 263. 260. 266. 274. 270. 269. 275. 280. 274 19541 19542 235. 240. 239. 238. 239, 244. 241. 242. 236. 242. 244. 246. 239. 243 261. 265. 264. 259. 266. 269. 268. 266. 266. 272. 272. 264. 272. 274 19543 266. 265. 276. 281. 274. 277. 281. 285. 287. 284. 289. 290. 284. 278 19544 265. 26S. 262. 269. 273. 276. 274. 271. 284. 278. 280. 275. 270. 278 19545 244. 247. 253. 253. 254. 255. 2S7. 259. 260. 261. 260. 263. 262. 265 19546 269. 274. 278. 274. 283. 288. 286. 285. 288. 292. 295. 293. 286. 294 19547 249. 2S2. 249, 251. 258. 259. 254. 257. 263. 262. 259. 263. 267. 265 19548 261. 260. 261. 266. 268. 267. 264. 266. 269. 268. 268. 266. 265. 268 ---------------------------------------------------------------------------------------------------------------- DAY . DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: ORAL IGAVAGE) PIIARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-6295.12) TABLE 14 (PAGE 6): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 111 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42a -------------------------------------------------------------------------------------------------------------- 19533 254. 252. 256. 259. 260. 257. 256. 261. 264. 259. 19534 302. 300. 299. 308. 308. 305. 302. 308. 307, 311. 19535 2S4. 254. 258. 257. 262. 263. 261. 262. 2S9. 259. 19536 262. 264. 264. 262. 259. 266. 266. 264. 262. 268. 19537 246. 247. 247. 253. 252. 250. 248. 255. 253. 254. 19S38 258. 258. 259. 259. 261. 260. 258. 2S6. 260. 260. 19539 256. 254. 252. 258. 258. 259. 257. 257. 257. 255. 19540 280. 281. 280. 282. 289. 281. 278. 284. 289. 286. 19541 242. 243. 244. 246. 243. 248, 244. 249. 247. 246. 19542 271. 278. 274. 267. 274. 276. 274. 269. 273. 276. 19543 285. 262. 289. 291. 294. 288. 293. 296. 291. 286. 19544 274. 276. 283. 289. 282. 284. 275. 290. 295. 211. 19545 268. 267. 264. 260. 264. 272. 272. 270. 270. 290. 19546 291. 299. 300. 298. 295. 299. 308. 305. 300. 274. 19547 269. 268. 273. 272. 274. 271. 271. 275. 275. 297. 19548 264. 266. 266. 264. 261. 260. 261. 264. 267. 266. -------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (C). a. Last value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 7): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP IV 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 8 9 10 11 12 13 -------------------------------------------------------------------------------------------------------------- 19549 220. 220. 219. 226. 229. 227. 228. 231. 233. 233. 230. 235. 237. 2 19550 239. 236. 247. 250. 251. 251. 253. 264. 260. 263. 256. 264. 270. - 26 19551 228. 230. 230. 234. 240. 242. 242. 252. 250. 249. 250. 2Sl. 261. 26 19552 230. 230. 23S. 242. 236. 232. 237. 240. 239. 231. 239. 243. 242. 2 19553 218. 216. 215. 221. 228. 226. 223. 224. 233. 230. 232. 230. 233. 2 19554 232. 236. 234. 239. 247. 248. 246. 250. 254. 254. 251. 258. 260. 2 19555 231. 234. 237. 234. 240, 249. 248. 248. 246. 250. 253. 258. 253. 2 19556 230. 229. 232. 240. 238. 233. 240. 246. 243. 240. 247. 253. 251. 24 19557 221. 225. 224. 223. 226. 230. 234, 236. 231. 238. 239. 243. 243. 2 19558 234. 239. 239. 244. 247. 250. 246. 249. 252. 256. 256. 258. 259. 2 19559 220. 227. 228. 230. 227. 235. 237. 234. 235. 239. 237. 233. 238. 2 19560 225. 229. 232. 230. 232. 238. 243. 246. 240. 246. 247. 252. 248. 24 19S61 220. 220. 222. 228. 232. 224. 218. 230. 237. 235. 233. 234. 242. 2 19562 235. 239. 242. 243. 239. 236. 241. 241. 240. 240. 245. 249. 247. 2 19S63 216. 220. 221. 225. 229. 230. 229. 234. 233. 235. 229. 233. 239. 24 19564 231. 230. 229. 230. 240. 240. 242. 242. 241. 247. 245. 250. 251. 2 -------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 -------------------------------------------------------------------------------------------------------------- 19549 195SO 19551 19552 19553 19554 248. 246. 248. 250. 2S2. 252. 248. 253. 254. 253. 251. 254. 249. 2 271. 267. 265. 271. 273. 275. 268. 274. 275. 272. 266. 272. 275. 2 265. 270. 272. 273. 273. 275. 279. 284. 280. 278. 279. 281. 274. 2 24S. 240. 244. 247. 246. 243. 244. 249. 249. 246. 248. 252. 247. 24 232. 238, 237. 236. 235. 240. 236. 238. 233. 234. 239. 238. 232. 2 266. 267. 263. 267. 271. 271. 266. 269. 271. 269. 267. 271. 271. 26 195ss 19556 19SS7 19558 195S9 19560 266. 263. 268. 272. 272. 270. 275. 276. 277. 273. 282. 284. 282. 2 258. 251. 260. 263. 266. 264. 264. 268. 261. 262. 272. 267. 261. 2 251. 247. 246. 2Sl. 253. 2S4. 252. 254. 256. 2S5. 257. 256. 249. 2 266. 266. 268. 271. 269. 276. 273. 273. 276. 277. 280. 282. 280. 2 242. 244. 24S. 242. 246. 250. 248. 244. 248. 247. 248. 242. 246. 24 257. 253. 255. 260. 260. 2S6. 259. 264. 262. 256. 260. 262. 257. 2 19561 242. 239. 236. 241. 244. 245. 241. 247. 249. 244. 242. 245. 247. 24 19562 258. 256. 259. 256. 264. 264. 267. 266. 262. 266. 266. 267. 262. 2 19563 243. 247. 246. 243. 242. 245. 248. 246. 247. 246. 246. 249. 249. 2 19564 255. 253. 254. 257. 260. 259. 258. 256. 261. 261. 259. 259. 254. 2 --------------------------------------------------------------------------------------------------------------- DAY - DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS ISPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PACE 8): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA ------------ ------------------------------------------------------------------------------------------------ PAT # DOSAGE GROUP IV 1.6 MG/KG/DAY ------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42a ------------------------------------------------------------------------------------------------------------- 19549 19550 19551 19552 19553 19554 256. 272. 280. 250. 237. 273. 255. 274. 284. 252. 228. 272. 252. 266. 284. 254. 231. 267. 258. 272. 284. 257. 235. 272. 259. 272. 280. 257. 231. 272. 263. 270. 278. 260. 226. 268. 266. 266. 283. 262. 227. 268. 264. 269. 283. 262. 232. 271. 269. 271. 283. 261. 233. 273. 272. 270. 282. 263. 230. 270. 19555 19556 19557 284. 257. 252. 280. 2S4. 252. 287. 261, 25S. 287. 261. 255. 290. 258. 2S9. 282. 250. 256. 288. 258. 2S2. 288. 258. 254. 286. 256, 255. 280. 2S3. 251. 19558 19559 19560 19561 19562 19563 19564 282. 248. 261. 246. 266. 254. 249. 281. 250. 259. 246. 263. 251. 256. 284. 249. 265. 242. 266. 253. 2S6. 288. 247. 266. 246. 268, 2SO. 254. 288. 252. 265. 249. 269. 2S3. 2S3. 290. 254. 260. 252. 271. 257. 250. 291. 255. 266. 244. 268. 260. 257. 294. 252. 264. 249. 268. 258. 2S6. 291. 254. 259. 246. 273. 260. 256. 290. 252. 258. 239. 263. 255. 250. -------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 9): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP V 3.2 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAY 1 2 3 4 5 6 7 a 9 10 11 12 13 1 --------------------------------------------------------------------------------------------------------------- 19565 19566 19567 217. 238. 234. 222. 234. 237. 214. 240. 239. 221. 246. 241. 225. 245. 24S. 226. 240. 240. 224. 243. 231. 222. 249. 236. 224. 251. 237. 224. 242. 232. 232. 240, 230. 229. 245. 234. 224. 22 252. -24 235. 23 19568 240. 246. 247. 249. 248. 247. 250. 247. 248. 250. 251. 256. 2S5. 25 19569 19570 19571 19572 19573 220. 221. 224. 226. 229. 236. 233. 238. 236. 234. 236. 238. 236. 23 228. 234. 234. 234. 233. 237. 238. 243. 242. 238. 243. 246. 247. 24 235. 230. 228. 243. 241. 245. 249. 252. 252. 248. 253. 255. 252. 25 227. 227. 226. 226. 228. 230. 231. 234. 233. 235. 238. 241. 240. 24 221. 224. 221. 221. 226. 231. 233. 231. 230. 236. 234. 233. 232. 23 19574 236. 237. 237. 244. 249. 249. 245. 254. 255. 255. 253. 254. 258. 26 19575 230. 233. 230. 237. 238. 245. 242. 248. 247. 248. 245. 245. 249. 25 19576 230. 233. 234. 236. 242. 237. 243. 249. 242. 238. 247. 248. 247. 24 19577 222. 224. 225. 226. 226. 229. 229. 231. 235. 236. 238. 240. 239. 24 19578 232. 232. 235. 242. 245. 244. 250. 254. 252. 252. 256. 2SO. 258. 25 19579 220. 221. 223. 224. 221. 224. 227. 225. 222. 224. 217. 215. 215. 22 19580 230. 230. 228. 238. 242. 240. 237. 237. 243. 242. 234. 241. 238. 23 --------------------------------------------------------------------------------------------------------------- DAY 17 18 19 20 21 22 23 24 25 26 27 28 29 3 --------------------------------------------------------------------------------------------------------------- 19565 230. 227. 236. 230. 232. 230. 227. 229. 232. 231. 227. 231. 236. 23 19566 255. 253. 246. 252. 257. 253. 241. 249. 251. 251. 243. 252. 249. 24 19567 234. 230. 227. 233. 235. 229. 226. 232. 230. 228. 223. 230. 231. 22 19568 258. 262. 258. 264. 265. 266. 262. 261. 263. 267. 264. 263. 264. 26 19569 237. 241. 241. 241. 243. 245. 241. 237. 246. 247. 242. 238. 240. 24 19570 247. 248. 251. 250. 254. 255. 249. 252. 249. 250. 254. 251. 251. 24 19571 256. 252. 257. 255. 258. 254. 257. 256. 258. 252. 256. 2SS. 250. 24 19572 243. 243. 242. 242. 242. 243. 241. 242. 241. 244. 245. 248. 245. 24 19573 235. 235. 234. 241. 242. 242. 238. 235. 238. 240. 241. 237. 237. 23 19574 258. 254. 258. 258. 256. 258. 258. 259. 252. 247. 253. 254. 250. 24 19575 247. 254. 255. 256. 256. 254. 255. 260. 261. 257. 255. 256. 256. 25 19576 254. 250. 253. 250. 252. 253. 259. 260. 259. 253. 261. 257. 257. 25 19577 241. 243. 242. 239. 241. 241. 238. 234. 232. 232. 228. 227. 222. 22 19578 256. 257. 261. 262. 261. 259. 262. 262. 260. 259. 264. 264. 258. 24 19579 212. 219. 215. 213. 221. 220. 220. 220. 218. 213. 215. 214. 213. 21 19580 239. 238. 238. 245. 244. 243. 239. 247. 243. 245. 241. 247. 244. 24 --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 14 (PAGE 10): BODY WEIGHTS - PRECOHABITATION - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP V 3.2 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAY 33 34 35 36 37 38 39 40 41 42a --------------------------------------------------------------------------------------------------------------- 19565 233. 234. 233. 233. 232. 233. 227. 231. 232. 225. 19566 246. 248. 236. 242. 249. 243. 236. 236. 244. 244. 19567 224. 224. 221. 222. 223. 226. 224. 222. 225. 223. 19568 264. 263. 267. 266. 267. 267. 265. 269. 268. 268. 19569 246. 245. 240. 244. 241. 241. 238. 239. 238. 234. 19570 252. 250. 247. 250. 249. 250. 247. 24S. 246. 249. 19571 244. 243. 252. 244. 244. 239. 243. 242. 241. 241. 19572 238. 235. 236. 239. 237. 234. 236. 243. 240. 245. 19573 238. 241. 243. 243. 238. 240. 237. 240. 236. 235. 19574 240. 245. 249. 251. 251. 248. 247. 250. 251. 240. 19575 249. 250. 245. 241. 246. 243. 245. 246. 243. 233. 19576 257. 257. 265. 268. 261. 261. 262. 264. 261. 255. 19577 227. 225. 227. 226. 227. 226. 224. 224. 224. 221. 19578 256. 252. 254. 257. 258. 251. 253. 254. 251. 249. 19579 211. 209. 208. 210. 214. 217. 214. 216, 212. 211. 19580 244. 242, 239. 240. 237. 235. 234. 238. 237. 231. --------------------------------------------------------------------------------------------------------------- DAY = DAY OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PAGE 1): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY ----------------- ----------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 1 -------------------------------------------------------------------------------------------------------------- 19501 p 19502 NP 19503 P 257. 263. 269. 276. 288. 294. NO CONFIRMED MATING DATE; 270. 297. DELIVERED 274. 300. ON DAY 278. 280. 304. 303. 66 OF STUDY 277. 307. 285. 306. 289. 129 308. 31 19504 P 295. 306. 308. 308. 311. 313. 317. 316. 318. 320. 32 19505 p 281. 294. 301. 301. 302. 308. 309. 306. 309. 316. 32 19506 P 266. 273. 279. 284. 287. 295. 297. 294. 301. 304. 31 19507 p 272. 284. 286. 290. 291. 298. 296. 289. 287. 296. 30 19508 P 291. 297. 306. 313. 314. 313. 315. 319. 318. 325. 33 19509 p 269. 273. 273. 284. 287. 293. 299. 299. 293. 302. 30 19510 P 259. 267. 278. 279. 290. 291. 301. 302. 302. 311. 31 19511 P 291. 299. 305. 307. 311. 311. 316. 322. 310. 321. 32 19512 P 306. 314. 312. 316. 323. 318. 324. 318. 322. 324. 32 19513 P 330. 329. 334. 340. 350. 349. 35S. 355. 353. 353. 36 19SI4 NP 290. 298. 302. 301. 311. 313. 316. 321. 321. 325. 32 19515 P 258. 267. 274. 274. 278. 283. 285. 286. 287. 295. 29 19SI6 P 282. 288. 298. 306. 302. 309. 311. 285. 311. 315. 32 -------------------------------------------------------------------------------------------------------------- DAY 13 14 is 16 17 18 19 20 21 CR -------------------------------------------------------------------------------------------------------------- 19501 p 311. 310. 315. 19502 NP 318. 314. 310. 19503 P 19SO4 P 19505 P 19506 P 19507 P 19508 P NO CONFIRMED MATING DATE; 342. 347. 348. 333. 337. 343. 325. 326. 33S. 312. 316. 304. 350. 349. 364. DELIVERED 357. 3S6. 340. 323. ON DAY 368. 374. 360. 335. 66 OF 385. 390. 376. 350. STUDY 396. 415. 397. 363. 418. 433. 412. 371. 430. 452. 431. 394. 19509 p 19510 p 322. 330. 324. 338. 328. 339. 333. 345. 363. 380. 399. 377. 19511 p 338. 345. 351. 19512 P 19513 P 336. 374. 341. 380. 344. 386. 354. 376. 388. 406. 416. 431. 19514 NP 335. 337. 342. 19515 p 309. 313. 316. 19516 P 334. 347. 350. 357. 375. 390. 412. 422. 446. -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY - DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOMINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PACE 2): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA --------------- ---------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP Il 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 1 -------------------------------------------------------------------------------------------------------------- 19517 NP 19518 P 19519 P 19520 277. 306. 276. 247. 276. 316. 280. MORIBUND 268. 270. 319. 320. 288. 287. SACRIFICED ON DAY 272. 322. 296. 0 OF 272. 329. 296. PRESUMED 271. 333. 302. GESTATION; 272. 338. 304. PREGNANCY 272. 334. 307. STATUS 273. 334. 316. COULD NOT 27 33 32 BE D 19521 P 292. 299. 308. 309. 303. 306. 315. 306. 313. 321. 32 19522 P 292. 295. 305. 305. 308. 318. 320. 320. 319. 328. 32 19523 P 288. 294. 304. 310. 309. 312. 315. 330. 317. 318. 33 19524 P 296. 304. 311. 319. 318. 327. 327. 323. 331. 336. 34 19525 P 259. 261. 271. 272. 274. 277. 278. 281. 281. 285. 28 19526 P 278. 280. 288. 267. 291. 291. 294. 301. 303. 306. 30 19527 P 281. 293. 294. 296. 300. 302. 310. 310. 303. 310. 34 19528 P 258. 27S. 285. 290. 300. 300. 300, 308. 315. 317. 31 19529 P 2S8. 265. 273. 268. 280. 282. 282. 284. 284. 289. 29 19530 P 271. 285. 293. 287. 296. 297. 312. 307. 309. 313. 31 19531 P 270. 270. 281. 280. 290. 292. 294. 296. 296. 298. 30 19532 P 281. 293. 298. 297. 296. 299. 306. 303. 309. 316. 31 -------------------------------------------------------------------------------------------------------------- DAY 13 14 is 16 17 is 19 20 21 GR) --------------------------------------------------------------------------------------------------------------- 19517 NP 279. 273. 281. 19518 P l9sl9 p 19520 19521 P 19522 P 354. 333. MORIBUND 336. 339. 3S7. 359. 370. 385. 392. 338. 339. 361. 379. 397. SACRIFICED ON DAY 0 OF PRESUMED GESTATION; 335. 339. 353. 370. 389. 346. 355. 412. 414. PREGNANCY 402. 424. 429. STATUS 413. 448. 433. COULD NOT BE DETERMINED 429. 19523 P 19524 P 19525 P 339. 351. 292. 346. 361. 297. 32S. 367. 303. 353. 373. 368. 393. 378. 411. 390. 430. 405. 450. 411. 470. 19526 P 318. 322. 324. 19527 P 328. 335. 342. 19528 P 19529 P 19530 P 330. 309. 333. 337. 308. 335. 329. 320. 314. 318. 327. 329. 344. 343. 360. 355. 380. 370. 400. 378. 420. 19531 P 328. 332. 333. 19532 P 332. 332. 347. --------------------------------------------------------------------------------------------------------------- P - PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY m DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PACE 3): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP 111 0.4 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 --------------------------------------------------------------------------------------------------------------- 19533 P 19534 P 19535 P 19536 P 19537 P 19538 P 19539 P 19540 P 19541 19542 P 19543 P 19544 P 19545 NP 267. 323. 253. 274. 2S7. 260. 262. 285. 234. 284. 304. 290. MATING 270. 273. 273. 325. 333. 329. 244. 260. 266. 275. 282. 265. 257. 266. 265. 267. 268. 274. 261. 271. 277. 291, 301. 302. MORIBUND SACRIFICED ON DAY 292. 294. 294. 316. 318. 319. 304. 314. 310. NOT CONFIRMED 275. 333. 267. 286. 268. 272. 280. 310. 0 OF 297. 323. 308. 274. 336. 270. 289. 269. 278. 284. 310. PRESUMED 302. 328. 306. 278. 342. 274. 289. 272. 279. 286. 315. GESTATION; 306. 327. 313. 276. 336. 268. 290. 257. 269. 279. 303. PREGNANCY 304. 331. 315. 281. 342. 278. 293. 264. 279. 292. 308. STATUS 303. 331. 327. 278. 345. 281. 295. 266. 280. 298. 319. COULD NOT 304. 333. 313. 282 341 281 296 267 285 296 320 BE DE 310 338 320 19546 P 296. 286. 