Document jm3m6aa4XyKjwOpj2E5VX4RJN

spf Sa {N773 2per MR 54374 UNITED STATES ENVIRONMENTAL PROTECTION AGENeY AR226_ 1093 WASHINGTON, D.C. 20460 AR 2E np Dear Interested Party: On March 28, 2002, EPA made available copies in PDF file format (389KB)ofthe Draft aHnwasazsraaermdtehAntststeiDsssimdieoincotunmOefonPftertifnlcEulPourdAoeodfcatenaneroioocrfAPcionildhlue(tPitFoonOxAPa)rpepAveenandrtiiInotsgnSoaannltdsp,Tagoperwea8p.aireEsdPecbAtyihotanhse2bR0iescokfomtehe document, "Production of PFOA and its Salts." This ovo as ow bees tified, nd cored copy of te ull doourset fs tached Please replace your earlier versionofthe draft assessment with this one, or print only the cover page, the Tableof Contents, and page 8 from this corrected file, and use them to replace those pages from the original file printout. An new revised text in bold is also attached efrorraytaousrheceotnvsehnoiweinncgetshoe page 8 deletions that you can see iimn msetdriikaetoeultyand where the corrections were made. We reget any confston or verre hs ey have caused Seecneniivfeee1a8SCyeoodpuyHboayvpefhheaonnAyerqamuee2ns0t21i0t-oh5ns64o-rA7sc6s3oc4ms,msemoernrtbtsy,sewomhaniieclrhaacinoegmeadhieneAnrsonsbieusfsst@menepnutem,egprolivecss1eoycfoohunewsiststhditeos fi Hplsdpn reviewed in the Assessment, Mary Dominiak by phone at 2o0r2i-f5y6o4u-8h1a0v4e, any by fdaixffaitcu2l0ti2e-s56o4p-e4n7i7n5g,thoersbeyfielmesa,ipllaetase contact Sine, sl 8 CChhaermliesalM,CoAnuterro,l DDiirveicstioorn 32 23m Atachments x552 25 Aem----r--aA--e UCeO1rNWTi AoINonNH 0O0 CRI o----, ERRATA SHEET Corrected 4/15/2002 Table 1. Reported Physicochemical Properties Commi] w|p ae | # VP | Sol-H20 g RECCOF [seas | | bic [1 1 7] RECO2H | 335-671 189C |om34m gnH |g] | RNEHCO2-- || s38e25a2s61|| 130C| sublimes| 1x10E5| 2g0eglsL <s RF bow sora | [we |||] H_(1 g fee acid /L Water) = 2.6 Free acid pKa is approximately 0.6 Sodium orSilversalts of PFOA decompose above 250 C to generate perfluoroolefins 2.0 Production of PFOA and its Salts PFOA is commerelally manuftcturedby two major altemative processes: 1) the Simons Electro- | Chemical Fluorination (ECF) process or 2) the telomerization process. IfnlutohreidEeCaFndpraonceosrsg,anaincefleeectdrsitcoccukorrfenotctisanpoaiscseadcitdhroorugahdaesroiluvtaiotniovfeanlh-yodertoaunschsyudrfiolfugoernniydle. | "The ECF process replaces the carbon-hydrogen bonds on moleculesofthe organic feedstock `owcictuhrcsarwbhoenn-fallulotrhienecabronbdosn,-hiyndarongiednenbtoincdals maraenrneeplracuesdedwittohmcaarkbeonP-FlOuSo.rinPeerbfolnudosr.inTahteionECF | (prPoFcOesFs)ypieelrdfsibueotrweoeonc3t0a-n4c5spuerlcfeontnystkr-aifguhRtoOcrShiEadi)ne,(anloornmgalw)itpherafvlauroiraobloectmainxotnuyrleoffluboyrpirdoeducts and impurities. The output of the ECF process is not a pure chemical, but instead a mixture of isomers and homologues including higher and lower straight-chain homologues; branched-chain perfluoroalkyl fluoridesofvarious chain lengths; straight-chain, branched, and cyclic perfluroalkanes and ethers; and other byproducts (3M Company, 20004). After disposal or recoveryofsome of the byproducts and impurities, the acid fluoridePOS is base hydrolyzed in | batch reactors to yield PFOA. The PFOA salts are synthesized by base neutralizationofthe acid to the salt in a separate reactor (3M Company, 20000). In the telomerization process, tetrafluoroethylene is reacted with other fluorine-bearing chemicals to yield fluorinated carboxylic acids. This process yields pure straight-chain acids with an even numberof carbon atoms. Distillation can be used to obtain pure components (ECT, `1m9i9x4t).ureCsoomfmpeerrcfiluaolripnraotdeudctcsommapnouufnadcstuwrietdhtehvreonugchartbhoentenluommbeerirzsat(iRoennnperro,ce2s0s01a)r.e generally 3M 3M ChaosmcphaanrayctiesrtihzeedlairtgsemstanmuafnaucftaucrteuroefrPaFndOAimapnordteitrs of PFOA and is sals in ammonium and sodium the United States. salts in 1997 at Tfehsesstehafnig5u0re0s,0m0a0ykogvepresriyaetaert,haentdotiatls pirmopdourcttaitoinonvaotlluemsesotfhaPn F1O00A,0s0i0nckegth(e3vMasCtommapjaonriyt,y2o0f003). 5 CONTAIN NO CBI Ea DRAFT HAZARD ASSESSMENT OF PERFLUOROOCTANOIC ACID AND ITS SALTS U.S. Environmental Protection Agency Office of Pollution Prevention and Toxics Risk Assessment Division February 20, 2002 (Corrected April 15, 2002) CONTAIN NO rai 3 PREFACE "aTshsiosciisaatedprweiltihmienxaproysausrseetsosmpeenrtfolufortohoecptoatneonitciaalcihdaz(aPrFdsOAt)o haundmaints hsealatlst.h Tahned mthaejoernivtiyroofnmtehent toxicology information is for ammonium perfluorooctanoic acid (APFO). This assessment includes a review of the studies that were available asof July 2001 A two-generation reproductive available in the springof 2002. toxicity Effects study were of APFO observed is in currently being conducted and two-generation reproductive will be toxicity studyof a related compound, perfluorooctane sulfonate. The results of the APFO study will be important to determine whether similar effects are observed. Corrected 41152002 Table of Contents Executive Summary 1 10 Chemical Identity 6 LI Physicochemical Properties 6 20 Production ofPFOA and its Salts (Corrected April 15,2002) 8 21 Uses of PFOA and its Salts 10 22 Environmental Fate n 22.1 Photolysis n 222 Volatility In 223 Biodegradation 2 224 Hydrolysis 12 225 Bioaccumulation 13 226 Soil Adsorption 14 23 Environmental Exposure 14 23.1 Combustion 14 232 Discharge to Water 14 233 Discharge to Land 15 234 Environmental Monitoring 15 24 Human Biomonitoring 16 30 Human Health Hazards 20 3.1. Metabolism and Pharmacokinetics 3.01 Halflifien Humans 2200 3.1.2 Absorption Studies inAnimals 21 3.13 Distribution Studies in Animals 2n 3.1.4 Metabolism Studies in Animals 25 3.15 Elimination Studies in Animals 2 32 Epidemiology Studies 29 32.1 Morality Study El 322 Hormone Study 2 323 Cholesterol Study 33 32.4 Study on EpisofdCearse (Morbidity) 36 33 Acute Toxicity Studies in Animals 38 33.1 Oral Studies 38 332 Inhalation Studies ES 333 Dermal Studies 39 334 Eye lritation Studies 335 Skin Imitation Studies 3399 3345 SMuubteahgrenoinciictTyoSxtiucdiiteysStudies in Animals a309 36 Developmental Toxicity Studies in Animals a8 37 Carcinogenicity Studies in Animals 52 5 37.1 Cancer Bioassays 52 372 Mode of Action Studies 5 372.1LiverTumors 5 3.7222 Leydig Cell Tumors sa 372.3 Mammary Gland Tumors 55 3.72.4 Pancreatic Tumors 55 40 Hazards o the Environment 55 41 Introduction 55 42 Acute Toxicity to Freshwater Species 57 50 References 0 ANNEX I - Robust Summaries " [2 Introduction EXECUTIVE SUMMARY pPerrofdluucoerdooscytnatnhoeitcicaaclildy o(rPFthOrAo)ugahndthietsdesaglrtasdaarteiofnulolyr mfleutoarbionlatiesdmoorfgoatnhiecrcfolmupooroucnhdesmitchaalt can be. products. PFOA is primarily used as a reactive intermediate, while its salts arc used as processing aids uses. In recent in the years, production of fluoropolymers and less than 600 metric tonsperyear fluoroelastomers and of PFOA and its sats in other surfactant have been manufactured in the United States or imported. Mostofthe toxicology studies have been conducted this report. with the ammonium salt of perfluorooctanoic acid, which is referred to as APFO in Environmental Fate and Effects PFOA is persistent in the environment. It has very low volatility and vapor pressure. It does not hydrolyze, photolyze or biodegrade under environmental conditions. Several wildlife species have been sampled around the world to determine levels of PFOA. PFOA has rarely been found in fish sampled from the U.S. certain European countries, the. North Pacific Ocean and Antarctic locations, or in fish-cating bird samples collected from the U.S.. including Midway atoll, the Baltic and Mediterrancan Seas, and Japanese and Korean coasts. PFOA was found in few mink livers from Massachusetts aa concentration range of <18 10 108 ng/g, dry wt, but not found in mink from Lousiana, SouthCarolinaand Illinois. PFOA concentrations in river otter livers from Washington and Oregon States were less than the quantification limitof 36 ng/g, wet wt. PEOA was not detected at quantifiable concentrations in oysters collected in the Chesapeake Bay and Gulfof Mexico of the U.S. coast "The concentrationsof PFOA in surface war, sediments, clams, and fish collected from two locations upstream and five locations downstreamofthe 3M manufacturing facility at Decatur AL have been determined. Ofthe five downstream sampling locations, the two closest to the facility had PFOA surface water concentrations significantly greater than the two upstream sites (means of 1900ug/L and 1024 ug/L); the nearest three locationshadsediment concentrations. significantly greater than the upstream sites (wet wt. means 1855 ug/ke. 892 uke, 238 ugg). "The average fish whole body PFOA concentration for the two upstream locations was 11.7 ug/kg (wet wt), while that for the five downstream locations was 106.4 ug/ke. The average PFOA concentration in clams at the two upstream locationswas 4.38 ug/kg, while the average for the five downstream locations was 8.42 ug/ke. Based on available data, APFO does not appear to bioaccumulate in fish. Ina study of fathead minnows, the calculated BCE for APFO was 1.8, 1 Several species were tested to assess the acute toxicity of APFO; these included the fathead minnow (Pimephalespromelas), bluegill sunfish (Lepomis machrochirus), water flea (Daphnia magna), and a green algae (Selenasirum capricornutum). Comparisonsof the different studies are problematic for several reasons. The studies were conducted with different test substances. Generally the ammoniumsaltor the tetrabutylammonium salt was tested. Purity of the test `material is a major concern and was not sufficiently characterized in these tesis. In some tests it appeared that 100% test chemical was used, for others a chemical of lesser purity (approximately 2710 85%) was used. Water, a solvent (isopropanoolr) a combination of both was used in other tests, for no obvious stated reason. Finally, only nominal test chemical concentrations were reported; the actual concentrations were not reported. Twelve tests were conducted with fathead minnows; 96-h LCS0 values (based on mortality) ranged from 70 to 843 mg/L. Iti unclear why this range is so wide. Assuming these studies arc valid, and dueto the limitations discussed above, these toxicity values indicate low toxicity. The two acute mg/L). values for bluegill sunfish also indicate low toxicity (96-h LCS0s of >420, and 569 Nrainngeeadcfutreomte3st9s1w0e>r1e0c0o0ndmugc/tLe.d wTihtehldoawpehrnivdasluaensdar4e8-ihndEicCaSti0vevoalfumesod(ebraasetde otonxiicmitmyo,bibluitztahteion) wide range makes interpretation difficult. Seven tests were conducted with green algac; 96-h ECS0 values (based on growth rate, cell density, cell counts, and dry weighs) ranged from 1.2 10 >66 mg/L (the ErS0 cell density value of 1,000 mg/L is excluded from this discussion). The lower value indicates high to moderate toxicity, based on the acute criteria. The lower value would also be indicativeofmoderate toxicity, based on the chronic moderate criterion (0.110 mg/L). A 14-d EC50 value of43 mg/L, based on cell counts, for green algae was also calculated in one study. This is indicative of low chronic toxicity, based on the chroniccriterion (10 mg/L). Green algae appeared to be the most sensitive test species in the 44% APFO test sample, daphnids were the next most sensitive, and fathead minnows were the least sensitive. Human Health Effects and Biomonitoring Littl information is available concerning the pharmacokineticsof APFO in humans. A preliminary studyofretired workers suggests simply that the serum half-life is between 1 and 3.5 years. These data provide evidenceof the potential to bioaccumulate PFOA in humans. In addition, this study provides preliminary evidence that the serum half-life maybe longer in females than in males. Animal studies have shown that APFO is well absorbed following oral and inhalation exposure, and 10 a lesser extent following dermal exposure. In rats and dogs, there are major gender differences in the distribution and elimination of APFO. APFO distributes primarily to the liver, 2 8 povlaarsym.aT,tadnodeksidnnoetyp,aarnitditoonatoletshseerliepxitdenftr,acottihoenrotriassduiepsosoef ttihsesubeo.dyAPinFcOludbiinngdsthteotestis and macromolecules in the tissues listed above. APFO is not metabolized and there is evidence of enterohepatic circulationof the compound. The urine i the major route of excretion ofAPFO in the female rat, while the urine and feces are both major routesofexcretion ofAPFO in male ra. In female rats, the half-life is 24h in the serum and 60 h in the liver; in male rats, the half-life is 105 h in the serum and 210 h in the liver. In beagle dogs, the plasma half-life is 254 h in females and S07 h in males. In rats, the elimination half-life is one day in females and 15 days in males. Female ratsappearto have a secretory mechanism that rapidly eliminates APFO; this secretory. mechanism is either lacking or relatively inactive in males. Other studies in rats have shown that testosterone exerts an inhibitory effect on renal excretion ofAPFO. Hormonal changes during pregnancy do not appear to change the rateofelimination in rats. The gender difference observed in rats and dogs has not been observed in primates and humans. "There are limited data on PFOA serum levels in workers and the general population. Occupational data from plants in the U.S. and Belgium that manufacture or use PFOA indicate that mean serum levels in workers range from 0.84 10.6.4 ppm. The highest level reported in a worker in 1997 was 81.3 ppm. In non-occupational populations, serum PFOA levels were much lower. In both pooled blood bank samples and in individual samples in both adults and children, `mean PFOA levels ranged from 3 to 17 ppb. The highest serum PFOA level reported was in sample from a child (56 ppb). Epidemiological studies on the effects of PFOA in humans have been conducted on workers. `Two mortality studies, as well as studies examining effects on the liver, pancreas, endocrine system, and lipid metabolism, have been conducted to date. In addition, a morbidity study was also recently submitted. A retrospective cohort mortality study demonstrated a weak association with PFOA exposure and prostate cancer.A statistically significant association was observed in prostate cancer mortality as lengthofemployment increased. This result was not observed in a recent update to the study; however, the results cannot be directly compared because the exposure categories. were modified in the update. In a morbidity study, workers with the highest PFOA exposures for the longest durations sought care more often for prostate cancer reatment than workers with Tower exposures. Another study reported an increase in estradiol levels in workers with the highest PFOA serum levels; however, none of the other hormone levels analyzed indicated any adverse effects. Some ofthe same employees who participated in the hormone study also were included ina study of cholecystokinin (CCK) levels in employees. No positive association was noted between CCK. values and PFOA. The other available study examined cholesterol and other serum components in workers. There did not appear to be any significant differences among workersofdifferent exposure levels, except among obese workers (aspartate amino transferase and alanine amino transferase). However, PFOA was not measured directly, but indirectly as total serum fluorine. 3 `PTFheOrAe asreermuamnlyevleilmsigtrateiaotnesr 1t0hatnhe1s0e pstpumd.iesT,hbeurtefmoorse,t anloltoaftblhyestheersemsaulltlsnmuumsbtebreofinwtoerrpkreertesdwith carefully. aInndac2u5t0e-t5o0x0icimtgy/sktgudfioersfienmaanleism,alasn,dt<he10or0a0l mLgD/Sk0gvfaolrumeaslfeoranCdDfreamtaslweeWriest>a5r0r0atms.g/Tkhgerfoerwmaasles no mortality following inhalation exposure of 18.6 mg/L. for one hour in rats. The dermal LDSO in rabbits was determined to be greater than 2000 mg/kg. APFO is a primary ocular iitant in rabbits, while the data regarding potential skin iritancy are conflicting. APFO is not mutagenic. APFO did not induce mutation in ther. typhimurium or E. coli when tested either with or without mammalian activation. APFO did not induce chromosomal aberrations in vitro in human lymphocytes when tested with and without metabolic activation up to cytotoxic concentrations. aberrations in CHO cells in viAtrPo.FOInwtahse tfeisstteadstsawyi,ceAfPoFr Oitsianbdiulicteydtbooitnhdcuhcerocmhorsoommoaslomal aberrations and polyploidy in both the presence and absence of metabolic activation. In the second assay, no significant increases in chromosomal aberrations were observed without activation. However, when tested with metabolic activation, APFO induced significant increases in chromosomal aberrations and in polyploidy. APFO was negative in a cel ransformation assay in CsH 10T,; mouse embryo fibroblasts and in the in vivo mouse micronucleus assay. Subchronic studies in rats and mice with 28 and 90-daysof exposure have demonstrated that the liver is the primary target organ and that males are far more sensitive than females. Dietary exposure to APFO for 90 days resulted in significant increases in liver weight and hepatocellular hypertrophy in female rats at 1000 ppm (76.5 mg/kg/day) and in male rats at doses as low as 100 ppm (5 mg/kg/day). Analysesofserum and liver levels of APFO showed a marked gender difference that accounts for the difference in sensitivity. In 90-daystudy with rhesus monkeys, exposure to doses of 30 mg/kg/day or higher resulted in death, lipid depletion in the adrenals, hypocellularityofthe bone marrow, and moderate atrophyof the lymphoid follicles in the spicen and lymph nodes. Unlike rodent studies, analysesofthe serum and liver levels did not reveal a gender difference in monkeys, but the sample size was very small (N=2). Chronic dietary. exposureofrats to 300 ppm APFO (14.2 and 16.1 mg/kg/day for males and females, respectively) for 2 years resulted in increased liver and kidney weights, hematological effects and liver lesions in males and females. In addition, testicular masses were observed in males at 300 ppm and ovarian tubular hyperplasia was observed in females after exposure to 30 ppm (1.6. mg/kg/day), the lowest dose tested. Prenatal developmental toxicity studies in rats resulted in death and reduced body weight in dams exposedto oral doses of 100 mg/kg/day or by inhalation to 25 mg/m' APFO. There was no evidenceofdevelopmental toxicity after oral exposure to doses as high as 150 mg/kg/day, while inhalation exposure to 25 mg/m' resulted in reduced fetal body weights. In a rabbit oral developmental toxicity study there was a significant increase in skeletal variations afer exposure 4 0 to 50 mg/kg/day APFO. highest dose tested. There was no evidenceof matemal toxicity at 50 mg/kg/day, the Areptrwood-ugcetnievreattoixoincirteyprsotduudcytoivfePtFoOxiScistyhoswteuddyhiisgchurmroernttallyitbyeoifngF1conpduupcsteadt.dosAestwaos-gleonwearsat1i.o6n mefgf/ekcgt/disayob.serTvheed.results of the APFO study will be important to determine whethear similar CianrdcuicnionggeLneicyidtiyg scteuldliaesdeinnoSmparsagiun et-hDeamwalleey r(aCtsDa)nrdatmsasmhmoawrtyhatfiAbProFaOdeinsowmeaaskilnytchaercfiemnaolgeesn.ic, f1o6.l1lomwgi/ngkgd/ideatayryinexfepmoasluesr)e.toTh3e00copmppmofuno2dr(yaeta3r0s0(epqpumi)vahlaensta(l0so1b4.e2enmgr/ekpogr/tdeadytionbmeales and carcinogenic toward the liver and pancreasofmale CD rats `tThheeimnedcuchtainoinsomf(st)umoofrAsPbFyOAPtuFmOoriisgednueesitos aisnnoont-gcelenaortloyxuincdmerescthoaondi.sAmv,aiilnavbollevidnagtaacitnidviactaitoen tohfat receptors and perturbationsofthe endocrine system. The liver carcinogenicity/toxicity of APFO appear to be related to inductionofperoxisome proliferation following binding to the peroxisome proliferation activation receap(tPPoArR a) in the liver. Available data suggest that ttohehoinrdmuocntailonoimfbLaelyandciegrceeslullttiunmgorfsro(mLaCcTt)ivaantidonmaofmmthaerPyPgAlRan:d naenodpilnadsumcstiboyn AofPtFhOe cmyatyocbherodmuec P4530 enzyme, aromatase. Preliminary data suggest that the pancreatic acinar cell tumors are related to an increase in serum level of the growth factor, cholecystokinin. Athsatthtehemteucmhoarnsisinmdsoucfecdarincirnatosgaerneicrealcetviaonntotfo AhuPmFanOs.havReenvoitewboeefnafvualillyaeblluecmiedactheadn,isittiiscadsastaumoefd oaltlhebrutdrtuwgosmaonddecshoefmiincdaulcsttihoantoifntdhuecelLutCeiTniizniangnihmoarlmsohnaes(lLeHd)a,w"odrokpsahmoipnepaangeolntioscmo"nacnldude that "GnRH agonism", are considered t0 be relevant to humans, and that the possibility of induction of Leydig cell adenoma in humans by specific agents with other modesofaction cannot be ruled out despite the rarity of LCT in humans. At present, there is no evidence that the induction of LCT by APFO is via the "dopamine agonism" or "GnRH agonism" modeofaction. It is recognized that there are quantitative differences in certain biological parameters between rats and humans. However, the principal cell control mechanisms appear similar, and the difference in carcinogenic response is probably quantitative. As binding to the PPARa appears to be the critical event leading to hormonal imbalance and APFO tumorigenesis, and the level of PPARa in human livers is lower than that in rodent liver, it appears that humans may be less sensitive than rodents in the development of LCT, mammary gland tumors, or liver neoplasms. s i 1.0 Chemical Identity Chemical Name: Perfluorooctanoic Acid Molecular formula: C8 H F15 02 Structural formula: F-CF2-CF2-CF2-CF2-CF2-CF2-CF2-C(=0)-X, "The free acid and some common derivatives have the following CAS numbers: "The perfluorooctanoate anion does not have a specific CAS number. Free Acid (X=OM+ M =H) [335-67-1) Ammonium Sali Sodium Salt Potassium Salt Silver Salt (X=OM+M=NH (X=OM+; M=Na) (X= OM; M=K) (X=0M=; M= Ag) [3825-261] 335-955) [2395-00-8 [335-933] Acid Fluoride X=F) [335-66-0) Methyl Ester Ethyl Ester (X=CH3) (X= CH2-CH3) 376272) [3108-245] Synonyms: 1-Octanoic acid, 2.2,3,3.4,4,58 58-6pe,nt6ad,e7ca,fl7uo,ro8PFOA 1.1 Physicochemical Properties For this report, perfluorooctanoic acid is consistently referred to as PFOA. Most of the toxicology studies have been conducted with the ammonium salt of perfluorooctanoic acid, which wil be referred to as APFO in this report. PFOA is a completely fluorinated organic acid. "The typical structure has a linear chainofeight carbon atoms produced by the telomerization of tetrafluoroethylene. The physical chemical properties noted below are for the free acid, unless otherwise stated. The data for the free acid, pentadecafluorooctanoic acid [335-67-1),i the. most complete. The reported vapor pressure of 10 mm Hg appears high, but is consistent with other perfluorinated compounds with similar boiling points. The free acid is expected to completely dissociate in water. Determinationofthe vapor pressureof APFQ is problematic. For APFO, the recently reported vapor pressure of < | x 10E-5 (3M Environmental Laboratory, 1993) seems t00 low foar material that sublimes as the ammonium salt. This study measured the water solubility of APFO to be > 10%. It was noted in an earlier study that concentrationsof 20 g/L "gelled" (3M. " 1 Ccaolmcpulaantye,d v1a97l9u),. -0T.9h,emipagrhttitniootn bcoeefafciccuireanttewdauserteoptohretemdeitnhothdesueseedarl(yHastnusdcihesaonfd5.LeoA1n9o7t9h)er "The formationof an emulsified layer between the octanol and water surface interface would `make determinationof log P difficult. "The available physicochemical properties for the PFOA free acid are: MW: 414 (Beilstein, 1975) MP: 45-50C (Beilstein, 1975) BP: 189 - 192 C/ 736 mm Hg (Beilstein, 1975) VP: 10 mm Hg @ 25C (approx.) (Exfluor MSDS) Sol. - Water: 3.4 g/L (telomeric [mp = 34 C ref. 0.01 - 0.02 mol/L~4 - 8 g/L) (MSDS from Merck, Fischer, and Chinameilan Internet sites) pKa: 2.5 (USEPA AR-226 473) PH (g/L): 2.6 (MSDS Merck) iDsuaenttiocitphaetseudrtfoacfeo-racmtmiuveltpirpolpeerltaiyeesrsofinPocFtOaAn,olawnadtteh,e tmesutcphroltiokceotlhofsore tohbeseOrEveCdDfomretPhFoOdS,, PFOA Therefore, an n-octanol/water partition coefficient cannot be determined. Water solubility has been reported for PFOA, but it is unclear whether these values are for a microdispersion of `mmiicceelllleess,artahthiegrhetrhacnontcreunetrsaotliuobnilsi.ty(.SiSmeivsteerra,l 1re9p9o2r;tsClnfootuertsh,at1P98F5O;AEdswalatrsdsse,lf1-9a9s7s)o.ciaItneaaqsueous solutions, micelles partion betwee the ai/r water interface on the surface. Decompositionofdifferent salts produces perfluoroheptene (loofmsetsal fluoride and carbon dioxide). This occurs at 320C for the sodium salt and at 250-290C (Beilstein 1975). The ammonium salt sublimes at 130C (USEPA AR-226 473). `The physicochemical properties of PFOA and its derivatives are summarized in Table 1. ; 3 Corrected 4/15/2002 `Table 1. Reported Physicochemical Properties cctowe| ora |[we | | [7] Compound| CAS REG # ve LogP RECCOF | 35648 | | dic [--| T--" RRFE-CCO2O- _[ 335-67: 189C [To3m2a0ggmLl| Hg|| Nir | 3825-261| 130C| sublimes| Ix10E-5 | gels <s RF H_(1 g free acid /L Water) = 26 Free acid pKa is approximately 0.6 Sodium or Silver sasof PFOA decompose above 250 C to generate perfluoroolefins. 