Document 0JO7bwN2EMzmVvpke29G8j87m

: AR126-0561 . Summary PFOS Rat Two-Generation Reproduction Study * Study Numbers: 3M T-6295.9, Argus 418-008 (in-life), FACT-TOX-012 (analytical). Compound& Lot: PFOS (Perfluorooctanesulfonate) ~ Lot 217, 98.4% pure (SMD Analytical Request 53030). CAS No. 2795-39-3 SRteupdroydTuicttlei:onCToomxbiicinteydSOtruadly(oGfaPvaFgOeS) FinerRtialtisty, Developmental and Perinatal/Postnatal Report Date: 10 June 1999 (in-life); 19 April 1999 (analytical). Study Year and GLP stats: 1997-1999 and GLP study Procedures: Groups of male and female rats (35/sex/group) were administered (daily oral intubation) PFOS at dose levels of 0,0.1,0.4, 1.6, and 3.2 mg/kg/day. Dosing started when the animals were = 70 days old. The male and female rats (Fy generation) were dosed for 42 days prior to mating. After mating, dosing continued in the mated females. throughout gestation. At day 10 of gestation, Caesarean section was done on 10 females/group to check for any effects in early gestation. The remaining 25 females/group were allow to deliver their liters. Dosing of the females continued during the 21-day lactation period. At weaning, 2 males and 2 females per litter were randomly selected for continuation on the study (Fy generation). Note: Because of post-natal effects at high doses (see below for details) only three groups of Fj animals continued the study, namely 0, 0.1 and 0.4 mg/kg/day. Daily dosing began in the Fy pups the day after weaning and continued throughout their gavrooiwdtahnctoespeaxruaadligmma.turTithye. aAnitm2a4lsdawyesreofalasgoettehseteFd,inrataswwaeterre-ftielslteeddMin-maapzasesiwveh.en they wlietrteer7ma0tedamyastionldg.s wCeohraebiatvaotiidoend.foDromsaitnigngcobnetgiannuewdhiennthtehefeFymaalneismtahlrsowueghrege9s0tadtaiyosn oalndd- 21-day lactation after delivery of F2 pups. The study ended upon weaning of the Fs pups. Results: No deaths occurred in any of the Fo animals and there were no compound arenldatfeedmcalliensicaalttshiegnhsi.ghDeorsteworeldaotseed lreevdeulsce(d3.w2e&igh1t.6bmogd/ykgw/ediagyh)t.s oRcecduurrceeddifnoboodth male consumption correlated with these body weight effects. The 0.4 mg/kg/day males also hada slight, but statistically significant, reduced body weight gain during the pre-mating period. Dosing of PFOS as high as 3.2 mg/kg/day had no effect on estrous cycling; all dmiaftfienrgenacnesd afteratinlyitdyopsaeralmeevtelerisn orfegtahredFtoorlaittsewrearveeruangaefsfefcotrecd.orpTohreareluwteear,eimnpolsaingantiifoincsa,nt viable embryos or nonviable embryos from the day 10 of gestation Caesarean section fUepmoalnesl.itering, post-natal survival was reduced at the higher two dose levels (16 & 3.2 mg/kg/day). The post-natal survival effect was manifested by increased number of Cons +m : / 04161 apparent sill-births and increased pup deaths during the first 24 to 72 hours after birth. Necropsy of dead pups revealed many had not nursed ~ no milk curd in the stomach. Reflex and physical development (surface righting, pinna unfolding, eye opening, acoustic startle reflex. air right reflex, pupil constriction) was not affected in the 0.1 & 0.4 mg/kg/day Fi pups. Some of these parameters were slightly delayed in the 1.6 mg/kg/day Fy pups and they were not evaluated at the high dose (3.2 mg/kg/day) as no pups survived beyond four days post-natal. Because of adverse effects in the higher two dose levels (1.6 & 3.2 mg/kg/day) Fi pups, reduced survival and/or reduced growth, the decision was made, in conjunction with the attending laboratory veterinarian, not to continue these two dose levels into the second generation. It had been determined that dose of 1.6 mg/kg/day or higher produced `compound toxicity. Continuance of these dose levels would not result in any additional useful information and would