300. 307. 310. 312. 317. 304. 298. 316. 326 19547 P 272. 285. 288. 292. 292. 296. 290. 289. 315. 294. 296 19548 NP 270. 276. 279. 280. 284. 283. 283. 267. 284. 288. 288 --------------------------------------------------------------------------------------------------------------- DAY 13 14 is 16 17 10 19 20 21 GPAI ---------------------------------------------------------------------------------------------------------------- 19533 P 19534 P 19535 P 294. 363. 303. 298. 367. 305. 298. 334. 312. 306. 378. 323. 380. 338. 400. 352. 405. 371. 438. 387. 445. 19536 P 19537 P 315. 281. 320. 284. 317. 285. 338. 353. 371. 397. 418. 429. 19538 P 301. 302. 291. 19539 P 310. 321. 319. 19540 P 331. 340. 335. 19541 19542 P MORIBUND 343. SACRIFICED ON DAY 0 OF PRESUMED 334. 339. GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19543 P 19544 P 353. 329. 361. 333. 364. 34S. 375. 396. 419. 430. 447, 458. 19545 NP MATING NOT CONFIRMED 19546 P 19547 P 334. 314. 339. 323. 333. 333. 350. 353. 371. 375. 377. 396. 19548 NP 296. 291. 286. 284. 284. 284. 285. 291. 294. ---------------------------------------------------------------------------------------------------------------- P - PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY - DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: ORAL ICAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PAGE 4): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA -------------------- ----------------------------------------------------------------------------------------- RAT # DOSAGE GROUP IV 1.6 MG/KG/DAY ------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 1 -------------------------------------------------------------------------------------------------------------- 19549 NP MATING NOT CONFIRMED 19550 P 19551 NP 276. 276. 279. 279. 278. 284. 290. 266. 281. 291. 29 MATING NOT CONFIRMED 19552 NP MATING NOT CONFIRMED 19553 P 19554 P 19555 NP 19556 NP 19557 245. 24S. 252. 2S4. 256. 253. 255. 256. 259. 261. 26 282. 275. 282. 278. 280. 282. 286. 282. 286. 288. 29 292. 293. 300. 299. 301. 300. 295. 296. 287. 293. 28 267. 268. 261. 264. 266. 264. 258. 262. 263. 261. 25 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19558 P 286. 287. 302. 30S. 303. 308. 299. 297. 309. 302. 30 19559 NP 260. 260. 264. 267. 265. 269. 268. 268, 271. 273. 27 19560 P 274. 278. 283. 286. 284. 291. 288. 286. 285. 291. 29 19561a 254. 260. 262. 259. 264. 265. 262. 268. 262. 266. 26 19562 P 272. 277. 282. 283. 283. 288. 292. 285. 287. 294. 30 19563 P 256. 263. 273. 267. 270. 266. 267. 266. 265. 264. 26 19564 P 258. 257. 264. 266. 268. 268. 273. 271. 274. 276. 27 -------------------------------------------------------------------------------------------------------------- DAY 13 14 15 16 17 18 19 20 21 GR, -------------------------------------------------------------------------------------------------------------- 19549 NP MATING NOT CONFIRMED 19550 P 19551 NP 311. MATING 314. 322. NOT CONFIRMED 19552 NP MATING NOT CONFIRMED 19553 P 286. 287. 292. 19554 P 307. 318. 323. 19555 NP 298. 296. 299. 19556 NP 19557 268. MORIBUND 263. 261. 268. 270. 265. 272. SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 276. 270. 19558 p 324. 337. 332. 19559 NP 19560 P 19561a 267. 309. 260. 259. 308. 262. 250. 319. 263. 252. 328. 2S4. 349. 252. 363. 2S8. 377. 258. 389. 257. 410. 19562 P 19563 P 316. 278. 326. 285. 328. 295. 350. 366. 392. 412. 438. 443. 19564 P 287. 292. 295. -------------------------------------------------------------------------------------------------------------- P = PREGNANT NP - NOT PREGNANT DAY - DAY OF PRESUMED GESTATION (VALUES EXCBUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (C). a. Necropsy and Caesarean-section observations were not recorded for rat 19561. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 15 (PAGE 5): MATERNAL BODY WEIGHTS - PRESUMED GESTATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- PAT N DOSAGE GROUP V 3.2 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAY 0 1 2 3 4 5 6 7 8 9 10 --------------------------------------------------------------------------------------------------------------- 19565 P 19566 P 19567 P 19568 P 19569 P 19570 P 19571 P 19572 P 19573 P 19574 P 19575 P 19576 NP 234. 224. 227. 228. 229. 230. 234. 219. 234, 240. 2SO 244. 252. 249. 251. 246. 244. 244. 245. 241. 249. 266 228. 226. 228. 230. 231. 234. 237. 236. 24S. 250. 2S4 2S9. 263. 264. 266. 266. 270. 270. 264. 270. 275. 276 249. 247. 248. 247. 244. 252. 252. 248. 245. 249. 262 257. 2S7. 258. 260. 253. 266. 265. 260. 265. 272. 275 245. 247. 249. 253. 2S6. 255. 259. 245. 245. 259. 268 2SO. 255. 252. 25S. 253. 257. 259. 259. 262. 261. 264 229. 225. 223. 227. 238. 244. 247. 250. 243. 248. 256 252. 253. 256. 261. 261. 261. 266. 245. 254. 262. 275 243. 239. 239. 228. 239. 242. 250. 256. 2SO. 252. 269 MATING NOT CONFIRMED 19577 P 19576 NP 230. 229. 230. 227. 219. 208. 208, 219. 217. 221. 230 MATING NOT CONFIRMED 19579 P 219. 216. 213. 211. 213. 227. 223. 217. 222. 229. 238 19580 p 237. 23S. 242. 235. 240. 244. 244. 236. 240. 245. 251 --------------------------------------------------------------------------------------------------------------- DAY 13 14 15 16 17 18 19 20 21 GRA --------------------------------------------------------------------------------------------------------------- 19565 P 19566 P 19567 P 19568 P 19569 p 19570 P 19571 P 19S72 P 19573 P 19574 P 19575 P 19576 NP 19577 P 19578 NP 259. 278, 269. 288. 264. 294. 288. 282. 268. 285. 294. MATING 247. MATING 264. 261. 286. 285. 278. 266. 291. 290. 266. 278. 302. 291. 293. 297. 290. 276. 273. 275. 288. 291. 292. 299. NOT CONFIRMED 245. 248. NOT CONFIRMED 296. 287. 304. 300. 2SB. 312. 306. 332. 317. 264. 315. 319. 346. 330. 274. 320. 334. 357. 349. 280. 341. 346. 369. 360. 286. 351. 349. 385. 376. 295. 19579 P 19seo p 240. 266. 248. 270. 250. 275. 2S8. 269. 208. 299. 304. 320. ---------------------------------------------------------------------------------------------------------------- P - PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAY = DAY OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 416-013- ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 1): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAYS 1- 8 8- 15 15- 22 22- 29 29- 36 36- 41 41- 42a --------------------------------------------------------------------------------------------------------------- 19501 19502 19503 19504 19505 19506 131. 114. 129. b 124. 92. 18. 131. 123. 128. 119. 107. 81. 19. 138. 145. 150. 128. 141. 100. 23. 144. 147. 152. 140. 141. 9s. 20. 144. b 140. b 143. 90. 22. 147. b 147. 133. 122. b 24. 19507 141. b b 146. 148. 97. 19. 19508 124. 131. 144. 129. 130. b 21. 19509 123. 128. 127. 121. 120. b 23. 19510 161. b b b 132. b 18. 19511 149. b 150. 150. 140. 99. 22, 19512 126. 124. 131. 133. 120. 93. 16. 19513 b 149. 167. b 158. 114. b 19514 156. 144. 140. 141. 131. 92. b 19515 120, 120. 138. 107. 106. 88. c 19516 130. 122. 123. 120. 135. 87. 14. -------------------------------------------------------------------------------------------------------------- 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. Value was incorrectly recorded and was excluded from group averages and statistical analyses. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 2): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP 11 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAYS I- a 8- 15 15- 22 22- 29 29- 36 36- 41 41- 42a -------------------------------------------------------------------------------------------------------------- 19517 127. 116. 119. 126. 109. 87. 11. 19518 143. 139. 137. 135. 129. 98. 21. 19519 128. 128. 130. 128. 125. 83. 16. 19520 122. 115. 125. 132. 132. 87. 15. 19521 148. 143. 149. 152. 143. 115. 16. 19522 19523 138. 126. 125. b 124. 87. 19. 120. b ISO. 119. 120. 83. 18. 19524 140. ISO. 149. 153. 127. 99. is. 19525 121. 119. 123. 108. 114. 73. 17. 19526 140. 148. 146. 148. 151. 97. la. 19527 145. 138. 144. 124. 121. 82. 23. 19528 137. 124. 128. 126. ill. 84. 16. 19529 146. 131. 127. 117. 112. so. 20. 19530 129. 135. 145. 140. 128. 94. 19. 19531 154. 142. IS3. 144. 143. 102. 20. 19532 133. 121. 133. 136. 135. 85. 14. -------------------------------------------------------------------------------------------------------------- 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. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER! T-6295.12) TABLE 16 (PAGE 3): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP 111 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAYS I- a 8- 15 15- 22 22- 29 29- 36 36- 41 41- 42a -------------------------------------------------------------------------------------------------------------- 19533 125. 108. 122. 113. 126. 88. 17. 19534 151. 131. 152. 141. 129. 94. 23. 19535 115. 103. 122. 117. lie. 84, is. 19536 124. 123. 125. 122. 121. 79. 20. 19537 130, 118. 118. 109. 122. 80. 20. 19538 129. 115. 118. 117. 115. 87. 17. 19539 117. 105. 114. 110. 115. 79. 15. 19540 122. 122. 132. 133. 133. 87. is. 19541 112. 112. 117. 112. 127. 85. b 19542 144. 135. 132. c 123. 87. c 19543 149. 138. c ISO. 126. 101. c 19544 138. 135. 140. 133. 130. 97. 24. 19545 130. 123. 128. c 124. 94. 19. 19546 146. c 168. 162. 158. c 21. 19547 127. 124. 126. 134. 139. 96. 21. 19548 146. 131. 134. 123. 118. 78. 16. -------------------------------------------------------------------------------------------------------------- DAYS = DAYS OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Last value recorded before cohabitation. b. Value was incorrectly recorded and was excluded from group averages and statistical analyses. c. Spilled feed precluded the calculation of this value. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PPOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 4): FEED CONSUMPTION VALUES - PRECOHABITATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- DAYS 1- a 8- 15 IS- 22 22- 29 29- 36 36- 41 41- 42a --------------------------------------------------------------------------------------------------------------- 19549 195so 19551 19SS2 19553 19554 19555 127. 121. b 118. 124. 92. 18. 146. b 123. 107. 108. 75. is. 141. 142. 137. 126. 125. 74. 16. 121. 103. 108. 105. 104. 82. 17. 131. 145. 116. b 122. b 17. 141. 147. b b 139. 96. 18. b 123. 141. 138. 122. 89. 13. 19556 19557 19558 19SS9 19560 19561 19562 19563 127. 130. 135. 125. ill. 77. 15. 124. 124. 124. ill. 108. 72. c 144. 143. 140. 140. 140. 98. b 137. 122. b b 115. 92. b 132. 122. 115. 118. 121. 85. 12. 109. 118, 119. 116. Ill. 87. 10. 140. 119. 129. 123. 120. 84. 16. 143. 138. 137. 134. 142. 105. 21. 19564 120. ill. 109. 105. 105. 77. 9. --------------------------------------------------------------------------------------------------------------- DAYS - DAYS OF STUDY ALL WEIGHTS WERE RECORDED IN GRAMS (C). a. Last value recorded before cohabitation. b. Spilled feed precluded the calculation of this value. c. Value was incorrectly recorded and was excluded from group averages and statistical analyses. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 5): FEED CONSUMPTION VALUES - PRECOMABITATION - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP V 3.2 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- DAYS I- a 8- 15 15- 22 22- 29 29- 36 36- 41 41- 42a -------------------------------------------------------------------------------------------------------------- 19565 118. 109. 102. 104. 100. 68. 11. 19566 143. 119. 124. 106. 103. 78. 17. 19567 125. 101. 105. 101. 97. 72. 14. 19568 132. 123. 125. 117. 114. 81. Is. l9s69 19570 141. 117. 114. 100. 113. 67. 10, 139. 128. 144. 120. lie. 84. 17. 19571 140. 122. 115. 112. 95. 73. is. 19572 117. 118. Ill. 112. 93. 73. 19. 19573 132. 113. 110. 98. 117. 73. b 19574 157. b b 117. 113. b b 19575 144. 125. 114. 115. 97. 77. c 19576 141. 121. 116. 122. 126. 82. 9. 19577 126. 128. 103. 76. 90. 68. 10. 19578 13S. 123. ill. 116. 103. 77. is. 19579 128. 90. b 91. b 80. 13. 19580 127. 103. ill. 112. 106. 74. 13. -------------------------------------------------------------------------------------------------------------- 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. Value was incorrectly recorded and was excluded from group averages and statistical analyses. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 1): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- PAT # DOSAGE GROUP 1 0 IVEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 --------------------------------------------------------------------------------------------------------------- 19501 p 17. 20. 22. 22. 24. 23. 22. 24. 22. 23. 2 19502 NP 21. 22. 22. 23. 22. 23. 23. 21. 19. 24. 2 19503 P NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 P 23. 24. 22. 22. 27. 21. 24. 25. 24. 24. 2 19505 P 22. 21. 24. 23. 24. 25. 22. 22. 23. 26. 2 19506 p 24. 29. 28. 23. 25. 26. 23. 23. 27. 26. 1 19507 P 27. 32. 27. 21. 28. 25. 20. 17. 25. 24. 2 19508 P 21. 23. 26. 24. 23. 24. 24. 21. 25. 25. 2 19509 p 20. 22. 23. 23. 24. 27. 22. 21. 25. 25. 2 19510 P 16. 25. 23. 26. 45. 30. 27. 23, 27. 29. 2 19511 p 18. 23. 23. 24. 25. 23. 26. 22. 22. 22. 2 19512 P 24. 21. 23. 24. 21. 24. 19. 21. 21. 22. 1 19513 P 22, 21. 25. 29. 27. 28. 29. 24. 25. 28. 2 19514 NP 21. 13. 21. 24. 24. 26. 27. 24. 28. 28. 2 19515 p 22. 21. 22. 22. 23. 23. 24. 22. 24. 23. 2 19516 P 20. 26. 22. 24. 24. 24. 17, 24. 20. 23. 2 --------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 --------------------------------------------------------------------------------------------------------------- 19501 P 23. Is. 19502 NP 19. 16. 19503 P NO CONFIRMED KATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 P 25. 20. 25. 28. 27. 24. 33. 26. 19505 p 26. 23. 23. 29. 28. 28. 27. 24. 19506 p 21. 25. 22. 25. 28. 25. 22. 23. 19507 P 21. 17. 22. 27. 28. 27. 16. 23. 19508 p 23. 25. 19509 P 20. 25. 18. 20. 2S. 23. 24. 11. 19510 P 25. 18. 19511 P 21. 17. 19512 P 20. 17. 28. 24. 24. 23. 25. 17. 19513 P 26. 23. 19514 NP 26. 22. 19sis p 24. 20. 19516 P 27. 23. 24. 31. 26. 30. 26. 25. --------------------------------------------------------------------------------------------------------------- P - PREGNANT NP - NOT PREGNANT (VALUES DAYS - DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). EXCLUDED FROM AVERAGES) PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 2): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP 11 0.1 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 a - 9 9 - 10 10 --------------------------------------------------------------------------------------------------------------- 19517 NP 21. 12. 16. 19. 18. 15. is. 16. 16. 14. 1 19518 p 26. 24. 26. 23. 24. 28. 25. 2S. 25. 23. 2 l9sl9 p 19. 23. 15. 23. 21. 24. 27. 20. 25. 24. 2 19520 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINE 19521 P 2S. 23. 25. 19. 21. 22. 15. 24. 24. 25. 2 29522 P 17. 23. 20. 22. 23. 23. 24. 19. 25. 22. 2 19523 P 22. 21. 25. 23. 24. 24. -21. 22. 25. 25. 2 19S24 P 23. 24. 29. 27. 29. 28. 25. 26. 27. 32. 2 19525 P 16. 21. 19. 21, 19. 19. 22. 20. 19. Is. 2 19526 P 20. 2S. 23. 26. 23. 23. 28. 22. 24. 25. 2 19527 P 20. 19. 20. 21. Is. 24. 26. 18. 22. 19. 1 19528 P 25. 24. 20. 28. 26. 26. 30. 26. 29. 24. 3 19529 P 21. 23. 20. 26. 22. 21. 24. 18. 24. 21. 2 19530 P 22. 27. 23. 29. 25. 30. 24. 26, 28. 27. 2 19531 P 16. 21. 24. 28. 24. 24. 25. 22. 28. 26. 2 19532 P 20. 23. 20. 23. 23. 25. 22. 23. 27. 24. 2 --------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 --------------------------------------------------------------------------------------------------------------- 19517 NP 19518 p 19519 P 19S20 19521 P 19522 P 19523 P 19524 P 19525 P ii. 22. 21. MORIBUND 20. 21. 26. 24. 22. a a 27. 25. 32. 22. 26. 28. 25. SACRIFICED ON DAY 0 OF PRESUMED 17. 28. 29. 25. 22. Is. 21. 23. 26. 22. 27. 26. 28. 18. 28. 29. GESTATION; 26. 28. 30. 28. 22. PREGNANCY 26. 24. 28. 28. 12. STATUS 18. COULD NOT BE DETERMINE 20. 26. 19526 P 19527 P 19528 P 19529 P 19530 P 19531 P 25. 18. 19. a 21. 19. 4. 16. 16. 28. 26. 23. 20. 21. 27. 26. 25. 31. 24. 23. 23. 15. 22. 17. 19532 P 24. 21. --------------------------------------------------------------------------------------------------------------- P - PREGNANT DAYS - DAYS ALL WEIGHTS a. Spilled NP = NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES)OF PRESUMED GESTATION WERE RECORDED IN GRAMS (G). feed precluded the calculation of this value. PROTOCOL 418-013; ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 3): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RAT N DOSAGE GROUP 111 0.4 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 --------------------------------------------------------------------------------------------------------------- 19533 P 19. 