2.0 Production of PFOA and its Salts PFOA is commercially manufactured by two major aliemative processes: 1) the Simons ElectroChemical Fluorination (ECF) process or 2) the telomerization process. In the ECF process, an electric current is passed through a solutionofanhydrous hydrogen fluoride and an organic feedstockofoctanoic acidor aderivative. The ECF process replaces the. cidaernbtoinca-lhymdarnongeern ubsoenddstoomnamkoelePcFuOlSe.s oPfetrhfeluoorrgiannaiticofneoecdcsutorcskwwhietnh aclalrbtohen-cfalruboornin-ehybdornodgse,nin an bonds are replaced with carbon-fluorine bonds. The ECF process yields between 30-45 percent straight chain (normal) perfluorooctanonyl fluoride (PFOF), along with a variable mixture of byproducts and impurities. The outputof the ECF process is not a pure chemical, but instead a . `mixtureofisomers and homologues including higher and lower straight-chain homologues; branched-chain perfluoroalkyl fluorides of various chain lengths; sraight-chain, branched, and eyelic perfluroalkanes and ethers; and other byproducts (3M Company, 2000a). After disposal or recoveryofsome of the byproducts and impurities, the acid fluoride is base hydrolyzed in batch reactors salt in a 10 yield separate PFOA. reactor T(h3eMPCFoOmpAasnaylt,s 2ar0e0s0y)nthesized by base neutralization of the acid to the In the telomerization process, tetrafluoroethylene is reacted with other fluorine-bearing chemicals to yield fluorinated carboxylic acids. This process yields pure straight-chain acids w1i9t94h).anCeovmemnerncuimablerporfodcuacrtbsomnanautofmasc.tuDriesdtitllhartoiuognhcatnhebteeluosmeedritzoaotbitoaninprpoucreesscoarmepogneneenrtasll(yECT, mixturesofperfluorinated compounds with even carbon numbers (Renner, 2001) 3M Company is the largest manufacturer and impoorftPFeOrA and its salts in the United Stats. 3M has characterized its manufacture of PFOA and its ammonium and sodium salts in 1997 at less than 500,000 ke per year, and ts importation at less than 100,000 kg (3M Company, 2000). These figures may overstate the total production volume of PFOA since the vast majority of 5 1% PFOA is consumedi themanufuctureoftheammoniumor sodium salts.Moreprecise production volumesof PFOA and the ammonium and sodium salts have been reported to dUiSscElPosAurbey 3Mt,hisburtephoarvte been claimed as TSCA confidential business information, preventing Industry participants have characterized 3M as the dominant global producerof PFOA related chemicals, manufacturing approximately 85 percent or more of total worldwide volumesofthe ammonium salt of PFOA (FMG, 2001). USEPA has not located information that would ch(oawnshtinrcoahtdicbcetoetnhnipsnrcolviainidfmeo.drmCbauytririoennndtounsptroyo,dnuncpotorilosynmtveoraliaucfmolerbgliaenifioenrmcUahteSimoEincPaflAosr mCmaahnneuumffiaacccattluurUreeprddsamtotehtheSerystUthnaeintme3dM SctuartmelsaoirviempporordtuecdtiinonvovloulmuemsesaboofvPeF4,O5A25snkge)s. iFtuirtlhecrommomreer,iathlezraetiisonn.o informatioonn the total Since 1985, USEPA has received a totalofapproximately 25 notifications for PFOA-related chemicals that were not previously on the TSCA Chemical Inventory. Most ofthese notifications were from companies other than 3M. In most cases, the notifications qualified for tbhuetLgodwioVnoylume Exemption for new chemicals with a production volume less than 10 metric In terms of on-going production, 3M has not committed publicly to a complete phase-out of 3PMFOhAasaninddiPcFaOtAe-drhealtatiet d cphheasmiincgaoalusscietrhtaasinfoFrLPUFOORSAaDndBPrFaOSn-rsepelcaitaeldtychmemaicralsl. tHhoawecvoemra,in PFOA and its salts such as FC-26, FC-118 and FC-143, FX-1001 and others (3M Company, 2000). AisncilduedefrFormantchee, UGneirtmeadnSyt,atlesa,yO,EaCndDJaMpeanm.beTrhecrouenmtraiyesatshoatbreepsoormteodlpyrohdauvcetipornodnuoctnio-nOcEapCaDcity countries such as China. Following are companies that may manufacture PFOA and its salts. (3M Company, 2000b; Directory of World Chemical Producers, 1998; Dynax, 2000; Renner, 2001; SEMI, 2001): OECD . 3M Company (United States) . DuPont (United States) . Exfluor Research Corporation (United States) . PCR Inc. (United States) . CAitboafiSnpaec(iFarlatnyceC)hemicals (Germany) . HoCleDaycrnhieasontnt G(AeGkeirremmnaagnneyys))llschat (German) . 15 + EMintieCnhieSmpSAy.nt(hleisliys)S.p.A. (Italy) + Asahi Glass (Japan) + Daikin (Japan) + Dainippon (Japan) = Tohkem Products Corporation (Yapan) Non-OECD + Chenguang Research Institute of the Chemical Industry (China) Shanhai 3F New Materials Co., Ltd. (Ching) 2.1 Uses of PFOA and its Salts PFOA is used mainlyas a chemical intermediate, and its salts are used in emulsifier and surfactant applications According to 3M, the vast majorityofPFOA is consumed to make the ammonium or sodium salts. 3M also uses PFOA as areactive intermediate in the industrial synthesis ofa fluoroacrylic ester. The fluoroacrylic ester is used in an industrial coating application (3M Company, 20002). "The saltsof PFOA have additional uses, mostly in surfactant and emulsifier applications. These include the following: Processing aid in the industrial synthesis of fluoropolymers and fluoroelastomers such as polytetrafluoroethylene and polyvinylidene fluoride witha variety of industrial and consumer uses (3M Company, 2000a; DuPont, 2000; Daikin, 2001). + Post-polymerization processing aids in the stabilization of suspensions of fluoropolymers and fluoroelastomers prior to further industrial processing (3M Company, 2000a). Processing aid for factory-applied fluoropolymer coatings on fabrics, metal surfaces, and fabricated or molded parts (3M Company, 20000). Extraction agent inion-pairreversed-phased liquid chromatography (Petrits, 1999). Based on the physicochemical propertiesofthe salts of PFOA, they may also have other related surfactant or emulsifier uses as a photographic chemical or in the manufactureofelectronic components such as semiconductors. These same properties may lead industry to explore PFOA asa replacement chemical for PFOS in other applications in which PFOA is not currently used. 10 1e 22 Environmental Fate 2.2.1 Photolysis Direct photolysisof APFO was examined in two separate studies (Todd, 1979; Hatfield, 2001) and photodegradation was not observe in either study. In the Todd (1979) study, a solution of 50 mg/l APFO in 2.8 litersofdistilled water was exposed to simulated sunlight at 22:2 C. Spectral energy was characterized from 290-600 nm witha max output at ~360 nm. Direct photolysis of the test substance was not detected. However, the author noted that sample purity was not properly characterized which may have contributed to experimental error. In the Hatfield (2001) study, both direct and indirect photolysis were examined utilizing techniques based on EPA and OECD guidance documents. To determine the potential for direct photolysis, APFO was dissolved in pH 7 buffered water and exposed to simulated sunlight (Serano, 1999; Nubbe, 1995). For indirect photolysis, APFO was dissolved in 3 separate `matrices and exposed to simulated sunlight for periodsof time from 69.5 to 164 hours. These exposures tested how each matrix would affect the photodegradation ofAPFO. One matrix was a pH 7 buffered aqueous solution containing H202 as a well-characterized source ofOH radicals (Ogata, 1983; Lunak, 1992). This tested the propensity ofAPFO to undergo indirect photolysis "The second matrix contained Fe203 in water that has been shown to generate hydroxyl radicals via a Fenton-type reaction in the presence of natural and artificial sunlight (Kachanova, 1973; Behar, 1966). The third matrix contained & standard solutionof humic material. Neither direct nor indirect photolysis of APFO was observed based on loss ofstarting material. Predicted degradation products were not detected above their limitsofquantitation. There was no conclusive evidenceofdirect or indirect photolysis whose ratesofdegradation are highly dependent on the experimental conditions. Using the iron oxide (Fe203) photoinitiator matrix model, the APFO half-life was estimated to be greater than 349 days. 222 Volatility Impinger studies were performed to examine the volatility of APFO and PFOS. Solutions of APEO or PFOS containing ammonium acetate in water/1-propanol (50:50) or phase transfer agents, e.2., n-alkyldimethylbenzylammonium chloride (3M Environmental Laboratory, 1993) were blown with 280 litersofair a flow rate of I Limin. (3M Environmental Laboratory, 1993). The results indicate there is some lossofAPFO and POS, but mostofthe solutions retained over 80% or more of the fluorochemicals. The average retention was 92% for both APFO and PFOS. This indicates that there is loss from the solutions. However, some of the solutions, particularly the n-alkyldimethylbenzylammonium chloride solution, appearto retain all the fluorochemicals. These results were reviewed by Dr. Edwin Tuckofethre Chemistry Dept. at the Universityof Oklahoma (3M Environmental Laboratory, 1993). He concluded that n 17 itis very unlikely that these fluorochemicals were removed by bubblingairthrough water due to their very low vapor pressures. He suggested that a more plausible mechanism for loss from the solution phase is concentrationofthe surfactants in foam and loss from the bubbled solutions as foam or micro-droplet. In the second partof the experiment, air was passed over the fluorochemicals and bubbled through a train of impingers containing the ammonium acetate solution. It was expected that if any fluorochemicals were presenti the arthey would be transferred and retainedby the ammonium acetate solution. However, no fluorochemicals were present in cither the first or second impinger. The report concludes that the vapor pressureofboth compounds i less than 10 E07. According to these experiments, APFO and PFOS (potassium salt) have very low volatility and vapor pressure. Quantitative conclusions regarding rates of volatilization from water or Henry's Law constant are not possible. However, APFO and PFOS are capableoftransport out ofwater. Also, the lossof the fluorochemicals may have been as the free acids, not the salt forms. APFO sublimes at 130 C (sec Physicochemical Properties Section 1.1). There is no information on the validityof the test method for determining volatilityof the est substance. The study also lacks characterization of the purityof the test substance. 2.2.3 Biodegradation Using an acclimated sludge inoculum, the biodegradation of APFO was investigated using a shake culture study modeled after the Soap and Detergent Association's presumptive test for degradation (Reiner, 1978). Both thin-layer and liquid chromatography did not detect the: presenceofany metabolic products over the course of2 1/2 months indicating that PFOA does not readily undergo biodegradation. In a related study, 2.645 mg/L APFO was not measurably degraded in activated sludge inoculum (Pace Analytical, 2001). Test flasks were prepared using aamHiPneLrCa/l MsaSlDts smyesdteimu.m,S|evmerLalmeotthhaenroslt,udaineds cSoOnmduLctseedttlbeedtswleuednge1.97A7n-a1l9y8si7sawlsaosdciodnndoutctoebdsewrivteh APFO biodegradation using what probably were standard COD and BOD methods, however, the methods used in these studies were either insufficiently described (ic. no description of experimental protocols) or there were indicationsofa high degreeof experimental error. The r1e9s8u0l;ts3wMereC,omthpearnefyo,re1,9d8e5eb:mePdacuenrAenlailaybtliecably, t1h9e97s)u.bmitter (3M Company, 1977; 3M Company. 22.4 Hydrolysis `The 3M Environmental Laboratory (20012) performed a study ofthe hydrolysis ofPFOA. The study procedures were based on EPA's OPPTS Guideline Document 835.2110 (EPA 1998); although the procedures do not fulfil all the requirementsofthe guideline, they were more than n 18 adequate for these studies. Results were based on the observed concentrations of PFOA in buffered aqueous solutions as & functionoftime. The chosen analytical technique was high performance liquid chromatography with mass spectrometry detection (HPLC/MS). During the study, samples were prepared and examined at six different pH levels from 1.5 to 11.0over a period of 109 days. Experiments were performed at 50C and the results extrapolated to 25C. Data from twoofthe pH levels (3.0.and 11) failed to meet the data quality objective and were rejected. Also rejected were the data obiained for pH 1.5 because ion pairing led to artificially low concentrations for all the incubation periods. The result for the remaining PH levels (5.0, 7.0, and 9.0) indicated no clear dependenceofthe degradation rate of PFOA on PH. From the data pooled over the three pH levels, it was estimated that the hydrolytic half-life of PFOA at 25C is greater than 92 years, with the most likely value of 235 years. From the mean value and precisionof PFOA concentrations, it was estimated the hydrolytic half-life of PFOA to be greater than 97 years. 2.2.5 Bioaccumulation To determine the potential for bioaccumulation, Fathead minnows were exposed to 25 mg/l APFO for 13 days (Howell etal, 1995). After 13 days exposure, the fish were then removed from APFO contaminated water and analyzed for depuration over 15 days. Aer 192 and 312 hours exposure to APFO contaminated water, the average concentration of APFO in fish tissue was 44.7 and 46.7 pg/g wet weight (ww), respectively. At this point, APFO appeared to reach steady state. Twenty-four hours afier being transferred to clean water, the concentration of APFO decreased to 19.9 pglg ww and by 96 hours post-exposure, the concentration had decreased to approximately ug/g ww and remained relatively constant until test termination at 360 hours. The calculated BCE for APFO was 1.8. It shouldbenoted that questions have been raised about this study regarding the analytical techniques, high-test chemical concentration, and short test duration. Vraspir (1979) conducted a study to determine ifbluegill sunfish bioaccumulate fluorochemicals from the 3M Decatur plant. Two lots of 30 fish were used. One lot was exposed to Decatur plant effluentfor21 days and the other to river water only for 23 days. Exposed fish, both living and dead, as well as the control ish were homogenized and analyzed for fluorochemicals by GC, TLC. and GC/MS. There were no detectable amountsofAPFO in the ethyl acetate or toluene extractsofthe tissues. No fluorochemicals were detected in the river water exposed fish. However, interpretationof his study is problematic for several reasons. Effluent concentrations of subject fluorochemicals were not characterized and the specific protocol for exposureofthe fish was not found. There was also no information on analysisof the Tennessee River water or effluent used in the study. Additionally, it was not known if there was any opportunity for the depurationofthe fish prior to sacrifice. No explanation was attemptaesd to what was the cause 5 K) ofthe twelve dead fish in the effluent-exposed group. The study also did not differentiate: between the bioaccumulationof the test compound and the sorption onto the surfaceof the fish. 22.6 Soil Adsorption `The adsorption-desorptionofAPFO was studied in 25 mi solutions of 14C-labeled APFO in distilledwaterwith S g Brill sandy loam soil or 24 hours at a temperature of 16-19C. The study reported a Kdof0.21 and a Koc of 14 indicating that PFOA has high mobility in Brill sandy loam soil (Welsh 1978). The Koc value, however, is questionable du to the lack of accurate information on the purity of the 14C-labeled test substance (Boyd 19932,0). Moody and Field (1999) conducted sampling and analysis of samples taken from groundwater | 103 meters below the soil surface in close proximity to two fire-training areas with a history of aqueous film forming foam use. Perfluorooctanaocidc was detected at maximum concentrations ranging from 116 10 6750 ug/L at the two sites many years afte is use at those sites had been discontinued. These results suggest that APFO may have the potential to migrate through soils to relatively shallowgroundwaterwhere it persists 2.3 Environmental Exposure 23.1 Combustion For 1997, 3M estimated 1950 pounds of PFOA-compound (PFOA and related salts) stack releases at its Cottage Grove MN location and another 4500 Ibs. from Cottage Grove incinerated offsite (3M Company, 2000a,b). In 1998, 70% of the fluoride-containing wastes at 3M's Decatur location were incinerated off-site; incineration is now the primary disposal method for these materials (3M Company, 2000a,0). For 1999, DuPont estimated stack releasesof24,000 Ibs. APFO at ts Washington Works WV location, plus another 16,000 Ibs. from Washington Works incineratedoffsite (DuPont, 2000). Canadian research has stated that the thermolysisoffluoropolymers, e.g. Teflon, Kel-F. can liberate small quantitiesof polycarboxylic acids, which include PFOA (Ells et al., 2001). This information was insufficient to estimate potential yields. 23.2 Discharge to Water By analogy to PFOS, PFOA discharged to water may remain there, become adsorbed to particulate matter and sediment, and/orbeassimilated by organisms. For 1999, 3M estimated PFOA-compound water releasesof <30,000 Ibs. a its Decatur AL location, and <15,000 Ibs. at its Cottage Grove MN location (3M Company, 2000a,b). For 1999, DuPont estimated the 1" 20 following APFO water releases per location: Washington Works WY, 55,000 lbs; Parlin NJ, 300 Ibs. Spruance VA, 150 Ibs; Chambers Works NJ, 9500 bs. (DuPont, 2000). DuPont measured and modeled the following APFO concentrations at ts sites: Washington Works WV: 0.552 ug/l from a 1999 drinking water sample obtained from GE Plastics immediately downstream on the Ohio River. Modeled 1996 APFO-compound releases indicated an average annual PFOA concentrationof0.423 ug/l, with APFO concentrations likely to exceed 1 ug C-8/1 about 50% of the time during the year, and likely to exceed 10 ug APFO/L about 22%of the time during the year. 2.3.3 Discharge to Land 3M reported that land treatmentof sludge from wastewater treatment at their Decatur AL location ended in mid-1998; less than S00 Ibs. were disposed to land at that ite in 1997. Sludge from the Decatur site is now transported to an offsite landfil; sludge from 3M's Cottage Grove MN facility is sent to an industrial landfill (3M Company, 2000a,5). DuPont (2000) estimated 3.900 Ibs. of APFO sludge landfilled on site in 1999 at their Chambers Works NJ facility. DuPont estimated 2,600 Ibs. APFO transferred offsite to a hazardous waste landfill from their Washington Works WV facility. Prior operations resulted in ground- and surface water concentrationsof APFO monitored at three landfills operated by DuPont's Washington Works WV facility. Average surface water concentrations for two landfills were 1392 ug/L and 18.5 ug/L, respectively. A third landfill had a maximum concentrationof33 ug/L in the permitted outfall. Average groundwater concenirations for two landfills were 2537 ug/L and 8.83 ug/L, respectively. A third landfill had maximum groundwater concentration of 15 ug/L (DuPont, 2000). DuPont also reported the following APFO concentrations, measured January 2000, in three drinking water wells of the Lubeck Public Service District, downstream of DuPont's Washington Works WV site: 0.8 ug/L, 0.44 ug/L, and 0.313 ug/L (DuPont, 2000). As ofAugust 2000, the Lubeck Public Service District (LPSD) reported APFO concentrations of 0.2 ppb in drinking water at DuPont's Washington Works facility, and 0.2, 0.5, and 0.1 ppb in the three LPSD wells (LPSD, 2000). 2.3.4 Environmental Monitoring. 3M's Mult-City Study reported on PFOA concentrations from water, sludge, sediment, POTW effluent, and landfill leachate samples taken in six cities (3M, 2001). Four of the cities (Decatur AL, Mobile AL, Columbus GA, Pensacola FL) were "supply" cities that have manufacturing or industrial useoffluorochemicals; twoof the cites (Cleveland TN, Port St. 1s 21 Lucie FL) were "control"citiesthatdo not have significant fluorochemical activities. Across all cities, POTW effluent concentrations rangedfrom 0.040 to 2.42 ppb. The POTW sludge (dry wt) range was non-detect {0 244 ppb; the drinking water range was non-detect to 0.029 ppb; the landfill leachate range was non-detect 0 45.1 ppb; the surfuce water range was non-detect to 0.083; the sediment range was non-detect to 1.75 ppb (dry wt); and the quiet water range was non-detect (0 0.097 ppb. The "control" cities samples generally inhabited the lower endof the above ranges, except for the POTW effluent and sludge findings for Cleveland, which were intermediate in their ranges. Giesy reported that PFOA was rarely found in fish and fish-cating water birds. Fish were sampled from the U.S., certain European countries, the North Pacific Ocean, and Antarctic locations (Giesy, 2001a). Fish-eating bird samples were collected from the U.S., including Midway atoll, the Baltic and Mediterranean Seas, Japanese and Korean coasts (Giesy, 2001b) Giesy reported on PFOA in mink and river otter livers from the U.S. (Giesy, 2001c). PFOA was found in a few mink livers from Massachusetts at a concentration rangeof<18 to 108 ng/g, dry Wt. but not found in mink from Louisiana, South Carolina and Illinois. PFOA concentrations in river otter livers from Washington and Oregon States were less than the quantification limit of 36 ng/g wet wt. Giesy reported that PFOA was not detected at quantifiable concentrations in oysters collected in the Chesapeake Bay andGulf of Mexicoofthe U.S. coast (Giesy, 2001d). Giesy reported on the concentraotfiPoFnOAs in surface water, sediments, clams, and fish collected from locations upstream and downstreamof the 3M facility at Decatur AL (Giesy, 2001). Of the five downstream sampling locations, the two closest to the 3M facility had PFOA surface water concentrations significantly greater than the two upstream sites (means of 1900ug/L. and 1024 ug/L, vs. 0.008 (est.) and 0.028 ug/L); the nearest three locations had sediment concentrations significantly greater than the upstream sites (wet wi. means 1855 ug/ke, 892 ug/g, 238 ug/kg vs. 0.08(est.) and 0.09(est). Clam and fish samples were collected at two locations, one upstream and one downstream of the 3M facility. The average fish whole body PFOA concentration for the upstream location was 11.7 ug/kg (wet wt.), while that for the downstream location was 106.4 ug/kg. The average PFOA concentration in clams at the upstream location was 4.38 ug/kg; that for the downstream location was $.42 ug/ke. 2.4 Human Biomonitoring "Table 1 provides serum PFOA levels in both occupational cohorts and in the general population. "The highest levels reported to date in the general population are similar to some of the lowest 16 22 levels in workers exposed to PFOA occupationally. The data are currently limited to those discussed below. 