subject the animals to unnecessary stress. Death did not occur post-weaning in the Fy animals nor were there any compound related clinical signs. The male and female Fy rats in the 0.4 mg/kg/day group had somewhat reduced body weight and food consumption; at some points in time (weeks of growth) the differences from controls were statistically significant. The appearance of extemal evidence of sexual development (preputial separation in males & vaginal patency in . females) was not affected at either dose level. Also, the passive avoidance and water maze testing did not reveal any differences in regard to leaming, short-term retention nor long-term memory. Likewise, none of the mating or reproductive performance measurements (parameters) were adversely affected inthe 0.1 & 0.4 mg/kg/day F, male and female rats when they were mated and allowed to deliver litters. There were no significant differences in regard to Fa pup survival or pup weights at weaning, Conclusions: The results of the study indicated the following NOEL levels. The Fy maternal and paternal NOEL is 0.1 mg/kg/day. [higher dose levels had reduced body weight and food consumption] `The Fo reproductive NOEL is > 3.2 mg/kg/day. [no effect on reproductive performance even at highest dose level) The F; post-natal NOEL is 0.4 mg/kg/day. (higher dose levels had reduced pup viability and growth] `The Fy matemal and paternal NOEL is 0.1 mg/kg/day. (0.4 mg/kg/day animals had reduced body weight and food consumption] The Fy reproductive NOEL is > 0.4 mg/kg/day. [no effect on reproductive performance at either 0.1 or 0.4 mg/kg/day] `The F: post-natal NOEL is 0.4 mg/kg/day. [no important toxicological effect on pup survival or growth at either 0.1 or 0.4 mg/kg/day] 01162 . Compound Level Samples: At the terminal sacrifice of the Fy males (after mating was completed), serum and liver samples were collected from 5 males in each of the dose groups. Serum samples were collected from 5 Fy females in all the dose groups except 3.2 mg/kg/day group at the terminal sacrifice the day after weaning the F liters. Liver samples were collected from the same Fo females and liver samples were also collected from the weaned Fy pups of the same dams. Analysis of the samples for PFOS (3M Analytical Report TOX012) yielded the following mean PFOS concentrations, Male Fo Serum: control - 0.024 ppm, 0.1 mg/kg/day ~ 10.5 ppm, 0.4 mg/kg/day -- 45.4 ppm, 1.6 mg/kg/day -- 152 ppm, 3.2 mg/kg/day - 273 ppm. Male Fo Liver: control - 0.665 ppm, 0.1 mg/kg/day - 84.9 ppm, 0.4 mg/kg/day - 176 ppm, 1.6 mg/kg/day - 323 ppm, 3.2 mg/kg/day ~ 1357 ppm. Female Fo Serum: control ~ 0.037 ppm, 0.1 mg/kg/day -- 5.28 ppm, 0.4 mg/kg/da~y 18.9 ppm, 1.6 mg/kg/day - 82.0 ppm. Female Fo Liver: control ~0.171 ppm, 0.1 mg/kg/day ~14.8 ppm, 0.4 mg/kg/day - 58.0 ppm, 1.6 mg/kg/day ~ 134 ppm. Pup Fy Liver: control - 0.051 ppm, 0.1 mg/kg/day~ 6.19 ppm, 0.4 mg/kg/day ~ 57.6 ppm, 1.6 mg/kg/day - 70.4 ppm. Note: Serum and liver reflect PFOS concentrations of Fo dams and Fi pups at the end of 21-day lactation, i.e. when the samples were collected. Compound levels at parturition when reduced pup survival was observed could differ and those are being measured in a follow-up PK study. Data Evaluation: A well conducted GLP study following established testing guidelines (OECD 416). Study evaluated reproductive parameters and post-natal pup evelopment including neurological parameters. The study established valid NOEL values. 