17. 17. 20. 19. 19. 19. 19. 18. is. 2 19534 P 24. 27. 23. 24. 22. 26. a 21. 23. 25. 2 19S35 P 8. 17. 17. 21. 18. 20. a 20. 21. 22. 2 19536 P 16. 21. 21. 21. 21. 17. 21. 22. 19. 17. 2 19537 P 16. 16. 19. 21. 22. 20. a 18. 21. 20. 1 19538 P 19. 18. 19. 18. 17. 22. 16. is. 20. 19. 1 19539 P Is. 17. 20. 22. 20. 22. a 25. 22. 20. 2 19540 P 17. 24. 20. 24. 23. 25. a 21. 27. 24. 2 19541 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINE 19542 P 25. 20. 22. 21. 24. 24. 20. 21. 20. 23. 2 19543 P 27. 23. 25. 26, 23. 26. 24. 24. 25. 26. 2 19544 P 24. 28. 19. 25. 24. 22. a 24. 24. 30. 2 19545 NP MATING NOT CONFIRMED 19546 P 12. 22. 24. 27. 26. 30. a 25. 29. 26. 2 19547 P 21. 23. 23. 25. 21. 23. a 21. 22. 23. 2 19548 NP 16. 21. 20. 21. 20. 11. a 21. 23. 22. 2 --------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - 15 15 - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 --------------------------------------------------------------------------------------------------------------- 19533 P 19, 16. 22. 25. 25. 23. 22. 19. 19534 P 26. 11. 24. 17. 17. 25. 30. 23. 19535 P 22. 19. 19536 P 21. 10. 21. 20. 25. 26. 28. 24. 19537 P 20. 17. 19538 P 17. 9. 19539 P 22. 18. 19540 P 22. 14. 19541 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINE 19542 P 22. b 19543 P 23. 19. 39. 27. 28. 23. 28. 20. 19544 P 24. 21. 19545 NP MATING NOT CONFIRMED 19546 P 26. 20. 31. 21. 31. 34. 27. 21. 19547 P 22. 18. 19548 NP 20. 19. le. is. 20. Is. 20. 19. --------------------------------------------------------------------------------------------------------------- P . PREGNANT NP - NOT PREGNANT IVALUES EXCLUDED FROM AVERAGES) DAYS - DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value not recorded. b. Spilled feed precluded the calculation of this value. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PAGE 4): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA ---------------------------------------------------------------------------------------------------------------- PAT N DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 a - 9 9 - 10 10 ---------------------------------------------------------------------------------------------------------------- 19549 NP MATING NOT CONFIRMED 19550 P 19551 NP 14. 16. 16. 16. 18. 20. a 17. 23. 22. 23 MATING NOT CONFIRMED 19552 NP MATING NOT CONFIRMED 19553 P 15. 16. 20. 21. 18. 17. a 19. 22. 21. 23 19554 P b 17. 15. 19. 23. 22. a 23. 22. 21. 22 19555 NP 19. 20. is. 24. is. 16. a 17. 19. 15. 20 19556 NP 19. 11. 18. 16. 15. 11. 17. 18. Ia. 15. 1 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19558 P 14. 22. 22. 22. 20. 18. a 23. 25. 24. 2 19559 NP 15. 18. 18. 19. 20. 21. 21. 20. 20. 21. 2 19560 P 24. 23. 21. 22. 23. 18. 16. 21. 17. 22. 2 19561c Is. 20. le. 17. 20. 18. a 19. 20. 20. 2 19562 P is. 21. 19. 20. 21. 21. 17. 20. 23. 23. 2 19563 P 19. 23. 21. 21. 19. Is. a 18. 22. 22. 22 19564 P 13. 18. 16. 20. is. 20. a 17. 20. 20. 2 ---------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - IS IS - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 ---------------------------------------------------------------------------------------------------------------- 19549 NP MATING NOT CONFIRMED 19550 P 20. 16. 19551 NP MATING NOT CONFIRMED 19552 NP MATING NOT CONFIRMED 19553 P 20. 18. 19554 P 21. 18. 19555 NP 17. 17. 19556 NP 14. b 21. 20. 14. 19. 18. 15. 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19558 P 23. 15. 19559 NP 8. b 13. 14. 14. 17. 20. 17. 19560 p 17. 24. 28. 26. 27. 27. 29. 22. 19561c 18. 13. 19562 P 25. 21. 22. 33. 25. 28. 26. 21. 19563 P 22. 18. 19564 P 19. 15. ---------------------------------------------------------------------------------------------------------------- P - PREGNANT NP - NOT PREGNANT (VALUES EXCLUDED FROM AVERAGES) DAYS - DAYS OF PRESUMED GESTATION ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. value not recorded. b. Spilled feed precluded the calculation of this value. c. Necropsy and Caesarean-section observations were not recorded for rat 19561. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 17 (PACE 5): MATERNAL FEED CONSUMPTION VALUES - PRESUMED GESTATION - INDIVIDUAL DATA ---------------------------------------------------------------------------------------------------------------- RAT # DOSAGE GROUP V 3.2 MG/KG/DAY ---------------------------------------------------------------------------------------------------------------- PREGNANCY STATUS DAYS 0 - 1 1 - 2 2 - 3 3 - 4 4 - 5 5 - 6 6 - 7 7 - 8 8 - 9 9 - 10 10 ---------------------------------------------------------------------------------------------------------------- 19S65 P 6. 9. 12. 10. 15. 14. a 26. 21. 19. 21 19566 P 19. 17. 17. 13. 10. 13. 14. 14. 20. 22. 21 19567 P 10. is. 17. is. is. 21. 20. 19. 22. 22. 21 19568 P 13. 16. 17. 17. 17. 17. a 19. 20. 19. 21 19569 P 16. 14. is. 13. 19. 17. 15. 10. 24. 18. 21 19570 P 16. 15. 15. 14. 22. 22. 23. 14. 21. 19. 17 19571 P Is. 15. 19. 22. 19. le. 11. 16. 21. 22. 21 19572 P 19. 18. 18. 13. is. 17. 33. 14. 20. 18. 19 19573 P 4. a. b 19. 22. 20. a is. 20. 19. 21 19574 P 15. Is. 21. 23. 18. 21. a is. 20. 23. 24 19575 P 7. 12. b b 19. 20. a 13. 20. 23. 22 29576 NP MATING NOT CONFIRMED 19577 P 13. 14. 11. 6. 2. 6. a b 18. 20. 23 19S78 NP MATING NOT CONFIRMED 19579 P 9. 13. 6. 30. 27. 16. 10. 17. 16. 23. 19 19580 p 9. 16. 15. 13. 18. 15. a 15. 17. 17. 21 ---------------------------------------------------------------------------------------------------------------- DAYS 13 - 14 14 - IS IS - 16 16 - 17 17 - 18 18 - 19 19 - 20 20 - 21 ---------------------------------------------------------------------------------------------------------------- 19565 P 19566 p 19567 P 19568 P 19569 P 19570 P is. 14. 20. b 22. b 21. 17. 20. 23. 25. 26. 24. 23. 14. 17. 24. 22. 24. 17. 21. 11. 21. b 19571 P 19S72 P 21. 19. 2S. 27. 2S. 29. 24. 23. Is. 14. 25. 25. 31. 26. 23. 30. 19573 P 19574 P 22. 16. 20. 19. 19575 P 25. 20. 19S76 NP MATING NOT CONFIRMED 19577 P 18. 20. 23. 17. 22. 24. 23. 18. 19578 NP MATING NOT CONFIRMED 19579 P 17. 15. 19. 23. 24. Is. 15. 19. 19580 P is. is. ---------------------------------------------------------------------------------------------------------------- P - PREGNANT NP = NOT PREGNANT (VALUES DAYS = DAYS OF PRESUMED GESTATION EXCLUDED FROM AVERAGES) ALL WEIGHTS WERE RECORDED IN GRAMS (G). a. Value not recorded. b. Spilled feed precluded the calculation of this value. PROTOCOL 418 013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 16 (PAGE 1): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTION ON DAY 15 OF PRESUMED GESTATION -------------------------------------------------------------------------------------------------------------- VIABLE EMBRYOS ---------------- NONVIABLE EMBRYOS ------------------- IMPLANTATION SITES -------------------- CORPORA LUTEA --------------- RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 119501 5 9 14 1 0 1 6 9 is 9 10 19 19502 NOT PREGNANT 19508 9 7 16 0 0 0 9 7 16 9 7 16 19510 5 6 11 1 1 2 6 7 13 6 7 13 19511 a 11 19 0 0 0 8 11 19 11 12 23 19513 10 6 16 0 0 0 10 6 16 10 6 16 19514 NOT PREGNANT 19515 7 6 13 2 1 3 9 7 16 10 7 17 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19517 NOT PREGNANT 19522 8 7 15 0 0 0 8 7 15 9 a 17 1952S 4 2 6 a 1 1 4- 3 7 7 9 16 19526 5 5 10 0 0 0 5 5 10 10 9 19 19527 8 9 17 0 1 1 8 10 18 10 10 20 19530 4 7 11 4 0 4 8 7 is 9 9 is 19S31 8 7 is 1 0 1 9 7 16 9 11 20 19532 8 4 12 1 0 1 9 4 13 10 6 16 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP III 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19535 4 3 7 1 1 2 5 4 9 8 7 15 19537 0 3 3 1 2 3 1 5 6 4 a 12 19538 8 4 12 0 0 0 8 4 12 9 6 is 19539 7 6 13 2 0 2 9 6 15 9 9 18 19540 9 7 16 0 0 0 9 7 16 11 7 18 19542 8 8 16 0 0 0 8 a 16 9 10 19 19544 6 5 11 2 1 3 a 6 14 12 10 22 19547 s 11 16 0 0 0 5 11 16 6 12 18 -------------------------------------------------------------------------------------------------------------- PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN PATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE IS (PACE 2): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTION ON DAY 15 OF PRESUMED GESTATION --------------------------------------------------------------------------------------------------------------- VIABLE EMBRYOS ---------------- NONVIABLE EMBRYOS ------------------- IMPLANTATION SITES -------------------- CORPORA LUTEA --------------- RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RAT N HORN TOTAL HORN TOTAL HORN TOTAL OVARY TOTAL --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19550 9 5 14 0 0 0 9 5 14 9 10 19 19553 6 7 13 1 0 1 7 7 14 10 10 20 19554 9 s 14 0 1 1 9 6 15 10 7 17 19555 NOT PREGNANT 19558 7 6 13 3 0 3 10 6 16 10 8 is 19562 NECROPSY AND CAESAREAN-SECTION OBSERVATIONS WERE NOT RECORDED 19563 5 7 12 1 1 2 6 8 14 8 11 19 19564 7 4 11 0 1 1 7 5 12 13 13 26 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19565 2 4 6 0 0 0 2 4 6 6 7 13 19566 7 5 12 1 0 1 a 5 13 9 a 17 19567 8 5 13 0 0 0 8 5 13 9 8 17 19570 6 9 15 1 1 2 7 10 17 7 10 17 19573 5 9 14 0 1 1 5 10 is 5 10 15 19574 7 6 13 0 0 0 7 6 13 10 10 20 19575 7 6 13 0 4 4 7 10 17 8 11 19 19580 7 8 15 0 0 0 7 8 is 11 10 21 --------------------------------------------------------------------------------------------------------------- PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 18 (PAGE 3): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION -------------------------------------------------------------------------------------------------------------- VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES ------------------------ ---------------- ---------------- ---------------- ---------------- SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT R RAT N M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19503 NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 8 4 5 7 12 0 0 0 2 3 5 0 0 0 7 10 17 1950S a 9 4 13 17 0 0 0 0 2 2 0 0 0 4 15 19 19506 4 9 5 8 13 0 0 0 1 0 1 0 0 0 6 8 14 19507 7 7 9 5 14 0 0 0 0 1 1 0 0 0 9 6 15 19509 5 7 6 6 12 0 0 0 1 0 1 0 0 0 7 6 13 19512 7 7 4 10 14 0 0 0 0 0 0 0 0 0 4 10 14 19516 5 9 6 a 14 0 0 0 0 0 0 0 0 0 6 8 14 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19518 6 5 6 5 11 0 0 0 2 0 2 0 0 0 a 5 13 19519 9 7 9 7 16 0 0 0 0 0 0 0 0 0 9 7 16 19520 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 7 a 9 6 15 0 0 0 0 0 0 0 0 0 9 6 is 19523 3 7 8 2 10 0 0 0 1 1 2 1 1 2 10 4 14 19524 12 4 8 a 16 0 0 0 0 1 1 0 0 0 8 9 17 19528 3 2 1 4 5 0 0 0 1 2 3 0 0 0 2 6 a 19529 6 8 7 7 14 0 0 0 1 0 1 0 0 0 8 7 is -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 111 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19533 8 4 4 8 12 0 0 0 0 0 0 0 0 0 4 0 12 19S34 8 6 7 7 14 0 0 0 0 0 0 0 0 0 7 7 14 19536 5 13 6 12 18 0 0 0 0 0 0 0 0 0 6 12 16 19541 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19543 11 6 10 7 17 0 0 0 1 1 2 0 0 0 11 8 19 19545 MATING NOT CONFIRMED 19546 4 1 4 1 5 0 0 0 1 3 4 0 0 0 5 4 9 19548 NOT PREGNANT -------------------------------------------------------------------------------------------------------------- M = MALE PLACENTAE F = FEMALE APPEARED NORMAL UNLESS NOTED OTHERWISE. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 18 (PAGE 4): CAESAREAN-SECTIONING OBSERVATIONS - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION --------------------------------------------------------------------------------------------------------------- VIABLE FETUSES DEAD FETUSES EARLY RESORPTIONS LATE RESORPTIONS IMPLANTATION SITES ------------------------ ---------------- ---------------- ---------------- ---------------- SEX RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT RIGHT LEFT Ri RAT N M F HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL HORN TOTAL --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19549 MATING NOT CONFIRMED 19SSI MATING NOT CONFIRMED 19552 MATING NOT CONFIRMED 19556 NOT PREGNANT 19SS7 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19559 NOT PREGNANT 19560 5 7 9 3 12 0 0 0 0 0 0 0 0 0 9 3 12 1 19562 10 10 8 12 20 0 0 0 0 2 2 0 0 0 8 14 22 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19568 6 0 6 0 6 0 0 0 0 3 3 0 0 0 6 3 9 19569 6 7 9 4 13 0 0 0 0 0 0 0 0 0 9 4 13 1 19571 DELIVERED ON DAY 21 OF GESTATION 19572 6 6 7 5 12 0 0 0 1 2 3 0 0 0 8 7 15 19576 MATING NOT CONFIRMED 19577 1 3 4 0 4 0 0 0 1 0 1 0 0 0 5 0 5 19578 MATING NOT CONFIRMED 19579 DELIVERED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- M = MALE PLACENTAE F = FEMALE APPEARED NORMAL UNLESS NOTED OTHERWISE. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 1): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION --------------------------------------------------------------------------------------------------------------- NUMBER OF LIVE ---- CONCEPTUSES ------ EMBRYOS NONVIABLE --------------------------------------------------------------------------------------------------------------- RAT N TOTAL N N t --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19501 14 15 1 6.7 19502 NOT PREGNANT 19508 16 16 0 0.0 19sio 11 13 2 15.4 19511 19 19 0 0.0 19513 16 16 0 0.0 19514 NOT PREGNANT 19515 13 16 3 18.8 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP Il 0.1 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19Sl7 NOT PREGNANT 19522 15 is 0 0.0 19525 6 7 1 14.3 19526 10 10 0 0.0 19527 17 la 1 5.6 19530 11 15 4 26.7 19531 15 16 1 6.2 19532 12 13 1 7.7 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 111 0.4 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19535 7 9 2 22.2 19537 3 6 3 50.0 19538 12 12 0 0.0 19539 13 is 2 13.3 19540 16 16 0 0.0 19542 16 16 0 0.0 19544 11 14 3 21.4 19547 16 16 0 0.0 --------------------------------------------------------------------------------------------------------------- PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 2): LITTER OBSERVATIONS (CAESAREAN-DEBIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- PATS CAESAREAN-SECTIONED ON DAY 15 OF GESTATION --------------------------------------------------------------------------------------------------------------- NUMBER OF LIVE ---- CONCEPTUSES ------ EMBRYOS NONVIABLE --------------------------------------------------------------------------------------------------------------- RAT # TOTAL N N t --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19sso 14 14 0 0.0 19553 13 14 1 7.1 19554 14 is 1 6.7 19555 NOT PREGNANT 19558 13 16 3 18.8 19S61 NECROPSY AND CAESAREAN-SECTION OBSERVATIONS WERE NOT RECORDED 19563 12 14 2 14.3 19564 11 12 1 8.3 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19565 6 6 0 0.0 19566 12 13 1 7.7 19567 13 13 0 0.0 19570 15 17 2 11.8 19573 14 15 1 6.7 19574 13 13 0 0.0 19575 13 17 4 23.5 19580 15 is 0 0.0 -------------------------------------------------------------------------------------------------------------- PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 3): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- NUMBER OF LIVE AVERAGE FETAL ---- CONCEPTUSES ------ FETUSES BODY WEIGHT (G) RESORBED --------------------------------------------------------------------------------------------------------------- RAT 0 MALE FEMALE TOTAL MALE FEMALE TOTAL a N N t --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19503 NO CONFIRMED DATE OF MATING; DELIVERED ON DAY 66 OF STUDY 19504 8 4 12 4.75 4.44 4.64 17 5 29.4 19505 8 9 17 5.13 5.10 5.12 19 2 10.5 19SO6 4 9 13 5.66 5.48 5.54 14 1 7.1 19507 7 7 14 S.47 5.21 5.34 is 1 6.7 19509 5 7 12 5.34 5.14 5.22 13 1 7.7 19512 7 7 14 5.14 4.87 S.01 14 0 0.0 19516 s 9 14 5.17 4.87 4.98 14 0 0.0 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19518 6 s 11 4.80 4.52 4.67 13 2 15.4 19519 9 7 16 S.29 5.16 S.23 16 0 0.0 19520 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE 19521 7 8 is 5.12 4.79 4.94 is 0 0.0 19523 3 7 10 5.47 5.28 5.34 14 4 28.6 19524 12 4 16 5.51 5.11 5.41 17 1 5.9 19528 3 2 5 6.45 S.06 5.89 8 3 37.5 19529 6 8 14 5.87 5.54 5.68 is 1 6.7 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 111 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19533 8 4 12 5.30 4.89 5.16 12 0 0.0 19534 8 6 14 5.00 4.82 4.92 14 0 0.0 19S36 5 13 is 5.31 5.05 5.12 18 0 0.0 19541 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE 19543 11 6 17 5.18 4.72 5.02 19 2 10.5 19545 19546 MATING NOT CONFIRMED 4 1 5 5.18 4.70 5.09 9 4 44.4 19548 NOT PREGNANT -------------------------------------------------------------------------------------------------------------- a. TOTAL - SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES. PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 19 (PAGE 4): LITTER OBSERVATIONS (CAESAREAN-DELIVERED EMBRYOS OR FETUSES) - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- NUMBER OF LIVE AVERAGE FETAL ---- CONCEPTUSES ------ FETUSES BODY WEIGHT (G) RESORBED --------------------------------------------------------------------------------------------------------------- RAT N MALE FEMALE TOTAL MALE FEMALE TOTAL a N N I --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19S49 MATING NOT CONFIRMED 19551 MATING NOT CONFIRMED 19552 MATING NOT CONFIRMED 19556 NOT PREGNANT 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19559 NOT PREGNANT 19560 5 7 12 S.26 5.06 5.14 12 0 0.0 19562 10 10 20 4.97 4.84 4.91 22 2 9.1 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19568 6 0 6 4.50 ---- 4.50 9 3 33.3 19569 6 7 13 4.84 4.50 4.66 13 0 0.0 19571 DELIVERED ON DAY 21 OF GESTATION 19572 19576 6 6 12 4.78 4.57 4.68 15 MATING NOT CONFIRMED 3 20.0 19577 1 3 4 5.18 5.00 5.04 5 1 20.0 19578 MATING NOT CONFIRMED 19579 DELIVERED ON DAY 21 OF GESTATION --------------------------------------------------------------------------------------------------------------- a. TOTAL - SUM OF FETAL WEIGHTS/NUMBER OF LIVE FETUSES. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 1): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF PRESUMED GESTATION --------------------------------------------------------------------------------------------------------------- EMBRYO 1 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 le 19 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # CLs 19501 9/10 E A A A A A A A A A A A A A A 19502 NOT PREGNANT 19508 9/ 7 A A A A A A A A A A A A A A A A 19510 6/ 7 E A A A A A A A A A A E A 19511 11/12 A A A A A A A A/ A A A A A A A A A A A 19513 10/ 6 A A A A A A A A A A/ A A A A A A 19514 NOT PREGNANT 19515 10/ 7 E A A E A A A A A A A E A A A A --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19517 NOT PREGNANT 19522 9/ 8 A A A A A A A A A A A A A A A 19525 7/ 9 A A A A/ E A A 19526 10/ 9 A A A A A/ A A A A A 19527 10/10 A A A A A A A A A A E A A A A A A A 19S30 9/ 9 E A E A A E A E A A A A A A A 19531 9/11 A A A A A A A A E A A A A A A A 19532 10/ 6 A E A A A A A A A A A A A --------------------------------------------------------------------------------------------------------------- A - VIABLE EMBRYO E = NONVIABLE EMBRYO DENOTES POSITION OF CERVIX CLa - CORPORA LUTEA/OVARY PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PACE 2): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF PRESUMED GESTATION -------------------------------------------------------------------------------------------------------------- EMBRYO N 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 is 19 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 111 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- PAT # CLe 19535 S/ 7 A E A A A E A A A 19537 4/ 8 E/ E A A A s 19S38 9/ 6 A A A A A A A A A A A A 19539 9/ 9 A A A A E E A A A A A A A A A 19540 11/ 7 A A A A A A A A A A A A A A A A 19542 9/10 A A A A A A A A A A A A A A A A 19544 12/10 E A A A A A A E A A A A E A 19547 6/12 A A A A A A A A A A A A A A A A -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19550 9/10 A A A A A A A A A A A A A A 19S53 10/10 A A A A E A A/ A A A A A A A 19554 10/ 7 A A A A A A A A A A A A E A A 19555 NOT PREGNANT 19558 10/ 8 E E E A A A A A A A/ A A A A A A 19561a 19563 8/11 A E A A A A/ A A E A A A A A 19S64 13/13 A A A A A A A/ A E A A A -------------------------------------------------------------------------------------------------------------- A = VIABLE EMBRYO E - NONVIABLE EMBRYO DENOTES POSIT10N OF CERVIX CLs - CORPORA LUTEA/OVARY a. Necropsy and Caesarean-section observations were not recorded for rat 19561. PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 3): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 15 OF PRESUMED GESTATION --------------------------------------------------------------------------------------------------------------- EMBRYO # 1 2 3 4 5 6 7 8 9 10 11 22 13 14 15 16 17 18 19 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT 0 CLa 19565 6/ 7 A A/ A A A A 19566 9/ 8 E A A A A A A A A A A A A 19567 9/ 8 A A A A A A A A A A A A A 19570 7/10 A A A A A A E E A A A A A A A A A 19573 5/10 A A A A A/ A A E A A A A A A A 19574 10/10 A A A A A A A A A A A A A 19575 8/11 A A A A A A A A A E E A A E E A A 19580 11/10 A A A A A A A A A A A A A A A -------------------------------------------------------------------------------------------------------------- A - VIABLE EMBRYO E = NONVIABLE EMBRYO DENOTES POSITION OF CERVIX CLs - CORPORA LUTEA/OVARY PROTOCOL. 418-013: ORAI, (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABL.E 20 (PAGE 4): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA --------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION --------------------------------------------------------------------------------------------------------------- FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 is 19 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY --------------------------------------------------------------------------------------------------------------- RAT # CLa 19503 NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 19505 19506 10/13 4/16 7/ 8 FA 4.29 FA 5.04 E E FA 5.46 FA MA 4.79 MA 5.37 MA E FA 5.41 FA KA 4.76 t4A 5.47 MA MA 4.62 FA 4.69 MA MA / E 4.71 MA E 4.15 PA PA MA 5.10 FA 5.13 PA E MA 5.21 MA MA 4.01 MA 4.81 FA FA 4.84 FA 5.12 FA MA 5.21 MA S.31 FA E MA 5.36 FA FA 3.80 MA 5.38 MA 4.77 E FA 4.83 FA 4.78 FA FA 5.34 4.93 5.53 5.93 5.66 5.31 S.43 5.40 5.68 5.64 5.95 4.67 5.87 5.22 5.70 19507 13/12 MA FA MA MA MA MA PA FA FA E FA MA MA PA FA 5.48 5.42 5.34 5.53 5.47 5.64 5.06 5.06 5.28 5.32 5.32 5.49 5.18 5.13 19509 7/ 8 FA FA FA FA MA FA 'E MA MA FA FA MA MA 5.03 5.31 4.90 5.51 S.12 S.14 5.44 5.31 5.12 4.98 5.27 5.54 19512 4/11 MA FA FA FA MA FA MA MA MA MA PA FA FA MA 5.45 4.87 4.76 5.07 4.78 4.S4 4.82 5.29 5.21 4.98 5.30 5.00 4.58 5.47 19516 S/ 9 MA FA FA PA FA FA MA MA MA FA PA FA MA FA 5.34 4.85 4.87 5.03 5.46 4.92 4.84 5.38 5.06 3.84 5.13 4.91 5.23 4.85 --------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 11 0.1 MG/KG/DAY --------------------------------------------------------------------------------------------------------------- 19518 B/ 7 FA 4.21 E MA FA FA 4.70 4.70 4.52 E FA FA / MA MA MA MA MA 4.53 4.56 4.76 4.70 4.85 4.72 5.07 19519 19520 10/ 7 FA MA FA FA MA FA FA MA MA / FA MA MA MA FA MA MA 4.99 5.31 5.48 5.14 5.63 5.06 4.63 5.31 5.83 5.59 5.16 4.98 4.88 5.22 5.00 5.51 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 9/ 6 FA FA FA MA FA FA MA MA MA / FA FA MA FA MA MA 4.89 4.74 4.62 4.66 4.90 4.81 4.92 4.92 5.28 4.49 4.86 S.20 4.98 5.45 5.42 19523 10/ 4 E FA L FA FA FA FA FA MA MA / MA E FA L 4.80 5.52 5.42 S.23 5.57 4.81 5.58 5.32 5.52 5.62 19524 9/12 FA MA MA MA MA FA MA MA MA MA E MA MA MA FA MA FA S.16 S.37 5.51 5.64 5.54 S.11 5.70 S.55 5.00 5.49 5.73 S.32 5.56 5.16 5.72 S.00 19528 6/ 9 E MA / MA E E FA MA FA 6.86 6.59 4.83 5.89 S.30 19529 B/ 8 FA MA FA MA E PA FA MA FA MA FA MA PA MA FA 5.13 5.87 5.63 5.97 5.54 5.64 5.19 5.68 6.11 5.27 6.25 5.71 5.81 5.74 --------------------------------------------------------------------------------------------------------------- M - MALE F - FEMALE A - ALIVE E - EARLY RESORPTION L - LATE RESORPTION CLO - CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G). DENOTES POSITION OF CERVI 418-013:PAGE C-12 Protocol418-013 Page 12 Caesarea n-Sectionina Observations - Day 21 Presumed Gestation: All remaining female rats in each dosage group willbe Caesarean-sectioned on day 21 of presumed gestation.The gravid uterus willbe excised and weighed. Individual placentalweights willbe recorded. The fetuseswillbe removed from the uterus, weighed and placed inindividuaclontainers.The female rats(pregnant dams) willbe examined fornumber and distributioonf. Corpora Lutea. ImplantationSites. [Placentaethatappear abnormal (size,coloror shape) willbe noted inthe raw data]. Live and Dead Fetuses. (A livefetusisdefined as one thatresponds to stimulia; dead fetusisdefined as a term fetusthatdoes not respond to stimuliand thatisnot markedly autolyzed; dead fetusesdemonstrating marked to extreme autolysisare considered to be lateresorptions.) Earlyand Late Resorptions. (A conceptus isdefined as a lateresorptionifitisgrosslyevident that organogenesis has occurred;ifthisis not the case, the conceptus isdefined as an earlyresorption.) Fetal Observation-s: Caps and labeledtubes willbe weighed (combined weight,to the nearest .001 gram) before and afterretentionof pooled fetalsamples forsubsequent use in pharmacokinetic analyses. These weights willbe documented in the raw data,and copies of these weights willbe includedwith the packing listpriorto shipment. PlacentalSamr)les and Amniotic Fluid: Samples ofthe amnioticfluid(when possible)and the placentaof each fetuswillbe collectedi,ndividuallpyooled (perlitterf)r,ozen and stored(-700C or below) until shipment to the Sponsor foranalysis. 418-013: PAGE C-1 3 Protocol418-013 Page 13 Gmss ExternalAfterationSse,x, Body Weiohts and Identification: Fetuses willbe examined forsex and forgross externalalterations.Late resorptions and dead fetuseswillbe examined forgross externalalterationsto the extent possible. The individualbody weight of each fetuswillbe recorded. Only body weights of live fetuseswillbe used to determine littefretalbody weight averages. Representative photographs of fetalgross externalalterationswillbe taken. Blood Samples: Blood samples willbe collectedfrom each pup via decapitation,pooled (perlittera)nd transferredintoserum separatortubes. The samples willbe spun in a refrigerated centrifuge.The serum will-betransferredintopolypropylene tubes labeledwith the study number, animal identificatiodna,te of collections,tudy day and collection timepoint.Allsamples willbe immediately frozen on dry ice and maintained frozen (-70'C or below) untilshipment to the Sponsor foranalysis. LiverSamples: The liverofeach fetuswillbe collected,pooled (perlifterf)r,ozen and stored(-700C or below) untilshipment to the Sponsor foranalysis. Fetal Carcasses: The remaining carcass (and head) of each fetuswillbe collected,frozenand stored (-70*C or below) untilshipment to the Sponsor foranalysis. Shipi)ingInstructions: Allsamples willbe maintained frozen(-70'C or below) untilshipment foranalysis. Samples willbe shipped frozen on dry iceviaovernightmail. A packing liswtillbe includedwiththe 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: Rats willbe sacrificedby carbon dioxideasphyxiation. Live fetuses willbe sacrificedvia decapitation. 41 Ml 3:PAGE C-14 Protocol418-013 Page 14 NECROPSY: Gross lesions willbe retained in neutral buffered 10% formalin for possible future evaluation (a table of random unitswillbe used to selectone controlgroup ratfrom which alltissues examined at necropsy willbe retained,in order to provide control tissuesforany possible histopathologicalevaluations of gross lesions). Unless specificallcyited below, allother tissues willbe discarded. Scheduled Sacrifice: On eitherday 15 of presumed gestation or day 21 of presumed gestation,followingthe finalcollectionintervalfor urine and fecal samples and blood sample collection,female ratswillbe sacrificed,and a gross necropsy of the thoracic,abdominal and pelvic viscera willbe performed. The number and distributioonf implantationsiteswillbe recorded. A liversection (rightlaterallobe) and the milk-secretingglands from the axillaryt,horacic,abdominal and inguinalregions of each dam (leftside only)willbe collected,frozen and stored (-70*C or below) untilshipment to the Sponsor for analysis. Uteriof apparently nonpregnant ratswillbe stained with 10% ammonium confirm the absence of implantationsites(5). sulfideto Rats Found Dead or Moribund: Rats that die or are sacrificedbecause of moribund condition,abortion or premature deliverywillbe examined forthe cause of death or mohbund condition on the day the observation ismade. The ratswillbe examined forgross lesions. A liversection (right laterallobe) and the milk-secretingglands from the axillaryt,horacic,abdominal and inguinalregions of each dam (leftside only) willbe collected,frozen and stored (-70'C or below) untilshipment to the Sponsor foranalysis. Pregnancy statusand uterine contents of female ratswillbe recorded. Aborted fetuses and/or delivered pups willbe examined to the extent possible, using the same methods described forterm fetuses. Uteriof apparently nonpregnant rats willbe stained with 10% ammonium sulfideto confirm the absence of implantationsites('). Shit)t)ingInstructions: Allsamples willbe maintained frozen (-70'C or below) untilshipment foranalysis. Samples willbe shipped frozen on dry ice via overnight mail. A packing listwillbe included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipientand the Study Monitor willbe notifiedin advance of sample shipment. 418-013:PAGE C-15 Protocol418-013 Page 15 STATISTICAL EVALUATION: Averages and percentages willbe calculated. Littervalues willbe used where appropriate. Additionalprocedures and/or analyses may be performed, ifdeemed appropriate. DATA ACQUISITION, VERIFICATION AND STORAGE: Data willbe hand- and/or computer-recorded. Records willbe reviewed by the Study Directorand/or appropriate management 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 supplied to the Sponsor upon request. Preserved tissueswillbe stored at the Testing Facilityat no charge forone year aftermailing of the draftfinalreport,afterwhich time the Sponsor willbe contacted to determine the dispositionof these materials. RECORDS TO BE MAINTAINED: Protocol and Amendments. Test Article,Vehicle and/or Reagent Receipt, Preparation and Use. Animal Acquisition. Randomization Schedules. Mating History. Treatment (ifprescribed by 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. Caesarean-Sectioning and Fetal Observations. Gross Necmpsy Observations. Organ Weights. Photographs (ifrequired). Study Maintenance (room and environmental records). Feed, Water and Bedding Analyses. Packing and/or Shipment Lists. 4lMl3:PAGE -:@-16 Protocol418-013 Page 16 KEY PERSONNEL: Executive Director of Research: Mildred S. Christian, Ph.D., Fellow, ATS Director of 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 Member, InstitutionAanlimal Care and Use Committee: Dena C. Lebo, V.M.D. Directorof Operations and Compliance: Barbara J. Patterson,B.A. Consultant,VeterinaryPathology: W. Ray Brown, D.V.M., Ph.D., ACVP FINAL REPORT: A comprehensive draftfinalreportwillbe prepared on completion of the study and will be finalizedfollowingconsultationwith the Sponsor. The reportwillincludethe following: Summary and Conclusion. Experimental Design and Method. Evaluationof Test Results. Appendices: Figures,Summary and IndividualTables Summarizing the Above Data, Protocoland Associated Amendments and Deviations,Study Directoes GLP Compliance Statement, Reports of Supporting Data (ffappropriate)and QAU Statement. INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE STATEMENT: The procedures described inthisprotocolhave been reviewed by the Testing Facility's InstitutionAanlimal Care and Use Committee. Allprocedures described in thisprotocol that involvestudy animals willbe conducted in a manner to avoid or minimize discomfort,distressor pain to the animals. The Sponsor's signaturebelow documents the factthatinformationconcerning 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. 4,1,8-013:PAGE C-17 Protocol418-013 Page 17 REFERENCES: 1 Christian, M.S. and Voytek, P.E. (1982). In Vivo Reproductive and Mutagenicity Tests. Environmental Pmtection Agency, Washington, D.C. National Technical InformationService,U.S. Department of Commerce, SpringfieldV,A 22161. 2. Christian,M.S. (1984). Reproductive toxicitaynd teratologyevaluationsof naltrexone(Proceedings of Naltrexone Symposium, New York Academy of Sciences, November 7, 1983), J.Clin.Psychiat.45(9):7-10. 3. Lang, P.L. (1988). Embryo and Fetal Developmental Toxicity (Teratology) Control Data in the Charles River CD.*CDODBR Rat. Charles River Laboratories, Inc.,Wilmington, MA 01887-0630. (Data base provided by Argus Research Laboratories,Inc.) 4. Instituteof Laboratory Animal Resources (1996). Guide forthe Care and Use of Laboratory Animals. National Academy Press, Washington, D.C. 5. Salewski,E. (1964). F.rbemethode zum makroskopischen Nachweis von lmplantationsstelleanm Uterus der Ratte. Arch. Pathol.Exp. Pharmakol. 