3M has offered voluntary medical surveillance to workers at plants that produce or use perfluorinated compounds since 1976. Serum PFOA levels have been measured and reported since 1993. Prior to this time, only total organic fluorine was measured. The results of biomonitoring for PFOA have been reported for 3 plants: Cottage Grove, Minnesota; Decatur, Alabama; and Antwerp, Belgium. Surveillance years include 1993, 1995, 1997, 1998, and 2000, although not all of the plants offered surveillance in allof these years. The 1998 data reported for the Decatur plant consist ofa random sampleof employees; however, volunteers participated in all of the other sampling periods for allofthe plans. Mean serum PFOA levels have increased slightly at both the Cottage Grove and Decatur plants since 1993. Workers at the Cottage Grove plant, where PFOA exposures are highest, have the highest PFOA serum levels. The latest sample was in 1997, and the mean serum PFOA level was 6.4 ppm (range = 0.1 -- 81.3 ppm) (Olsen et al., 1998). Only 74 employees participated in the 1997 surveillance. The total numberofemployees working at the plant was not reported. Atthe Decatur plant, 263 of 500 employees participated in 2000 (Olsen et al, 2001d). The mean Serum PFOA level was 1.78 ppm. `This was slightly higher than the mean in 1998 (1.54 ppm). In 2000, 5 employees had serum levels greater than 5 ppm, the Biological Limit Value established by the 3M Exposure Guideline Committee. Cell operators had the largest increase in serum PEOA between 1998 and 2000. The highest level was in a chemical operator on the Scotchgard team (12.70 ppm). The mean level for the est of the members of the team was 5.06 ppm (range 5 -9 ppm). Otherjob categories did not exhibit such a large increase. 3M reports that this is due to increased PFOA production at the Decatur plant beginning in 1999. Serum PFOA levels at the Antwerp plant have been lower than at Decatur or Cottage Grove, and have decreased slightly since 1995 (Olsen et al., 2001). Participation in medical surveillance at the Antwerp plant was the highest it had ever been in 2000 (258 volunteers outof 340 workers). The mean serum PFOA level was 0.84, and the highest serum level reported was 7.04 ppm Three employees had levels greater than $ ppm. 3M's Specialty Materials Manufacturing Division laboratories, where employees perform fluorochemical research (Building 236), conducted voluntary biomonitoringof 45 employees in 2000 (Olsen et al, 20011). The mean PFOA serum level was 0.106 ppm (range 0.008 - 0.668 ppm). Data on PFOA levels in the general population are very limited. They are very recent and are only available on small cohorts. The mean serum PFOA levels are much lower in the general population than in workers exposed to PFOA. " 43 PCooomlpeadnbyl,oo1d99sa9m,pl1e9s99bf)r.omTUh.eS.hibglhoeosdt bpaonoklsedisndaimcpalteemreepaonrtPedFOwaAsl2e2velpspbo.f3Satmop1l7esppwber(e3M collected in 1998 and 1999. These data provide a very preliminary viewofthe PFOA levels that amraeyrebperepsreensteanttivienotfhethUe.SU..Sg.enpeorpaullpaotpiuolnatfioorn.seveHroawlerveears,onist:ca1n)nobtlbooedasdosnuomresdatrhe notthneesceelsesvaerlisly representative of the U.S. population, 2) manyofthe blood banks originally contacted for possible inclusion in the study declined to participate, 3) onlay small number of samples have aicntfuoarlmlaytiboenenaraenaavlayizleadblfeoroPnFtOheA,doanonrds.4) no other data such as age, sx. or other demographic P20r0e1li)m.inaBrlyodoadtasaomnpliendsifvirdoumal63b2loUo.dS.saadmuplltesblhoaovdedroencoernst,lyabgeeesn20r-e6p9o,rtweedr(eOlosbeinaienteadl,fr2o0m01sbix, American Red Cross blood banks located in: Los Angeles, CA; Minneapolis/St. Paul, MN; Charlotte, NC; Boston, MA; Portland, OR, and Hagerstown, MD. The mean serum PFOA level was 5.6 ppb. The range was <lowerlimitofquantitation (LLO=Q 1.92 or 2.11) to 52.3 ppb. Blood samples from U.S. children have also been analyzed for serum PFOA. A sampleof 599 children, ages 2-12 years old, participating in a study ofgroup A streptococeal infections, revealed a mean PFOA serum level of 5.6 ppb. The range was <LLOQ to 56.1 ppb. The LLOQ was 1.92 or 2.88. The samples were collected from equal numbersof male and female children residing in 23 states. The samples in bothof these studies were analyzed using high-pressure liquid chromatography/electrospray tandem mass spectrometry (HPLC/ESMSMS). These data are only preliminary and have not completed quality assurance procedures. In another study, the PFOA concentration was analyzed in human sera and liver samples (Olsen etal., 2001g). Thirty-one donor sampleswere obtained from 16 males and 15 females over an 18-month period from the International Institute for the Advancement of Medicine (LAM). The average ageof the male donors was 50 years (SD 15.6, range 5-69) and the average age of the female donors was 45 years (SD 18.5, range 13-74). The causesofdeath were intracranial hemorrhage (n = 16 or 52%), motor vehicle accident (n = 7or 23%), head trauma (n = 4 or 13%), brain tumor (n = 2 or 6%), drug overdose (n = | or 3%) and respiratory arrest (n = | or 3%). Both serum and liver tissue were obtained from 23 donors; 7 donors contributed liver tissue only and I donor contributed serum only. Serum samples were obtained from 5 miofblood; liver samples consisted of 10 oftissue. Samples were frozen at I1AM and shipped frozen to 3M for analysis. Samples were extracted using an ion-pairing extraction procedure and were quantitatively assayed using HPLC-ESMSMS and evaluated versus an unextracted curve. eExxttreancstiivoen mpraotcreidxursep.ikeSsetruudimesvawleurees fpoerrfPoFrOmeAdrtaonegveadlufartoemt<heLprOeQcis(i<o3n.a0n--)d 7a.c0cunrga/cmyLo.f the Assuming the midpoint value between zero and LOQ serum value for samples <LOQ, the mean serum PFOA level was 3.1 ng/mL with a geometric meanof 2.5 ng/mL. No liver to serum rations were provided because more than 90% of the individual liver samples were <LOQ. Serum PFOA levels in corporatestaff and managers at a 3M plant in St. Paul, Minnesota, where occupational exposure to PFOA should not have occurred, were reported (3M Company, 1999), 1s oo Fourof 31 employees had serum PFOA levels greater than the detection limit of 10 ppb. The `mean for these employees was 12.5 ppb. Table 1. SERUM PFOA LEVELS IN HUMAN POPULATIONS Arithmetic Mean Geometric | 95% | Mean Confidence | Interval Cottage Grove Plant 1997 (n=74) 64 0.1-81.3 1995 (n = 80) 68 1993 (n=111 50 0.0-114.1 0.0-80.0 Decatur Plant 2000 (n = 263) 1.78 004-1270 | L13 0.99 -1.30 1998 (n =126) 1.54 1997 (n = 84) 1.57 1995 (n= 90) 1.46 0.02-6.76 0.90 not reported | * not reported | * 072-112 . . 2000(n = 258) 0.84 0.01 -7.04 033 0.27-0.40 2000(n = 45) 0.0- 00.6868 | 0.053 [Source------ JArithmeticMean [Range `Commercial sources of (n ee = 35 lots) American Red Cross blood banks, 2000 56 427-523 0.0 - 03.0776 | ER le E : 2$ 3.0 Human Health Hazards 3.1. Metabolism and Pharmacokinetics 3.1.1 Half-life in Humans In order to determine the half-life of PFOA, a group of retirees (n = 20) volunteered to participate in a 5-year half-life study in which serum samples willbedrawn every 6 months (Burris et al., 2000). The only other data available on the half-life of PFOA is from a 1980 study in which it was estimated to be approximately | year; however, this analysis was based on total organic fluorine in blood serum. Twenty-seven retirees, age 55 to 74 years, volunteered to participate i this half-life study. PFOA levels in this group ranged from 0.1 to 3.1 ppm. Mostof the retirees were employed at the Decatur, Alabama plant for an averageof28 years. The number of years since retirement varied greatly among the participants. The average length of time between retirement and the startofthe study was 30 months (2.5 years) but ranged from S to 130 months (~ 5 to 10 years) "There were 3 collection periods during which serum PFOA samples were collected and analyzed: November 1998, June 1999, and November 1999. Cottage Grove employees, where PFOA exposure was much higher, have only participated in 2 sampling periods; therefore, they were not included in this analysis. Half-lives were calculated using a one-compartment model. A log-linear relationship (slope = c~oknes(t2a.n3t03w))aswadsetuesremidnteod,esutsiimnagtethteherehlalaft-iloinfse.hipT:htezha=lf0-.l6i9f3e/wkqa.s cOanllcuylattheodsearfeetrirteheesewlhiominfiatttihoen linear one compartmental model (+ 0.6) for PFOA were included in the analyses. 1f3 data points were not available for anyofthe subjects and if there was a lack of fit to the model, that retiree was not included in the analysis. Twenty participants met these requirements. The median serum half-oflPiFfOeA was 344 days, with a range of 109 to 1308 days. The two highest half-life calculations were for the 2 female retirees who participated in this study (654 and 1308 days). It should be noted that the difference in PFOA serum levels between retirees was quite large (0.1 - 3.1 ppm). It was not specifically sated in the report; however, based on a statement in the report, it is assumed that the 2 femaleretireesdid not have the highest PFOA serum levels For mostofthe participants not included in the analysis, the second measurement was higher than the first. Therefore, the data did not fit the model and they were excluded. Although this may justify not including those participants in the analysis, it is an indicationof the many limitationsofthe data. Its stated in the report that neither age nor numberofmonths retired was associated with the serum PFOA half-life calculations; however, this statement is not supported with any data in the report. In addition, no individual data were provided in the report and the relationship between numobfyeearrs exposed in the workplace and PFOA levels and half-life were excluded. Also, elimination of PFOA occurs via urine and feces; however, these. 2 26 measurements were not collected. Therefore, it cannotbedetermined whether the half-life suggested by the preliminary results reported here represents a true elimination half-life from the body. Finally, the efoffcoentcinuted non-occupational, low-level exposure on the half-life is unknown. The data presented above provide a very rough estimate of the plasmahal life ofPFOA. It does not provide an elimination rate. In addition, these data do not provide any information about the distribution of PFOA in the body. Without the individual data or supporting information, the statement that time between retirement and entry into the study does not affect the half-life calculation is highly suspect. One would expect age, lengthofexposure, and time elapse since occupational exposure to affect PFOA serum levels. Since these data were not provided in the report and since only 3 data points have been calculated to date, one can only estimate that the half-lifeof PFOA is between | and 3.5 years. These data provide evidenceof the potential to bioaccumulate PFOA in humans. In addition, these preliminary data suggest that gender plays a role in thehal life. 3.1.2 Absorption Studies in Animals APFO is well absorbed following oral and inhalation exposure, and to a lesser extent following dermal exposure. In rats, an averageof 749 ug or 37%of the fluorine in the administered dose was recovered in the urine within 4.5 hr after PFOA dose (by stomach intubation 2 mlofan aqueous solution containing 2 mg PFOA) (Ophaug and Singer, 1980). The quantityof nonionic fluorine recovered in the urine increased to 61%of the dose at hr, 76% at 24 hr, and 89% at 96 hr. mAaflteearratssi,nagtleleoarsatl9d3o9s%eofofth"e"Cto-tPalFOcAar(bmone-a1n4dwosaes,a1b1s.o0rbmegd/kagt)24inhsooulrusti(oGnitbosognroaunpdsoJfothhnrsoene, 1979). The half-life for eliminationoftotal carbon1-4 from plasma was 4.8 days. Following APFO head-only inhalation exposure in male rats (6 hriday, 5 days/wk for 2 wk to 0, 1,8 or 84 mg/m3) concentofroragatnoiflouonridse in the blood showed a dose relationship with initial levels of 108 ppm in rats treated at 84 mg/m3 (Kennedy et al., 1986). Immediately after the tenth exposure period, the mean organofluoride blood levels were 13 ppm, 47 ppm, and 108 ppm in the 1, 8, and 84 mg/m3 dose groups. Subchronic dermal APFO treatment in rats and rabbits (10 applications, 5 doses, 2 rest days, 5 doses) with either 0, 20, 200, or 2000 mg/kg resulted in elevated blood organofluorine levels which increased in a dose-related manner (Kennedy. 1985). Oab'sMoarlplteioynoanfdPEFbbOiAnsin(1m9a8l1e)acnodndfuecmtaeldeararbbaintgse. fiPnFdiOnAg s(t1u0d0ymwgh/ikcgh, i1n0d0ic0amtegs/kseig,niafnidca2n0t0d0ermal mu/kg in saline slurry) was applied to approximately 40%ofthe shaved trunk of the animals, which were then fitted with a plastic collar, and the trunk was wrapped with impervious plastic 2 27 sheeting. The exposure period was 24 hr, 5 days/week over 14 days. Mortality was 100% (4/4) in the 2000 mg/kg group, 75% (3/4) in the 1000 mg/kg group and 0% (0/4) in the 100 mg/kg group. In the past, Chemolite workers have been exposed to large dermal dosesof PFOA. It appears that dermal exposure may have playead significant role in the absorption of PFOA in these workers. Upon recognition that PFOA could be absorbed dermally, work practices were changed and engineering controls were adopted that reduced dermal exposures (Gilliland, 1992). A t-butyl ammonium salt of perfluorooctanoate in the formoftreated fabric and as a liquid formulation was applied dermally to rabbits (Johnson, 199Sb). Liver samples were analyzed at 28 days post dose for total organic fluorine. The results from treated animals were the same as control values. All total organic values were below the practical quantitation limit. Serum levels were also below the practical quantitation limits of the analysis for samples collected at day 1 and 2 after administration of the mixture or the treated fabric. From the pharmacokinetic study (Johnson, 1995a), it would be unlikely that any extent of absorption could have been detected in this study. 3.1.3 Distribution Studies in Animals oPfFtOheAbdoidsyt.ribIuttdesoepsrinmoatriplayrttiotitohnetolivtehre, lpilpaisdmfar,acatnidonkoirdnaedyi,poasnedttiossauel,esbsuetrdeoxetesntb,inodthetrotissues macromolecules in the tissues. There is evidenceofenterohepatic circulationof the compound. Major sex-related differences in the dispositionofPFOA have been observed. Serum and liver concentrations of PFOA were determined in thesus monkeys in a 90 day oral toxicity study (Griffith and Long, 1980). In monkeys at the 3 mg/kg/day dose, mean serum PFOA was 50 ppm in males and 58 ppm in females. At the same dose, males had 3 ppm and females 7 ppm in liver samples. At 10 mg/kg/day doses, male monkeys had a mean serum PFOAof 63 ppm and females 75 ppm. Liver levels were 9 and 10 ppm for males and females, respectively. Ophaug and Singer (1980) measured ionic fluoride and total fluorine in the serum of female rats following the administrationofPFOA by stomach intubation (2 miof an aqueous solution containing 2 mg PFOA). Serum from rats 4.5 hr afte the administration of PFOA had a nonionic fluorine level 13.6 ppm and virtually all of this was bound to components in the serum and not ultrfilterable. Despite the large increase in nonionic fluorine in the serum, the ionic. fluoride level remained very low (0.03 ppm). Prior o intubation of PFOA, the ionic and `nonionic fluorine levels in serum were 0.032 and0.07 ppm, respectively. The nonionic fluorine level in the serum decreased to 11.2 ppm at hr, 0.35 ppm at 24 hr, and 0.08 ppm at 96 hr. The authors conclude that PFOA is rapidly absorbed from the gastrointestinal tract and rapidly cleared from the serum. 2 28 `Twenty-four hours after oral administration of APFO (2 mg APFO in 2 mi aqueous solution by stomach intubation), female rats had a mean serum nonionic fluorine levelof 0.35 ppm, while mwaaslbroautsndhatdo aa smiemailnarseerxtuemntnoinnitohneicplfalsumorainoeflmeavleleoafn4d4.fe0mpalpemr(atHsan(h9i7j.a5rv%ibeotuanld,).1982). APFO In male and female rats administered 14C-PFOA in propylene glycolwater (9.4 umole, ip). the concentration of 14C-PFOA-derived radioactivity in the blood was higher and climinated more slowly in males (t1/29 days, malesvs 4 hr, females, Vanden Heuvel et al., 1991). In the male rat, the liver had the highest PFOA concentration (21%ofdose at 2 hr, 2%ofdose at 28 days) followed by the plasma and kidney. Far lower PFOA concentrations were found in the hear, esti, fat, and gastrocnemius muscle. In females at 2 hr post dose, the highest concentrationsofPFOA were found in the plasma followed by the kidney, liver and ovaries in that order. The average 112 for elimination of PFOA from the liver in male rats was 11 days compared to an average of9 days for extrahepatic tissues. In females, the average t1/2 for tissue elimination was approximately 3 hr. Vanden Heuvel etal. (1991) investigated the disposition of PFOA in perfused male rat liver. Approximately 11% of the cumulative dose of '*C-PFOA infused (0.08 umol/min x 48 min, 3.84 umol total) was extracted by the liver durinag firs pass. In addition, the cumulative percent of PFOA extracted by the liver at2min (33%) was substantially greater than that seen after 48 min (119%) indicating that first-pass hepatic uptake of PFOA may be saturable. Ylinen et al. (1990) studied the difference between male and female Wistar ats in the distribution and accumulation of PFOA aftear single and subchronic administration. The single: dose of PFOA (50 mg/kg in propylene glycol-water mixture, 1:1, vol. 0.25 m/100g) was administered intraperitoneally to 10 week old rats (20 male, 20 female). Subchronic administration ofPFOA consisted of 3, 10, and 30 mg/kg/day by gavage (in 0.9% NaCl, 0.5 m/100g) to newly weaned rats (18 male, I8 female). After the single dose, samples were collected for PFOA determination 12, 24-168 (at 24 h intervals), 244 and 336 hours aftr the administration, and in the subchronic test on the 28th day. The serum was collected by cardiac puncture; after decapitation the brain and at necropsy samples from the liver, kidney, lung, spleen, ovary, tesis, and adipose tissue were collected and frozen. The biological half-life of PFOA in the serum and tissues was determined from the linear relationship between time and PFOA concentration in the semilogarithmic plot. In the single-dose study, concentration of PFOA in the serum and tissues was higher in males than females at all time periods. Twelve ours afte the administration of PFOA about 10% ofthe dose was found in the serum of females, whereas about40% was in the serum of males. After 14days about 3.5%ofthe dose: remained in the serum. In females, PFOA concentration in the serum, liver, and kidney occurred in adiscontinuous fashion, indicating distinct phases. The half-life in the serum was 24 and 105 nin the females and males, respectively. In the females, a half-lifeof60h was estimated in the liver during the first week. In the males, thehal-life in liver was 210 h. Although PFOA was retained by the liver, it was not found in the lipid fraction. In the kidney, the hal-life was 145 h and 130 hin females and males, respectively. In the spleen, the half-life was 73 hand 170h in 3 2 females and males, respectively. PFOA was also found in brain tissue. PFOA was not detectable in adipose tissue. In the subchronic study, samples taken on the 28th day indicated significantly higher PFOA concentrations in the serum and tissues of males versus females in all three dose levels.Aft subchronic, as well as single-dose administration, PFOA was mainly distributed in the serumof ats. High concentrationsofPFOA were also found in the liver, kidney, and lung of males and females. At the high dose level (30 mg/kg/day), females and `males exhibited, respectively, serum concentrations of 13.92 and 51.65 ug/ml, liver concentrationsof6.64 and 49.77 ug/g, kidney concentrations of 12.54 and 39.81 ug/g, spleen concentrations of 1.59 and 4.10 ug/g, lung concentrationsof 0.75 and 23.71 ug/g, and brain concentrationsof 0.044 and 0.710 ug/g. The ovary contained 1.16 ug/g and the tests contained 7.22 ugle. A significant positive correlation existed between the administered dose and the concentrationof PFOA in the liver, kidney. spleen, and lung of females. On the contrary, no significant correlation between the administered dose and the concentration of PFOA was observed in the males, as 10 mg/kg/day produced higher PFOA concentrations in the serum and organs than 30 mg/kg/day. However, in males, the concentration i the spleen, testis, and brain correlated positively with the concentration in the serum. Vanden Heuvel et al. (1992) demonstrated that PFOA covalently binds to proteins in the liver, plasma, and testes of rats in vivo. Carbon-I4-labeled PFOA was administered to six-week old male Harlan Sprague-Dawley rats in propylene glycol/water (1:1, v/v; | ml/kg) ata doseof9.4. umolkg, ip. No time-dependent changes in either absolute or relative concentrations of covalently bound PFOA-derived 14C were found at2 h, 1and 4 days post-treatment. Covalently bound PFOA was represented by 0.1 to 0.3%ofthe tissue 14C content. The absolute concentration of covalently bound PFOA was significantly higher in the plasma than in th liver. The testes had the highest relative concentration of PFOA-derived radioactivity covalently bound. In in vitro tests, covalent binding of 14C-PFOA to a constant concentration of albumin (8 uM) increased in2 linear fashion with increasing PFOA concentration. The covalent binding. of PFOA to hemoglobin in vitro was diminished by the additionofcysteine but not methionine, suggesting that protein sulfhydryl groups may be involved. Hanhijarvi et al. (1987) compared the disposition of PFOA between male and female Wistar rats during subchronic administration. PFOA was administered by gavage to 48 newly-weaned animals at 0, 3, 10, and 30 mg/kg (in 0.9% NaCl, 0.5m/100g) for 28 consecutive days. Urine was collected on the 7th and 28th day of the study (discussed below). At the endof the study, blood was collected via cardiac puncture. At each dose level, the mean PFOA concentrations in the plasmaofthe male rats were significantly higher than those ofthe female rat. The mean plasma PFOA concentrations for the male rats were 48.6+-26.5 ug/ml (dosed at 3 mg/kg), 83.14 24.7 ug/ml (10mg/kg), and 53.4+-11.2 ug/ml (30 mg/kg). The corresponding figures for female rats were 2.43+-5.96 ug/ml, 11.3+-8.59 ug/ml, and 9.06+-8 80 ug/ml in the same order. The PFOA concentrations in the plasmaof the male animals suggested that the binding sites of PFOA may become saturated at the chronic daily dose level of 30 mg/kg. Although the plasma PFOA dciofnfceernetnrcaetsiobnestwweereen tsihgensifeixceasntwleyrheioghbesrerivnetdheatmnaelcerorpatssy,.no significant histopathological x 30 "The disposition of PFOA was studied in male Wistar rats afer castration and estradiol administration as well a in intact males and females (Ylinen et al., 1989). The male rats (N=20) were castrated at the ageof 28 days and after 5 weeks were used in the tests.Half of the operated and 10 intact males were administered estradiol valerate subcutaneously 500 g/kg every second day during 14 days before the test. Blood samples were collected by cardiac pwuansctcuornes.idAetratbhelyenhidgohferth(e17t-es4t0(t9imehsr)),tthhaencionnctehnetsreatriuomnooffotPhFeOrAgrionuptsh.e sTehreurmeowfaisntnaoct males statistically significant difference in the serum concentrations between the other groups. PFOA was similarly bound to the proteins in the serum of males and females. Johnson ct al. (1984) investigated the effectof feeding cholestyramine to rats on the fecal elimination of APFO. Since APFO exists as an anion at physiologic pH, it would be expected to `complex with cholestyramine in vivo. Ten Male Charles River CD rats (12 weeks old, 300-342 g) were administered ammonium 14C-perfluorooctanoate (2.1 mg/ml) dissolved in 0.9% NaCl as achsoilnegslteyirnatmrianveenionufseeddo.seU(r2inmeaatnd,faevceersasgaemAplPeFsOwedroeseco1l3lemcgt/ekdg)at. inFtievrevarlastsfowrer14e dgaiyvse,na4t % wthheicmheatnimpeetrhceenatnaigmeaolfs AwePrFeOsadcorsifeieceldimainndatleidveirnstahmepfleecseswoefrechcoollelsetctyerda.miAnet-t1r4edaateyds rpaotsst(d4o3se2,+ 5.5) was 9.8-fold the mean percentageof dose eliminated in feces by untreated rats (4.4+-1.0). Excretion in urinewas 41% for treated rats and 67% for untreated rats. Carbon-14 present in the liver represented 12.1+-2.1 ug eq/g and 22.3+-6.2 ug eq/g in treated and untreated rats, respectively (4% and 8%ofdose, respectively). In plasma, the levels were 5.1+-1.7 ug eq/ml and 14.7--6.8 ug eq/ml in treated and untreated rats, respectively. In red blood cells, the levels were 1.8--0.7 ug eq/ml and 4.2+-2.4 ug eq/ml in treated and untreated rats, respectively. The high concentration of 14C-APFO in liver at 2 weeks afer dosing and the fact that cholestyramine treatment enhances fecal eliminationofcarbon-14 nearly 10-fold suggests that there is enterohepatic circulation of APFO. "The disposition of PFOA (etrabutyl ammonium salt perfluorooctanoic acid) in female rabbits has been reported (Johnson, 1995a). Individual rabbits were given intravenous doses at 0, 4, 16, and 24 mg/kg and appeared normal throughout the study (the animal treated at the 40 mg/kg dose leveldied within 5 minutesofdosing). Serum samples were analyzed for total organic fluorine at 2, 4, 6, 8, 12, 24, and 48 hours post dose. At 2 hrs, serum organic fluorine levels in the 0,4, 16, and 24 mg/kg dosed rabbits were 1.25 ppm, 4.09 ppm, 14.9 ppm, and 41.0 ppm, respectively. Therewas a rapid decrease in serum leveloftotal organic fluorine with time, nondetectable at 48 hr. The biological half-life was on the order of4 hours. The total organic fluorine in whole liver at 48 hr post dose for control animals4, mg/kg, 16 mg/kg, and 24 mg/kg intravenous doses were 20 ug, 43 ug, 66 ug, and 54 ug. 3.1.4 Metabolism Studies in Animals Vanden Heuvel et al. (1991) investigated the in vivo metabolismofPFOA in rats administered HC-PFOA (9.4 umol/kg, i... Pooled daily urine samples (0-4 days post-treatment) and bile 2 3 extracts analyzed by HPLC contained a single radioactive peak eluting identically to the parent compound. Tissues were taken from ras treated 4, 14, and 28 days previously with 14C-PFOA to determine the presenceofPFOA-containing lipid conjugates. Only the parent compound was present in rat tissues; no PFOA-containing hybrid lipids were detected. Fluoride concentrations in plasma and urine before and afier PFOA treatment were unchanged, indicating that PFOA does not undergo defluorination in vivo. `Ophaug and Singer (1980) also found no change in ionic fluoride level in the serum or urine following oral administrationofPFOA to female rats. Ylinen et al. (1989) found no evidence of phase Il metabolism of PFOA following a single intraperotoneal PFOA dose (50 mg/kg) in male. and female rats. 3.15 Elimination Studies in Animals "The urine is the major routeofexcretion ofPFOA in the female rat, while the urine and feces are both major rooufetxcreetison of PFOA in male rats (Vanden Heuvel etal. 1991). Male and female rats were administered ""C-PFOA in propylene glycol/water (9.4 umol/kg, ip.). Female rats eliminated PFOA-derived radioactivity rapidly in the urine with 91% of the dose being excreted in the first 24 hr, while male rats excreted only 6%ofthe dose in that time period. Negligible radioactivity was recovered in the feces of female rats. In male rats during the 28-day collection period the cumulative excretion of PFOA-derived 14C in urine and feces was 36.4% and 35.1%, respectively. The female rat retained less than 10%of the administered dose after 24 hr, while the male rats retained 30% of the administered dose afer 28 days. The whole-body elimination half-life in females was less than one day, and in males it was 15 days. In renalligated rats injected ip. with 14C-PFOA, approximately 0.3%of the PFOA-derived radioactivity was excreted in the bile after 6 hr (Vanden Heuvel et al, 1991). No sex-related difference in the biliary excretion ofPFOA was observed when the kidneys were ligated. Johnson and Gibson (1980) observed a sex difference in extent and rateof excretion oftotal carbon-14 between male and female rats aftar single iv dose (mean dose: female, 16.7 mg/kg: male 13.1 mg/kg) of 14C-PFOA. Female rats excreted essentially all of the dose via urine in 24 hours while at the same time period male rats excreted only 20 percentof the dose; male rats excreted 83% via urine and 5.4% via feces by 36 days post dose. No radioactivity was detected in tissues of female rats at 17 days post dose; male rats had 2.8%ofthe dose in liver and 1.1% in plasma at 36 days post dose with lower levels (< 0.5%ofthe dose) in other organs. Ophaug and Singer (1980) investigated the metabolic fate of PFOA in female Holtzman rats Animals weighing approximately 250 g were administered by stomach intubation 2 miof an aqueous solution containing 2 mg PFOA. The animals were then placed in metabolism cages and provided rat chow and tap water for 4.5, 8, 24,or 52.5 hr. In addition, four rats were placed hirn. meAttatbhoeliesnmdocfatgehseaenxdpefreidmeanltaolw pfelruoiroiddeth(e<0ur.i5nep,pmfe)cedsieatnadnsdedriustmilwleedrewactolelrecftoerda. peWriitohdionf94.65 hr after PFOA dose, an average of 749 ug or 37% of the fluorine in the administered dose was 2% 32 recovered in the urine. The quantityofnonionic fluorine recovered in the urine increased to 61%of the dose at hr, 76% at 24 hr, and 89% at 96 hr. Urinary excretion of ionic fluoride in the PFOA dosed animals was not significantly different than that of the control animals. Fecal excretion of nonionic fluorine was 4.5%ofthe administered dose at 52.5 hr and 14.3% at 96 hr. "The urine from undosed animals contained no detectable nonionic fluorine. "The urinary excretion of APFO in rats was investigated by Hanhijarvi et al. (1982). Four male and six female Holtsman rats were administered 2 mg APFO in 2 ml aqueaus solution by stomach intubation. Seven female rats were administered 2 ml distilled wateras controls. The animals were then placed in metabolism cages with rat chow and tap water. Urine was collected until animals were sacrificed at 24 h by cardiac puncture. Serum was collected. lonic fluoride and total fluorine contentof serum and urine was determined, and nonionic fluorine was calculated as the difference. For clearance studiesofAPFO and inulin, the ats were anesthetized with Inactin. The femoral artery was cannulated for continuous infusion of 5% mannitol in isotonic saline and the femoral artery was cannulated for drawing blood samples. "The urinarybladder was also cannulated for serial collectionsofurine. Intravenous priming doses of 5.2-5.6 mg [1-14C] ammonium perfluorooctanoate (sp act 0.5 uCi/mg) and 8.8 ug. witiated inulin (methoxy-3H, sp act 114 uCi/mg) were given to each animal. The radiolabled inulin and APFO in 5% mannitol in isotonic saline was then infused at a rate of 0.21 ml/min. An additional 0.42-0.63 mg/hr 14C-APFO and 9.6 ug/hr rtiated inulin was infused during the experiments. When the urine and serum collections for the clearance study were complete, probenecid was administered (65-68 mg/kg, ip) and additional clearance tests were performed. In the cumulative excretion study, rats were dosed iv with a mixture ofradiolabeled APFO (1020%) and unlabeled APFO (80-90%). Five percent mannitol in isotonic saline was infused at a rate of 0.081 ml/min and urine specimens were collected over 30-min intervals. The effect of probenecid was assessed by administering 65-68 mg/kgipat least 30 min prior to the: administration of APFO. Twenty-four hours after oral administration of APFO, female rats had excreted 76+-2.7% of the dose in the urine and had a mean serum nonionic fluorine level of 0.35+-0.11 ppm, while male rats had excreted only 9.2+-3.5% of the dose and had a mean serum nonionic fluorine levelof44.0+-1.7 ppm. APFO was bound to a similar extent in the plasma of male and female rats (97.5+-0.25% bound). The clearance studies demonstrated major differences between the sexes in ats. The APFO clearance in female rats was several times greater than the inulin clearance. Administrationofprobenecid, which strongly inhibits the renal active secretionoforganic acids, reduced APFO/inulin clearance ratio in females from 14.5 to 0.46. APFO clearance was reduced from 5.8 t0 0.11 ml/min/100g. Net APFO excretion was reduced from 4.6 to 0.13 ug/min/100g. In male rats, however, the APFO/inulin clearance ratio and the net excretion ofAPFO were virtually unaffected by probenecid. In the males, APFO clearance was 0.17 ml/min 100g, APFO/inulin clearance ratio was 0.22, and net APFO excretion was 0.17 ug/min/mg. In the cumulative excretion studies, female rats excreted 76%of the APFO dose, while males excreted only 7.o8ft%he dose overa 7-hr period. Probenecid administration `modified the cumulative excretion curve for males only slightly. However, in females probenecid markedly reduced PFO elimination to 11.8%. It is concluded that the female rat possesses an active secretory mechanism which rapidly eliminates APFO from the body. This 7 33 Secretory mechanism is lacking or is relatively inactive in male rats and accounts for the greater toxicity of APFO in male rat. Hanhijarvi et al. (1987) compared the urinary elimination of PFOA between male and female Wistar rats during subchronic administration. PFOA was administered by gavage to 48 newlyweaned animals at 0, 3, 10, and 30 mg/kg (in 0.9% NaCl, 0.5mi/100g) for 28 consecutive days. Urine was collected on the 7th and 28th dayofthe study. At the end of the study, blood was collected via cardiac puncture. At necropsy, tissue specimens for histopathologic examination were collected from the controls and from the group receiving 30 mg/kg/day PFOA. On the seventh day of the study period, the female rats in lowest dose group (3 mg/kg/day) exhibited significantly greater urinary PFOA excretion than the males (3.12+-0.30 vs 1.50+-0.57 mg/24hr/kg). Unlike the female rats, on the 7th dayofthe study all three groupsofmale rats excreted significantly less PFOA than their daily dose of PFOA, which suggested that the males had not reached a steady state by seven days. On the 28th day, the males excreted an amount of PFOA equal to theirdaily dose. Hanhijarvi etal. (1988) investigated the excretion Kinetics of PFOA in the beagle dog. Six laboratory bred beagle dogs (3 male, 3 female) were anesthetized with methoxyflurane and catheters were placed in both ureters afte laparototomy and cystotomy. The animals were given an intravenous dose of 30 mg/kg of PFOA followed by continuous infusion with 5% mannitol solution at 1.7 mUimin. Urine was collected at 10 minute intervals for 60 min. A S ml blood samplewascollected in the middle of each urine sampling period. Probenicid (30 mg/kg i.v.) was then administered, and urine and blood samples were again collected as before. Renal clearanceof PFOA was calculatedforthe before and after probenecid injection periods. Four additional dogs (male, 2 female) were given 30 mg/kg PFOA intravenously. These dogs were kept in metabolism cages, and blood samples were collected intermittently for 30 days. From these dogs, plasma PFOA half-lives were determined. There was no difference between the. renal clearances of the male and female dogs either before or after probenecid. Renal clearance rate was approximately 0.03 mUmin/kg. Probenecid significantly reduced the PFOA clearance in both sexes, indicating an active secretion mechanism for PFOA. The plasma half-lives of PFOA were longer in the male dogs (473 h and S41 h) than in the female dogs (202 h and 305 h) "The urinary excretion of PFOA was studied in male Wistar ats afer castration and estradiol awdemrienicsatsrtartatieodn aats twheellagaesoif2i8ntadcatymsaalneds aafntderfSemwaeleekss(Ywleirneenuscetdl.i,n t1h9e89t)e.sts.TheHamlafloef rtahtes (N=20) eopveerraytseedcaonndd 1d0ayindtuarctinmgal1e4sdwaeysrebeafdomrienitshteetreestd. esUtrriandieolwavsalceorlalteecstuebdciuntmaneetoaubsollyis5m00caugge/skg during 96 hr after a single intraperotoneal PFOA dose (50 mg/kg). Blood samples were collected by cardiac puncture. Castration and administrationofestradiol o the male rats had a significant stimulatory effect on the urinary excretion of PFOA. During the first 24 hours, female rats excreted 72+-5% (N=6)of the dose, whereas the intact males excreted only 9+-4% (aNm=o6u)n.tsAsfitmeirltahreteosftreamdailoelst(re6a1t+m-en1t9,%baonthd t6h8e-i1nt4a%ct,arnedspceacsttirvaetleyd).maTlhees ceaxsctrreatteeddPmaFlOeAs wiinthout ES 3h teshterafdeimoalletsreaantdmetnhteeexsctrreotgeednPtFreOatAedinmaulreisn.e Afatsttehretehnadnotfhethienttaecsttm(a9l6ehsr)(,5t0h+e-1co3n%c)e,ntbruattileosnsotfhan PFOA in the serumofintact males was considerably higher (17-40 times) than in the serum of other groups. There was no statistically significant difference in the serum concentrations between the other groups. PFOA was similarly bound by the proteins in the serum of males and females. Vanden Heuvel et al. (1992a) investigated whether androgens or estrogens are involved in the marked sex-differences in the urinary excretion of PFOA. Castrationofmales greatly increased (> 1-fold) the elimination of ""-PFOA (9.4 umol/ke, ip.) in urine, demonstrating thata factor produced by the testis is responsible for the slow eliminationof PFOA in male rats. Castration plus 17B-estradiol had no further effect on PFOA elimination whereas castration plus testosterone replacementa the physiological level reduced PFOA elimination to the same level as rats with intact testis. Thus, in male ras, lestosterone exerts an inhibitory effect on renal excretion of PFOA. In female rats, neither ovaricctomy nor ovariectomy pius testosterone affected the urinary excretion of PFOA, demonstrating that the inhibitory effectoftestosterone on PFOA renal excretion is a male-specific response. Probenecid, which inhibits the renal transport system, decreased the high rate of PFOA renal excretion in castrated males but had no effect on male rats with intact esti. Hormonal changes during pregnancy do not appear to cause a change in the rateof elimination ofcarbon-14 afer oral administration ofa single dose of ammonium '""-PFOA (Gibson and Johnson, 1983). At8 or 9 days afier conception, four pregnant rats and 2 nonpregnant female ats were dosed (mean dose, 15 mg/kg) and individual urine samples were collected at 12, 24, 36, and 48 hours post dose and analyzed for carbon-14 content. Essentially all of the carbon-14 was eliminated via urine within 24 hours for both groupsofats. Feedingofcholestyramine to rats enhanced the fecal eliminationofAPFO (Johnson et al. (1984). Male rats were administered ammonium [14]perfluorooctanoate (2.1 mg/ml) dissolved in 0.9% NaCl as a single intravenous dose (2 mirat, average APFO dose 13 mg/kg). At 14 days post dose, the mean percentage of APFO dose eliminated in the fecesofcholestyramine-treated rats (43.2+-5.5) was 9.8-fold the mean percentageofdose eliminated in feces by untreated rats (3.4+1.0). Excretion in urinewas41% for treated rats and 67% for untreated rats 32 Epidemiology Studies 3.2.1 Mortality Study A retrospective cohort mortality study was performed on employees at the Cottage Grove, Minnesota plant which produces APFO (Gilliland and Mandel, 1993). A this plant, APFO permopdluocyteidonatwathselpilmaitnedfotroathleeCashte6mimcoanlthDisvibseitonw.eenThJeancuoahroyrt1c9o4n7siasntdedDeocfewmobrekerrs19w83h.o hDeaadtbheen certificatesofallof the workers were obtained to determine causeofdeath. There was almost 2 3 `wcaosmpclaetteegofroilzleodw-buapse(d99o.n5%th)eiorfjaollbofhistthoreiesst.udyIfptahretiycihpaadntbs.eenTheempelxopyoesdurfeorstaatulseaostf t1hemownotrkherisn. the Chemical Division, they were considered exposed. All others were considered to be not exposed to PFOA. The numberofmonths employe in the Chemical Division provided the cumulative exposure measurements. Of the 3537 (2788 men and 749 women) employees who apanrdti1c4i8paotfedthien t3h4i8s smteundy,wo3r9k8e(d34in8tmheenChaenmdic5a0lwDoimveisni)onw,earneddtehceeraesfeodr.e, Ewleerveencoonfstihdeer5e0d women exposed 10 PFOA. `Standardized Mortality Ratios (SMRs), adjusted for age, sex, and race were calculated and compared to U.S. and Minnesota white death rates for men. For women, only state rates were apveariiloadblsoe.fdTuhraetSioMnRosf feomrpmlaolyemsenwetr(e5,st1r0a,tiafniedd2f0orye3arlsa)t.ency periods (10, 15, and 20 years) and 3 For all female employees, the SMRs for all causes and for all cancers were less than 1. The only elevated (although not significant) SMR was for lymphopoictic cancer, and was based on only 3 deaths. When exposure status was considered, SMRs for all causesofdeath and for all cancers were significantly lower than expected, based on the U.S. rates, for both the Chemical Division workers and the other employees of the plant. In al male workers at the plant, the SMRs were close to 1 for mostof the causesofdeath when compared to both the U.S. and the Minnesota death rates. When latency and duration of employment were considered, there were no elevated SMRs. When employe deaths in the Chemical Division were compared to Minnesota death rates, the SMR for prostate cancer for workers in the Chemical Division was 2.03 (95% CI 5 - 4.59). This was based on 4 deaths (1.97 expected). There was also a statistically significant association with length of employment in the Chemical Division and prostate cancer mortality. Based on the results of proportional hazard models, the relative risk foar 1-year increase in employment in the Chemical Division was 1.13 (95% C1101 10 1.27). Itrose to 3.3 (95% CI 1.02 -10.6)for workers employed in the Chemical Division for 10 years when compared to the other employees in the plant. The SMR for workers not employed in the Chemical Division was less than expected for prostate cancer (58) An updateofthis study was conducted to include the death experienceof employees through 1997 (Alexander, 2001a). The cohort consistedof 3992 workers. The eligibility requirement was increased to | yearofemployment at the Cottage Grove plant, and the exposure categories were changed to be more specific. Workers were placed into 3 exposure groupsbasedon job history information: definite PFOA exposure (n = 492. jobs where cell generation, drying, shipping and packaging of PFOA occurred throughout the historyof the plant); probable PFOA exposure (n = 1685, other chemical division jobs where exposure to PFOA was possible but with cThoewmeircoarltdriavnissiieonnt jeoxbpso).sures); and not exposed to fluorochemicals (n = 1815, primarily non- 30 36 In his new cohort, 607 deaths were identified: 46 of these deaths were in the PFOA exposure. group, 267 in the probable exposure group, and 294 in the non-exposed group. When all ehmigphleoryfeoersalwleorfetchoemcpaaurseedsotfodtehaetshtaatneamloyrzteadl.itNyornateeos,fSthMeRSMweRrewleerssetshtaatnisItiocralolnylsyigsnliifgihctalnyt at P= .05. The highest SMR reported was for bladder cancer (SMR = 1.31, 95% C1 = 0.42- 3.05). Five deaths were observed (3.83 expected). A few SMRs were elevated for employees in the definite PFOA exposure group: 2 deaths from cancerofthe large intestine (SMR = 1.67), 1 from pancreatic cancer (SMR = 1.34), and I from prostate cancer (SMR = 1.30). In addition, employees in the definite PFOA exposure group were 2.5 times more likely to die fom cerebrovascular discase (5 deaths observed, 1.94 expected; 95% CI = 0.84-6.03). In the probable exposure group, 3 SMR should be noted: cancer of the testis and other male genital organs (SMR = 2.75, 95% CI = 0.07 ~ 15.3); pancreatic cancer (SMR = 1.24, 95% C1 = 0.45 2.70); and malignant melanoma of the skin (SMR = 1.42, 95% C1 = 0.17 - 5.11). Only 1, 6, and 2 cases were observed, respectively. The SMR for prostate cancer in this group was 0.86 (0-9). `There were no notable excesses in SMRs in the non-exposed group, except for cancerofthe bladder and other urinary organs. Four cases were observed and only 1.89 were expected (95% C1=0.58 - 5.40). Its difficult to interpret the results ofthe prostate cancer deaths between the first study and the update because the exposure categories were modified in the update. Only 1 death was reported inthe definite exposure group and $ were observed in the probable exposure group. Alofthese deaths would have been placed in the chemical plant employees exposure group in the first study. The numberofyears that these employees worked at the plant and/or were exposed to PFOA was not reported. This is important because even | prostate cancer death in the definite PFOA exposure group resulted in an elevated SMR for the group. Therefore, ifanyof the: employees' exposures were misclassified, the resultsofthe analysis could be altered significantly. "The excess mortality in cerebrovascular disease noted in employees in the definite exposure group was further analyzed based on number of yearsof employment at the plant. Three of the 5 deaths occurred in workers who were employed in jobs with definite PFOA exposure for more than 5 years but< 10 years (SMR = 15.03, 95% CI = 3.02 - 43.91). The other 2 occurred in employees with less than 1 yearofdefinite exposure. The SMR was 6.9 (95% C1 = 1.39 20.24) for employees with greater than yearsofdefinite PFOA exposure. In order to confirm trehgaiotnhaelrmeosrulttaslirteygarratdeisngwecreereubsreodvafsocrultaher dreifseeraesnecweeproepunloattiaonn.artTifhaectorefsudletastdhidcenrottifcihcaantegec.oding, `When these deaths were further analyzed by cumulative exposure (time-weighted according to ewixtphos9uryeeacrastoefgodreyf)i,niwtoerPkeFrOsAwietxhp2o7suyreearwseorfe e3x.3potsiumrees imnoprreoblaikbelleyPtoFdOiAeoefxcpeorscebdrojvoabsscourlatrhose. 3 37 disease than the general population. A dose-response relationship was not observed with years of exposure. "The slight excess in bladder cancer in the cohort as a whole should be noted, especially given the bladder cancer mortality experience at 3M's Decatur plant, which produces mostly PFOS. Bladder cancer mortality was 4 times higher in workers with high PFOS exposure jobs at 3M's Decatur, Alabama plant than the general population (SMR = 4.81, 95% CI = 0.99 14.06) (grAoluepx.andSeerr,u2m00P1Fb)O.ATlherveelesdienawthosrkweerrsearreeplorotweedr,aatntdheallDeocfatthuremploacntc,urwrheedrienPthFeOhAigihs euxspeodsuarse:an elastomer in fluoropolymer production or is producedas a by-product, than at Cottage Grove; `however, the manufacture ofPFOA began at Decatur in 199. Therefore, PFOA exposures will likely increase at this plant. It is not clear whether POA, PFOS or some other chemical may be responsible for the bladder cancer deaths observed at these plants; however, follow up should continue in an effort to shed some light on this observation. is difficult to compare the results of the first and second mortality studies at the Cottage Grove: plant since the exposure categories were modified. Although the authors claim that the newer exposure categories are more accurate, it is still likely that exposure misclassification occurred. Without measured exposures (serum PFOA levels), it is difficult to judge the reliability of the exposure categories that were defined. In the second study, the chemical plant employes were sub-divided into PFOA-exposed groups, and the film plant employees essentially remained in the "non-exposed" group. This was an effort to more accurately classify exposures; however, these: new categories do not take into account durationof exposure or lengthof employment. Another limitation to this study is that 17 death certificates were not located for deceased employees and therefore were not included in the study. The inclusion or exclusionofthese deaths could greatly change the analyses for the causesofdeath that had a small numberofcases. Follow up of worker mortality at Cottage Grove (and Decatur) needs to continue. Although there were `more than 200 additional deaths included in this analysis, itis a small number and the cohort is still relatively young. Given the resultsofstudies on fluorochemicals in both animals and humans, further analysis is warranted. 