01163 . Laboratory Report FACT-TOX-012 cSoemrpulmetaendd),LsiveerrumSaamnpdlelsi:verAstamthpeletserwmeirnaelcsoalclreicftiecde fofrotmheSFmyamlaelseisn (eaafctehromfatthiengdowsaes 3gr.o2upmsg./kgS/edrauymgsraomupplaetstwheerteercmoilnlaelctseadcrfirfoicme thFeodfaeymaalfetesriwneaalnl itnheg dtoheseFigrloiutpersse(xdcaeypt21 woferleacataltsioonc)o.llLeicvteerd sfarmopmlethsewweeraenceodllFeyctpeudpfsr(olmivtehressamapmleeFsywfeermealpeoso.leLdivbeyrlsitatmerp)loefs the same dams. rMeidlukcCedurtd0 8Sapmupplsesp:erOlinttedr."a4Aytotfhilsacttiamteiosn,toamllaclihttcerosn,taesntssta(tmeidliknctuhred)prsoatmocpolle,s,weporoeled mbgy/lkigtt/erd,ayw)eraendcohlilgehctdeodsfer(o3m.24 mogr/5kgl/idtaerys).fAromvtahlriedeatdeodsaenaglryotuipcsal--cmoentthroodl,folromwidloksceu(r0d.1 `matrix has not been developed and these samples have not been analyzed. 01164 . S, PRMEDICS Slmiiinisely Argus Division _ 90S Sheshy Drive, Bldg. A| Horsham, PA 15044 April 13, 2000 Marvin T. Case, D.V.M., Ph.D. 3M Toxicology Services 3M Center, Building 220-2E-02 St. Paul, Minnesota 55144-1000 Telephone: Telefax: (612) 733-5180 (612) 733-1773 RE: Final Report 418-008 - Combined Oral (Gavage) Fertility, Developmental and Perinatal/Postnatal Reproduction Toxicity of PFOS in Rats Sponsor's Study Number: 6295.9 Dear Dr. Case: reper, Enclosed is one copy of the final report Amendment 1 to the above-referenced If you have any questions, please do not hesitate to contact me. Sincerely, RGY:kgp Attach. Raymon: . York, Ph.D, DABT Associated Director of Research and Study Director 01465 : irvr, MEDICA _-- Argus50R5esSehaerechhyLDarbiovrea,toBruieisd,inIgncA. TelTeeplheofHnaoexr::sh((a22m11,55)) P4444A331.-858507841740 REPORT AMENDMENT | 13 APRIL 2000 PROTOCOL 418-008 SPONSOR'S STUDY NUMBER: 6295.9 PERINACTOAMLB/IPNOESDTNOARTAALL(RGEAPVRAOGDEU)CFTEIROTINLTIOTYX,ICDIETVYESLTOUPDMYENOTFAPLFOASNIDN RATS FINAL REPORT 1. Page 1:9, paragraph 4 has been revised to: rPartesgansasnicgyneodcctourcroehdabiinta2t2io(n95i.n6t%h)e,021(V(e8h4ic.l0e%)), a0n.1d a2n4d(09.64.0m)go/fktgh/eda23y,d2o5saagnedg2r5oufpesm,ale arcesrpoescstitvheelyt.hreAelldopsraeggenagnrtoudpasm.sVidaebliilvietryeadnlditgers.owtThhoefgetshteastieocnoninddgeexnewraasticoonmpoafrfasbplrieng T(Fh2erpeupwse)retonwoetaonxiicnoglowgeircealallysiomupnoarftfaenctteddifbfyertehnecehsifghreosmt tdhoesacgoenttreosltegdr,o0u.p4 mvgal/ukegs/.dayN.o clinical the test aorrtincelceraosphsiygohbasser0v.a4timogn/skgi/ndtahye.F2 generation pups were attributable to dosages of 2. Page 1-10, paragraph 4 has been revised to: eTfhfeecFtsI ognenmeartaitniognorrefperrotidluicttyiovcecuNrOreEd.LiTs hgereNaOteErLthfaonravidaobsilaigteyoafnd0.g4rmogw/tkhg/indatyh;e Fn2o egfefneecrtastoionnpoufpfsspurrivnigviaslaolrsogr0o.w4tmhg/aktgt/hdeayh.ighTehsetrdeowseargee nteostteodx,ic0o.l4omggic/aklgl/ydiamyp.ortant 3. Page V-8, paragraph 3 has been revised to: dTohseagvieabtielsitteydo,f0t.h4emgs/ekcgo/nddayg.eneSrmaatliloninocfrfesapsreisnign(tFh2epnuupsm)bewraosfudnaafmfsecwtietdhasttitlhlebohringpheuspts taonxdicionlotghiecanlulmybiemrpoorftpanutpbdeecaatuhsse:aft1e)rtDhLe v2aliunetshwee0r.e4 nmogt/ksgig/ndiafyicdaonstlaygedigfrfeoruepntwferroemnot DthLe 2co1ntwreorleggrroeuaptevratlhuaesn;o2r)ctohmepaavrearbalgeeftoorthleivcelointttreorsligzreosudpelviavleureesd; aannddsu3)rvnieviitnhgerttohe fvriaobmilitthyeocronltarctoaltgironouipndviacleusefso.r the 0.4 mg/kg/day dosage group remarkably differed 01156 4. V-8, paragraph 4 has been revised to: FOAL RERORT AVENONsHEoaRrnTts| `Phuigphebsotddyosweaiggehttessotfed,t0h.e4semcgo/nkdg/gdeanye.raStmiaonllofrfesdpurcitnigon(sF2inppuupps)bwoedryewueniagfhftescitnedthaet the 0.1 samnadll0.d4ifmfge/rkegn/cedsaybedtowseaegne tghreopupusp wbeordeynwoetigthoxtiscfoolrogtihcealcloyntirmoploratnadnt0.b1emcgau/skeg:/daI)ythe dosage groups were liter sizesofthe 0.1 nmogt/kstga/tdisatyicdaollsyasgiegngirfoiucapnotnanDdLsweIretharsosuogcihat4,edaswictohmtphaerleadrgweirthlitvhee control group values, and the DL 4; 2) the small reductions random selection of in pup body weights pinuptshefo0r.4cmognt/ikngu/eddayobdsoesravgaetigornosuopn pwreerceulalsisnogc,iaastecdowmiptahraedmiwniitmhatlhley cloanrtgreorllgivreoulipttevralsuiezes,atanbidrtthhe(rDaLn1d)oamnsdeloenctDioLn 4of pupsfor continued observation on DL 4; and p<D.01, respectively) present on DLs the statistically significant reductions (p<0.05 7 and 14, as compared to the control group values, were transient and disappeared by DL 21. 