247:367. PROTOCOL APPROVA FOR THE TESTING FACILITY Alan M. Hoberman, Ph.D., DABT Directorof Research @7 @ alai@i@ ayr@%nd G. Yoc@oqrP0 @ LY Associate Dire Study Director D.,DABT esearch arbara J. Pattersu,,fI A. hairpersorillnst fnnaa-llAnimal Care and i FOR THE SPONSOR 7-,r., Marvin T. Case, D.V.M., Ph.D. Study Monitor 41 Ml 3:PAGE C-14 Protocol418-013 Page 18 9 Date 09 - &()LI-99 Date (3 Date /o Date 418-013:PAGE C-19 SCHEMATIC ATTACHMENTI OF STUDY DESIGN AND STUDY SCHEDULE ATTACHMENT 1 418-013:PAGE C-20 Protocol418-013 Page 1 of 2 STUDY SCHEMATIC PHARMACOKINETIC STUDR S@qOf Dosage Prematng Period (42Da@) Female Rats Cohabb@on Period (5dap@ Enodf Dosage Presumed Gestation Period Caesarean- Sedoning' Day21of Presumed Caesarean- Gestation Sedoninbg Day15of Day20of Presumed Presumed Gestaton Gestation Dosage Period. a. For additionadletailsee "Tests,Analyses and Measurements" sectionofthe protocol. b. Evaluationofcorporalutea,implantatiosnitesand viableand nonviableembryos. C. Fetalevaluations(alfletuses-externalexaminations). A7rACHMENT 1 41"l 3:PAGE C-21 Protocol418-013 Page 2 of 2 SCHEDULEs 10 NOV 98 16 NOV 98 -21 JAN 99 27 DEC 98 PM -01 JAN 99 AM 28 DEC 98 -01 JAN 99 12 JAN 99 -16 JAN 99 18 JAN 99 -22 JAN 99 26 MAY 99 Animals Arrive-AcclimationBegins. Dosage Period-Female Rats(42days priorto cohabitatiounntilday 14 or20 ofpresumed gestation). CohabitationPeriod. Day 0 of Presumed Gestation. Caesarean-SectioningPeriod(Day 15 of presumed gestation). Caesarean-SectioningPeriod(Day 21 of presumed gestation). DraftFinalReport. a. The studyinitiatidoante istheday the Study Directorsignsthe protocol. 4,1,8-013:PAGE C-22 ATTACHMENT 2 MATERIAL SAFETY DATA SHEET 418-013:PAGE C-23 IOATERIAL SAFM DATA SHM 3H 3H Cont*r St. Paul, Minnesota 551"-1004 I-BOD-364-3577 or (612) 737-6601 (24 hours) Copyright, 1M , Minnesota Mining and Manufacturing Company. AU rights reserved. Copying and/or downloading of this information for the purpose of properly utflivi g 3H products is allowed provided that: 1) th* information is oopied in full with no changes unless prior agreement is obtained from 3M, and 2) neither the copy nor the original is resold or otherwise distributed with the intention of earning a profit thereon. DIVISION: 3H CHEMICALS TWE MAKE: FC-95 FLUORAD Brand Fluorachanical Surfactant ID NUMBER/U.P.C.: 98-0207-0103-7 00-5113S-CROS4-1 93-0207-0104-S 98-0211-0988-5 00-SI135-09362-7 98-0211-3916-1 ZF-0002-10"-l ISSUED: January 29, 1998 SUPERSEDES: November OS, 1907 DOCUMENT: 10-3796-9 00-61135-09OSS-8 00-SI135-02311-2 ----------------------------------------------------------------------------- 1. INGREDIENT C.A.S. NO. PERCENT ----------------------------------------------------------------------------- POTASSIUM PERFLUORDALKYL SULFONATE ...... 279S-39-3 82 POTASSIUM PERFLUOROALKYL SULFONATE ...... 3871-99-6 3 POTASSIUM PERFLUORGALKYL SULFONATE ...... 29420-49.3 3 POTASSIUM PERFLUOROALKYL SULFONATE ...... 60270-55-S 2 POTASSIUM PERFLUOROALKYL SULFONATE ...... 3672-2S.1 I . 86 -a -7 .6 -3 ----------------------------------------------------------------------------2. PHYSICAL DATA ----------------------------------------------------------------------------- BOILING POINT: ...............N./.A VAPOR PRESSURE: ..............N./.A VAPOR DENSITY: ...............W./A. EVAPORATION RATE: .............N/.A SOLUBILITY IN HATER: ..........s.light SPECIFIC GRAVITY: .............c.a. 0.6 Water-1 (Bulk) PERCENT VOLATILE: .............0.% PH: ........................7..-.a. (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 418-013:PAGE C-24 low: FC.95 FLUOM Brand FluDrOChORLOal Surfactan't January 29, 1998 PAGE 2 ......................................a............................ 3. FIRE AND EXPLOSION K42MD IDATA ..............a -.-.-.-.----------------------------------------------- FLASH POINT: ...............N.on.e.. FLAMMABLE LIMITS - LEL: ...a-aa... MIA FLAMMABLE LIMITS - UEL: ...... MIA AUTDIGNITION TEMPERATURE: ......MIA EXTINGUISHING MEDIA: water, Carbon dioxide, Dry chemical, Foam SPECIAL FIRE FIGHTING PROCEDURES: Wear full protectiveclothing,includinghalm'tI self-contained, positive pressure or pressure demand breathingapparatus, bunker coat and pants, bands around arms, waist and logs, face mask, and protectivecoveringfor exposed areas of the head. UNUSUAL FIRE AND EXPLOSION K4ZARDS: See hazardousDecompositionsection for products of combustion. ----------------------------------------------------------------------4. REACTIVITY DATA ----------------------------------------------------------------------- STABILITY: Stable INCOMPATIBILITY - MATERIALS/CONDITIONS TO AVOID: Not applicable. HAZARDOUS POLYMERIZATION: hazardous polymerization will not occur. HAZARDOUS DECOMPOSITION PRODUCTS: Carbon Monoxide and Carbon Dioxide, Oxides of Sulfur, Hydrogen Fluoride,Toxic Vapors, Gases or Particulates. ----------------------------------------------------------------------S. ENVIRONMENTAL INFORMATION ----------------------------------------------------------------------- SPILL RESPONSE: Observe precautionsfrom other sections. Vacuum, use wet sweeping compound or water to avoid dusting. CAUTIC)NIA vacuum cleaner could be an ignitionsource. Clean up residuewith water. Place in an approved metal container. Seal the container. RECOMMENDED DISPOSAL: Do not release to waterways or sewer. Do not use in productsor processesthat could result in aquaticconcentrationsgreater than 1/10 of the lowestECSO or LCSO concentration. Incineratein an industrialor commercialfacilityin the presenceof a combustible material. Combustionproductswill includeHF. Disposal alternative: Disposeof waste productin a facilitypermittedto ----------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 418-013:PAGE C-25 MM: Fr,-eFsLLN)W Brand FluoroChadeal Surfactant january 29, 1998 PAGE 3 ------------------------------------------------------------------------ S. ENVIRONNOffAL INFORMATION (continued) ------------------------------------------------------------------------ acceptchemicamlost*. ENVIROMIMAL DATA: 96-Mr. Aquatic Fish LCSO, Fathead Kinnow(Pinsphaloo prouslas)w38 mg/l, oluegillguntish(Lepmis mcrochirus)-40 mg/j, RainbowTrout(Salmo gairdneri)-limg/l; 48-Mr. ECSO, Daphnis Magna - 50 ag/l; COD-.004 g/g; 80020- Nil. REGULATDRY INFORMATION: Volatile Organic Compounds: N/A. VOC Less M20 & Exempt Solvents: N/A. Since regulationsvary, consult applicableregulationsor authorities before disposal. U.S. EPA Mazardous Waste Number - None (Not U.S. EPA hazardous). This productcomplieswith the chemicalregistrationrequirementsof TSCA, EINECS, CDSL, AICS, MITI and Korea. EPCRA HAZARD CLASS: FIRE K42ARD: No PRESSURE: No REACTIvrTY: No ACUTE: Yes CHRONIC: Yes -----------------------------------------------------------------------6. SUGGESTED FIRST AID -----------------------ft----------------------------------------------------- EYE CONTACT: Immediatelyflush eyes with large amountsof water for at least 15 minutes.Get immediate medical attention. SKIN CONTACT: Immediatelyflush skin with large amounts of water. Remove contaminatedclothing.If irritationpersists,call a physician.Wash contaminatedclothing before rouse. INHALATION: If signs/symptomsoccur, remove personto fresh air. If signs/symptomscontinue, call a physician. IF SWALLOWED: Drink two glasses of water. Call a physician. .................................................................... 7. PRECAUTIONARY INFORMATION ------------------------------------------------------------------------ EYE PROTECTION: Avoid eye contact. Wear vented goggles. -----------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 418-013:PAGE @@---26 MM: FC-95 FLTRWAD Brand fluarochanicalSurtactant January 29, 199B PAGE 4 ----------------------------------------------------------------------7. PRECAURIONAIRWYORMATION (continued) --------------------------------------------w------------------------ SKIN PROTECTION: Avoid skin contact. Weer appropriategloves when handling this material. A pair of glovesmade from the followingmterial(s) are recommended: butyl rubber. Us* one or more of the following personalprotectionItems as necessaryto preventskin contact: head covering,coveralls. Protectivegarments(otherthan gloves)should be made of either of the followingmaterials: polyethylen*/polyvinylidencehloride(Saranex). RECOMMENDED VENTILATION: Use with appropriatelocal exhaustventilation. Use in a wellventilatedarea. Providesufficientventilationto maintain emissionsbelow recommendedexposurelimits. If exhaust ventilation is not adequate,use appropriaterespiratoryprotection. RESPIRATORY PROTECTION: Avoid breathingof dust. Select one of the followingNIOSM approved respiratorsbased on airborneconcentrationof contaminantsand in accordancewith OSHA regulations: half-maskdust and aist respirator, half-masksuppliedair respirator,full-facedust and mist respirator, full-facesuppliedair respirator. PREVENTION OF ACCIDENTAL INGESTION: Do not eat, drink or smoke when using this product.Wash exposed areas thoroughlywith soap and water. Wash hands after handling and before eating. RECOMMENDED STORAGE: Keep containerdry. Keep container closed when not in use. FIRE AND EXPLOSION AVOIDANCE: Nonflammable. OTHER PRECAUTIONARY INFDRMATION: No smoking:Smoking while using this product can result in contaminationof the tobacco and/or smoke and lead to the formation of the hazardousdecompositionproducts mentionedin section 4 of this MSDS. Hmrs KAZMD RATINGS: HEALTH: 2 FLAMMABILRTY: 0 REACTIVITY: 0 PERSDNAL PROTECTION: X (See precautions, section 7.) EXPOSURE LimrTs RNGREDIENT VALUE UNIT TYPE AUTH SKIN* ------------------------------------------------------------------------ POTASSIUM PERFLUDROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUDROALKYL SULFONATE ... 0.1 MG/M3 TWA 3M y POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MG/K3 TWA 3M y POTASSIUM PERFLUOROALKYL SULFONATE ... 0.1 MG/M3- TWA 3M y ------------------------------------------------------------------------ Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 4"B1-013:PAGE C-27 mDa: rc-esFLuoM BrandFlwrocheai=l ISurfactant January 29, 1998 EXPOSURE LIMITS (continued) PAGE 5 INGREDIENT VALUE UNIT TYPE AUTH SKIN* .................................................................. POTASSIUM PERFLUOROALKYL SULF M TE ... 0.1 mG/m3 THA 3M y - SKIN NOTATION: Listed substances indicatedwith 'Y' under SKIN refer to the potentialcontributionto the overallexposureby the cutaneousroute Including mucous membrane and *ye, either by airborne or, more particularly, by directcontactwith the substance. Vehiclescan alterskin absorption. SOURCE OF EXPOSURE LIMIT DATA: - 3M: 3M Recommended Exposure Guidelines -----------------------------------------------------------------------a. HEALTH HAZARD DATA ------------------------------------------------------------------------ EYE CONTACT: Ki3.dEye Irritation:signs/symptomscan include redness,swelling, pain, and tearing. SKIN CONTACT: Mild SkinIrritation(afterprolongedor repeatedcontact): signs/symptomscan include redness, swelling, and itching. May be absorbedthrough the skin and persist in the body for an extended time. INHALATION: May be harmfulif inhaled. May be absorbed by inhalation and persist in the body for an extended time. Single overexposure,above recomended guidelines, may cause: Irritation(upper respiratory):signs/symptomscan include SWAL:oreness of the n03e and throat, coughing and sneezing. OWED: Ingestionis not a likely route of exposureto this product. Illnessmay result from a single swallowingof a moderatequantityof this material. May be harmful if swallowed. f4UTAGENICITY: Mutagenicityassays indicate the productis not autagenic. .................................................................... Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately 418-013:PAGE C-28 peDS.- FC-es nmno January 29, 1999 Brand Fluorodwsioal Surfactant PACE 6 .............................................w ................... S. HEALTH HAZARD DATA (continued) ------------------------------------------------------------------------ REPRODLICTIVE/DEVELOPMENTAL TOXINS: Not toratogenicin the rat at oral doses below maternallytoxic levels. OTHER HEALTH HAZARD INFORMATION: This productis not known to contain any substancesregulatedunder California PropositionGS. A Product Toxicity Summary Sheet is available. -----------------------------------------------------------------------SECTION CHANGE DATES ------------------------------------------------------------------------ HEADING SECTION CHANGED SINCE November OS, 1997 ISSUE -----------------------------------------------------------------------Abbreviations:N/D - Not Determined N/A - Not Applicable CA - Approximately ------------------------------------------------------------------------ The informationin this Material Safety Data Shoot (MSDS)is believedto be correct as of the date issued. 3M K4KES NO WARRANTIES, EXPRESSED OR IMPLIED, INCLUDING, BUT NOT LIMITED TO, ANY IMPLIED WARRANTY OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE OR COURSE DF PERFORMANCE OR USAGE OF TRADE. User is responsible for determining whether the 3M productis fit for a particularpurposeand suitablefor user's methodof us* or application. Giventhe varietyof factorsthat can affect the use and applicationof a 3H product,some of which are uniquelywithin the user's knowledgeand control,it is essentialthat the user evaluatethe 3M product to determinewhetherit is fit for a particularpurposeand suitable for user's methodof use or application. 3M providesinformationin electronic fare as a serviceto its customers. Due to the remote possibilitythat electronictransfermay have resulted in errors,omissionsor alterationsin this information,3M makes no representationsa& to its completenessor accuracy. In addition, informationobtainedfrom a database any not be as currentas the informationin the HSDS available directlyfrom 3M. 418-013:PAGE C-29 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION PROCEDURE 418-013:PAGE C-30 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION Pmtocol 418-013 Version:4l8-Ol3-fO5NOV 98) Page 1 of 3 PROCEDURE TestArticle: PFOS Vehicle: 0.5% Tween4D 80 inR.O. DeionizedWater A. Purpose: The purpose ofthisprocedureistoprovidea method forthe preparation ofdosage suspensionsofPFOS and the vehiclefororaladministratiotno ratson Argus Study 418-013. B. General Information: 1 . Allsuspensioncontainerswillbe labeledand colorcoded. Each labelwill specifytheprotocolnumber, testarticliedentificatiAorng,us batch number, concentrationd,osage level,preparatiodnate,expiratiodnate and storageconditions. 2a. Suspensions willbe prepared: X . Daily Weekly - For days of use 2b. Vehiclewillbe prepared: - Daily X Weekly - For- daysofuse 3. Suspensions willbe preparedata finaldosage volume of5 mukg. 4. Safety: X Gloves,labcoat,goggles orsafetyglassesand faceshield X Dust-MistRespirator - Half-FaceRespirator - Full-FaceRespirator/PositiPvreessureHood - Tyvek Suit/Apron 5. Dosage suspensionsadjustedforFree base and % Purity. Yes X No (Calculationbsased on 100%) Free Base Purity 6. Sampling requirements:Citedinprotocol. 7. Storage:Citedinprotocol. 418-013-.PAGE C 31 ATTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION PMtoCol 418-013 Version:418-013-LO5NOV 98) Page 2 of3 PROCEDURE NOTE: Test articlewillbe prepared as a serialdilutionfrom 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, 5*C, add the required amount of TweendD 80 and mix untiluniform (See TEST ARTICLE CALCULATIONS). D. Test ArticleSuspension Preparation: 1 . To prepare the 0.64 mg/mL, Group V suspension, add the required amount oftestarticl(eSee TEST ARTICLE CALCULATIONS) intoan appropriatelysized,labeled container.OS ad to the requiredamount with vehicleand heat the mixture to BOOC 50C forapproximately 30 minutes or untilthe TA/S dissolves. 2. Once the testarticlehas 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 0.32 mg/mL, Group IV suspension, remove the required amount of stock suspension (Group V) (See TEST ARTICLE CALCULATIONS), OS ad with the vehicleand mix. 4. To prepare the 0.08 mg/mL, Group IIIsuspension, remove the required amount ofstock suspension (Group IV) (See TEST ARTICLE CALCULATIONS), OS ad with the vehicleand mix. 5. 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. 