3.2.2 Hormone Study Endocrine effects have been associated with PFOA exposure in animals; therefore, 2 crosssectional studies were conducted on employees of a plant producing PFOA (Olsen, ct al., 19984). Medical surveillance, hormone testing and PFOA serum levels were obtained for volunteer workers in 1993 (n = 111) and 1995 (n = 80). Sixty-cight employees were common to both sampling periods. In 1993, the rangeofPFOA was 0-80 ppm (although 80 ppm was the limit of detection that year, so it could have been higher) and 0-115 ppm in 1995 using thermospray mass csopretcistorlo,pdheohtyodmreoterpyiaasnsdaryo.stEelreovneenshuloframtoen(eDsHwEeArSe)a,sessatyraeddioflr,oFmStHh,e s1e7rguammsmaamp.les. They were: hydroxyprogesterone (17-HP), free testosterone, total testosterone, LH, prolactin, thyroidstimulating hormone (TSH) and sex hormone-binding globulin (SHBG). 2 38 Employees were placed into 4 exposure categories based on their serum PFOA levels: 0-1 ppm, 1-< 10 ppm, 10- < 30 ppm, and >30 ppm. Statistical methods used to compare PFOA levels and hormone values included: multivariable regression analysis, ANOVA, and Pearson correlation coefficients. PFOA was not highly correlated with anyof the hormones or with the following covariates: age. alcohol consumption, BMI, or cigarettes. Most ofthe employees had PFOA serum levels less than 10 ppm. In 1993, only 12 employees had serum levels > 10 ppm, and 15 in 1995. However, these levels ranged from approximately 10 ppm to over 114 ppm. There were only 4 employees in the >30 ppm PFOA group in 1993 and only 5 in 1995. Therefore, it is likely that there was not enough power to detect differences in eitherofthe highest categories. The mean age of the employees in the highest exposure category was the lowest in both 1993 and 1995 (33.3 years and 38.2 years, respectively). Although not significantly different from the other categories, BMI was slightly higher in the highest PFOA category. Estradiol was highly correlated with BMI (r= 41,p <.001 in 1993, and r= 30,p < 01 in 1995). In 1995, all 5 employees with PFOA levels > 30 ppm had BMs > 28, although ths effect was not observed in 1993. Estradiol levels in the >30 ppm group in both years were 10% higher than the other PFOA groups; however, thedifference was not statistically significant. The authors postulate that the study may not have been sensitive enough to detect an association between PFOA and estradiol because measured serum PFOA levels were likely below the observable effect levels suggested in animal studies (55 ppm PFOA in the CD rat). Only 3 employees in this study had PFOA serum levels this high. They also suggest that the higher estradiol levels in the highest exposure category could suggesta threshold relationship between PFOA and estradiol Free testosterone was highly correlated with age in both 1993 and 1995. The authors did not report a negative association between PFOA serum levels and testosterone. There were no statistically significant trends noted for PFOA and either bound or free testosterone. However, 17-HP. a precursoroftestosterone, was highest in the >30 ppm PFOA group in both 1993 and 1995. In 1995, PFOA was significantly associated with 17-HP in regression models adjusted for opfososniebleemcpolnofoyuenede(rdsa.ta wHeorweevneort,prtohveiaduetdhoirnstshteatreeptorhta)t.thiTsheasrseocwiearteionnowsaisgnbiafisceadntonastshoeciraetsiuolntss between PEOA and cortisol, DHEAS, FSH, LH, and SHBG. "There are several design issues that should be noted when evaluating the results of this study. First, although there were 2 study years (1993 and 1995), the populations were not independent. Sity-cight employees participated in both years. Second, there were 31 fewer employees who participated in the study employees in cither year in 1995, thus reducing the powerof the study. with serum PFOA levels greater than 10 ppm. TThheirrde,wtehreecraolssso-very few sectional design ofthe study does not allow for analysis of temporalityof an association. Since the hall-lifeof PFOA is at least 1 year, the authors suggest that it is possible that there may be some biological accommodation to the effects of PFOA. Fourth, only one sample was taken for 3 39 ach hormone for each of the study years. In order to get more accurate measurements for some. ofthe hormones, pooled blood taken in a short time period should have been used for cach participant. Fifth, some of the associations that were measured in this study were done based on the results oafn earlier paper that linked PFOA with increased estradiol and decreased testosterone levels. However, total serum organic fluorine was measured in that study instead of PFOA, making it difficult to compare the results Finally, there may have been some measurement errorofsome of the confounding variables. In 1997, voluntary medical surveillance was again offered to employees (Olsen, et al., 1998). In this sampling period, cholecystokinin (CCK) levels were analyzed in 74 employees to determineifthey were positively associated with serum PFOA levels. CCK levels were observed because research has suggested that pancreas acinar cell adenomas seen in rats exposed to PFOA may be the result of increased CCK levels. Seventeenofthe subjects were common to all three sampling periods (1993, 1995, and 1997). The same statistical methods were used in this study period as used in 1993 and 1995, and the four PFOA exposure categories were also the The mean PFOA serum level in employees participating in the 1997 study period was 6.4 ppm (range 0.1 - 81.3 ppm). The mean CCK value was 28.5 pg/ml (range 8.8 - 86.7 pg/mi). The highest CCK values were reported in the 2 exposure categories less than 10 ppm. The means were 50% higher in these 2 categories than in the categories greater than 10 ppm (p = 06). `When adjusted for potential confounders, multivariable regression models indicated a weak negative association between CCK and PFOA; however, the data were not included in the report "The following explanations may indicate why this study failed to find a positive association between PFOA and CCK values = Itis possible that the hepatocarcinogenic effofpeercoxitsosme proliferators in rodents do not act the same biochemically in humans. + The scrum PFOA levels observed in workers may have been 00 low to detect an effect. Effects in animals were observed at higher doses than mostof the serum levels found in + CwoCrKkerresceptors may be different between rats and humans. Therefore, the monkey may + TbheeainmvoorlevaepmpernoptroifatCeCaKnimianlthmeodineiltiatotisontuodryptrhoempoatnicorneoatfipcaenfcfreecattsiocfcPanFcOeAr iisn humans. controversial. + Tsihneceraatcimnaayr cneoltl bmealaingnaapnpcrioepsr,iaitnedmuoceddelbyinctahrecisntougdeynosfinparnactsr,eaatriecrcaarnecienrhiunmhaunmsans, "The same methodological issues that applied tothe study in 1993 and 1995 applyto this portion. ofthe study as well 3 Wo 323 Cholesterol Study Based on animal testing which reported that animals exposed to PFOA develop hepatomegaly adentdearlmtienreaitfiosnismiinlalriepfidfemcettsabaroclipsrme,saentcrionssw-osrekcetirosnaelx,poocsceudpattoiPoFnOalA.stuIdnyawaPsFOpeArfporromdeucdttioon Vfaicpiolpirtoy,tei1n1s5,waonrdkehrepsawtiecreensztuydmieesd (tGoidlelitlearnmdianendwhMeatnhdeerl,se1r9u9m6)P.FFOorAtya-fefiegchttedwotrhkeeircshowlhesotewreorl,e efixvpeoesmepdltooyPeeFsOwAhforoeimth1e9r8v5o-l1u9n8t9eewreedreorinwcelruedeadskinedthteo sptaurdiyci(pa9t6e%, wpaerrteiciipnactliuodnedraien).theSixtytuhneeixrpjoosbeddesgcrroiuptpi.onT.heHsoeweemvpelro,yweehsenwesrceruamssluevmeelds wtoerheavaenalltyzledo,r ihowaPsFOnoAteedxptohasturtheisbagsreoduponof cwaotrekgeorrisesh,adotPaFlOsAerluemvefllsuomruicnehwgarseautseredthfaoncltahsesigfeynewroarlkpeorpsulianttioone.xpoTshuerreefgorroeu,psi.nstead ofjob "mTeoataslurseedr,umdufcluooritnhee cwoasst uosfeadnaalsyazisnugrrtohgeatseammpeleass.ureBlfoorodPFsOaAm.pleSsewreurmePaFnOalAyzweadsfonrottotal swaenrsuammfilnuaorsiene(,SsGePrTu)m, gglaumtammay!glouxtaalmoayc!ettriacnstfrearnassaem(inGaGsTe),(ScGhoOlTe)s,tesroelr,umlogwl-duetnasmiytyplyirpuovpircoteins (caLtDeLg)o,riaensodfhtiogtha-ldesnesriutym lMiupropirnoetelienvsel(sH:D<LI).ppAml,l o1f-3thpepmpa,rt>i3ci-pa1n0tsppwme,re>1pl0a-ce1d5ipntpom,fivae:nd > 15 pHaplmf.ofTthhee rwaonrgkeeorfs ftehlel sientroutmhefl>uo|ri-n3e vpaplumescwataesgo0ry0, w2h6ilpep2m3(hmaedasne3r.u3mplpemv)e.ls <AppI rpopxmimaantdel1y1 had levels > 10 ppm. "uTnhievraeriwaetreeannaolyssiegsniffoircacnhtoldeisftfeerroeln,ceLsDbLe,twaenednHeDxLp.osuIrnethceatmeuglotriiveasriwahteenanaanlaylsiysz,edthuesrienwgas nota aslicgnoihfoilcacnotnassusmopctiiaotnio,nabgee,twBeMeIn,taotnadl sciegrauremttfelusomroikneinagn.d cThhoelresetwereorleonroLsDtaLtiastfiecrallaydjsuisgtniinfgicfaonrt differences among the exposure categoriesof toal serum fluorine for SGOT, SGPT, and GGT. oHboewseevewro,rkienrcsre(aBseMsIin=S35GOkgT/ma?n)d SGPT occurred with increasing total serum fluorine levels in Since PFOA was not measured dirctly and there is no exposure information provided on the ienmfpolromayteieosn.(egT. hleenagutthhoofrscsmtpalteoytmheatnnto/exadpvoesrusree)c,litnhiecarlesouulttcsoofmetsheresltautdeyd tporoPvFidOeAleimxiptoesdure have bfeoernmeorbesmeprlvoeydeiens.theIsneaedmdpitliooyne,etsh;ehroawnegveoerf,eist uislnsotrecploeratretdhaftorthtehreelhivaesrbeeneznymfoelslowwe-ruepfoafirly woniedesafomrplmeanwyasofttahkeenefxrpoosmucraecchaetemgpolroiyese,e,intdhiicsaitsinngotvasruirapbriilsiitnyg.in1tthweoruesludltsb.e mGuicvhenmtohrateohnellypful 0 have several samples taken over ime to ensure their reliability. It also would have been tihnotesreewsthinogwteorceomorpiagrinealtlhye trehsouulgthsotfnotthe0wboerkeexrpsowsehdo0wesreee kifntohwernetwoebreeeaxnpyosdiefdfetroePncFeOsAamtoong 3s ul the employees in these groups. There were moreofthe "unexposed" employees (n = 65) participating in the study than those who worked in PFOA production (n = 48). 3.2.4 Study on Episodes of Care (Morbidity) In order to gain additional insight into the effectsoffluorochemical exposure on workers" health, an "episodeof care" analysis was undertaken at the Decatur plant to screen for morbidity outcomes that maybeassociated with long-term, high exposure to fluorochemicals. An "episode. of care" i a series of health care services provided from the start ofa particular disease or condition until solution or resolutionofthat problem. Episodes ofcare were identified in employees" health claims records using Clinical Care Groups (CCG) software. All inpatient and outpatient visits to health care providers, procedures, ancillary services and prescription drugs used in the diagnosis, treatment, and managementofover 400 diseases or conditions were tracked Episodesofcare were analyzed for 652 chemical employees and 659 film plant employees who worked at the Decatur plant for at least | year between January 1, 1993 and December31, 1998. Based on work history records, employes were placed into different comparison groups: Group A consistedofall film and chemical plant workers; Group B had employees who only worked in ther the film or chemical plant; Group C consistedofemployees who worked in jobs with high POSF exposures; and Group D had employees who worked in high exposures in the chemical plant for 10 years or more prior o the onsetofthe study. Film plant employees were considered to have little or no fluorochemical exposure, while chemical plant employees were assumed to have the highest exposures. Ratiosofobserved to expected episodesof care were calculated for each plant. Expected numbers were based on 3M's employee population experience using indirect standardization techniques.A ratio of the chemical plant's observed to expected experience divided by the film plant's observed to expected experience was calculated to provide a relative risk ratio for cach episode of care (RREPC). 95% confidence intervals were calculated for each RREPC. Episodes ofcare that wereofgreatest interest were those which had been reported in animal or epidemiologic literature on PFOS and PFOA: liver and bladder cancer, endocrine disorders involving the thyroid gland and lipid metabolism, disordersofthe liver and biliary tract, and reproductive disorders. "The only increased riskofepisodes for these conditions ofa prior interest were for neoplasms of the male reproductive system and for the overall categoryofcancers and benign growths (which tihneclouvdeerdalclacnacnecreorftchateegmoarlyefroerparlold4ucctoimvpeasryissteomn).groTuhpse.re Twhaes rainskirnactrieoawseadsrgirsekaotfeesptiisnotdheesgfroorup of employees with the highest and longest exposures to fluorochemicals (RREpC = 1.6, 95% CI = 12-211). Increased riskofepisodes in long-time, high-exposure employees also was reported for male reproductive cancers (RREPC = 9.7, 95% C1 = 1.1 - 458). It should be noted that the confidence interval is very wide for male reproductive cancers and the sub-categoryof prostate: 36 wl cancer. Five episoofdcearse were observed for reproductive cancers in chemical plant eobmspelrovyeedesin(1f.i8lmexppleacntteedm)p,loofyweheisc(h3.44 wexepreectperdo)s.tatTehciasncfeirnsd.ingOnsheouelpdisboedenootfepdrobsetcaatuesceanancer was ewxhceenssthienrperowsetraeteonclaync2erexmporotsaulrietycawtaegsoroibesser(vcehdemiinctahlepCloatnttaegmeplGoryoeveespalanndtfmiolrmtaplliatnyt study employees). The update of the study sub-divided the chemical plant employees and did not confirm this finding when exposures were divided into definitely exposed and probably exposed employees. `There was an increased riskofepisodes for neoplasmsofthe gastrointestinal tract in the high exposure exposure group group (RREPC (RREPC = = 1.8, 2.9, 95% 95% C1 C1 = = 1.21.7 3.0) and the 5.2). Most long-term employment, high of the episodes were atributable to benign colonic polyps. Similar numbersofepisodes were reported in film and chemical plant employees. In the entire cohort, only 1 episode of care was reported for liver cancer (0.6 expected) and 1 for bladder cancer (1.5 expected). Both occurred in film plant employees. Only 2 casesofcirrhosis of the liver were observed (0.9 expected), both in the chemical plant. There was a greater isk of lower urinary tract infections in chemical plant employees, but they were mostly due to recurring episodesof care by the same employees. It i difficult to dra any conclusions about these observations, given the small numberof episodes reported. Chemical plant employees in the high exposure, long-term employment group were 2 Y% times more likely to seek care for disordersofthe biliary tract than their counterparts in the film plant (RREPC = 2.6, 95% CI = 1.2 - 5.5). Eighteen episodesofcare were observed in chemical plant employees and 14 in film plant workers. The sub-categories that influenced this observation were episodesof cholelithiasis with acute cholecystitis and cholelithiasis with chronic or unspecified cholecystitis. Mostofthe observed cases ocurred in chemical plant employees. Risk ratiosofepisodesof care for endocrine disorders, which included sub-categories of thyroid disease, diabetes, hyperlipidemia, and other endocrine or nutritional disorders, were not elevated in the comparison groups. Conditions which were not identifieda priori but which excluded the null hypothesis in the 95% confidence interval for the high exposure. long-term employment `group included: disorders ofthe pancreas, cystits, and lower urinary tract infections. "The results ofthis study only should be used for hypothesis generation. Although the episode of care design allowed fora direct comparisonofworkers with similar demographics but different exposures, there are many limitations to ths design. The limitations include: 1) episodesof care amraeyrenpootrtneedc,esnsoatridliyseiansdeiciantceihdeingche,ri2s)k tofhie dnatca airedodifeffdiincsuelcatsteeo,i3n)temrparneytobfectahueseriasklraartgieosRRfoErpC eanpailsyosdiesswoafscalriemihtaedd tvoer6yyweiardse,c5ontfhiedeunticleizianttieornvoalfsh,etahletrhebcyarpersoevrivdiicnegsumnastyabrlefelercetsullotc,al4)metdhiecal practice patiems, 6) individualsmaybe counted more than once in the database because they can n 43 be categorized under larger or smaller disease classifications, 7) episodesof care may include the same individual several times, ) not all employees were included in the database, such as those on long-term disability 9) the analysis may be limited by the Software used, which may misclassify episodesof care, 10) the software may assign 2 different diagnoses to the same episode, and 11) certain services, such as lab procedures may not have been reported in the. database. 3.3 Acute Toxicity Studies in Animals 33.1 Oral Studies "The acute oral toxicity ofAPFO was tested in male and female rats in three studies. Death occurred at concentrations 2464 mg/kg (Intemat'l Res and Dev Corp., 1978). Abnorm findings upon necropsy (kidney, stomach, uterus) were observed (Glaze, 1997) at 500 mg/kg (higher concentrations were not tested). Clinical signsof toxicity observed in these three studies included the following: red-stained face, stained urogenital area, wet urogenital arca, hypoactivity, hunched posture, staggered gait, excessive salivation, ptosis, piloercction, decreased limb tone, ataxia, comeal opacity, and hypothermic to touch. In one study (Intemat'l Res and Dev Corp., 1978), the oral LDSO values for Charles River CD rats were 680 mg/kg (399 ~ 1157 mg/kg 95% confidence limit) for males; 430 mg/kg (295 - 626 mg/kg 95% confidence limit) for females; and $40 mg/kg (389 -- 749 mg/kg 95% confidence limit) for males and females. The remaining two studies provided LDSO valuesof (1) >500. `mek for male Crl:CD(SD)BR rats, and 250-500 mg/kg for female Crl:CD(SD)BR rats (Gilaza,1997); and (2) <1000 mg/kg for male and female Sherman-Wistar rats (3M Company, 1976). 332 Inhalation Studies "The acute inhalation toxicityof APFO was ested in male and female Sprague-Dawley rats, at a dose level of 18.6 mg/L. (nominal concentration), and exposure duration of one hour. Signs of toxicity, during, and up to 14 days afer the exposure period, included the following: excessive salivation, excessive lacrimation, decreased activity, labored breathing, gasping, closed eyes, mucoid nasal discharge, irregular breathing, red nasal discharge, yellow staining of the anogenital fur, dry and moist rales, red material around the eyes, and body tremors. Upon necropsy, lung discoloration was observe in a higher than normal incidence ofrats (8/10). Based on the study results, the test substance was not fatal {0 ats at a nominal exposure concentration of 18.6 mg/L and exposure durationofone hour (Bio/dynamics, Inc. 1979). 3 4H 333 Dermal Studies "The acute dermal toxicityof APFO was tested in male and female Hra(NZW)SPF rabbits, ata dose level of 2000 mg/kg, and a 24-hour exposure period. All animals appeared normal and exhibited body weight gain throughout the study, with the exception of one male that lost weight during the first week. Dermal imitation consisted oflight to moderate erythema, edema, and atonia; slight desquamation; coriaceousness; and fissuring. No visible lesions were observed upon necropsy. The dermal LDSO in rabbits was determined to be greater than 2000 mg/kg (Glaza, 1995). 33.4 Eye lrritation Studies "The eye imitation potential of APFO was tested in albino rabbits, ata dose levelof 0.1 gram. In two of three studies, APFO was determined to be 2 primary ocular irritant. In the studies in which APFO was found to be a primary ocular irritant, APFO was lef in contact with the eye for 7 days. then rinsed, or not rinsed. Imitation scores varied during the observation period. Imitation scores of the conjunctiva, irs, and comea ranged from 2 --4 in one study (Biosearch, Inc. 1976) and from 2 ~10 in the other study (3M Company, 1976a). In both studies, irritation remained evident for the durationofthe observation period (7-days post-exposure). In the study in which APFO was determined to be a non-iritant (Gabriel), the test substance was left in contact with the eye for S or 30 seconds, and then the eyes were rinsed. In this study, positive scores were reported for conjunctiva irritation for up to 7-days post-exposure, so the author's negative conclusion for ocular imitancy is problematic. 33.5 Skin Irritation Studies "The skin irritation potentialofAPFO was tested in albino rabbits in two studies, at a dose level 070.5 grams, under occluded test conditions. In one study (Riker Laboratories, Inc. 1983), APFO produced irreversible tissue damage in female rabbits, following a 3-minute, 1-hour, and 4ehour contact period. Moderate erythema and edema, as well as chemical bum, eschar, and necrosis, were observed following all three contact periods. An endpoint was not achieved in this study due to extreme irritation following each contact period. In the second study (Gabriel). APFO was reported as a non-irritantofskin after an exposure period of 24 or 72 hours, based on primary irritation scoresofzero. 3.4 Mutagenicity Studies APFO was tested twice (Lawlor, 1995; 1996) for is ability to induce mutation in the Salmonella E. coli/mammalian-microsome reverse mutation assay. The tests were performed both with and without metabolic activation. A single positive response seen at one dose level in S. typhimurium TAI37 when tested without metabolic activation was not reproducible. APFO did not induce mutation in either S. typhimurium or E. coli when tested either with or without mammalian activation. 3 ws aAnPdFwOitdhiodutnomtetianbdoulcieccahcrtoimvaotsioomnaulpatboercryattoitoonxsicincovnicternotirnathiuomnasn(Mluyrmlp,ho1c9y9t6es; wNhOeTnOtXes,te2dw00i0t),h APFO was tested twice for its ability to induce chromosomal aberrations in CHO eels in vitro. pInretsheencfiersatnadssaabys,eAncPeFoOfmientdaubcoeldibcoatchticvhatrioomn.osIonmtahleasbeecrornatdiaosnssaay,ndnoposliygpnliofiicdayntininbcortehastehse in cmehtraobmoolsiocmaacltivaabteirorna,tiAoPnsFwOeriendoubcseedrvsiegdniwfiitchaonuttiancctrievaasteisoni.n Hcohwreovmeors,omwahleanbetersrtaetdiownistahnd in polyploidy (Muri, 1996b). eAmPbFrOyowafisbrtoebsltaesdtsi.n aThceellcetlratnrsafnosrfmoartmiaotniaonndwcaystodteotxeircmiityneadssaasybcootnhdcuocltoendyitnrCa;nsHfo1r0mTayt,iomnouansde tfeoscitetdriannsiftohremrattihoencpooltoenntyiaolr.fTohceraesswaaysmneotheovdisde(nGcaeorfy t&raNneslfsoornm,at1i9o8n1)a,t anyof the dose levels SAiPgnFiOficwaants itnecsrteedastewsicinemiinctrohnuicnlveiivoanmdouwsaes cmoincsriodneurceldeunsegaastsaiyv.e uAndPeFrOthdeidconnodtitiinodnuscoeftahniys assay (Muri, 19962), 3.5 Subchronic Toxicity Studies in Animals Two unpublished 28-day feding studies were performed at Industrial Bio-Test Laboratories, Ine. (Metrick and Marias, 1977 andChristopher and Marias, 1977). In both rats and mice the liver was the target organ. In rats, males had more pronounced hepatotoxicity and histopathologic effects than females. Ina 28-day studyof ChR-CD albino rats, cight randomly assigned groups of five males and five females were studied (Metrick and Marias, 1977). After rats were allowed to acclimate foar week in individual cages they then received similar feed containing 0, 30, 100, 300, 1000, 3000, 8180,g0r00a,msorf3or0,m0a0l0esppanmdA7P6FgOrafmosr 2fo8rdfaeymsa.leAs.t tThheebeagniinmnailnsgowefrethoebssteurdvyedthdeaailnyimaanldsbaovdeyraged weights and food consumption were recorded weekly. Animals that died during the study were examinedforgross pathology, as were surviving animals at 28 days. It is stated that the study included a complete examination of gross pathology and a complete setoftissues and organs were examined, but the specific list is not supplied. Livers were weighed to determine relative organ weight then stained for histopathologic examination. Awlelreanniomaplrseminattuhree1d0e,a0t0h0saonrdot3h0e,r0c0l0i-npipcam gsirgonuspsoftdoixeidcbietfyorien tthhe oetnhderofgtrhoeupfsi.rstBwoedeyk.weTihgehrte gains were reduced in the groups receiving 1000 or more ppm. Slight reductions in body weight Rgaeidnucweerdefaolosdoionbtsaekrevweadsinobmsaelrevseedxipnorsaetsd fteod301000p0ppmpamnodrmhailgehseranindadfoesmea-lreeslfaetded10m0apnnpemr.. " ue Relative liver weights were increased in males fed 30 ppm or more and females fed 300 ppm or more. Gross pathological exam did not reveal treatment-related effects in kidneys or other organs besides livers. Focal to multifocal cytoplasmic enlargement of hepatocytes was noted in animals fed 300 ppm, and multifocal to diffuse enlargement of hepatocytes was noted in animals fed 1000 ppm or higher. These effects were more pronounced in males (Metrick and Marias. 