5. Page V-9: Revisions to paragraph 4 on page V-8 caused it to extend onto page V-9. These revisions were NOEL for the second made to clarify the interpretation of the generation offspring (F2 generation). observations reported and the is LLfi ince Raymond G. York, P/D., DABT ~ Date Associate and Study DDiirreeccttoorr of Research Hane]Lemp bosiiGlpa.ro Nancy{. Gonglievski Date Quality Assurance Manager 01157 418-008:PAGE |-9 REVISED PAGE feed consumption dosage group. values were reduced during lactation in the 0.4 mg/kg/day Dosages of the test article as day of vaginal patency in the high as 0.4 mg/kg/day did F1 generation female rats. not affect the average There were no bfioonlgo-gtiecralmlyreitmepnotritoannotrdriefsfeproennsceesinihnibtihteiovnalinuetshefoFr1lgeeanneirnga,tisohnorfte-mtaelrem rraettse,ntaison, evaluated paradigm. by performance in a passive avoidance or watermaze performance aDnodsafgeretislitoyfptahreatmeesttearrsticelveaalsuahtiegdhiansth0e.4F1mgg/eknge/rdaatyiodnidfenmoatlaeffreactts.any mating All necropsy observations in unrelated to the test article. the F1 generation female rats were considered 2P5refgenmaanlceyroatcscuarsrseidgnined22to(c9o5h.a6b%i)t,at2i1on(8in4.t0h%e)Oa(nVdeh2ic4le()9,6.00.)1 oafntdhe0.243,mg2/5kagn/dday dinodseaxgweagsrocuopmsp,arreasbpelcetiavcerlyo.ss Atlhleptrhergeneandtosdaagmesgdreoulpisv.eredVialbititleirts.y aTnhdeggreoswttahtioofn the the second highest gdeonseargaetitoenstoefdf,sp0r.i4ngmg(/Fk2g/pduapys.) tTohweeraenwienrgewneoretoaxliscooluongaicfaflelcyted by oibmspeorrvtaanttiodnisffienrtehneceFs2frgeonmetrhaeticoonntpruolpsgrwoeurpevaatltureisb.utaNbloectloindicoaslaogrenseocfrothpesytest article as high as 0.4 mg/kg/day Cr1168 Cc. Conclusions 418-008:PAGE I-10 REVISED PAGE On the basis of these data, the abservable-effect-level (NOEL) Fo generation maternal and paternal no- of PFOS is 0.1 mg/kg/day (0.4 mg/kg/day and higher dosages caused reductions in body weight gain and reduced feed consumption values). The Fo generation reproductive NOEL is greater than 3.2 mg/kg/day; no effects `ionntmhaetFi1ng,gefneertrialittiy oonr oefsftsrporuisngcyiscl0i.n4g mogc/cukrgr/edda.y The (1.6 NmgO/EkgL/dfoaryviaanbidlity and growth higher dosages caused preimplantation loss and reductions in litter size, pup viability, growth and survival). The (0.4 mFg1/gkegn/edraaytidoonsamagteercnaaulseadndrepdautcetmiaonlsNinOEboLdoyfwPeiFgOhSt is 0.1mg/kg/day gain and reduced feed consumption values). The F1 generation reproductive NOEL is greater than a dosage of 0.4 mg/kg/day; no effects on mating or fertility occurred. The NOEL for viability and growth in toxicologically tihmepoFr2tagnetneerffaetcitosnoonffpsuprpinsgurivsivaallsoor0.g4romwgt/hkga/tdatyh.e There highest were no dosage tested, 0.4 mg/kg/day. NAPS - "of NI( 3 / Be Mildred S. Christian, Ph.D., Fellow, ATS Date Execytive Director of Research M. Hoberman, Ph.D., DABT Date Director of Research ime KAfe saa08:00 Rayond G. York, PD., DABT Date Associate Directorof Reearch and Study Director C00114259 418-008:PAGE V-8 REVISED PAGE B.8. Necropsy Observations (Summary - Table E18; Individual Data - TableE34) All necropsy observations in the F1 generation female rats were considered unrelated to the test article because: 1) the observations occurred in rats in the control group; and 2) the observation commonly occurs in this strain of rat. These observations included rough and/or pitted cortex of the kidneys with calculi in the Kidney and/or urinary bladder of two