41Ml 3:PAGE C-32 A'TTACHMENT 3 TEST ARTICLE AND VEHICLE PREPARATION Protocol418-013 Version*418-013 (05 NOV 98 Page 3 of3 PROCEDURE 6. To preparethe 0 mg/mL, Group Isuspension,add requiredamount of vehicletoan appropriatelsyized,labeledcontainer(See TEST ARTICLE CALCULATIONS) and mix. Writtenby: Approved by: - U Clarification:X No QS Initials/Date Date: 0,9 (See attachedclarificatifoonrm.) q-1 418-013:PAGE C-3.Jo PPJil4EDIC.A Argus ResearchLaboratoriesi.m. 905 Shechv Drive.Buildin.pA horsham. PA 19044 Telephone:(215)443-97 10 Telefax:(215)443-8587 PROTOCOL 418-013 Oral(Gavage)PharmacokinetiSctudyofPFOS inRats SPONSOR'S STUDY NUMBER: T-6295.12 Amendment 1 - 6 January1999 Caesarean-SectioniOnbgservation-sDay 21 Presumed Gestation(pages12 and 13 oftheprotocola)nd Necrops (page 14 oftheprotocol): At Caesarean-sectioinngon day 21 ofpresumed gestationl,ungand liver sampleswilble collectefdromthreefetusesperliftferromfivelittepresrdosage group.These collectiownislolccuraftewreighingand examinatiofnorsex and grossexternaallterations. One halfofthelungsand one laterallobeofthelivewrilble individualrleytained inneutrabluffered10% formaliinnscintillatviioanls.Priortofixatiosne,veral (minimumoftwo)section(sapproximatel1ymm thicko)fthishalfofthelungs and thislobeofthelivewrilble removed usinga scalpealnd wilble retaineidn McDowell-Trump'Fsixativ(estoreudnderrefrigerationsncintillatvioanlsf)or futurevaluationT.he otherhalfofthelungsand theotherlaterallobeofthe livewrilble flashfrozeninliquinditrogeannd storedinheat-sealablpeouches on dryice.The procedureforcollectianngd flashfreezintghesetissuewsilble availablientheraw data.The liveorfeach oftheremainingfetusesp,lusany remaininglivefrromthethreeselectedfetusesw,ilble collectedp,ooled(per littefrr)o,zenand stored(-700Corbelow)untislhipmenttotheSponsorfor analysiass describedon page 13 oftheprotocol. The lungand livesrectionisnMcDowell-Trump'sFixativaend thelungand liver samplesinneutrablufferefdormaliwnillbe shipped(oncoldpacksand ambient conditionrse,spectivelfyo)r,possiblfeutureevaluatiobny electromnicroscopyor lighmticroscopyr,espectiveltyo,: 418-013:PAGE C-34 Jeanne deward PathologyAssociatesInternational 4915 D,ProspectusDrive Durham, NorthCarolina27713 Telephone: (919)544-5257 Telefax: (919)544-3218 Protocol418-013 Amendment 1 Page 2 The frozensamples willbe shipped(on dryice)toKrisJ.Hansen, Ph.D.,atthe address citedinthe protocolforpossiblefuturebiochemicalevaluation. Allrecipientwsillbe notifieidnadvance ofsample shipment. Reason forChanae: The Sponsor has requestedthatthese additionaslamples be retainedfor continuedevaluationofthe reasonsforreduced pup survivaolbserved in previousstudies. Alan M. Hoberman, Ph.D.,DABT Directorof Research -- ------- 99 Date Raymond G. York,Ph.1'a@E3ABT Date AssociateDirectorof Research Study Director Dena C. Lebo, V.M.D. Date 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 418-013:PAGE D-1 DEVIATION FROM THE PROTOCOL AND THE STANDARD OPERATING PROCEDURES OF THE TESTING FACILITY Eightyvirgifnemale ratswere randomly assignedtothestudyand receivedthetestarticlfeorforty-twdoays priortocohabitationr,ather than 120 virginfemalesassignedtostudy,dosed, mated and then assignedas the populationforthestudy. Female ratswithconfirmeddate ofmatingwere randomly selectedfor gestationday 15 (DG 15)Caesarean-sectioningA.llremainingrats (withand withoutconfirmeddatesofmating)were assignedto DG 21 Caesarean-sectioningR.ats withno confirmeddate ofmating were Caesarean-sectionedon theirestimatedDG 21. These deviationsdidnotadverselyaffecthe outcome orinterpreation ofthestudybecause sufficienrtatswere availablteocollecthe pharmacokineticdata. Alldeviatni@ofnsare cumented inthe raw data. R ond G. Yor D, DABT AssociateDirectorof search and Study Director Date TEMPERATURE APPENDIX E AND RELATIVE HUMIDITY REPORTS ARGUS 418-013:PAGE E-'. Temperature and RelativeHumidity Report Location:Room 04 Protocol Number.- 418-013 Range of Dates: 1O-Nov-1998 15:00 to 23-Nov-1 998 16:49 TargetRange: Species:Rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber ofData Points: Temperature 64OF to 79*F 14 312.75 312 RelativeHumidity 30% to70% 14 312.75 312 Mean (SD): 69.1 Maximum: 76.4 Median: 68.3 Minimum: 66.8 Number ofPointsinRange (%): 312 Number ofPointsHigh (%): 0 Number ofPointsLow (%): 0 Report Generated:03-Feb-1999at13:44 (2.5) 58.5 64.4 58.6 53.6 (100.0) 312 (0-0) 0 (0-0) 0 (1.7) (100.0) (0.0) (0.0) COMMENTS: REVIEWED BY: DATE: 4111-79 Cumulativeby Location(vD4.01.97) ARGUS 418-013:PAGE E-2 Temperature and RelativeHumidity Report Location:Room 27 Protocol Number: 418-013 Range of Dates: 23-Nov-1998 15:49 to 22-Jan-1999 16:00 TargetRange: Species:Rat TotalNumber ofDays: TotalNumber ofHours: TotalNumber of DataPoints: Temperature 64*F to79OF 61 1439.74 1442 RelativeHumidity 30% to70% 61 1439.74 1442 Mean (SD): Maximum: Median: Minimum: Number ofPointsinRange (%): Number ofPointsHigh (%): Number ofPointsLow (%): 70.7 (0.7) 54.1 72.6 69.8 70.8 56.1 69.0 14.9 1442 (100.0) 1418 0 (0-0) 0 0 (0.0) 24 8.2) (98.3) (0-0) (1.7) Report Generated:03-Feb-1999at13:48 COMMENTS: REVIEWED BY: DATE: ?13 Cumulativeby Location(vO4.01.97) ARGUS 418-013:PAGE E-3 RelativeHumidity DeviationsReport Location:Room 27 Protocol Number. 418-013 Range of Dates: 23-Nov-1 998 15:49 to 22-Jan-I999 16:00 Humidity Target Range: Species: Rat 30% to 70% Date 04-Jan-1999 05-Jan-I999 06-Jan-1999 06-Jan-I999 06-Jan-I999 06-Jan-I999 06-Jan-1999 06-Jan-1999 D6-Jan-1999 06-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 11-Jan-1999 14-Jan-1999 rime 16:00 23:00 00:00 01:00 02:00 03:00 04:00 05:00 06:00 07:00 11:00 12:00 13:00 14:00 15:00 16:00 00:00 R.H. 29.1 L 26.1 L 23.7 L 23.3 L 20.1 L 18.4 L 17.6 L 16.9 L 14.9 L 27.0 L 27.9 L 29.1 L 26.1 L 28.6 L 26.9 L 28.7 L 29.6 L Date 14-Jan-I999 14-Jan-I999 14-Jan-I999 14-Jan-1999 14-Jan-I999 14-Jan-I999 14-Jan-1999 Time 02:00 03:00 04:00 05:00 06:00 07:00 08:00 R.H. 28.6 L 27.8 L 27.5 L 26.3 L 27.0 L 26.3 L 25.8 L H Value out of range -High L Value out of range - Low R.H. = RelativeHumidity Report Generated: 03-Feb-1999 at 13:57 1 V These deviations did not adversely affect the outcome or interpretation of the study. The followingdeviation(si)mpacted on the outcome ofthe study as described: Study Director Date: e-A -99 Deviationsby Location(vD4.01.97) APPENDIX F HISTORICAL CONTROL DATA 418-013:PAGE F-1 SUMMARY OF REPRODUCTIVE INDICES CD RAT PERIOD JUNE 1996 -JUNE 1998 NUMBER OF STUDIES 105 NUMBER OF RATS: TESTED 2233 PREGNANT 2065 FOUND DEAD 4 ABORTED 0 DELIVERED 0 NUMBER OF RATS PREGNANT AT CAESAREAN-SECTIONING 2052 NUMBER OF RATS WITH SINGLE CONCEPTUS LrTTEk* LIVE 6 RESORBED I ABORTED 0 % PREGNANT AVERAGE # CORPORA LUTEA AVERAGE # IMPLANTATIONS AVERAGE LITTER SIZE AVERAGE # LIVE FETUSES AVERAGE # DEAD FETUSES AVERAGE # RESORPTIONS AVERAGE # EARLY RESORPTIONS AVERAGE # LATE RESORPTIONS MEAN or% 92-8 16.9 15.3 14.5 0.1 0.7 0.7 0.0 RANGE/STUDY MEAN or% (64.0-100) (14.0-21.0) (12.9-18-0) (11.8-16.9) (0-1.4) (0-2.0) (D-1.9) (0-0.1) 418-013:PAGE F-2 SUMMARY OF REPRODUCTIVE INDICES CD RAT MEAN or % AVERAGE % DAMS W" ANY RESORPTIONS 46.1 AVERAGE % DAMS WITH ALL CONCEPTUSES RESORSED 0.1 AVERAGE % DAMS WrrH ONE OR MORE LIVE FETUSES gg.a AVERAGE SEX RATIO. (% MALESILrrrER) 50.2 AVERAGE FETAL BODY WEIGHT (G) 3.4a AVERAGE FOR MALES (G) 3.5a AVERAGE FOR FEMALES (G) 3.39 AVERAGE % DEAD OR RESORBED CONCEPTUSES/LffTER 4.7 RANGEISTUDY MEAN or% (o-aT.5) (0-4.5) (95.4-100) (42.a-57.0) (3.10-3.78) (3.17-3.90) (2.98-3.67) (0-12.4) 418-013:PAGE F-3 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT PERIOD STUDIES RATS TESTED RATS PREGNANT RATS DIED RATS ABORTED RATS DELIVERED RATS WITH 100% JUNE RESORPTION 1996 -JUNE 1998 124 2668 2,M* S" 0 431 3 EXTERNAL MEAN RANGE/STUDY OBSERVATIONS N% N % Red substancearound nose and/ormouth 2 0.07 0-1 (0-12.5) White substancepresent inanteriocrhamber of eyes 1 0.04 0-1 (0-4.2) Red perivaginaslubstance 1 0.04 0-1 (D-4.0) GROSS LESIONS ESOPHAGUS Tom( attributteodan intubatioanccident) LUNGS Dark red(attributteDdan intubabonaccident) THORACIC CAVITY Fillewdithlighrtedor cloudyredfluid THYMUS Large AXILLA Mass present Lymph nodes enlarged on lefstide 1 0.04 0-1 (0-4.0) 1 0.04 0-1 (D-4.0) 2 0.07 D-1 (0-4.0) 1 0.04 D-1 (0-4.0) 1 0.04 D-1 (0-4.0) 1 0.04 0-1 (0-4.2) Pregnancystatusforone dam couldnotbe determined One was a moribundsacrificaend one was attributteodan intubabonaccident 418-013:PAGE F-4 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT GROSS LESIONS N LIVER Adhesionbetween left laterallobeand lefltateral abdominalwall 1 Rightlateraalnd rrvedian lobesprotrudedthrough diaphragm 1 STOMACWINTESTINE Stomach and Intestines distendedwithgas 1 Cocum containeda black substance 1 Stomach containeda dark red,granulasrubstance 1 BACK Spine,protrusioonf bone on dorsalowmcic portion 1 KIDNEY(S) Pelvis,slighttmoderate dilatiownithorwithout fluid 16 Pelvism,arked/extreme drabon 7 Mottled 1 Large 2 smwi 1 Cortexpitted 1 Left,pelviscontained yellowfluid 1 Bilateraple.lvicsontained numerous calculi 1 RighL indentation 1 Palo 1 Left,smau;righkenlarged cortexwithnumerous pinpoinytellowareas and viscousyellowftuidin pelvis 1 ADRENAL GLAND(S) Large 1 MEAN RANGE /STUDY % N % 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 C)-l (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.2) 0.04 0-1 (D-4.0) 0.60 0-2 (0-12.5) 0.26 D-1 (0-4.0) 0.04 0-1 (0-4.0) 0.07 0-1 (0-4-0) 0.04 0-1 (04-0) 0.04 0-1 (0-3.4) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-12.5) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 418-013:PAGE P-5 SUMMARY OF MATERNAL NECROPSY OBSERVATIONS CD RAT GROSS LESIONS N ABDOMINAL CAVITY Filledwithlighrtedfluid 1 SPLEEN Two whiteareason serosalsurface 1 Large 2 BLADDER Wall thickand contained one calculus 1 URETERS Distendedwithclearfluid 2 VAGINA/CERVIX Cervixcontaineda darkred. gelatinous ubstance 1 Cervbc containedgreen, vl%wus fluid 1 Vagina containedbrown. viscousfluid 1 UTERUS Containedred-brownfluid 1 Righthom, absent 1 Right hom, threadlike 1 Lefthom, lumen absenton cervicalend;ovarianend. distendedwithclearfluid 1 Righthom, clearmasses containing elatinous substance present 1 Portionof lefutterinheom prolapsed 1 OVARIES Right containeda red, fluid-filbluerdsa[cyst 1 MEAN % 0.04 RANGE /STUDY N % 0-1 (G-4.0) 0.04 0-1 (G-4.0) 0.07 G-2 (0-8-0) O.G4 0-1 (G-4.0) 0.07 0-1 (G-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-2.1) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-2.1) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 0.04 0-1 (0-4.0) 418-013:PAGE F-6 SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS CD RAT PERIOD JUNE 1996-JUNE 1998 # STUDIES INCLUDED # LrTrERS EXAMINED # LIVE FETUSES EXAMINED 84 1652 23689 ALTERATION HEAD Exencephaly L F Hematoma L F Microcapha@y L F EYE(S) Eye bulgesdepressed L F EyoUds open L F Microphthalmia L F EARS Low set L F SNOUT Short L F TONGUE Absent L F Protrudes L F FusedtD lowermargin L oforalopening F N% 4 0.24 4 0.02 1 0.06 1 0.00 1 0.06 1 0.00 RANGE N 0-1 D-1 0-1 0-1 0-1 0-1 /STUDY % (0-4.2) (0-0.3) (0-2.2) (0-0.2) (0-4.0) (0-0.3) 9 0.54 9 0.04 2 0.12 2 0.01 1 0.06 1 0.00 0-2 (0-12-5) 0-2 (0-0-9) 0-1 (0-4.2) 0-1 (0-0.3) 0-1 (0-4.0) G-1 (0-0.3) 1 0.06 1 0.00 G-1 (0-12.5) 0-1 (D-0.9) 2 0.12 2 0.01 D-1 (0-4.2) 0-1 (0-0.3) 1 0.06 1 0.00 1 0.06 1 0.00 1 0.06 1 0.00 0-1 (0-4.0) 0-1 (0-0.3) 0-1 (0-4.2) 0-1 (0-0.3) 0.1 (0-4.8) 0-1 (0-0.4) L:LITTER INCIDENCE F:FETAL INCIDENCE 418-013:PAGE F-7 SUMMARY OF FETAL GROSS EXTERNAL ALTERATIONS CD RAT ALTERATION JAWS Micrognathia Agnathia Cleft BODY Edema Umbilicalhemia Spina bifida Hematoma Conjoinedtwin HINDLIMBS Thirdlimbpresent ANUS No openingpresent TAIL Threadlike Agenesis Short Consbieted N% RANGE /STUDY N % L 2 0.12 0-1 (G-4.5) F 2 0.01 0-1 (0-0.3) L 1 0.06 0-1 (0-12.5) F 1 0.00 0-1 (0-0-9) L 1 0.06 G-1 (0-4.8) F 1 0.00 0-1 (0-0.4) L 2 0.12 0-1 (0-4.3) F 2 0.01 0-1 (0-0.3) L 5 0.30 0-1 (D-4.3) F 5 0.02 0-1 (0-0.3) L 1 0.06 0-1 (0-4.2) F 1 0.00 0-1 (0-0.3) L 2 0.12 0-1 (0-4.0) F 2 0.01 0-1 (0-0.3) L 1 0.06 0-1 (0-4.0) F 1 0.00 0-1 (0-0.3) L 1 0.06 0-1 (0-14.3) F 1 0.00 0-1 (0-0.9) L 1 0.06 D-1 (0-2.6) F 1 0.00 0-1 (0-0.2) L 5 0.30 0-1 (0-12.5) F 5 0.02 0-1 (D-0-9) L 2 0.12 0-1 (0-4.5) F 2 0.01 0-1 (0-0.3) L 1 0.06 0-1 (0-2.4) F 1 0.00 0-1 (0-0.2) L 1 0.06 0-1 (0-2.4) F 1 0.00 0-1 (0-0.2) L:LITTER INCIDENCE F:FETAL INCIDENCE 418-013:PAGE F-8 SUMMARY OF FETAL SOFT TISSUE ALTERATIONS CD RAT PERIOD JUNE 1996 - JUNE 1998 # STUDIES INCLUDED 36 # LffTERS EXAMINED 830 FETUSES EXAMINED 5753 ALTERATION BRAIN Lateralvenbicies, L moderatedilation F Lateravlentricles, L marked dilation F Thirdventricle, L marked dilation F Lateraalnd thirdvent- riclesI,rTegularly L shaped F lrregularsthyaped L F EYES Mk:rophthalniia L F TONGUE Absent L F JAW Micrognathia L F HEART Septaldefect L F VESSELS lnnominate,absent L F RANGE/STUDY N% N % 1 0.12 0-1 (0-4.3) 1 0.02 0-1 (0-0.6) 2 0.24 0-1 (04.3) 2 00..03 0-1 (".6) 1 12 0-1 (0-4.2) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (04.2) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (04.2) 1 0.02 0-1 (0-0.6) 3 0.36 0-1 (0-4.3) 3 0.05 0-1 (0-0.7) 1 0.12 0-1 (04.0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 6 0.72 0-2 (0-8.3) 6 0.10 0-2 (0-1.2) L:LITTER INCIDENCE F:FETAL INCIDENCE 418-013:PAGE F-9 summARY OF FETAL SOFT TISSUE ALTERATIONS CD RAT ALTERATION VESSELS (CONT.) lnnominatea,riseson L left F Umbilicaalrtery. descendstoleftof L urinaryblomer F Situsinversus L F Aorta,descends to L right F PuWnonary artery. descends toright L behindaorta F LUNGS Rightapicalc,ardiac and diaphragmatic L lobw appearas one F intermediatkebe. L absent F BODY Edema L F ABDOMINAL CAVITY Situsinversuosf liver, intestinesst,mach. spleen,pancreasand L kidneys F KIDNEY(S) Pelviss.lighdtilation L F PeMs. moderatedilatioLn F ADRENAL(S) Dark red L F L:LITTER INCIDENCE F:FETAL INCIDENCE RANGE/STUDY N% N % 1 0.12 0-1 (0-4.0) 1 0.02 0-1 (0-0.6) 15 1.81 0-2 (0-8.7) 16 0.28 0-2 (0-1.4) 1 0.12 0-1 (0-4-2) 1 0.02 0-1 (0-0-6) 1 0.12 0-1 (D-4-0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4-0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4-0) I o.02 0-1 (0-0.6) 1 0.12 0-1 (0-4-0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4-0) 1 0.02 0-1 (0-0.6) 1 0.12 0-1 (0-4-0) 1 0.02 0-1 (0-0.6) 2 0.24 0-2 (0-8.7) 2 0.03 0-2 (0-1.3) 1 0.12 0-1 (0-4.3) 1 0.02 0-1 (0-0.6) 2 0.24 0-1 (0-4.3) 2 0.03 0-1 (0-0.6) 418-013:PAGE F-10 SUMMARY OF FETAL SKELETAL ALTERATIONS CD RAT PERIOD STUDIES INCLUDED LITTERS EXAMINED FETUSES EXAMINED JUNE 1996 -JUNE 1998 36 816 6151 ALTERATION SKULL Frontal(s)i:ncompleteloyr not ossified Paristal(si)n:completeloyr notossified Nasal(s):shcxt Sphenoid: incompletely ossified Maxillasand Prenia)dllae: short Skull:incompleteloyr not ossified RANGE/STUDY N% N% L 2 0.24 0-1 (0-4.2) F 2 0.03 0-1 (0-0-6) L 2 0.24 0-1 (0-4.2) F 2 0.03 0-1 (0-0.6) L 3 0.37 0-1 (0-4.2) F 3 0.05 0-1 (0-0.6) L 1 0.12 0-1 (0-4-2) F 1 0.02 0-1 (0-0.6) L 3 0.37 0-1 (0-4.2) F 3 0.05 0-1 (0-0.6) L 1 0.12 0-1 (0-4.2) F 2 0.03 0-2 (0-1-2) VERTEBRAE Thoracic:Cenbum, bifid L 67 8.21 F 74 1.20 : Contra.unilateral L 6 0.74 ossification F : Centrum, incompletely L 6 0.10 3 0.37 or notossified F 4 0.06 : Centra,fused L 1 0.12 F 1 0.02 : Arch,small L 1 0.12 F 1 0.02 0-5 (0-20.8) 0-6 (0-3.4) 0-2 (0-8-0) C@.2(0-1.0) 0-2 (D-8.3) 0-3 (0-1.8) D-1 (0-4.2) 0-1 (0-0.5) 0-1 (0-4.0) 0-1 (0-0.6) Lumbar Centrum,bifid Centrum. incompletely or notossified Arches,Incompletelyor notossified Centra,unilateral ossificabon L 1 0.12 F 1 0.02 L 2 0.24 F 3 0.05 L 12 1.47 F 17 0.28 L 2 0.24 F 2 0.03 0-1 (0-4.3) 0-1 (0-0.6) D-2 (0-8.3) 0-3 (0-1.8) 0-3 (0-12.0) 0-4(0-2.1) 0-1 (0-4.2) 0-1 (0-0.6) L LrrTER INCIDENCE F: FETAL INCIDENCE 18-013:PAGE F-11 SUMMARY OF FETAL SKELETAL ALTERATIONS CD RAT ALTERATION RIBS CervicaRlib(s)present One ormore,wavy One ormore,incompletely ossifie(dhypoplastico)r,not ossified Extraribs,i4th,unilateral Extraribs.14th,bRaterW RANGEISTUDY N% N % L 23 2.82 F 26 0.42 L 32 3.92 F 52 0.84 0.4 (D-17.4) 0-5 (0-2.9) 0-4 (0-33.3) 0-8 (0-5-1) L 22 2.70 F 36 0.58 L 4 0.49 F 4 0.06 L 1 0.12 F 1 0.02 0-2 (0-9-1) 0-5 (0-2.6) 0.4 (0-16-0) 0-4(0-2.2) 0-1 (0-4.0) 0-1(0-0.6) STERNEBRAE One ormore Incompletely ossifieodr notossified Fused Asymmetric SCAPULAE Bent L 109 13.36 0-9 (0-36.0) F 150 2.44 0-12 (0-6.7) L 1 0.12 0-1 (0-4.2) F 1 0.02 0-1 (0-0.6) L 1 0.12 0-1 (0-42) F 1 0.02 0-1 (0-0-5) L 1 0.12 0-1 (0-4.2) F 1 0.02 0-1 (0-0.5) PELVIS Pubis(esa)ndforlschium(a): L 116 14.22 0-7 (0-30.4) incompleteloyrnotossified F 180 2.93 0-16 (0-9.5) Pubis(es)i:ncompletely L 96 11.76 0-7 (0-30.4) ossified F 147 2.39 0-16 (0-9.5) Pubis(es)n:otossified L 3 0.37 0-1 (0-4.2) F 3 0.05 0-1 (0-0.6) lschium(a)i:ncompleteloyrnot L 39 4.78 0-4 (0-16.7) ossified F 58 0.94 0-9 (0-4.7) FORELIMB(S) Radiusand Ulna:Bent L 1 0.12 0-1 (0-4.2) F 1 0.02 0-1 (0-0.6) L:LITTER INCIDENCE F:FETAL INCIDENCE 418-013:PAGE F-12 SUMMARY OF FETAL OSSIFICATION SITES. SKELETAL AVERAGES CD RAT (CAESAREAN-SECTIONED DAY 20 GESTATION) PERIOD: # STUDIES # LffTERS # FETUSES JUNE INCLUDED EXAMINED EXAMINED 1996 -JUNE 1998 35 793 5961 SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (pairs) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS (Calculateads average perlimb) CARPALS METACAP.PALS DIGITS PHALANGES HINDPAVVS (Calculatedas average per limb) TARSALS METATARSALS DIGITS PHALANGES FETUS/LfrTER MEAN RANGEISTUDY 0.86 (0.69-0.98) 7.00 13.04 5.95 3.00 4.82 13.03 (13.00-13.15) (5.85-5.99) (2.96-3.01) (4.35-5.16) (13.00-13.09) I.W (0.98-1.01) 3.61 (3.26-3.75) 0.99 (0.94-1.00) 0.00 3.,54 5.00 5.D4 (3.33-3.74) (4.90-5.27) 0.00 3.99 (3.93-4.03) 5.00 4.98 (4.82-5.08) 418-013:PAGE F-13 FETAL OSSIFICATION SITES SKELETAL AVERAGES CD RAT (CAESAREAN-SECTIONED DAY 21 GESTATION) STUDY # LITTERS EXAMINED # FETUSES EXAMINED SKELETAL AVERAGES HYOID VERTEBRAE CERVICAL THORACIC LUMBAR SACRAL CAUDAL RIBS (pairs) STERNUM MANUBRIUM STERNAL CENTERS XIPHOID FOREPAWS (Calculateads averageperlimb) CARPALS METACARPALS DIGITS PHALANGES HINDPAWS (Calculateads average perlimb) TARSALS METATARSALS DIGITS PHALANGES 580 23 190 0.96 7.00 13.06 5.94 3.00 7.23 13.05 1.00 3.97 1.00 0.00 3.99 5.00 7.53 0.02 4.62 5.00 5.78 STATEMENT APPENDIX G OF THE STUDY DIRECTOR 0 PUMMICA 418-013:PAGE G-1 Argus ResearchLaboratoriesI,nc. 905 Sheehy Drive,BididingA Horsham, PA 19044 Telephone:(215)443-8710 Telefax(:215)443-8597 PROTOCOL 418-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS SPONSOR'S STUDY NUMBER: T-6295.12 STATEMENT OF THE STUDY DIRECTOR This finalreportaccuratelyreflecttshe raw data obtainedduringthe performance ofthe study. No deviationsfrom the U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations;FinalRule',the Japanese Ministryof Healthand Welfare (MHW) Good LaboratoryPracticeStandard forSafety Studieson Drugs" and the European Economic Community (EEC) Council decisionon 28 July 1989 on the acceptance by the European Economic Community ofan OECD decisiorilrecommendatioonn compliance withprinciples ofgood laboratorpyracticecoccurredthataffectedthe qualityor integritoyfthe study. Ll0@; qmond G. Y Ass-oCiateDirec r and Study Direct ov-o@wo-,g D.,DABT Research Date a. U.S. Food and Drug Administration.Good LaboratoryPractice Regulations;FinalRule. 