1977). In a 28-day studyofCharles River-CD albino mice, cight randomly assigned groups of five males and five females were studied (Christopher and Marisa, 1977). After mice were allowed to acclimate for 8 days in individual cages they then received similar feed containing 0, 30, 100, 300, 1000, 3000, 10,000, or 30,000 ppmof APFO for 28 days. At the beginningofthe study the animals averaged 88 grams for males and 76 grams for females. The animals were observed daily and body weights and food consumption were recorded weekly. Animals that died during the study were examined for gross pathology, as were surviving animals at 28 days. Itis stated the study included a complete examination ofgross pathology anda representative setoftissues and organs were examined, but the specific list is not supplied. Livers were weighed to determine relative organ weight then stained for histopathologic examination. All animals in the 1000-ppm and higher groups died before the endofda9y.The entire 300-ppm group died within 26 days except 1 male. One animal in eachofthe 30 and 100-ppm groups died prematurely. Clinical signs were observed in mice exposed to 100 ppm and higher doses of PFOA. At100 ppm some animals exhibited cyanosis on days 10.and 11oftesting, but appeared normal throughout the restof the study. Animals feed 300 ppm exhibited roughed fur and muscular weakness as well as signsof cyanosis after 9 daysof treatment. Animals fed 1000 ppm exhibited similar effects after 6 days and those receiving 3000 ppm or greater doses exhibited effects afer4 days. All mice fed APFO lost weight. Reductions in body weight gain were followed by weight losses in mice fed 30, 100, or 300 ppm. A dose-related pattern was seen in the depressed body weights Relative and absolute liver weights were increased in mice fed 30 ppm or more APFO. Gross pathological examinationofkidneysorother organs besides livers is not discussed. Treatmentrelated changes were observed in the livers among all APFO treated animals including enlargement andor discoloration of 1 or more liver lobes. Histopathologic examinationofall APFO treated mice revealed diffuse cytoplasmic enlargement of hepatocytes throughout the liver (Dpeagnenleorbautlaironhyapnedrt/roropnheyc)roascicsoomfhpeapnaiteodcybtyefsocaanldtofomcualltbiifloecadlucctytporpolliafsemriactivoancuwoelrees.also noted in mice within all groups (Christopher and Marias, 1977), "Three 90-day subchronic toxicity studies have been conducted. One was conducted in rats w(Gaoslcdoenntdhuaclt,ed19i7n84m)a,loenreatwsa(sPacloanzdzuoclot,ed19i9n3r)h.esus monkeys (Goldenthal, 1978b) and the third a +7 In the monkey study, Goldenthal (1978b) administered rhesus monkeys (2/sex/group) dosesof 0, 3,10, 30or 100 mg/kg/day perfluorooctanoic acid (FC-143) in 0.5% Methocel7 by gavage for 7 days/week for 90 days. All doses were given in a constant volume; individual daily doses were based upon the weekly body weights. Animals were observed twice daily for general physical appearance and behavior and pharmacotoxic signs. General physical examinations were performed during the control period and monthly during the study period. Individual body weights were recorded weekly. Blood and urine samples were collected once during the control period and at | and 3 monthsofthe study for hematology, clinical chemistry and urinalysis. Monkeys were fasted overnight prior to the collection ofblood and urine samples. Organs and tissues from animals that were sacrificed at the endofthe study and from animals that died during the treatment period were weighed, examined for gross pathology and samples taken for histopathology. Histopathology was performed on the following organs from all monkeys in the control and treatment groups: adrenals, aorta, bone, brain, esophagus, eyes, gallbladder, heart (with skin, mceosreonntaerryicvelsysemlpsh),ndoudoe,derneutmro,phialreyunmg,ejaeljulnyummp,hcneocduem,,mcaomlomna,rryecgtluamn,d,kindenrevyes,(wliivtehr, lung, muscle), spleen, pancreas, prostate/uterus, ibjunction (bone marrow), salivary gland, lumbar spinal cord, pituitary, stomach, testes/ovaries, thyroid, parathyroid, thymus, trachea, tonsil, tongue, urinary bladder, vagina, identifying tattoo, and any tissues(s) with lesions. All monkeys in the 100-mg/kg/day groups died during the study. The first death occurred during week 2; all animals were dead by week S. Signs and symptoms which first appeared during week I included anorexia, frothy emesis which was sometimes brown in color, pale face and gums, swollen face and eyes, slight to severe decreased activity, prostration and body trembling. Three monkeys from the 30-mg/kg/day group died during the study; one male died during week 7 and the two females died during weeks 12.and 13. Beginning in week 4, all four animals showed slight to moderate and sometimes-severe decreased activity. One monkey had emesis and ataxia, swollen face, eyes and vulva, as well as pallorofthe face and gums. Beginning in week 6, two monkeys had black stools and one monkey had slight to moderate dehydration and ptosis of the eyelids. No monkeys in the 3 or 10 mg/kg/day groups died during the study. Animals in the 3mg/kg/day-dose group occasionally had soft stools or moderate to marked diarrhea; frothy emesis was also occasionally noted in this group. One monkey in the 10 mg/kg/day group was anorexic during week 4, haad pale and swollen face in week 7 andhadblack stools for several days in week 12. The other animals in the 10-mg/kg/day groups did not show any unusual signs or symptoms. Changes in body weight were similar to the controls for animals from the 3 and 10 mg/kg/day dose groups. Monkeys from the 30 and 100 mg/kg/day groups lost body weight afer week I. Atthe endof the study, this loss was statistically significant for the one surviving male in the 30mg/kg/day group (2.30 kg vs 3.78 kg for the control). 2 w8 Hematology values at the endofthe 1 and 3 months of treatment were similar for the control and the 3 and 10 mg/kg/day groups. At 30 mg/kg/day, the surviving male had decreased numbers of erythrocytes, decreased hemoglobin, decreased hematocrit, and increased platelets. Prothrombin time and activated prothrombin time were also increased. These increases were apparent at | `month but were much more marked at three months, Following one month of treatment, glicose was significantly elevated in the 3-mg/keg/day group (117 v5 89 mg/100 ml in the control). The authors of the report autibute this to a single high value for male #7366 who had a value of 131. The other three monkeys in the 3-mg/kg/day g`grroouuppss wheardele1v0e4lsaonfd11122,3-1m0g5/,1a0n0dm1,20r-emspge/c1t0iv0elmly., aGftleurcoonsee mleovnetlhsoifn tthreea1t0meanntd. 3A0t mthgr/ekeg/day `months of treatment, glucose levels were 81, 96, 88, and 66-mg/100 ml in the control, 3, 10 and 30 mg/kg/day groups respectively. "There was a decrease in alkaline phosphatase levels in the 30-mg/kg/day group (365 vs 597 IU/L in the control)at one month, which persisted in the one surviving male (360 vs 81 1U/1 in the control) at 3 months. Alkaline phosphatase levels in the 3- and 10 mg/kg/day groups at three: months were 783 and 743 U/l showing a dose-related trend toward decreased levels. SGOT levels were reduced in the 30-mg/ke/day groups at one month (59 vs 29 IU in the control) and in the one surviving male at 3 months (88 vs 45 U/l in the control). SGPT was elevated in both the 10 and 30 mg/kg/day dose groups at | month; the levels were 13, 34, and 44 IU/L in the control, 10 and 30 mg/kg/day groups, respectively. SGOT levels in the 10-mg/kg/day group were comparable to the controlsat 3 months (34 vs 31 1U/i the control) but were still elevated in the one surviving male in the 30-mg/kg/day dose group (46 IU). Cholesterol in the one surviving male in the 30 mg/kg/day group was elevated (240 vs 165 amcgo/n1t0r0omlIl)evaenldotoftal.2p1 ro/t1e0in0 amnldaanldbutomtianl ailnbtuhmsinanwiamsal2.w0e0revsreadcuocnetdr.olTolteavlelporfot4e.i6n2wa/s1050.5m2lvs "There were no treatment related changes in urinalysis studies at any time period studied. There were no macroscopic lesions noted at gross necropsyofany animals which died during the study or which were sacrificed at the endofthe treatment period. "The following changes in absolute and relative organ weight changes were noted: absolute and relative weightofthe hearts in females from the 10 mg/kg/day group were decreased; absolute brain weightof females from this same group were also decreased and relative group mean weightof the pituitary in males from the 3 mg/kg/day group was increased. The significance of these weight changes is difficultot assess, as they were not accompanied by morphologic changes. " 49 One male and two females from the 30 mg/kg/day group and all animals from the 100 mg/kg/day `group had marked diffuse lipid depletion in the adrenals. All males and females from the 30 and 100 mg/kg/day groups also had slight to moderate hypocellulariyof the bone marrow and. moderate atrophy of lymphoid follicles in the spleen. One female from the 30-mg/kg/day group and all animals in the 100-mg/kg/day group had moderate atrophy of the lymphoid follicles in the lymph nodes. No other compound related lesions were seen in at the 30 and 100 mg/kg/day groups. No treatment related lesions were seen in the organs of animals from the 3 and 10 mg/kg/day groups. "The levels of PFOA in the serum and liver are presented below. Dose Serum (ppm) Liver (ppm) Liver total (ug) Males Females ~~ Males Females [I 005 007 3s 6s 37 ETI ND ND 04 9 ND 07 7 ND 10 0 ND ND 125 80 30 145 ND 60 125 10 ND ND 100 325 Males Females 3s 250 350 ND ND 600 ND ND 750 8000 7500 4000 9000 6000 20000 Inthe first rat study, Goldenthal (19782) administered CD rats (S/sex/group) dietary levels of 0, 10, 30, 100, 300, and 1000 ppm perfluorooctanoic acid. These dose levels are approximately equivalent (0 0.56, 1.72, 5.64, 17.9, and 63.5 mg/kg/day in males, and 0.74, 2.3, 7.7, 22.36 and 76.47 mg/kg/day in females. Animals were housed individually in wire mesh cages and had free a`mcocretaslsittyo. fDoeotdaailneddweaxtaerm.inaAtniiomnaslwsewreerpeeorbfsoerrmveeddotnwcieceadwaeielky.foIrndsiivgindsouafl tbooxdicyitwyeiagnhdtfaonrd food consumption were recorded weekly during the pretest and treatment periods. Blood and urine samples were collected during the pretest period and at | and 3 monthsof the study for hematology and clinical chemistry and urinalysis. At week 13, sex and group, frozen and shipped to the sponsor for analysis, pooled serum samples. Organs and tissues from animals that were sacrificed al the end of the study and from two females that died during the treatment period were weighed, examined for gross pathology and samples taken for histopathology. Histopathology was performed on the following organs from rats from the control, 100, 300, and 1000 ppm dose groups: brain with cervical cord, lumbar spinal cord, peripheral nerve, eyes, pituitary, thyroid with parathyroid, adrenals, lung, heart with coronary vessels, aorta, spleen. (medsueondteenruimc,ljyemjpuhnunmo,dei,letuhmy)mucsol,obn,onpeanwcirtehasm,alrirveorw, K(isdtneerynsu,m)u,risnaalirvyabrlyagdldaenrd,,tessmteasl, oivnatreisetsi,nes ptrhoest1a0tea,ndute3r0u-s,ppsmkidno(sme agmrmouaprsywegrlaenadl),soaneyxatimsiunee(ds)miwcirtohsgcroopsiscallelsyioansn.d Lliivveerrssafmrpolmesratfsrofmoamll dose groups were frozen and sent10 the sponsor for analysis w" so One female in the 100 and one female in the 300-ppm group died during collection of blood. These deaths were not considered to be treatment related. All ther animals survived until scheduled sacrifice. There was a significant reduction in mean body weight in males in the 1000-ppm group (362 vs 46 2 in the control group). Food consumption was reduced in males in the 100, 300 and 1000-ppm groups, but the differences were not statistically significant. Males in the 30, 100, 300 and 1000-ppm groups had significantly reduced numbers of erythrocytes at the endofthe treatment period. The values were 7.95, 7.05, 7.16, 6.72, and 6.94 in the control, 30, 100, 300 and 1000-ppm groups, respectively. Males had reduced leukocyte values compared to the controls in all dose groups, but were statistically significant at the 300 ppm group only; leukocyte values were 10.64, 8.88, 9.33, 9.35, 7.63, and 8.06 in the control, 10, 30, 100, 300 and 1000 ppm groups, respectively. A similar phenomenon was seen with hemoglobin values, which were reduced at al, dose levels but were significant at the 10-ppm dose level only. Hemoglobin values were 16.2, 14.7, 15.0, 15.4, 14.9, 13.1 in the control, 10, 30, 100, 300 and 1000 ppm groups, respectively. There was no similar effect upon the hematological parameters of female rats in the study. Males at the 30, 100, 300, and 1000-ppm dose levels had increased glucose levels (mg/100 mi), which were statistically significant at all but the 100-ppm dose level. Reported glucose levels were 121,120, 136, 134, 143 and 135 mg/100 mi forthe0, 10, 30 100, 300 and 1000 ppm groups, respectively. B.U.N. levels were elevated in males at the 100, 300, and 1000 ppm dose: rleesveplesc;timveelayn, vcaolmupeasraetd9100d1a6y.s2wmeer/e10200.4m,l2f3o.r9tahnedco3n5t.r1olsm.g/A1l0k0almilnefoprhtohsephtahtraeesedowsaesgerloeuvpast,ed in males in the 100, 300, and 1000-ppm groups; the levels were 147, 204 and 212 [UA for the three groups, respectively, compared to 104 1U/1 for the controls. Females showed no similar changes in biochemical measurements, Neither males nor females showed any treatment related changes in urinalysis parameters although females from all groups showed a higher frequencyofoccult blood in the urine than did males. "The only gross necropsy observation was noted in males a the 1000-ppm dose level. These animals had enlarged livers that showed varying degrees of surface discoloration. Neither females from the 1000-ppm dose level nor males or females from the lower dose levels showed such effects. Both absolute and relative liver weighs were significantly increased in males in the 30, 300 and 1000-ppm groups and in one femal in the 1000-ppm group. Compound-related liver lesions occurred in all male rats in the 100, 300 and 1000-ppm groups. These lesions consisted of focal to multifocal, very slight-to-slight hypertrophyof hepatocytes in centrilobular to midzonal regionsof the affected liver lobules. Insome instances these lesions were accompanied by an 4s sl increased amount of yellowish-brown pigment resembling lipofuscin in the cytoplasm of hepatocytes and occasionally in sinusoidal lining cell. The incidence and severity of the lesions was more pronounced among male rats at the 1000-ppm dietary level, A comparisonof the serum levels of PFOA is shown below. The greater toxicity observed in the males than in the females is due to the gender difference in elimination as demonstrated by the differences in serum PFOA levels. Dose PFOA in Serum (ppm) Males Females 0 0 0 10 21 ND 30 34 0.15 100 36 ND 300 38 025 000 49 065 ND = Not Determined. In the second rat study, Palazzolo (1993) administered 45-55 male Sprague-Dawley rats per group, doses of 1, 10, 30, or 100 ppm (approximate mean compound consumption at week 13 of 0.05, 0.47, 1.44, and 4.97 mg/kg/day) APFOadlibitum in the dict for 13 weeks. Two control groups (a nonpair-fed control group and a control group pair-fed to the 100 ppm dose group) were also exposed during that period. Following the 13-week exposure period, 10 animals per `group were fed basal diet for an additional 8-weeks post-treatment and observed for any signs of recovery. All est diets were assayed and evaluated for test material homogeneity and stability. All animals were observed twice daily for mortality, moribundity, and general clinical signs of toxicity. Body weights were recorded once before exposures began, weekly during the treatment Period, and then on the day ofnecropsy. Food consumption was recorded weekly for all dosedgroups, including the nonpair-fed control groups daily for the pair-fed animals, and then weekly for all of the animals retained for the recovery phaseof the study. A total of 15 animals per dosed-group were sacrificed following 4, 7, or 13 weeksof treatment; 10 animals per dosed group were sacrificed after 13 weeks of treatment and following weeks of non-treatment. Serum samples collected from 10 animals per dosed-group at each scheduled sacrifice during treatment and from $ animals per dose-group during recovery were analyzed for estradiol, total testosterone, luteinizing hormones, and for test material residue. The levelofpalmitoyl CoA oxidase, an indicator ofperoxisome proliferation, was analyzedfroma sectionof iver that was obtained from animals per dosed-group at each scheduled sacrifice. The following organs sfermoimnaalll vaensiimclael,spartoestaacthe,sccohaegduullaetdinsagcrgilfaincde,waenrdeuwreetihgrhae.d:Tbhreaifno,llloivweirn,gltuinsgssu,etseisntsth(eosnee)s,ame animals were preserved in 10% phosphate-buffered formalin and examined macroscopically: extemal surfaceof the body, all orifices, the cranial cavity, the extemal surfacesofthe brain and spinal cord, the nasal cavity and paranasal sinuses; the thoracic, abdominal, and pelvic cavities 6 [2 and viscera; and also examined microscopically: any observed lesions, brain, live, lungs, testes (one), seminal vesicle, prostate, coagulating gland, and urethra. [n addition, the following tissues were preserved in glutaraldehyde for electron microscopic examination: brain, liver, lungs, testes (one), seminal vesicle, and prostate. In the analysisofthe data, animals in groups exposed 10 1, 10, 30, and 100 ppm APFO were compared to the control animals in the nonpair-fed group, while the data from the pair-fed control animals were compared to animals exposed 0100 ppm APFO. All test diets were considered to be homogeneous and stable under the experimental conditions. All animals survived to scheduled sacrifice, with the exception ofone animal in the 100-ppm dosed-group that was sacrificed on week 4 due to severe neck sores unrelated to treatment. Twice-daily examinations ofall animals were unremarkable. At 100 ppm, significant reductions in body weights were seen compared to the pair-fed control group during week | and the nonpair-fed control group during weeks 1-13 (i.. throughout treatment). During recovery, however, no reductions in body weights were apparent. Body weight data in the other dosed-groups were comparable to conirols. At 100 ppm, mean body weight gains weresignificantlyhigher than the pair-fed control group during week | and significantly lower than the nonpair-fed control group during weeks 1-13. At 10 and 30 ppm, mean body weight gains were significantly lower than the nonpair-fed control group at week 2. These differences in body weight gains were not observed during the recovery period.Significantdifferences in food consumption were observed at 100 ppm during weeks | and 2 only, when compared to the nonpair-fed control group; no other significant differences in food consumption were noted. There were no significant differences among the groups for anyof the hormones evaluated in the serum. Likewise, serum analysis of test material residue showed no increase in serum APFO levels over the course of treatment. Statistically significant higher hepatic palmistry CoA oxidase activity was observed at:30 and 100 ppm; however, this effect returned to control levels by the endof the recovery period. At 10 ppm, statistically significant higher levelsof hepatic palmitoyl CoA oxidase. activity were observed at week S only. Mean enzyme activities were highest during week 8 for animals exposed to 10, 30, and 100 ppm. All dosed groups exhibited significant increases in absolute and relative liver weights and hepatocellular hypertrophy were observed at weeks 4, 7, and 13, compared to the pair-fed control group. The authors suggested that these changes might be associated with peroxisome proliferation, especially since increases in hepatic palmitoyl CoA. oxidase activity were also observed at this dose level during treatment. During recovery. however, none of the liver effcts were observed, indicating that these treatment-related liver effects were reversible. Therefore, under the conditionsofthis study, NOAEL of 1.0 ppm (0.05 mg/kg/day) and a LOAEL of 10 ppm (0.47 mg/kg/day) are indicated based on reductions in body weight and body weight gain, and on increases in absolute and relative liver weights with hepatocellular hypertrophy. a <3 3.6 Developmental Toxicity Studies in Animals Three prenatal developmental toxicity studies ofAPFO have been conducted, one inhalation and two oral studies "The firstofthese studies was an oral developmental toxicity study in rats (Gorner, 1981). Based on the results ofa range-finding study, an upper dose level of 150 mg/kg/day was set for the definitive study in which five groups of 22 time-mated Sprague-Dawley rats were administered 0,005, 1.5, 5, and 150 mg/kg/day APFO in distilled water by gavage on gestation days (GD) 615. Doses were adjusted according to body weight. Dams were monitored on GD 3-20 for clinical signsof toxicity. Individual body weights were recorded on GD 3, 6,9, 12, 15, and 20. Animals were sacrificed on GD 20 by cervical dislocation and the ovaries, uteri, and contents were examined for the numberofcorpora lutea, numberofviable and non-viable fetuses, number ofresorption sites, and numberof implantation sites. Fetuses were weighed and sexed and subjected to external gross necropsy. Approximately one-thirdofthe fetuses were fixed in Bouin's solution and examined for visceral abnormalities by free-hand sectioning. The remaining fetuses were subjected to skeletal examination using alizarin red. Signsofmaternal toxicity consistedofstatistically significant reductions in mean maternal body weights on GD 9, 12, and 15 at the high-dose group of 150 mg/kg/day. Mean maternal body weight on GD 20 continued to remain lower than controls, although the difference was not statistically significant. Other signs of matemal toxicity that occurred only at the high-dose `group included ataxia and death in three rat dams. Noothereffects were reported. `AdministrationofAPFO during gestation did not appear to affect the ovaries or reproductive: tractof the dams. Under the conditionsofthe study, a NOAEL of5mg/kg/day and a LOAEL of 150 mg/kg/day for matemal toxicity were indicated. A significantly higher incidence in fetuses with one missing stemebrae was observed at the highdose group of 150 mg/kg/day: however this skeletal variation also occurred in the controls and the other three dose groups (at similar incidence but lower than the high-dose group) and therefore was not considered to be treatment-related. No significant differences between treated and control groups were noted for other developmental parameters that included the mean number ofmales and females, total and dead fetuses, the mean number ofresorption sites, implantation sites, corpora lutea and mean fetus weights. Likewise, fetal lens finding initially described as a variety of abnormal morphological changes localized to the areaof the embryonal nucleus, was later determined to be an artifactof the free-hand sectioning technique and therefore not considered to be treatment-related. Under the conditionsofthe