control group rats. One of these rats also had a tan granular material in the renal cortex and thickened walls of dosage the urinary group dam bladder. that died Necropsy observations for the 0.4 as the result of an intubation error mg/kg/day were described previously. B.9. Natural Delivery and Litter Observations (Summaries - Tables E19 and E20; Individual Data- Tables E35 through E38) Pregnancy occurred in 22 (95.6%), 21 (84.0%) and 24 (96.0%) of the 23, 25 and 25 female rats assigned to cohabitation in the 0 (Vehicle), 0.1 and 0.4 mg/kg/day dosage groups, respectively. All pregnant dams delivered litters. The gestation index (the percentage of pregnant rats with live offspring) was comparable across the three dosage groups. The viability of the second generation offspring (F2 pups) was unaffected at the highest dosage tested, 0.4 mg/kg/day. Small increases in the number of dams with stillbom pups and in the number of pup deaths after DL 2 in the 0.4 mg/kg/day dosage group were not toxicologically important because: 1) the values were not significantly different from the control group values; 2) the average for live litter sizes delivered and surviving to DL 21 were greater than or comparable to the control group values; and 3) neither the viability or lactation indices for the group values, 0.4 mg/kg/day dosage group remarkably differed from the control Pup body weights of the second generation offspring (F2 pups) were unaffected at the highest dosage tested, 0.4 mg/kg/day. Small reductions in pup body weights in the 0.1 and 0.4 mg/kg/day dosage groups were not toxicologically important because: 1) the small differences between the pup body weights for the control and 0.1 mg/kg/day dosage groups were not statistically significant and were group on associated with DLs1 through 4, the as larger live compared litter with sizes of the 0.1 mg/kg/day dosage the control group values, and the random selection of pups for continued observations on DL 4; 2) the small reductions in pup body weights in the 0.4 mg/kg/day dosage group were associated with a minimally larger live litter size at birth (DL 1) and on DL 4 preculling, as compared with the control group values, and the random selection of pups for continued observation on DL 4; the statistically significant reductions 04370 418-008:PAGE V-9 REVISED PAGE (P<0.05 and p=0.01, respectively) present on DLs 7 and 14, as compared to the control group values, were transient and disappeared by DL 21. Administration of the test article at dosages as high as 0.4 mg/kg/day did not adversely affect any other parameter evaluated at natural delivery or during the 21-day lactation period (duration of gestation, averages for implantations and live litter sizes, numbers of dams with all pups dying during lactation, viability and lactation indices, surviving pups per liter and pup sex ratios). B.10. Clinical Observations from Birth to Day 21 Postpartum and Necropsy Observations (Summaries - Tables E21 and E22; Individual Data - Tables E39 and E40) No clinical or necropsy observations were attributable to dosages of the test article as high as 0.4 mg/kg/day because: 1) the incidences were not dosagedcleipniecnadleonbts;eravnadt/ioorns2)itnhceluodbesderovnaeticoonntorcoclugrrroeudpinpuopnltyhoatnewaosr tnwoot npeusptsi.ngTohrese nursing, had decreased motor activity and was cold to the touch, one pup in each of the control and 0.4 mg/kg/day dosage groups that had a missing tip of tail and one 0.4 mg/kg/day dosage group pup that had an umbilical hema. No milk in stomach occurred in 2, 4 and 4 pups that were found dead in the three respective dosage groups. One control 0.1 mg/kg/day dosage group pup had a group raised pup tan had area hydrocephaly and one on the median and left lateral lobes of the liver at necropsy on DL 21. 001171