21 CFR Part58. b. Japanese Ministryof Health and Welfare (1988). Good Laboratory Practice Standard for Safety Studies on Drugs, MHW Ordinance Number 21, March 26, 1997. C. European Economic Community (1989). Council decision on 28 Juiy 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: Legislation3.2(No.L 315; 28 October):1-17. APPENDIX H QUALITY ASSURANCE UNIT FINAL REPORT STATEMENT iePiR-UVMDICA 418-013:PAGE H-1 Argus Research Laboratories.Inc. 905 Sheehy Drive.BuildingA Horshmn, PA 19044 Telephone: (215)443-8710 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 Protocol418-013: Oral (G,avage)PharmacokineticStudy of PFOS inRats Sponsors Study Number T-6295.12 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 July1989 on the acceptance by the European Economic Community of an OECD decision/recommendationon compliance withprinciples ofgood laboratorypracticeon 18 OCT 98. Criticaplhases of thisstudywere inspectedsixtimes;study information 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 RegulationsJ,apanese MinistryofHealthand Welfare(MHW); Good Laboratory PracticeStandard forSafetyStudieson Drugs, and European Economic Community (1989)councildecisionon 28 July 1989 on the acceptance by the European Economic Community ofan OECD decision/recommendationon compliance withprincipleosfgood laboratorypracticebetween 25 APR 99 and 26 MAY 99, and forrevisionsrequestedby the Sponsor 17 JUN and 18 JUN 99 and forfinalizatioonn 24 JUN 99. 418-013:PAGE H-2 This study was conducted accordingto U.S. Food and Drug Administratio(nFDA) Good LaboratoryPracticeRegulations,Japanese Ministry of Healthand Welfare (MHW); Good LaboratoryPracticeStandard forSafety Studieson Drugs, and European Economic Community (1989)councildecision on 28 July 1989 on the acceptance by the European Economic Community ofan OECD decisionlrecommendationon compliance withprincipleosfgood laboratorypractice. Nancy J. ongliewgsi Date QualityAssurance Manager isaA. Zaborowski, B.S. Date Senior QualityAssurance Associate and PrincipalAuditor TABLE 1 CRITICAL PHASES INSPECTED 418-013:PAGE H-3 Test ArticlPerer)aration Date of inspection:19 NOV 98 Date resultsreportedtothe Study Directorand Management: 20 NOV 98 Test ArticlAedministration- Gava-ge Date of inspection:19 NOV 98 Date resultsreportedto the Study Directorand Management: 20 NOV 98 Cohabitation Date ofInspection:28 DEC 98 Date resultsreportedto the Study Directorand Management: 31 DEC 98 Blood Collection Date ofinspection:04 JAN 99 Date resultsreportedtothe Study Directorand Management: 04 JAN 99 Urineand Fecal Collection Date of inspection:04 JAN 99 Date resultsreportedtothe Study Directorand Management: 04 JAN 99 Caesarea n-Sectioning Date of inspection:13 JAN 99 Date resultsreportedto the Study Directorand Management: 19 JAN 99 418-013:PAGEH-4 TABLE 2 RAW DATA AUDIT(S) The followisntgudyinformatiaonnd rawdatawere audited from18 MAR 99 to25 MAR 99: Protocol. Protocoalmendment. Listofpersonnealnd computeroperatocrodes. Errorcodesand codesforclinicsailgnobservations. Animalreceiprta,ndomizatiopnh,ysicaelxaminatioannd acclimation. In-litfreansactiroencord. Feed consumption. Cohabitation. Caesarean-sectioning. Maternaglrossobservations. Fetalgrossobservations. Necropsy. Graviduterinweeights. Placentawleights. Tissuepackinglists. Malebreedercolonyrecords. Generalcomments. Studymaintenancerecords. Temperaturaend relatihvuemiditryeports. Feed and wateranalyses. Editrequests. Randomizations. Dosagevolumes. Deviations. Datareviewpages. Bloodand livecrollectidoantaand packinglists. Maternaplackinglistfsorliver/glands. Fetalliverautnigssupeackinglists. Pooledliveprl,acental,ivearnd lungpackinglists. Fecal/uricnoellectidoantaand packinglists. Embryo/placenctoallectiaonnd packinglists. Fetalcarcasspackinglists. Amniotifcluicdollectiaonnd packinglists. The resultosfthisaudiwtere reportetdotheStudyDirectoarnd Management on 26 MAR 99. 418-013:PAGE H-5 The followingstudy informatioannd raw data were audited on 25 MAR 99: Vehicle/Controalrticlreeceiptp,reparationand use. Test article/substancreceiptp,reparationand use. Test article/substancpeacking lists. Test article/substancaenalysis. The resultsofthisauditwere reportedtothe Study Directorand Management on 29 MAR 99. PROTOCOL 418-013: ORAL IGAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PAGE 5): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION -------------------------------------------------------------------------------------------------------------- FETUS N 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 17 is 19 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP 111 0.4 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- RAT # CLe 19533 4/12 MA MA FA MA / FA FA MA MA MA MA MA FA 5.19 5.23 4.92 5.42 4.80 5.01 5.13 5.09 5.57 5.40 5.35 4.82 19534 13/15 FA MA MA MA MA 14A FA / FA FA MA MA FA FA MA 19536 19541 7/12 4.08 4.80 MA FA 5.57 5.15 MORIBUND 4.80 4.79 FA FA 5.13 4.97 SACRIFICED 5.20 5.06 MA FA 5.89 5.36 ON DAY 0 5.24 4.78 5.00 5.12 4.87 4.60 5.22 5.32 FA FA FA FA MA FA FA PA 4.88 5.22 5.08 4.93 5.16 4.82 4.74 5.23 OF PRESUMED GESTATION; PREGNANCY STATUS MA 4.46 COULD FA 5.12 NOT FA MA 4.97 5.46 BE DETERMINED 19543 11/10 19545 E FA MA FA 4.83 5.07 4.60 14ATING NOT CONFIRMED MA 5.10 MA 5.26 FA 4.66 KA 5.08 MA 5.19 FA 4.47 MA / FA 5.51 4.60 E MA MA MA MA MA FA 4.74 5.41 5.48 5.38 4.71 5.1 19546 14/11 MA KA FA E MA / t4A E E E 5.04 5.56 4.70 5.30 4.84 19548 NOT PREGNANT -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP IV 1.6 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19549 KATING NOT CONFIRMED 19551 MATING NOT CONFIRMED 19552 19556 MATING NOT CONFIRMED NOT PREGNANT 19557 19559 MORIBUND SACRIFICED NOT PREGNANT ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19560 12/ 7 FA FA MA MA MA FA FA PA MA / MA FA FA 5.17 5.13 5.20 5.43 5.44 5.12 5.27 4.70 5.13 5.08 4.85 5.15 19562 8/14 MA KA FA FA FA PA MA FA / FA FA E MA E MA FA MA FA MA FA S.50 5.70 4.75 5.27 5.28 4.61 4.84 4.78 4.97 4.74 4.79 4.95 4.93 4.94 4.37 4.75 4.7 -------------------------------------------------------------------------------------------------------------- M - MALE F - FEMALE A = ALIVE E - EARLY RESORPTION L = LATE RESORPTION CLS - CORPORA LUTFA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G). DENOTES POSITION OF CERV PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 20 (PACE 6): EMBRYONAL VITAL STATUS OR FETAL SEX, VITAL STATUS AND BODY WEIGHT - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- PATS CAESAREAN-SECTIONED ON DAY 21 OF PRESUMED GESTATION -------------------------------------------------------------------------------------------------------------- FETUS 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 is 19 -------------------------------------------------------------------------------------------------------------- DOSAGE GROUP V 3.2 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- RAT N CLa 19568 8/11 t4A 14A MA MA MA MA / E E E 4.12 3.73 4.16 5.19 4.98 4.84 19569 10/ 6 FA MA FA MA FA FA MA MA FA MA FA FA MA 4.33 4.85 3.43 5.12 4.78 4.33 4.81 4.86 4.66 5.12 5.34 4.61 4.30 19571 DELIVERED ON DAY 21 OF GESTATION 19572 19576 S/ 9 FA- MA FA MA MA 4.51 4.S6 4.55 5.02 4.96 MATING NOT CONFIRMED E MA PA PA 4.31 4.76 4.39 E FA 4.67 E PA MA MA 4.55 4.67 5.15 19577 19578 B/ 5 FA 4.98 MATING E PA MA FA 5.15 5.18 4.87 NOT CONFIRMED 19579 DELIVERED ON DAY 21 OF GESTATION -------------------------------------------------------------------------------------------------------------- M - MALE F = FEMALE A - ALIVE E = EARLY RESORPTION L = LATE RESORPTION CLS m CORPORA LUTEA/OVARY FETAL BODY WEIGHTS WERE RECORDED IN GRAMS (G). DENOTES POSITION OF CERV PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 21 (PAGE 1): PLACENTAL WEIGHTS - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION -------------------------------------------------------------------------------------------------------------- FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 18 19 -------------------------------------------------------------------------------------------------------------- RAT # MEAN DOSAGE GROUP 1 0 (VEHICLE) MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19503 NO CONFIRMED MATING DATE; DELIVERED ON DAY 66 OF STUDY 19504 FA E MA E MA MA MA / E MA E MA PA MA E FA MA FA O.SB 0.52 0.47 0.59 0.59 0.64 0.45 0.61 0.60 0.69 0.65 0.64 0.50 19505 FA FA MA FA MA FA MA E FA MA MA FA MA MA MA E FA FA FA 0.44 0.53 0.48 0.52 0.47 0.48 0.43 0.40 0.37 0.47 0.38 0.41 0.41 0.48 0.41 0.38 0.46 0.4 19506 E FA MA FA MA MA FA FA FA MA FA FA FA FA 0.54 0.63 0.48 0.63 0.41 0.61 0.55 0.53 0.52 0.65 0.46 0.53 0.4S 0.51 19507 MA FA MA MA MA MA FA FA FA E FA MA MA FA FA 0.41 0.41 0.44 0.38 0.39 0.38 0.44 0.32 0.35 0.38 0.43 0.37 0.44 0.44 0.52 19509 FA FA FA FA MA PA E MA MA FA FA MA MA 0.53 0.46 0.53 0.56 0.53 0.55 0.45 0.57 0.54 0.57 0.54 0.55 0.47 19512 MA FA FA FA MA FA MA MA MA MA FA FA FA MA 0.49 0.56 0.54 0.54 0.54 0.53 0.36 0.46 0.47 0.51 0.40 0.48 0.51 0.47 0.53 19516 MA FA FA FA FA FA MA MA MA FA FA FA MA FA 0.51 0.60 0.46 0.49 0.63 0.63 0.47 0.44 0.48 0.57 0.33 0.61 0.42 0.59 0.42 -------------------------------------------------------------------------------------------------------------- RAT N MEAN DOSAGE GROUP Il 0.1 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19518 19519 19520 0.51 0.45 FA 0.49 E MA FA FA 0.53 0.55 0.54 E FA FA / MA MA MA MA MA 0.53 0.42 0.66 0.44 0.41 0.46 0.54 FA MA FA FA MA PA FA MA MA / FA MA MA MA FA t4A MA 0.43 0.48 0.47 0.47 0.47 0.42 0.38 0.46 0.43 0.44 0.50 0.45 0.55 0.38 0.4S 0.49 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINED 19521 0.45 FA FA FA MA FA FA MA MA MA / FA FA MA FA MA MA 0.44 0.46 0.40 0.44 0.49 0.49 0.40 0.42 0.46 0.37 0.47 0.41 0.48 0.42 0.50 19523 0.53 E FA L FA FA FA FA FA MA MA / MA E FA L 0.49 0.55 0.56 0.44 0.55 0.55 0.56 0.49 0.52 0.59 19524 19528 0.53 FA 0.60 E MA MA 0.52 0.58 MA / MA MA MA 0.67 0.47 E E FA MA 0.55 0.52 FA MA MA MA O.SB 0.46 FA MA 0.46 E MA MA MA FA MA FA 0.67 0.54 0.53 0.40 0.49 0.42 0.66 1.08 0.53 0.63 0.52 O.S4 19529 FA MA FA KA E FA FA MA FA MA FA MA FA MA FA 0.62 0.93 0.77 0.54 0.63 0.56 0.67 0.69 0.49 0.51 0.50 0.59 0.68 0.53 0.58 -------------------------------------------------------------------------------------------------------------- M - MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION CLa = CORPORA LUTEA/OVARY PLACENTAL WEIGHTS WERE RECORDED IN GRAMS (G). DENOTES POSITION OF CERV PROTOCOL 410-013: ORAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 21 (PAGE 2) : PLACENTAL WEIGHTS - INDIVIDUAL DATA ------------------------------------------------------------------------------------------------------------- RATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION ------------------------------------------------------------------------------------------------------------- FETUS # 1 2 3 4 5 6 7 8 9 10 11 12 13 14 is 16 17 18 1 ------------------------------------------------------------------------------------------------------------- RAT # MEAN DOSAGE GROUP 111 0.4 MG/KG/DAY ------------------------------------------------------------------------------------------------------------- 19533 0.45 MA MA FA MA / FA FA MA MA MA MA MA PA 0.52 0.38 0.34 0.44 0.42 0.46 0.40 0.43 0.47 0.58 0.50 0.47 19534 0.57 FA MA MA MA MA MA FA / FA FA MA MA FA PA MA 0.49 0.58 0.50 0.51 0.60 0.61 0.62 0.50 0.61 0.53 0.65 0.61 0.53 0.64 19536 19541 0.46 MA FA FA FA MA FA FA FA FA FA MA FA FA FA 14A FA FA MA 0.45 0.48 0.50 0.54 0.53 0.40 0.38 0.54 0.42 0.39 0.49 0.47 0.45 0.47 0.33 0.48 0.46 0.47 MORIBUND SACRIFICED ON DAY 0 OF PRESUMED GESTATION; PREGNANCY STATUS COULD NOT BE DETERMINE 19543 19545 0.41 E FA MA FA MA 0.50 0.39 0.44 0.37 MATING NOT CONFIRMED MA 0.49 FA 0.38 MA 0.42 MA 0.43 FA 0.41 MA / FA 0.42 0.28 E MA MA MA MA MA F 0.38 0.43 0.43 0.40 0.39 0. 19546 MA MA PA E MA / MA E E E 0.51 0.44 0.47 0.36 0.49 0.79 19548 NOT PREGNANT ------------------------------------------------------------------------------------------------------------- RAT # MEAN DOSAGE GROUP IV 1.6 MG/KG/DAY ------------------------------------------------------------------------------------------------------------- 19549 MATING NOT CONFIRMED 19551 MATING NOT CONFIRMED 19552 MATING NOT CONFIRMED 19556 NOT PREGNANT 19557 MORIBUND SACRIFICED ON DAY 43 OF STUDY (DAY 2 OF COHABITATION) 19559 NOT PREGNANT 19560 FA FA MA MA MA FA FA FA MA MA FA FA 0.54 0.48 0.54 0.59 0.67 0.74 0.54 0.50 0.53 0.39 0.56 0.44 0.50 19562 MA MA FA FA FA FA MA FA / FA FA E MA E MA FA MA FA MA F 0.44 0.46 0.54 0.43 0.43 0.41 0.45 0.49 0.54 0.53 0.38 0.57 0.39 0.34 0.39 0.43 0.45 0. ------------------------------------------------------------------------------------------------------------- M = MALE F = FEMALE A = ALIVE E = EARLY RESORPTION L = LATE RESORPTION DENOTES POSITION OF CER CLS - CORPORA LUTEA/OVARY PLACENTAL WEIGHTS WERE RECORDED IN GRAMS (G). PROTOCOL 418-013: OPAL (GAVAGE) PHARMACOKINETIC STUDY OF PFOS IN RATS (SPONSOR'S STUDY NUMBER: T-6295.12) TABLE 21 (PACE 3) ! PLACENTAL WEIGHTS - INDIVIDUAL DATA -------------------------------------------------------------------------------------------------------------- PATS CAESAREAN-SECTIONED ON DAY 21 OF GESTATION -------------------------------------------------------------------------------------------------------------- FETUS 1 2 3 4 5 6 7 a 9 10 11 12 13 14 15 16 17 is 19 -------------------------------------------------------------------------------------------------------------- PAT # MEAN DOSAGE GROUP V 3.2 MG/KG/DAY -------------------------------------------------------------------------------------------------------------- 19568 MA MA MA MA MA MA / E E E 0.62 0.71 0.54 0.76 0.59 0.59 0.54 19569 19571 0.50 FA MA 0.54 0.45 DELIVERED FA MA FA FA MA 0.47 O.SS 0.51 0.44 0.48 ON DAY 21 OF GESTATION MA 0.50 FA 0.58 MA 0.42 FA O.S6 FA 0.53 MA 0.48 19572 19576 0.42 FA MA FA MA MA 0.38 0.38 0.39 0.58 0.56 HATING NOT CONFIRMED E MA FA FA 0.39 0.42 0.38 E FA 0.51 E FA MA MA 0.36 0.42 0.30 19577 19578 0.67 FA 0.69 MATING E FA MA 0.68 0.68 NOT CONFIRMED FA 0.63 19579 DELIVERED ON DAY 21 OF GESTATION -------------------------------------------------------------------------------------------------------------- M = MALE F - FEMALE A = ALIVE E = EARLY RESORPTION L - LATE RESORPTION CLa - CORPORA LUTEA/OVARY PLACENTAL WEIGHTS WERE RECORDED IN GRAMS (G). DENOTES POSITION OF CERV APPENDIX C PROTOCOL AND AMENDMENT 418-013: PAGE C-1 PRIi@@ICLA,. Argus Research Laboratories, Inc. 905 Sheehy Drive, BuildingA Homham, PA 19044 Telephone: (215) 443-8710 Telefax: (215) 443-8587 PROTOCOL 418-013 SPONSOR'S STUDY NUMBER: T-6295.12 STUDY TITLE: Oral(Gavage) PharmacokineticStudy ofPFOS inRats PURPOSE: The purpose ofthisstudyistoevaluatethe pharrnacokineticosfPFOS inFo generationdams and Fl generationfetusesfollowinPgFOS treatmentof Crl:CDOBR VAF/Plusg female ratsduringprematingand gestation. TESTING FACILITY: Argus Research LaboratoriesI,nc. 905 Sheehy Drive,BuildingA Horsham, Pennsylvania 19044-1297 Telephone: (215)443-8710 Telefax: (215)443-8587 STUDY DIRECTOR: Raymond G. York,Ph.D.,DABT AisociateDirectoorf Research SPONSOR: 3M CorporateToxicology 3M Center,Building220-2E-02 St.Paul,Minnesota 55144-1000 STUDY MONITOR: Marvin T. 