study, a NOAEL for developmental toxicity of 150 mg/kg/day (highest dose group) was indicated. A second oral prenatal developmental toxicity study was conducted in rabbits (Gortner, 1982). Based on the resuls ofa range-finding study, an upper dose level of 50 mg/kg/day was set for the definitive study in which four groups of 18 pregnant New Zealand White rabbits were administered 0, 1.5, 5, and 50 mg/kg/day APFO in distilled water by gavage on gestation days a Sh (GD) 6-18. Pregnancy was established in each sexually mature female by iv. injection of pituitary lutenizing hormone in order to induce ovulation, followed by artificial insemination wasitdhay0.05 omflgoefsptaotoilone.d sAemcoennstcaonltledcotsede fvroolmummealoef1rabmbIi/tksg; twhaesdaadyomifnisitnesreemdi.natIinodnivwiadusaldebsoigdnyated weights were measured on GD 3, 6,9, 12, 15, 18, and 29. The does were observed daily on GD 3-29 for abnormal clinical signs. On GD 29, the does were euthanized and the ovaries, uterus `and contents examined for the numberofcorpora lutea, live and dead fetuses, resorptions and implantation sites. Fetuses were examined for gross abnormalities and placed in a 37C ivinsccuebraatloarnfdosarke2le4t-ahloaubrnsourmravliivtailesc.hecAk. blPouopdsswaemrpelesuwbasseqtuaeknetnlyfreoumthsainxidzoeedsapnrdioerxatomidnoesidnfgorand twheerneosnenGt Dto t18heasnpdo2n9s;ora floirvearnaslaymspils.e wTahisstiankfeonrmfartoimonthweassaumneavaaniilmaabllseoant GthDe t2i9m.eoAfltlissamples review. Signs of matemal toxicity consistedofstatistically significant transient reductions in body weight gain on GD 6-9 when compared to controls; body weight gains retumed to control levels on GD 12-29. AdministrationofAPFO during gestation did not appear to affect the ovaries or reproductive tract contents of the does. No clinical or other treatment-related signs were reported. Under the conditions of the study, a NOAELof0 mg/kg/day, the highest dose tested, for matemal toxicity was indicated. mNaolessiganinfdicfaenmtaldeisf,fedreenacdesorwelrvee nfoettuesdesb,eatnwdeefnetcalonwterioglhstsa.ndLtirkeeawtiesdeg,rothueprse fwoerrtehenonusmigbneirfiocafnt differences reported for the numberofresorption and implantation sites, corpora lutea, the conception incidence, abortion rat, or the 24-hour mortality incidence of the fetuses. Gross dneevcerlooppsmyeanntdaslketloextiaclitvyisccoenrsaislteedxaomifanadtoisoen-srewlearteeduinnrcermeaarskeaibnlaes.keTlehteaolnlvayrisaitginono,fextra ribs or 13" rib, with statistical significance at the high-dose group (38% at 50 mg/kg/day, 30% at 5 mg/day. 20% at 1.5 mg/kg/day, and 16 % at 0 mgrky/dey). A statistically significant increase in 13" ribs-spurred occurred in the mid-dosegroup of 5 mg/kg/day; however, the biological significanceofthis effect is uncertain since in both the high- and low-dose groups, this effect oThcecruerfroerde,atutnhdeesratmhee croatnediatnidonwsaosfntohte ssttautdiys,tiacalLlOy AsiEgnLiffiocrandtelvyedliofpfemreennttaflrtoomxiccointtyrooflsS.O mg/kg/day (highest dose group) was indicated. Staplesct al. (1984) also conducted a developmental toxicity studyofAPFO. The study design consistedofan inhalation and an oral portion, each with two trialsor experiments. The first trial twhaesstehceotnedratrtioall,ogtyhepodratmiosnowfetrheealsltouwdey,d itnowlihtitecrhatnhdetdhaempsupwserweerseacsraicfriicfeidceodnoGnDda2y1;35w-hpiolsetin partum. Fraotrs tpheerignrhoaulpateixonpopsoertditoonoAfPtFhOe sbtyudwyh,otlhee-tbwoodytrviaalpsocroninshiasltaetdioonf0120,pr0e.g1n,an1,t1S0p,raangdue2-5Damwgle/ym,6' hfiorustrsa/nddays,ecoonnGdDtri6a-l1s5,.resIpnetchtievoerlayl,pwoerrteoandomfitnihstsetrueddy,0 2a5ndan1d0012mgS/pkrga/gduaey-DAaPwFleOyirnactsofmoroitlheby gavage on GD 6-15. For both routesof administration, females were mated on an s-necded bbaysibsodayndwewihgehnt tahnednausmsbigenreodftmo agtreodupfsembaylerostwataisonbriendo,rdtehreyofwrearnek.ranFkienadllwyi,thtiwnobardedeidtiinongadlays `roups (six dams per group) were added to cach trial that was pair-ed to the 10 and 25 mg/m' groups. 2F1oratnhdeotbesreatrovleodgydapiolrytifoornoafbntohremsatludcyli(nircaall soingea)s,. dOamnsGwDer2e1,wetiheghdeadmosnwGerDe s1,ac6r,i9f,ice13d,b1y6, and ctehrevriecaplroddiuscltoicvaetison aanotdfeeuxaacsmhinaendimfaorl awnaysgervoaslsuaatbendo.rmaTlhietioevsa,rileisv,eruwteeriugshtasndwecroentreenctosrwdeedreand examined for the numberofcorpora lutea, live and dead fetuses, resorptions and implantation vsiitsecs.eraPl,upasnd(lsvkeelaetnadl daletaedr)atwioenrse. cTohuentheeda,dwseoifaglhledcoanntdrosleaxnedd ahnigdhe-xdaosmeidnegdrofuopr efxetteumseasl,were examined for visceral alterations 2s well as macro- and microscopic evaluation of the eyes tFroiarltorinael tuwpo,toiGn Dwhi21c.h tThweodadmaysswbeerfeoraelothweeedxpteocltietde,datyhoefppraorcteudruirteiowna,seatchhe sdaammewaassthhotusfoerd in an individual cage. The date of parturition was noted and designated Day | PP. Dams were wweerieghseadcriafnidcede.xamPiunpesdwfeorrecclionuinctaelds,igwnesigohnedD,ayasnd1,e7x,am1i4n,eadndfo2r2ePxPt.emOalnaDltaeyrat2i3onPs.P aEllacdhampsup Twhaes esyuebsseoqfuetnhetlpyuwpseiwgehreedaalnsdo eixnsapmeicnteedd foonrDadavyesrs1e5 calnindic1a7l PsiPgnfsootnheDaiynhsal4a,t7i,on14p,oratnidon2a2nPdP.on Days 27 and 31 PP for the gavage portionofthe study. Pups were sacrificed on Day 35 PP and examined for visceral and skeletal alterations. Inhalation Exposure "Trial One: Treatment-related clinical signs of matemal toxicity for ral one (ieratology) occurred at 10 and 2u5nkmegm/pmt'aappnedacroannscies,teadnodflewtehtaragbydionmefonusr,dcahmrosmaotdtahceryeonrdrhofetah,echerxopmoosruhrienpoerrtihoeda,(haigghe-neral c1o3n,caenndtra17t)i.onFogordoucpononsluym).ptiTohnreweaosutsiognfif1i2cdanatmlsy rdeideudcdeudraitngbottrhea1t0m.eanntdat2525mgm/gm/'m;'h(oownevGeDr,1n2o, sbiogdnyifwiceaingthtdiwffeerreenaclessowoebrseernvoetdedatbtehtewseeecnontcreanttreadtaionnds,pawiir-tfhedstagtriosutipcsa.l sSiiggnniiffiiccaanncteraetduthcetihoingsh-in acotntcheenhtirgaht-icoonnocnelnyt.ratLiioknewgirsoeu,p.staUtinsdteicraltlhyesciognnidfiitciaonntsoinfctrheeasesstuidny,meaaNnOlAivEerLwaenigdhLtsOwAeErLe sfeoern `maternal toxicity of1 and 10 mg/m', respectively, was indicated. EF) 6 No effects were observed on the maintenanceof pregnancy or the incidenceofresorptions. Mean fetal body weights were significantly decreased in the 25-mg/m' groups and in the control `group pair-fed 25 mg/m'.A detailed microscopic visceral and eye examination of the fetuses did not reveal any treatment.related effects; however in the control group that was pair-fed 25 `wmags/mo'b,searvsetadt.istUicnadlleyrstihgenicfoincdainttiionncsroefatshede isntcuiddy,enaceNofOfAetEuLseasndwiLthOApaErtLialfloyr odsesvieflioepdmsetnetranlebrae toxicity of 10 and 25 mg/m', respectively, was indicated. Trial Two: Clinical signsofmaternal toxicity seen at 10 and 25 mg/m' were similar in type and incidence as tthhoasnecodnetsrcorlisb,eadlftohroutrgihaltohnee.difMfaetreenmcaelwabsodnyotwestiagthitstigcaailnlydusirginnigfitcraenta.tmeInntaadtdi2t5iomn,g/2mo'utwoafs 1l2e.ss dams died during treatment at 25 mg/m'. No other treatment-related effects were reported, nor were any adverse effects noted for anyofthe measurementsofreproductive performance. Under trheespceocntdivietliyo,nsowefrteheindsitcuadtye,d.a NOAEL and LOAEL for maternal toxicity of 1 and 10 mg/m', Signsofdevelopmental toxicity in this group consistedofstatistically significant reductions in pup body weight on Day 1 PP (6.1 gat 25 mg/m' vs. 6.8 gin controls). On Day4s and 22 PP, pup body weights continued to remain lower than control, although the difference was not mstga/tims'ticvasl.ly50s.i1gniinficcoannttro(lsD)a. 4yNoPP:sig9n.i7figcaatnt2e5ffmegct/smw'evrse.1r0e.p3oritnecdonftorlollosw;inDgayex2te2rnPaPl: e4x9a.m0ignaatti2o5n ofthe pups or with ophthalmoscopic examination of the eyes. Under the conditionsof the study, a NOAEL and LOAEL for developmental toxicity of 10 and 25 mg/m', respectively, were: indicated. Oral Exposure "Trial One: `Three outof25 dams died during treatment of 100 mg/kg APFO during gestation (one death on GD 113 two on GD 12). Clinical signs of matemal toxicity in the dams that died were similar to those seen with inhalation exposure. Food consumption and body weights were reduced in treated animals compared to controls. No adverse signsof toxicity were noted for any of the reproductive parameters such as maintenanceofpregnancy or incidenceofresorptions. Likewise, no significant differences between treated and control groups were noted for fetal weights, or in the incidencesof malformations and variations; nor were there any effects noted following microscopic examination of the eyes. si s7 "Trial Two: Similar observationsfor clinical signs were noted for the dams as in rial one. Likewise, no adverse effects on reproductive performance or in any of the fetal observations were noted. 3.7 Carcinogenicity Studies in Animals 3.7.1 Cancer Bioassays "The carcinogenic potential of APFO has been investigated in a two-year feeding study in rats (3M. 1987). In this study, groups of 50 male and 50 female Sprague-Dawley (Crl:CD BR) rats were fed diets containing 0, 30 or 300 ppm FC-143 for two years. Groups of 15 additional rats per sex were fed 0, or 300 ppm FC-143 and evaluated at the one-year interim sacrifice. The mean actual test article consumption was: males, 1.3 and 14.2 mg/kg/day; females, 1.6 and 16.1 `mg/kg/day for the low and high-dose groups, respectively. "There was a dose-related decrease in body weight gain in the male rats and to a lesser exten, in the female rats as compared to the controls; the decreases were statistically significant in the high-dose groups of both sexes. The body weight changes are treatment related since feed consumption was actually increased (rather than decreased). There were no differences in mortality between the treated and untreated groups; the survival rates at the end of 104 weeks for the control, low-, and high-dose groups were: male, 70%, 72% and 88%; females, 50%, 48% and 58%. The only clinical sign observed was a dose-related increase in ataxia in the female rats; the incidences in the control,low-and high-dose groups were: 4%, 18% and 30%. Significant decreases in red blood cell counts, hemoglobin concentrations and hematocrit values were observed in the high-dose male and female rats as compared to control values. Clinical chemistry changes indicaoftiivveer toxicity included increases in alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (AP) in both treated male `groups from 3-18 months, but only in the high-dose males at 24 months. Increases in relative liver and kidney weights were noted in both high-dose male and female rats. Significant nonnoplastic lesions were seen primarily in the liver and testis; there were increases in the incidence of liver masses, hyperplastic nodules and foci, and in testicular masses in the highdose male group. Other liver toxic effects include dose-related increases in the incidence of dbioftfhusmealheepaantdomfeegmaalloecyttroesaitsed, gcryosutposi;d dtehgeesneeirnactrieoans,esanwderpeorsttaaltimsotincoanlluyclsieganrifcieclalntinifniltthreathioinghi-n dose males. A statistically significant, dose-related increase in the incidenceof ovarian tubular hyperplasia was found in female rats; the incidenceof this lesion in the control, low-, and highdose groups was 0%, 14%, and 32%, respectively. Based on these toxic effects, the high dose. selected in this study appears to have reached the Maximum Tolerated Dose (MTD). Based on decreased body weight gain, increased liver and kidney weights and toxicity in the hematological and hepatic systems, the LOAEL for male and female rat is 300 ppm. [Based on increases in the incidenceofataxia (a clinical sign) and ovarian tbular hyperplasia (which is reversible), the LOAEL for female rats is 30 ppm.] ES sg Atthe termination of the study,a slight increase in the incidenceofvarious neoplasms (tumors of the liver, testis, thyroid, adrenal and mammary glands, etc.) was seen in the treated animals. Among them, the increased incidences of testicular (Leydig) cell adenomas in the high-dose: male rats, andofmammary fibroadenoma in both groups of female rats were statistically significant (P< 0.05) as compared to the concurrent controls. The incidenceofthe Leydig cell tumors (LCT) in the control, low- and high-dose males was 0%, 4% and 14%, respectively: the respective incidencesof mammary fibroadenoma in the female groups were 22%, 42% and 48% "The increases are also statistically significant as compared to the historical control incidences (LCT, 0.82%; mammary fibroadenoma, 19.0%) observed in 1,340 male and 1,329 female Sprague-Dawley control rats used in 17 carcinogenicity studies (Chandra ct al, 1992). The spontaneous incidence of LCT in 2-yearold Sprague-Dawley rats in other studies was reported to be approximately $%(cited in: Clegg etal, 1997). Therefore, under the conditionsofthis study. APFO is carcinogenic in Sprague-Dawley rats, inducing Leydig cell tumors in the male rats and mammary fibroadenomas in the female rats Ina follow-up 2-year dietary study (300 ppm) in male Sprague-Dawley (CD) rats, APFO was fhoouwnedvetro,idnedtuacilesloivnerthteumstourdsyadnedsipganncarnedattiucmaocrinianrccideelntceumwoerrseinnoatddrietpioorntetdo (LCeoyodkigctcealll.,t1u9m9o4r)s.; APFO has also been shown to promote liver carcinogenesis in rodents (Abdellateital,1991; Nilsson etal., 1991). 3.7.2 Mode of Action Studies `The mechanisms of toxicological/carcinogenic actionof APFO are not clearly understood. Short-term genotoxicity assays suggest that APFO is not a DNA-reactive compound; it is nonmutagenic in the Ames test using five strofaSalimonnelsla typhimurium, or in an assay with oSfactcuhmaorrsombyyceAsPcFeOrevisisdiuaee 1(0G.r3ifnfoitnh-gaenndotLooxnigc,m1e9c80h)a.nisAvma,ilianbvloelvdiantgaaicntdiivcaattieonthoaft rtehceepitnodrusctainodn perturbations of the endocrine system. 37.2.1 Liver Tumors It has been well documented that APFO is a potent peroxisome proliferator, inducing peroxisome proliferation in the liveofrats and mice (e.g, Ikeda et al, 1985; Pastoor et al., 1987; Sohlenius etal. 1992). A sex-related difference in the inductionofliver peroxisome. proliferation exists in rats (Kawashima et al., 1989), but not in mice (Sohlenius et al., 1992) "The higher inductionof liver peroxisome proliferation in male rats was shown to be strongly dependent on the sex hormone testosterone (Kawashima et al, 1989). Like many other peroxisome proliferators, APFO has also been shown to cause hepatomegaly (an carly biomarker of peroxisome proliferator hepatocarcinogenesis) in rats (Takagi, et al., 1992; Cook, 1994) and mice (Kennedy, 1987), and induce oxidative DNA damage in liver of rats (Takagi et al., 1991). "The totality of these data appears to suggest that the liver toxicity and carcinogenicity of APFO `may be related to induction of peroxisome proliferation. Meanwhile, estrogen has been shown to 5 <9 promote hepatocarcinogenesis in rats (Yager and Yager, 1980; Cameron et al., 1982); an hiencpraetaoscearinciensatgroegnesnisleivnelrstsafter APFO exposure (discussed below) may also play a role in 3.7.2.2 Leydig Cell Tumors A large numberof non-genotoxic compoundsofdiverse chemical structures have been reported to induce Leydig cell tumors (LCT) in rats, mice, or dogs. A reviewofthe available information `ognroLupCsTbainsdeudcotinonthieniramniomdaelssolfeadctaiwoonrk(Cslheogpgpcatneall.,to19c9l7a)s.sifTyhtehecsoemcmoomnpotuhnedmse iinntothseemveonde of action for most compounds is tht these compounds affect the hormonal contro ofLeydig cel growth by disrupting the hypothalamic-pituitary-testicular axis at various points that result in increasing the serum levelsofluteinizing hormone (LH). It has been postulated that in addition to stimulating the production of testosterone, LH may also play a mitogenic role in the Leydig. cserlolsw;ihasstuismtualianteidnigncmreedaisaetoinrscisruccuhlaatsin1gGLF-H1,leTvGeFls-fa,ndlecuhkrootnriicensetsimaunldatviaorniooufsLferyedeirgadciecllaslsbcyan lead to LCT development (rev. in: Clegg et al., 1997). A seriesofstudies have been conducted to investigate the mechanism oftumor formation in `amla.le19S96p)r.agNueo-Dsiagwnliefiyca(nCtDi)ncrraetasseexspionsLedHtoweArPeFsOee(nCionotkheetraals.,a1f9e9r2;trBeiaetgmeelntetofalA.,P1F9O95;atLiu et vSiagrniiofuiscadnotslye lienvcerlesasfeod a1n4ddatyess,toHstoewreovneerl,evseelsruwmeraendsitgensitfiiccualnatrlyledveeclrseoafseeds.traIdtiwolasweproestulated that the elevated estradiol levels may cause Leydig cell hyperplasia and tumor formation by acting as a mitogen and/or enhancing growth factor secretion; the transforming growth factor a (TGF a), which binds to the epidermal growth factor (EGF) receptor and stimulated cell eprxopleirfiermaetnitosn,hafovreisnshtoawncne,thhaatsAbPeFenOdientcercetaesdedinthLeeyldeiveglcsoelfless(tTreaedridosl ebtyali.n,d1u9c9i0n)g. cyStuobcsheqruoemnet P450 XIX (aromatase), which converts testosterone to estradiol. Peroxisome proliferators are known to induce B-oxidation and cytochrome P-450 monooxygenases by binding to the rpeecreoptxoirss)o.meIptrsolbiefleireavtieodntahcattiAvaPtFioOn rinedicceesacpy(ttPocPohArrRome; Pa4s5u0bfXaImiXly(oafrosmtaetraosied)hboyrbmionndeing to and activating the PPAR AAlPthOouginhtshiegn1i4fidcaayn-tsitnucdrieeass,eist ainppLeHarswetrhaetniontcrseeaesneiinnSLpHralgeuveel-sDacawnlneoytrbaesrauflteed torueattomebnet of involved (in addition to increased estradiol level) in the induction of LCT by APFO. In these studies, significant increase in hepatic aromatase (which converts testosterone to estradiol) aacbtsicvritvieeds aisnstohceiattreedatweidtrhasd.ecTreesatsoestdesreonreu,mwtheisctohstisersoynnethleevseilzsedanadndinscerceraesteedd ebsytrtahdeioLlelyedvieglscwlelre,. oisrdreergutloatmeadinbtyaiLnHa;deteqsutaotsetetreosnteosatnedroLneHpfloarsmmaa lcelvoesles,d-rleodoupcefdeetdebsatcokstseyrosnteemleivneltshe(cHaPusTedaxbiys In increased aromatase activity) are expected to lead o increased LH levels through the negative si 60 sfteueddibeasockfmcehcehmainciaslms.foIr hwahsicbheetnhepopirnotpeodseodutmtohdateoinfcarcetaiseosnicnalLlsHfmoraeylenvoattaedlwLaHy,s abnedsetheantin all caroempseonmseaotfiotnhemaexyplhaanvaetioocncsurfroredfatiolirnegsttooredehtoemcteocshtaansgiessainndLiHnaplpevreolpsri(aCtleetgigmeitnaglo,f1s9a9m7)p.ling 3.7.2.3 Mammary Gland Tumors Estradiol has also been and the overexpression shown to ofTGF a stimulate has been the secretion suggested as ofTGF by one possible mammary epithelial factor in producing cells snuesotpalianseida c(eLllipertolailf.,er1a9t8i7o)n.ofHemnacmem,airtiys ptousmsoirblceeltlshaantdhethAePFsOub-sienqduuecnetddeclveevlaotpimoenonft oefstradiol DleavwellseymaryatsalisnoabddeirteisopnotnosiLblCeTf(orditshceusdseevdelaboopvmee)n.toInffmacat,mmthaisriys cfoinbsriosatdeentnowmiatsh itnheSpmreacghuaenism brayts.whIit chhasspboenetnadneemoounssmtraamtmeadrtyhatntehoepleaarslmysawpepreeardaenvceeloapneddhiinghagsipnogntfaenmeaoluesSipnrcaigdueencDeaowfley mexapmomsuarreytogleannddotguemnooruss iensturnotgreeantaedn,dapgrionlgacfteinmaalseaSrpersaulgtuoef-Daanwalcecyelreartastiisngdueeffteoctioncnrenaosremdal, age-related perturbationsof the estrous cycle in this strainofrat (Cutts and Noble, 1964; Chapin etal. 1996) 3.7.2.4 Pancreatic Tumors TothheermpeecrhoaxniissommebyprwolhiifcehraAtoPrsFOalsiondpurcoedducpeanpcarnecarteiactiacciancairnacerlcleltlumtourmsoriss uinnkrnatosw.nA.vaAilanbulmebdeartaof s(uCgCgKes)tltehvaetltsheecopanndcarreyattiochaecpiantairc ccehlolletsutmaosriss (arCeooreklaetteadl.t,o1a9n94i;ncOrbeoasuerninetsael.r,um1c99h7o)l.ecCysCtKokiisnian gparnocwrtehatfiacctaocrintahartcehlalssbienernatssh(oLwonngtnoecstkiemru,la1t9e87n).ormHaol,weavdeaprt,ivdea,taanodn ntehoeprloalsetoifc gCrCoKwthinof pancreatic tumor formation are conflicting. 4.0 Hazards to the Environment 4.1 Introduction `mTahedeaqmuoatriecdtiofxfiicciultty bayndsehvaezraarldoprfobAlPeFmOs dtioscauqusasteidcboerlogwa.nisTmhsesweaspraossbelsesmesd.coTmhpilsictaatsekdwtahse task ofFurdtehteerrmmoirnei,ngtihfestehepreocboltoexmisciltiymitteesdtstwheerceonvfaildiednacnedthcaotulcdoubled used in the assessment be placed on the toxicity test values, and the inherent tthouxsiciintytuarndlohawzearreddotfheAcPoFnOfidteonacqeuaotficcoonrcglaunsiisomnss.that could be drawn in assessing ss 6 G1e)nAervaalrliye,tytohefdaimffmeorneintumAPsFalOtSorwtihtehtveatrryaibngutdyelsaimgnmaotniiounms saanltdwlaotsntuemstbeedr.sThweereexatecsttecdo.mposition and identification of impurities, which may affect toxicity, in each lot number used is not known. 2f)roAm vaaprpireotxyiomfatteesltyin1g9l7a4b-o1r9a9t6o.rieTshicsonsdituucatteidontsheerAvePdFtOo itonxcirceiatsyesotvuedriaelslotvesetrvaarpiearbiiolidotyfatnidmethus made inter-laboratory comparisons more difficult. m3)ajPourrictoynocftehr.e tPeusrtietdymwaatsernioat,sourffpiecriceennttlytceshtarmaactteerriiazleadnidnptehrecseenttesotst.heIrnmsatoemreialt(esst)s, iwt aapspeaared. utsheadt. 10P0ur%ittyeostfctheestmimcaatlerwiaals duoseesd;afinfeoctthteorxsicaitcyheamnidcaslhoofulledsbseertapkuernitiynt(oapapcrcooxuinmtawtehleynp5o%ss)ibwlae,s by expressing toxicity on the same purity basis m4)anWaytoefrt,haentiosxoipcirtoypatnesotls,soflovrennto,oobrviaocuosmbinidniactaitoendorfeabsootnh. wSeorleveunstesdawrietmhitxheedtewsittmhattheeritaelstin `umsaetderiinaltetsotsmawkheeriet tmhiescciobnlceewntirtahttihoenstoesfttdhielutteisotnmwaattereiralbeafroereextthreetmeesltyislboewguann.d aSovlevreyntssmaalrle: oafmotuhensteosfttuedsitesmawtheyriaalsmoluvsetntbewaasdduesdedtoorthweatsesetvcehnamfboeurnsd. to1:bweansecneostsacrlye.ar Ifnrofmactt,he3sMummaries tshuamtmoafrtihzeedtesetascuhbstetsatncaen.d"stTahtuesd,"iDnattahomseaytesntostwahcecrureat1e0ly0%retleasttemtatoexriialcowfiattshyenottesutsesda,mptlhee with vtoaxliuceistyonvaalu1es00h%adtetsotbceheamdijcuasltebdastiost,a5k0etihnattotahcecoteusntts ctoheulpdebrceenctomspoalrveendt.(s), and to express the c5)hetmniaclallthceosnecetnotxriactiityontsesatrseoinnlsytenaodmianlawlaytsesrt ecchoemmimceanldceodncseontthraattioonnes cwaenreacucsuerda.telMyedaestuerremdinteest dtheetearctmuianledestthecthtehmeicnaolmicnoanlcecnotnrcaetnitornattioownhsiacrhe otnhleyt,esftororegxaanmipslmes5ar0e%exopfotsheed.meIafsiutriesd `comnecaesnutrreamteinotnsso, fthcehteomxiiccaitlycvoanlcueenstrwaitlilohnasvheotuoldachcaovredibnegelnybteakaednjourstmeaddbeya5v0ai%l.ablAe.nalTyhteinc,al rvoelcaotvielirtyy)rattheastcmoiuglhdthlaovweerbetehen adcettuearlmiconnecde,natrnadtpiohnyssitcoowchhiecmihctahleptresotceosrsgeasn(ies.mg.s wheyrdreoleyxspioss,ed could have been taken into account. Nominals may be used when measured concentrations are taken and the relationshoifp both is known. tIhneosredetersttolipmriotcaetieodnswiatnhdatnoyassosrutomeftehnavtitrhoenmneonmtianlalhaczoanrcdenrtervaiteiwonist wwaesreneacne"sasdaerqyutaoteig"nore: teoxxpirceistsyiaornoe fbatsheedmoenasweulrle-dcstteastblcihsehmeidcvaalluceonsc(elntorwatiiso>n1s.00Cmrgi/teLr:iamfeodraisusmesosrimngoddeegrarteeeoisfa>c1:ut1e00 `mg/L: high is <1 mg/L). 6 62 42 Acute Toxicity to Freshwater Species `Semvienrnaolws(pPeicimeesphwaelreestpersotmeedltaos)as,sbelsusegtihlelascuuntfeisthox(iLcietpyoomfisAmPaFcOh;rotchheisreusi)nc,lwuadteedrthfelefaa(tDhaepadhnia madajgunsat)e,d atond1a00gr%eteenstalcghaem(iSceallenaansdtrtuesmtcraepsrulitcsorarneutpurme)s.enTtehdeitnoxTiacbilteyste2st(oerngdapnoiiznetds bhyavteestbeen Substance) and 3 (organized by test species). Each value is related toa testing facility and reference. Twelve ranged tests from were 70 to conducted 843 mg/L. with Itis fathead unclear minnows; 96-h why this range LCS0 values (based on mortality) is so wide. Assuming these studies arc vtawloida,caunted dvaulcuetso ftohre blliumeigtialtliosnusnfdiisshcuaslsseodiandbiocvaet,etlhoewsetotxoixciictiyty(9v6a-lhueLsCi5nd0iscoatfe>l4o2w0,toaxincdit5y6.9The mg/L). rNainngeeadcfutreomte3st9stwoe>r1e0c0o0ndmugc/tLe.d Twihtehldoawpehrnivdalsuaensdar4e8-ihndEicCaSt0ivevoalfumeosd(ebraasteed toonxiicmimtyo,bibluitztahtei:on) wide range makes interpretation difficult dSeenvseintyt,ecsetsllwceoruentcso,nadnudcdtredywwietihghgtrse)enraanlggaeed;f9r6o-mh 1E.C2S100 >va6l6uesmg(/baLse(tdhoenExgSr0owctehllrdaeten,sicteyllvalue toofxi1c.i0t0y0, bmags/eLd iosnexthceluadceudtefcrroitmertiha.isdTihsceuslsoiwoenr).valTuheewolouwledravlsaolubeinidnidciactaetsivheiogfh mtoodmeordaetreate tmoex/iLci,tyb,absaesdeodnocneltlhecocuhnrtosn,icformogdreereantaelgcraietewraisona(ls0o.c1a1lc0ulmagt/eLd).in oAne14s-tdudEy.CST0hivsaliuseoinfdi4ca3tive othfelmoowscthrseonnsiictitvoexitceistty,spbeacsieeds oinntthhee4c4hr%onAiPc FcrOitetersitonsa(m1p0lmeg,/Ld)a.phnGirdesewneariegateheanpepxetarmeodstto be sensitive, and fathead minnows were the least sensitive. 