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 418-013:PAGE C-2 Protocol418-013 Page 2 REGULATORY CITATIONS: U.S.Food and Drug Administrati(o1n994).InternationCaolnferenceon Harmonisation; Guidelineon detectionoftoxicittyoreproductionformedicinalproducts.Federal RegisterS,eptember 22, 1994,Vol.59,No. 183. U.S.Food and Drug AdministrationG.ood LaboratoryPracticeRegulations;FinalRule. 21 CFR Part58. Japanese Ministryof Healthand Welfare(1997). Good LaboratoryPracticeStandard forSafetyStudieson Drugs,MHW Ordinance Number 21, March 26, 1997. European Economic Community (1989).Councildecisionon 28 July 1989 on the acceptance by the European Economic Community ofan OECD decisionlrecommendation on compliance withprincipleosfgood laboratorpyractice.OfficiaJloumalof theEuropean Communities:Legislation3.2(No. L315; 28 October):1-17. REGULATORY COMPLIANCE: Thisstudywillbe conducted incompliancewiththe Good LaboratoryPractice(GLP) regulationcsitedabove. Allchanges orrevisionosfthisprotocolshallbe documented, signedby the Study Directorand the Sponsor,dated and maintainedwiththe protocol. The QualityAssurance Unit(OAU) willauditthe protocolt,heraw dataand thereport, and willinspectcriticpahlases ofthe studyinaccordance withthe Standard Operating ProceduresofArgus Research LaboratoriesI,nc. The finarleportwillincludea statementsigned by the Study Directorthatthe report accuratelryeflecttshe raw data obtainedduringthe performanceofthe studyand that allapplicableGLP regulationwsere followedinthe conductofthe study.Should significadnetviationsfrom GLP regulationosccur,each willbe describedindetail, togetherwithhow the deviationmight affecthe qualitoyrintegritoyfthe study. SCHEMATIC OF STUDY DESIGN AND STUDY SCHEDULE: See ATTACHMENT 1 tothe protocol. 41"13: PAGE @'o-3 Protocol418-013 Page 3 TEST ARTICLE AND VEHICLE: Identification: Test Article: Name: Physical Description: LoVBatch Number SpecificGravity: Purity: ExpirationDate: PFOS (Synonym: FC-95). Light-coloredpowder. 217. -0.6. 98.9%. May 2000. Informationon the identityc,omposition,strengthand purityof the testarticleison file with the Sponsor. Vehicle: 0.5% Tween@) 80 inReverse Osmosis Membrane Processed Deionized Water (R.O. Deionized Water). Supplierand lotidentificatioofnTweens 80 to be documented in the raw data. Neitherthe Sponsor nor the Study Directorisaware of any potentialcontaminants likely to be present inthe vehiclethatwould interferweith the resultsof thisstudy. Therefore, no analyses otherthan those mentioned in thispmtocol willbe conducted. Safety Precautions: Gloves, mask, appmpriate eye pmtection and a uniform/labcoat are to be wom during formulationpreparationand administration.The MaterialSafety Data Sheet (MSDS) is attachedto the pmtocol (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. 418-013:PAGE C-4 Protocol41M13 Page 4 Shipments shouldincludeinformatiocnoncerningstorageconditionsand shipping cartonsshouldbe labeledappropriatelyT.he recipiensthould be notifieidnadvance of shipment. FORMULATION: Freauency of Preparation: Formulations(suspensions)willbe prepared dailyatthe Testing FacilityD.ata verifying the stabilitoyf the testarticlein the vehiclefor48 hours under the conditionsof administrationare on filewith the Sponsor. Detailedpreparationprocedures are attached tothisprotocol(ATTACHMENT 3). Adiustment forPurity: The testarticlweillbe considered 100% pure forthe purpose of dosage calculations. Testina FacilityReserve Samt)les: The Sponsor willreserve a sample (1 g) ofeach lotofthe bulktestarticluesed during the course ofthisstudy. The Testing Facilitwyillreserve a sample (5 mL) of each lotof the vehiclecomponents used during the course of thisstudy. Samples willbe stored under the previouslycitedconditions. ANALYSES: Samples additionalto those described below may be taken ifdeemed necessary during the course of the study. Bulk Test ArticleSampling: No analyses of the bulk testarticlewillbe conducted during the course of thisstudy. Informationon the stabilitoyf the bulk testarticleison filewith the Sponsor. Analyses of Prepared Formulations: Homogeneity and stabilitoyf prepared formulationsison filewith the Sponsor. However, recordswillbe maintained to document how the testarticlfeormulationswere prepared. 418-013:PAGEC-5 Protocol41 "l 3 Page 5 ConcentrationofTest 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 (-700C or below) and discarded at the Testing Facility upon request of the Sponsor. Shippin-q Instructions: Samples to be analyzed willbe shipped (frozen on dry ice)to: Kris J. Hansen, Ph.D. 3M Environmental Technology and 935 Bush Avenue Building 2-3E-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 formulations willbe discarded at the Testing Facility.All remaining bulk test articlewillbe retumed to the Study Monitor at the previously cited address upon completion of allwork with the test article. TEST SYSTEM: Species/Strain and Reason for Selection: The Crl:CD@BR VAF/Plus@ (Sprague@-Dawley) ratwas selected as the Test System because: 1) thisstrainof ratwas used inthe reproductive and developmental toxicity studies;2) historicadlata and experience existat the Testing FaciliWI-3)a;nd 3) the test articleispharmacologically active inthe species and strain. Number: Initiaplopulation acclimated: Population selected forstudy: 120 virginfemale rats. 80 mated female rats(16 per dosage group). 418-013:PAGE C-6 Protocol418-013 Page 6 Body Weiaht and Age: Female ratswillbe ordered to have body weights of 200 g to 225 g each at receipt, at which time they willbe expected to be at least60 days of age. Actual body weights will be recorded the day afterreceiptand willbe documented inthe raw data. The weight range willbe included inthe finalreport. Sex: Female ratswillbe given the testarticle.Male ratsof the same source and strainwill be used only as breeders and are not considered partof the Test System. Source: Charles River Laboratories,Inc. The ratswillbe shipped in fifterecdartons by airfreightand/or truckfrom Charles River Laboratories,Inc.,to the Testing Facility. Identification: Rats are permanently identifieudsing MonelO 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. ANIMAL HUSBANDRY: Allcage sizesand housing conditionsare in compliance with the Guide forthe Care and Use of Laboratory Animals"'). Housinci: Female ratswillbe individuallhyoused in stainlesssteel,wire-boftomed cages, except during the cohabitationperiod and during collectionintervalsforurineand fecal samples. During cohabitation,each pairof ratswillbe housed inthe male rat'scage. During collectionintervalsforurineand fecalsamples, the female ratswillbe housed individualliyn metabolism cages. No nesting materialswillbe supplied because the female ratswillbe sacrificedbefore parturitioinsexpected. 41"13: PAGE C-7 Protoco4l1"l 3 Page 7 Room Air,Temperature and Humidily: The animal room is independently supplied with at least ten changes per hour of 100% fresh airthat has been passed through 99.97% HEPA filter(sAiroClean@ room). Room temperature willbe maintained at 64OF (180C) to 790F (260C) and monitored constantly.Room humiditywillalso be monitored constantlyand maintained at 30% to 70%. Light: An automatically controlled 12-hour light:12-hour dark fluorescent lightcycle willbe maintained. Each dark periodwillbegin at 1900 hours EST. Diet: Rats willbe given CertifiedRodent Diet@ #5002 (PMI NutritionInternationala)vailable ad libitumfrom individuafleeders. Water: Water willbe availablead libitumfrom individualbottlesattached to the cages or from an automatic watering access system. Allwater willbe from a localsource and passed through a reverse osmosis membran' e before use. Chlorinewillbe added to the processed water as a bacteriostatp;rocessed water isexpected to contain no more than 1.2 ppm chlorineat the time of analysis.Water isanalyzed monthly for possible bacterialcontamination and twiceannually forpossiblechemical contamination. Contaminants: Neitherthe Sponsor nor the Study Directorisaware of any potentialcontaminants likely to be present in the certifiedietor the drinkingwater at levelsthatwould interferewith the resultsof thisstudy. Therefore,no analyses otherthan those routinelyperformed by the feed supplieror those mentioned in thisprotocolwillbe conducted. RANDOMIZATION AND COHABITATION: Upon arrivalm,ale and female ratswillbe assigned to individualhousing on the basis of computer-generated random units.Afteracclimation,virginfemale ratswillbe selected forstudy on the basis of physicalappearance and body weights recorded during acclimation.The female ratswillbe assigned to dosage groups based on computergenerated (weight-ordered)randomization procedures. 418-013:PAGE C-8 Protocol418-013 Page 8 Withineach dosage group,consecutiveorderwillbe used toassignfemale ratsto cohabitatiownithbreedermale rats,one male ratper female rat.The cohabitation periodwillconsistofa maximum offivedays. Female ratswithspermatozoa observed ina smear ofthe vaginalcontentsand/ora copulatoryplugobserved insituwillbe consideredtobe atday 0 ofpresumed gestationand assignedtoindividuahlousing. A tableofrandom unitsora computer-generatedrandomizationprocedurewillbe used toselecteightfemale ratsper dosage group forCaesarean-sectionexaminationson day 15 ofpresumed gestation.The remainingfemale ratsineach dosage group willbe examined on day 21 ofpresumed gestation. ADMINISTRATION: Route and Reason forChoice: The oral(gavage)routewas selectedforuse because: 1)thiswas the mute of administratioinnthe developmentaland reproductivteoxicologsytudies;and 2)itisone ofthe possibleroutesofhuman exposure. Method and Freguency: Female ratswillbe giventhe testarticloervehicleonce dailybeginning42 days priorto cohabitatiotnhrougheitherday 14 ofpresumed gestationor day 20 ofpresumed gestation.Dosages willbe adjusteddailyforbody weightchanges and givenat approximatelythe same time each day. RationaleforDosage Selection: Dosages were selectedon the basisofa previousstudyconductedwiththe testarticle (ArgusResearch LaboratoriesI,nc.,Protocol418-008). Dosage Levels,Concentrationsand Volumes: Number of Dosage Female Grou:)-Rats Dosage (mglkgidsy) 1 11 If@111 16 0 (Vehide) 16 0.1 16 0.4 Concentration -(mg/mL) 0 0.02 0.08 Dosag volum: (mL/kg) 5 5 5 ArgusBatch Number B-418-013-A(Day.Month.Year) B-418-013-8(Day.Month.Year) 8418-013-C4Day.Month.Year) IV 16 1.6 v 16 3.2 0.32 5 B-418-013-D(Day.Month.Year) 0.64 5 B-41"13-E(Day.Month.Year)-ii The testarticlweillbe consideredloc)%pureforthepurposeofdosage calculawm. 418-013:PAGE C-9 Protocol418-013 Page 9 TESTS, ANALYSES AND MEASUREMENTS: Viabill : AllPeriods: At leasttwice daily. ClinicalObservations and/or General Appearance: AcclimationPeriod: At leastonce. Dosage Period: Twice daily.Priorto administrationand once approximately one hour postdosage. Postdosage Period: Once priorto sacrifice. Clinicalobservationsmay be recorded more frequentlythan citedabove, ifdeemed appropriateby the Study Directorand/or Study Monitor. Body Wei(:hts: AcclimationPeriod: At leastonce. Dosage Period: Sacrifice: Daily. Terminal weight. Feed Consumption Values (recordedand tabulated): Acclimation Period: At leastonce. Dosage Period: Weekly to cohabitationand dailyduring presumed gestation. Feed consumption values may be recorded more frequentlyifitis necessary to replenishthe feed. These intervalswillnot be tabulated. Matina Performance: Mating willbe evaluated dailyduring the cohabitationperiod and confirmed by observationof spermatozoa ina smear of the vaginalcontents and/or a copulatory plug observed in situ. 4lMl3:PAGE C-10 Protowl 4 1M 13 Page 10 PharTnacokineticSamt)leCollection: Urine and Fecal Samples: Female ratswillbe housed individuallyin metabolism cages forcollectionof urine and fecal samples forthe followingintervals:one day priorto initiatioonf cohabitationto the followingmorning and days 6 to 7, 14 to 15 and 20 to 21 of presumed gestation. Following each 24-hour collectioninterval,samples willbe collected intocentrifuge tubes, placed on dry ice and stored frozen (-700C or below) untilshipment for analysis. In the event that a dam begins to deliverbefore completion of urine and fecal sample collection(day 20 to day 21 of presumed gestation),the dam willbe removed from the metabolism cage and placed in a nesting box with sufficientbedding untilsacrifice. Blood Samples: Blood samples willbe collectedfrom each of the female ratsfollowing removal from metabolism caging (priorto administration)on each of the followingdays: on the day cohabitation isinitiate(dpriorto cohabitation)and on days 7 and 15 of presumed gestation,as well as day 21 of presumed gestation. The time of blood collectionwillbe recorded inthe raw data. On alldays of collectionexcept days 15 and 21 of presumed gestation (dams at their terminal collectioninterval)b,lood samples (approximately 1 mL each) willbe collected from the orbitalsinus. Ifnecessary, whole blood may be collectedfrom an alternate site;ifso, the alternatesitewillbe documented inthe raw data. On day 21 of presumed gestation,blood samples (approximately 4 mL each) willbe collectedvia the inferiorvena cava. Ifblood collectionon day 15 of presumed gestation willbe a final bleed forthe respective dam (i.e.t,he dam willbe Caesarean-sectioned that day), the sample willbe approximately 4 mL and willbe collected via the inferiorvena cava. Blood willbe collectedand transferred intoserum separator tubes. The samples willbe spun ina refrigeratedcentrifuge. The serum willbe transferred intopolypropylene tubes labeled with the study number, animal identificationd,ate of collection,study day and collectiontimepoint. Allsamples willbe immediately frozen on dry ice and maintained frozen (-700C or below) untilshipment to the Sponsor for analysis. Shir)pingInstructions: Allsamples willbe maintained frozen (-700C or below) untilshipment for analysis. Samples willbe shipped frozen on dry ice via overnight mail. A packing listwillbe included with the samples and sent to Kris J. Hansen, Ph.D., at the previously cited address. Both the recipientand the Study Monitor willbe notifiedin advance of sample shipment. 418-013: PAGE C -11 ProtoCol 418-013 Page 11 Caesarean-Sectionin-a Observations - Day 15 Presumed Gestation: Eight randomly selected female rats per dosage group willbe Caesarean-sectioned on day 15 of presumed gestation.The gravid uterus willbe excised and weighed. Placentae thatappear abnormal (size,coloror shape) willbe noted inthe raw data. The female rats(pregnant dams) willbe examined fornumber and distributioonf: Corpora Lutea. ImplantationSites. Viable and Nonviable Embryos. (A viableembryo isoval or crescent shaped, pink,firmand enclosed in an amnioticsac fillewdith clearfluid.A nonviable embryo isamorphous, small,pale pink to tan or deep red to black,softand enclosed inan amniotic sac fillewdith clear,cloudy or opaque fluid.) Sample Collection: Caps and labeledtubes willbe weighed (combined weight,to the nearest .001 gram) before and afterretentionof pooled embryonic samples forsubsequent use in pharmacokinetic analyses. These weights willbe documented inthe raw data,and copies of these weights willbe includedwith the packing lisptriorto shipment. Samples ofthe amnioticfluidof each viableembryo willbe collected,pooled (perlitter), frozen and stored (-70'C or below) untilshipment tothe Sponsor foranalysis. Each viableembryo willbe removed from the uterus with the attached placenta,pooled (perlifterf)r,ozen and stored (-70"C or below) untilshipment to the Sponsor. Shii)pinaInstructions: Allsamples willbe maintained frozen(-700C or below) untilshipment foranalysis. Samples willbe shipped frozen on dry icevia overnightmail. A packing liswtillbe includedwith the samples and sent to Kris J. Hansen, Ph.D.,at the previouslycited address. Both the recipientand the Study Monitor willbe notifiedin advance of sample shipment.