5 [2 [ablSummeary of AcuteEcologicalToxicity Dafor tAPFaO (groupedby test subsance) Fest Organism a mL) TestSample: APFO ammonium salt. [Fatheadminnow (Pimephalespromelas) ENLCS0 [766 [3M Company.1980s | 99661LCLCS500 [7[40Wardetal1.9,5 aegis (Lepomismachroching) b[PeenLLCCsSo0_|>[456290 T[o3wM CCoommppaannyy,, 11997758 Wate es (Daphnia mgr) a i | eves [wo 1052Environmental Laboratory, | | (Green age (Selena capricomutiom) [Bact (Photobacterumphosphoreun) Inetiated sade `est Sample: APFO Fatheadminnow (Pimephalespromelas) f[ePnGrEcCsSo J[i31o0o0 [WWardaetaln1d995 elt| [[3X00-mmiinn ECCSSO[$[ [17300M 3MEnvirCoomnpamney.n1t9a8lL7aaboratory, [TamiaNOEC [1000 19961 [3M Company, 1950 [30-minEC50 1000 [3VCompany.19874 | ] [P 96hELCsH0 L[5s4C04S3[031M CCoommppaannyy.. 11998754 1 FTaetshteaSdammipnlneo:w (APiPmFepOhalaems mproomneilausm)salti[n65h0L%CwSaOt[er500 pebNoiC 0 J[EenvmirioSsysotenms., nInec.., 119990290|s te nes (phi magne) lh ECs0 [ 19900 19900 s oH prams nt ep J pre `est Sample: APFO ammonium salt in 80% water [FWaattheeademainDnaopw(Pimmepahganle)spromelas)___E[CS9)6h[[210W.aWCanrSdde0taetlal_,.119_999660[2 s94| (Greensigac (Sclenasiam capricarmam)~~ 6[PGREECCsS00 [[3>[9666W6 a[Wnanddcat.al 1199996605 Fr Pahtrn ppc pOAECSD [01996E5 nna ab. | pores [3 [139M9En6vironmental Laboratory, | [Retvated hdge [min ECS0_ [663 1996E4nvironmental Laboratory, {Test Sample: APFO in 50% isopropanol 199% [ri ------ | | 19960 1995 Fristner Proslepie)| PVECS [S1995 mee 1995 Waterfle Daplniamagna) ______J_ s {[E WardH etalE 199C 5 S0| Waterfla(Daphniamagna) [Green sigse(Setenastrumcapricormuum) SHECSO [39 DehECS0 2.1 [|WWaanrddetetaall.,11999055 | [Green algae (Selenasirumcapricormumum) 96HECS0__6 [Wardetal,1995 | "VTahleuseesvwaelresamdajyubsetein0dcornespirsetseenntt d1u0e0{70d3ifcerenitndgireetdsiteenstted. 0 6S OrganTani Summary of loin Toil DtVaoir AVEO Grouped Ryefseprieees Fant inno ineptpromt [PERTCE0("0mg/_L)[|Compa. 19780| BeonLC5r 0 poe r" o 1 SMCCoommpp9 aannyy.15 9785 7]| | p[9De6ehANLLCCOsSR>0E l>oo5et00 J|[WoEanvrcidreohSyestemimos,nIbneocc.., 1990. p-- EN FNhotCt3017 [FLeRsoWlitlbsuraLpabroroanto.rie1s78ne 55 | fe cso [ir Wibny Libnre 1a95 Cp ------1 Fontes Fa Company, 1978 Penrcso [seoTM [3M Company, 1978 eros api may ERECS 207 Examen Los 98 [-- | [4E8KCS0__[>[31MEn0vir0onm0entTMal Laboratory, 1982 fiEsCESCs)0 25807 [oEnnvdiocsaln. c06. 19908 prece p5 erE [oaWilbury Laboratories, Inc., 3% [3M Company, 1954 [dayNOEC[227M Company. 954 | fihhEECCs S0 [$17 [WWaandnedvaal. 11999985 1 Green gas (Seas capr[OiE EcCS0ar[396mu[Wamrd cal 19965. BEehCESCS)) 56066 [[WTA Weatialr.r19Ld96a5 bora9.95 [dy ECS 457 [eatwawy, 981 fonECS) po Wardetal, 1995 b Be en hJ[3i6aE e7 v[C cWasrdeS seta,l,) o119o9955 6 us(tabacippm [ioimnnECC5s0[70 F9 o Compa8 ny 0] Bmo inECsM0lE_v[o5l0Labomion 5960 [50minEC50 [390 [3M Environmental Laboratory. 1996c [Activated studge Pmt he [30-min ECS0 [30-minECS0_|> 500 | [50E5m0 i6n647 "Values were adjusted 10 represent 100% active ingredient. - "These values may be inconsistent due to different diets tested. "Tested substance was APFO ammonium salt. "Tested substance was APFO nes wes ALE son lS wee Tested substance was APFO ammonium salt in 80% water. i Test Sample: APFO (44%) in 27.9%water and 27.2% isopropanol [3M Company, 1980b [3M Company, 1987d. 3[M3M EEnnvviirroonnmmeennttaall LLaabboroartaortyo,ry. 1199990b6.4| a 67 5.0 References 3M Company. 1976a. Primary Eye Iiation Study-Rabbits 3M Company. 1976b. Acute Oral Toxicity in Rats-T-1585. 3M Company. 1977. Ready BiodegradofaFtCi-1o4n3 (BODICOD/TOC). Environmental Laboratory. St. Paul, MN. 3FlMuoCroocmhpeamnicyalInFtCem-a1t4i3onAacluRteesOeraarlchToaxnidciDteyv(eLlDospom)eSnttuCdoyrpionrRaattiso.n.Stu1d9y78Nbo.. F1l3u7o-r0a9d1, 3M Company. 1979. Technical Report Summary- Final Comprehensive Report: FC-143. (USEPA AR-226 528) 3M Company. 1980c. Ready Biodegradation of FC-143(BOD/COD). Lab Request No. 56255. Environmental Laboratory. St. Paul, MN. 3M Company. 1981. Water, Acetone and Toluene Solubility Estimates. Environmental Laboratory. St. Paul, MN. 3M Company. 1984. Chronic toxicity to freshwater invertebrates. 3M Company. Environmental L1a9b8o5raa.to9r6y-,hSotu.rPaauclu,teMsNt.aticLtaobxRiecqituyetsotfNatuhmeabdermiC1n0n06o.--w FX-1001. Februar2y. 3NoM.CC1o0m0p6a.nyE.nvi1r9o85nbm.entRaeladLayboBriaotdoergyr.adaStt.ioPnauol,f FMXN-.10F0e1br(uBaOrDy/C14O.D). Lab Request 3M Company. 1987. Activated Sludge Respiration Inhibition Test. Environmental Laboratory; Lab Request Number E1282. St. Paul, MN. S3uMlfoCnoamtep:anCuyr.re1n9t99sau.m"mTahreySocfiehnucmeoanfsOerrga,anhiecalFthl,uoarnodchteomxiisctolroyg"yadnadta"".PerFfelburouraoroyctSa,ne1999. (8EHQ-0299-373). 3M Company. 1999b. SEHQ-0699-373. Supplement. May 26, 1999 3PeMrfCluoomrpoaonctya.ne2s0u0l0fao.niVcolAucnitdaarnydUVsaerainoudsESxaplotsFuorremsI.nf3orMmaCtoiomnpParnoyfilseubfmorission to USEPA, dated April 27, 2000. @ 68 `3AMcidCoamnpdaSnaylt.s. 2300M0bC.oVmoplaunntyasruyUbmsiessainodn tEoxpUoSsEuPreA,IndfaotremdatJiuonneP8r,of2i0l0e0.for Perfluorooctanoic h3uMtpC:o/mwpwaenwy3e,M.2c00o0m/.aboAubto3umt/w3oMrlWdowrilded/wriedlee:a3seMhtPmhla.sing Out SomeofIts Specialty Materials. 3M Company. 20012. Environmental Monitoring - Multi-City Study, 3M Environmental Laboratory, June 25. In U.S. EPA Administrative Record AR226-1030A. 3M Environmental Laboratory. 1990a. Microbics Microtox Toxicity Test. St. Paul, Minnesota. 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S3tManEdnavridr~onTmeesnttCaolnLtarboolraRteofreyr.en2c0e01#bT.CRC-h9ar9a0c3t0e-ri0z3a0t.ioPnhaSsteu:dySoolfubPilFiOtyAD(eltoter#m3i3n2a)t,ioPnr.im3aMry Laboratories, St. Paul, MN. cAabrdceilnloagtefn,eAsi.sG.byrepaetr,flVu.o,roToacptearn,oHi.cSa.ciadn,da Rpoebrerofxriosiodm,eMp.rol1i9f9e1r.atoTrh.eTmoxoidcuolla.tiAopnplo.fPahtarlmiavecrol. 111: 530-537 FAilneaxlanRdeepro,rtB..HD.iv2i0s0i1oan. ofMoErntvailriotnymsetnutdayloafnwdorOkcecurpsaetmiponlaolyeHdeaalttht,heSc3hMooClototfaPgueblGircovHeealftachi,lity. Universityof Minnesota, April 26, 2001. 61 Alexander, B.H. 2001. Mortality studyofworkers employed at the 3M Decatur facility. Final RUenpiovretr.sitDyiovfisMioinnonefsoEtnav,irAopnrmieln2t6a,l2a0n0d1Occupational Health, School of Public Health, Behar, B. Stein, G. Science 1966, Vol. 154, p.1012. BSyesatcehm,.S. 31M99C5ao.mpIannhyib,itEonrvyirEfofnemcetnotfalL-L1a3b4or9a2totroyM,iSctraPbaiucls,'MMNi,crLoatboRxeTMqTuoexsitcnituymbAenarlNy2z1er69. July 26. BEenavcihr,onSm. en1t9a9lb.LabIonrhaitboirtyo,ryStE.ffPeacutlo,fMLN-,13L4a9b2RoenquAecsttivNa2t1e6d9S,luJdugley.26.3M Company Biegel, LB, Hurt, M.E., Frame, S.R., O'Connor, J.C., and J.C. Cook. 2001. Mechanisms of extrahepatic tumor induction by peroxisome proliferators in male CD rats. Toxicological Sciences. 60: 44-55. Bio/dynamics Inc. 1979. An Acute Inhalation StudyofT-2305 CoC in the Rat. 3M Company, St. Paul, MN, Project No. 78-7184. Biosearch, Inc. 1976. Primary Eye Irritation Study -- Rabbits. Philadelphia, PA. 3M Company. St. Paul, MN. Boeri, R., Magazu, 1, Ward, T. 1995a. Acute toxicity of L-13492 to the Daphnid, Daphnia Magna. TR. Wilbury Laboratories, Inc., 3M Company Lab Request number N2332, July 13. PBoiemreip,hRa.,lesMparogmaelzausJ.,,TWRar.d,WilT.bu1r9y95Lba.borAactuortieesT,oxIincci.,ty3oMfLC-o1m3p4a9n2y10Lathbe RFeaqtuheesatd nMuimnbneorw, N2332, July 13. Boeri, R., Magazu, J, Ward, T. 1995. Growth and reproduction Toxicity Test with L-13492 and the Freshwater Alga, Selenastrum capricormutum. T.R. Wilbury Laboratories, Inc., 3M Company Lab Request number N2332. August 3. PBiomeerp,hRa.l,esKopwraolmseklia,s.P.T,R.WaWridl,buTr.y1L9a9b5od.ratAorciuetse,TInco., x3MioCfcoNmi2p8a0tn3y-y,2La0btRheeqFuaetshteandumMbienrnow, N2803-2. November 16, PBoiemreip,hRa.,lKeoswparlomseklia,s.P.T,-WRa.rdW,ilbT.ury19L9aSbeo.ratAocruitees,TIonxci.c,i3tyMoCfoNm2p8a03n-y4LtaobthReeqFuaetshteanduMmibenrnow, N2803-4. Novembe2r1 o 70 NBo2e8r0i,3-R2.,anKdowhaelsFkrie,shPw.,ateWrarAdl,gaeT,. S1e99e5n.asGirroowichapaincodRrneuptruomd.ucTt-iRo.nWTioxlibcuitryLTaebsotrawtiotrhies, Inc. 3M Company Lab Request numberN2803.2.November2. Boeri, R., Kowalski, P., Ward, T. 1996a. Growth and Reproduction Toxicity Test with N2803- 4CaonmdpatnheyFLriebshRweaqtueersAtlngau,mbSeelreNn2a8s0t3r-u5m.caMpiacorrcnuhtum. T.R. Wilbury Laboratories, Inc., 3M 2MBoaegrnia,.R., TKRo.waWlistkiu,ryP.L,abWoarradt,oriTe.s, 1I9nc9.6b3.MAcCuotmepTaoxniyciLtayboRfeNq2u8e0s3t-n4utmobethre ND2a8p0h3n-i5d., DMaaprhcnhia mBoaegrni,aR.T,RK.owaWlislkbiu,rPy.LabWoarradt,erTi.s,1I9n9c6.e.3MAcCutoemTpoaxniyciLtayobfReNq2u8e0s3t-N2utmobteherND2a8p0h3n-i2d., DMaaprhcnhia 25. Boyd, S. 1993a. ReviewofTechnical Report Summary: Adsorption of FC 95 and FC 143 in Soil. Michigan State University. May 19. Boyd, S.A. 1993b. Reviewof Technical Notebook. Soil Thin Layer Chromatography. Number 48277, p30. Michigan State University. CsBueorvmrepirsaa,lnyJM.Muo;roOclhseenn,icGa;lsS.impIsntoenr,imCR;epMaonrdte#l1,,JC. o(r2p0o0r0a)teDeOtcecrumpiantaitoinoanlofMseedircuimnehaDlfe-lpiaversmofen3t,M Cameron, R.G. Imaida, K., Tsuda, H. and Ito, N. 1982. Promotive effectsofsteroids and bile acids on hepatocarcinogenesis initiated by diethylnitrosamine. Cancer Res. 42:2426-2428. CarlforsJ,. etal. Colloid Interface Sci. 103, 332 -336.(1985) dChaanlderar,atMs..,ArRiclhe.y,ToMx.iGo.1e.olan6d6:J4o9hn6s-o5n0,2 D.E. 1992. Spontaneous neoplasms in aged sprague- CShyamppion,siRu.Em.,ovSetrevvieenws:, eJ.nT.d,oHcurignheesmo,dCu.lLa.t,ioKneolcfef,epW.rRo.d,ucHteisons., FRu.nAd..aAnpdplD.asTtooxn,icGa..P.24119-9167.. MCahjeomrguBarreda,ki2h0v00o.ughChinemAgLuFaErdFiIrnec.,FPigrhetsisngRFeloeaamseTeRce:hnCohleogmyg.uaJrudneI3n0c,or3p0o0r0a.ted Announces CMhircies,toFpihnearl,RBe.poarntd MInadruistsr,iaAlJ.Bio-1T97e7s.t L2a8bo-rDaatyorOireasl, ITnocx.icSittuyyStNuod.y 8w5i3t2h-F1C0-65154,3 3inMARlebfienroence No. T-1742C0C, Lot 269. o at Chlyepegrgp,lEa.sDi.,a aCndooakd,enJo.Cm. aCfhoarpmiant,ioRn.:Em,eFcohsatneri,smPsM.anDd.raenldevDaanscteonto,hGu.mPa.ns1.99R7e.prLoedyudci,g cTeolxlicol. 11107121 CLOGP(v4.71) ~ Calculationofhydrophobicity as Log P(o/w). 2001. Daylight Chemical information Systems, Inc. Cinodouckt,io1n.C.b,yHupretr,oxMisEo,meFprraomlei,frSaRt.orasnindCBri:egCeDl,BLR.(B.CD1)99r4a.t.MTeocxhiaconliosgmissto,f1e4x:t3r0a1h,epaabtsitcratcutm#or 1169. Cbeuhtasvi,or1.ofatnudmoNrosb.leC,anRc.Le.r R1e96s4..24Es:t1r1o1n6e--1i1n2d3u.ced mammary tumors in the rat I. Induction and PDearkfionr,m2a0n0c1e. CoNaotbiunhgsi"koinTsPauidnat,&DaCiokaitniInngduIsntdruisetsryLiMda.ga"zFilnueo,roJpuonley2m0e0r1E,mpu.l5s6i-o6n6.for High- SDeCrPvi,ce1s9.98D.allDaisr,ecTtXo.ryof World Chemical Producers: 1998 Edition. Chemical Information UDuSPEoPnAt,.2d0a0t0e.d JVuonleun2t3,ar2y00U0s.e and Exposure Information Profile. DuPont submission to 3DyMnAaF,F2F00A0g.enLtes,tedratferdomAuEgduusatrd2,K2l0e0i0n.er (Dynax) o Charlie Auer (USEPA), Re: Phase-Out of Edwards, PIB etal. LANGMUIR 13(10), 2663 2669 (1997) 7E8G.0&3GonBihoantochmaibcisltAyqoufateigceTsoaxnidcoglroogwythLaabnordastuorrvyi.val1o97f8.iyThoef feaftfhecetasdomficonnntoiwnu(oPiumseepxhpaolseusre to promelas). Report #BW-78-6-175. Research report submitied to 3M Company, St. Paul, MN. aEsllaispDo.tenAt.i,aSl. sAo.urMcaeboufryh,alJo.gWen.atMeadrtoirngaannidc Da.ciCd.s Gi.n tMhueiernv2i0r0o1n.meTnht.ermNoaltyusries: o4f12f,lupopr.o3p2o1-l3y2m4e.rs AEllgnaalbaArsaswayy,TMe.sTt.Me1t9h8o1d.. 3RMepToercthnNiucmalbeRrep0o0r6t.SPurmomjeacrtyN,uMmubleiir-9P9h7as0e03E0x0p0o0s.urOec/tRoebcoevre1r6y. PEnivmierpohSaylsetsepmrso,mIenlc.as.19H9a02m.ptSotant,icNHA.cutSetuTdoyxincuimtybeorf FX-1003 10 was 9014-3. the Fathead Minnow, o 12 HEnavmiprtooSny,stNeHm.s,EInnvc.ir1o9S9y0sbt.emStsastitcudAycuntuembToexri9c0i1t3y-o3.f FX-1003 to the Daphnid, Daphnia magna. ES&T, 2000. Cheryl Moody and Jennifer Field, "Perfluorinated Surfactants and the Environmental ImplicationsofTheir Use in Fire-Fighting Foams" in Environmental Science & Technology, Vol. 34, Issue 18, p. 3864-3870. PFlMasGt,ic2s0I0n1d.ustVrye.rbaFlMcGo/mEmPenAtsmebeytitnhge,FMlaurocrhop7o,l2y0m0e1r Manufacturers Groupofthe Society of the Gabriel, Karl. Summary of: Primary Skin Iitation Study ~ Rabbits. Performed by: Bioscarch Submitted to 3M Company, 3M Center, St. Paul, MN. Gabriel, Karl. Submitted to Summary of: 3M Company, 3PMriCmeanrtyerE,ySet.IrPraiutlat,iMonN.Study -- Rabbits. Performed by: Biosearch. G1a0rr%y,ClVo.nFa.,l aCnedlRL.Li.neNfeolrsotnhe. Te1s98t1.CheAmnicAaslsTa-yo29f4C2elCloCT.ra3nsMfoCromamtpiaonnya,nSdt.CyPtaoutlo,xMicNi.ty in C3H GAemimsyo,nJi.Pu.,mDPre.rfl1u9o9r5o.oc3tManoraetqeu:esStteadtiecxFpiesrht Toevsetr".vieGweoisfy"EBciootaocxciucmoulloagtyi,vIencP.ropMearrtciehs2o0f. GiesyJ. P. andK. Kannan, 2001a. Accumulationofperfluorooctanesulfonateand related fluorochemicals in fish tissues. Prepared for 3M, St. Paul MN. June 20. In U.S. EPA Administrative Record AR226-1030A GcaiteisnygJw.aPt.eranbdirdKs.. KParnenpaanre,d20fo0r1b3.M,PeStr.flPuaourlooMcNi.aneJsuunlefo20n.atIenaUn.dS.reElaPtAedAfdlmuionrioscthreamtiicvaelsReicnofridshAR226-1030A Gicsy J. P. and K. Kannan, 2001c. Accumulationof perfluorooctanesulfonate and related fluorochemicals in mink and river otters. Prepared for 3M, St. Paul MN. June 20. In U.S. EPA Administrative Record AR226-1030A GiesyJ. P. and K. Kannan, 2001d. Perfluorooctanesulfonate and related fluorochemicals in oyster, Crassostrea virginica, from the Gulfof Mexico and Chesapeake Bay. Prepared for 3M, St. Paul MN. June 20. In U.S. EPA Administrative Record AR226-1030A GiesJy. P. andJ. L. Newsted, 2001. Selected fluorochemicals in theDecatur,Alabama area. Prepared for 3M, St. Paul MN, Project 178401. June. In U.S. EPA Administrative Record AR226-1030A @ Gibson, S.J., and Johnson, J.D. 1979. Absorptionof FC-143-14C In Rats After a Single Oral Dose. Riker Laboratories, Inc., Subsidiary of 3M, St. Paul, Minnesota. TGoitbaslonC,ar5b.1o,n-a1n4d iJnohMnasoln,an.dD.Fe1m9a8l0e. REaxsteAntfaenard RSoiuntgleeoIfVExDcorseetioofnFaCn-d14T3is-s1u4eC.DisRtirkibeurtion of Laboratories, Inc., Subsidiary of 3M, St. Paul, Minnesota. Gibson, S.J., and Johnson, J.D. 1983. Extent and Route of ExcretionofTotal Carbon-14 in Pregnant Rats Aftera Single Oral Doseof Ammonium 14 C-Perfluorooctanoate. Riker Laboratories, Inc., Subsidiary of3M, St. Paul, Minnesota. GUinlilveerts,iJtya.mesM.ar1c99h3.. 3Vhrequesied xpert review of "Bioaccumulation Studies", Comell GDiolcltiolraandl,tFh.s1i9,92D.iviFsliuoonroocfhEenmviicraolnsmaenndtaHluannadnOcHceuaplatthioSntaulicHesalintha,nUOncicvueprasittiyoonaflMCionhnoersto.ta, Gilliland, F.D. and Mandel, 1.5. 1993. Mortality among employees ofa perfluorooctanoic acid production plant. JOM. 35(9): 950-954. SiGip6lo8lpirloantdi,nsF,DaannddcMhaonldeeslt,roJlS. s19t9u6d.y oSfoecrcuumpaPteiroflnuaolrloyocetxapnoosiecdacmiedn.andAhmepJatiincd eMnezdym2e9s:,560- Glaza, S. 1995. Acute dermal toxicity study of T-6342 in rabbits. Corning Hazelton, Inc. Madison, WI. Project ID: HWI 50800374. 3M Company. St. Paul, MN. Glaza, SM. 1997. Acute Oral Toxicty Study ofT-6669 in Rats Corning Hazleton Inc. CHW 61001760. January 10. Sponsored by 3M, St. Paul, Minnesota. Goldenthal, E.I 1978a. Ninety Day Subacute Rat Toxicity Study. Final Report Prepared for 3M, MStinPnaeuslo,tMai,nnNeosvoetma,bebry 6I,nt1e9r7n5ational Research and Development Corporation, St. Paul, Goldenthal, E.L 1978b. Ninety Day Subacute Rhesus Monkey Toxicity Study. Final Report Prepared for 3M, St Paul, Minnesota, by International Research and Development Corporation, St. Paul, Minnesota, November 10, 1978. Gortner, E.G. 1981. Oral Teratology StudyofT-2998CoC in Rats. Safety Evaluation Laboratory and Riker Laboratories, Inc. Experiment Number: 0681TR0110, December 1981. GLoarbotreart,orEy.Ga.nd19R8i2.kerOrLaabloTraetroartioelso,gyIncS.tuEdxyopefriTm-e3n1t41NCumobCeirn:R0a6b8biTt.BSOaf,etyFeEvbarluuaartyio1n952 7H Griffith, F.D., and Long J.E. 1980. Animal toxicity studies with ammonium perfluorooctanoate. Am. Ind. Hyg. 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MimopaocdteCyd.bAy. fairned-Jf.igAh.tinFgiealcdt,ivi1t9y9.9.EnDveitreornm.inScait.ioTneochfnpoelr.fl1u9o99r,oc3a3r,b2o8x0y0l-2a8t0ie6ns.groundwater MHauzlret,oHn. In1c9.9,5.ViMeuntnaag,eVnAi.cit3yMtesCtoomnpaTn-6y3.42St.inPaaunl,inMvNi.vo mouse micronucleus assay. Corning Marl, H. 1996a. Mutagenicity test on T-6564 in an in vivo mouse micronucleus assay. Study number 17750-0455. 3M Company, St. Paul, MN. n 71 Muri, H. 1996b. Mutagenicity Test on T-6364 Measuring Chromosomal Aberrations in Chinese Hamster Ovary (CHO) Cells: with a Confirmatory Assay with Multiple Harvest. Final Report. Coming Hazleton Inc. CHV Study No.: 17750-0-437CO. September 16. Murl, Blood LH.ym1p9h9o6c2.ytMeustaWgietnhicaiCtoynTfeisrtmaotnoTr-y6A3s42s,ayMWeaistuhrMiunlgtiCphlreoHmaorsveosmtas.l Aberrations in Whole Coming-Hazelton, Inc. (CHV). Vienna, VA. CHV Study No.: 17073-0-449CO. Muri, H. 1996d. Mutagenicity Test on T-6342, Measuring Chromosomal Aberrations in Chinese Hamster Ovary (CHO) Hazelton, Inc. (CHV). Cells: witha Vienna, VA. CCoHnVfiSrtmuadtyorNyo.A:ss1a7y07w3i-t0h-M4u3l7tCiOp.le Harvests. Coming- hNielpsastoonc,arR.c,inBogeeniiBc.tjy,oPefrepa,et,rVo.x,isEormixeopnr,olKi.fearnadtorRsa. mCheelmC.,-.Bi1o99l1.. On the mechanoiftshme Interact. 78:235-250. CNuOlTtuOrXed.Pe2r0i0p0h.eraElvaHluuamtainonoLfymtphheocAybtileist.yofNOT-T7O5X24PtroojIencdtuNceumCbherrom2o92s0o6m2e. AbHeerrrtaotgieonnbsosinch, The Netherlands. NOTOX. 2001. Assessmentof Contact Hypersensitivity to T-7524 in the Albino Guinea Pig (Maximisation-Test). NOTOX Project number 292027. Hertogenbosch, The Netherlands. Nubbe, M. E; Adams, V. D.; Moore, W. M. The Direct and Sensitized Photo- oxidation of Hexachlorocyclopentadiene, Wat. Res. 1995, Vol. 29, No. 5, ppl287- 1293 Obourn, 1.D., Frame. S.R., Bell, RH. J., Longnecker, D.S., Elliot, G.S. and Cook, J.C. 1997. Mechanisms for the pancreatic oncogenic effectsof the peroxisome proliferator Wyeth-14,643. Toxicol. Appl. 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Identification June 25, 2001 Olsen, GW, Burris, JM, Lundberg, JK, Hansen, KJ, Mandel, JH, Zobel, LR. 2001c. Identificationoffluorohemicals in sera of American Red Cross adult blood donors. Interim report. June 25, 2001. 2O0l0s1edn., GDWe,scrLiopgtainv,e PsWu,mmSairmyposfons,erCuAm, fBluurorrso,chJeMn,icBaulrlleewve,lsMaMm,onLgunedmbpelrgo,yeJeK,paMratnicdiepla,ntJsH.of the year 2000 Decatur fluorochemical medical surveillance program. Final Report. March 19, 2001. OMlasnedne,l,GWJH,.Sc20h0m1iec.kleDre,scMr,ipTtiievreenssu,mJmMa,ryLoogfasn,erPuWm,flBuuorrriosc,heImMi,caBlurlelveewl,s MamMo,ngLuenmdpbleoryg,eeJK, participants of the year 2000 Antwerp fluorochemical medical surveillance program. Final Report. March 19, 2001. fOllsueonr,ocGheWm,icMaaldlseevne,lsDaCm,onBugrr2i3s6, JbMui,ldMianngdeemlp,lJoHy.ee2s0.01F1i.nalDeRsecproirptt.iveMasrucmhma1r9,y2o0f01serum OFllsueonr,ocGhWem,icHaalnsseinn,HCulmeamnenT,isLsuAe,. BFuirnrails,RJepMo,rtM.anEdpeild,emJiH.olo2g0y0,1g2.2I0d-e3nWti-f0i5c,atiMoendiocfal Department, 3M Company, St. Paul, MN 55144. O"Malley, Study with KT.-D2.,61a8nCdoECbbiennAsl,bKi.nLo.Ra19b8b1i.ts.ReRpiekaetr Application 28 Day Percutaneous Laboratories, St. Paul, MN Absorption `Ophaug, RH. andL. Singer. 1980. Metabolic HandlingofPerfluorooctanoic Acid in Rats. Proc Soc Exp Biol Med. 163:19-23 Pace Analytical. 1997. Ready Biodegradationof EC-126(BOD/COD). 3M Company Lab Request No. E1282. Minneapolis, MN. May 29. BPaacseedAnCahleymtiisctarli.es2.0013. MThCeom1p8a-nDayyRAeeqrueosbti,c CBoinotdreagcrtadAnaatliyotniScatludPyroojfecPterIfDl;uoCroAo0c9t7an,esulfonylMinneapolis, MN. February 23, 3 1 PPaelrafzlzuoolroo,ocMt.aJn.oa1t9e93(.CATShiNrot.ee3n8-2W5e-e2k6-D1i)etianrMyaTloexiRcaitts.y SFtiundayl wRietphorTt-.51L8a0b,orAamtomroynPiruojmect Identification HWI 6329-100. Hazleton Wisconsin, Inc. sPtausdtioeosro,fTa.Pm.,moLenei,uKm.Pp.erPfelrruio,roMo.cAt.anaonatdeG-iilnldiusc,edP.hJ.ep1a9t87o.meBgiaolcyhaenmidcpaelraonxdismoomrephporolloigfiercaatlion. Exp. Mol. Pathol. 47:98-109. Perit, 1999. determination K. of pPoleatritusn,deertiavl.ati"zleodn-apmaiirnoreavceirdsseuds-ipnhgaspeerlfilquuoirdicnhatreodmactarobgorxayplhiyc for acids as fon pairing agent" in J. Chromatography A, Vol. $33, 1999, pp. 147-155. R3eiMneCro,mEpAa.ny,19E7n8v.irFoantmeoenftaFlluLoarboocrhaetomriyc.alsJuilnyth19e.Environment. Project Number 970612613 Reiner, E.A. 1981. 3M Company Environmental Laboratory, St. Paul, Minnesota, Dec. 7. Renner, 2001. "Growing Concern Over Perfluorinated Chemicals" in Environmental Science and Technology. Vol. 35, Issue 7, pp. 154A-160A, April 1, 2001 PRiekrecrutLaanbcooruastoArbiseso,rpItnci.,oSnaSfteutdyy wEivtahluTat-i2o6n1LCabooCrationrAy.lbi1n97o9.RabRbeiptesa.tStA.ppPlaiucla,tMiionnne2s8o-tDaa.y Experiment Number: 09790AB04SS Riker Laboratories Inc., Safety T3371 in Albino Rabbits. St. Evaluation Paul, MN. Laboratory. Experiment 1983. Primary #0883EB0079. Skin Iitation Test with Scrano, L.; Bufo, S.A; Perucei, P.; Meallier,P.; Mansour, M. Photolysisand Hydrolysis of Rimsulfuron, Pestic. Sci. 1999, Vol, pp.955-961. Simister, E. etal. J. Chem. Soc., Faraday Trans. $8(20), 3033-41 (1992) hSeophalteinciupse,rAo.xK.i,soAmnedeprrosloinfe,raKt.ioannadndDerPeileatrerdepJ,a.r1a9m9e2t.erTshseheoffwecntososfexp-errefllautoerdodoicftfaenroeincceasciind moince. Biochem. J. 265:779-783. TStearpalteosg,eRn.iEc.,PoBtuerngtieaslso,fB.AAm.moanndiuKemmPse,rfWl.uDo.rooc19t8a4n,oatTehe(AEPmFbrOy)oi-nFetthealRaTto.xiFcuintyd.anAdppl. Tox. 4, 429-440. T10aktahgeip,eAr.,oxSiasio,mKe.,prUolmimfeermatuorrsa,,pTe.,rflHuaosreogocatwaan,oiRc.aacnidd aKnudropkearwfal,uoYr.ode1c9a91n.oicShaocritd-,tcearumseesxposure significant increasesof 8-hydroxydeoxyguanosine in liver DNAofrats. Cancer Lett. 57:535-60. n 80 "cTaarklaygib.ioA.marSkaei,rKf.o,r Uhempmaetomcuarrcai,noTg.,enHeassiesgianwdau,ceRd. bayndpeKruorxoiksaowmae,pYr.oli1f9e9r2a.torHse.pJa. tEonmveigroanl.y is an Toxicol. Pathol. 11: 145-149. Tterearndssf,orKm.iJn.,g Rgormomwtehrtfsa,ctFo.rG-.a ianndLeDyodrirgincgetllond,ur1iHn.g t1h9e90d.evIemlmoupnmoeinsttoofchtehemiractaltesdtest.ecMtoiloenc.ofCell Endocrinol. 69:RI-R6. TR. Wilbury Laboratories, Inc. 1995. Growth and Reproduction Toxicity Test with N2803-3 and the Freshwater Alga, Selenastrum capricornutum. Marblehead, MA. Study number B93-TH TPiRm.epWhiallbeusrpyrLoambeolraast.orMiaesr,blIenhc.ea1d9,96MaA..AcSuttuedytonxuicmibteyrof8N912-8T0H3.-3 to the Fathead Minnow, TR. Wilbury Laboratories, Inc. 1996b. Acute Toxicity ofN2803-3 to the Daphnid, Daphnia magna. Marblehead, MA. Study number 892-TH. Todd, J.W. 1979. FC-143 Photolysis Study Using Simulated Sunlight. Project 9776750202, 3M Company Technical Report No. 002. February 2. USEPA 1998 Fate, Transport and Transformation Test Guidelines, OPPTS 835.5270 Indirect Photolysis Screening Test; EPAT12-C-98-099; United States Environmental Protection Agency, USS. Government Printing Office: Washington, DC, 1998, pp1-22. USEPA 1998. Fate, Transport and Transformation Test Guidelines: 835.2110: Hydrolysis as a FunctionofpH; EPA7I2-C-98-057; United States Environmental Protection Agency, U.S. Government Printing Office: Washington, DC. pVearnfdlueonroHoecutvaneoli,e aLPc.,idDainvimasl,e.rWa.t,s:SoImnhmiebristo,ryR.,eaffnecdtoPfetteersstoons,teRroEne.. 1J9.92B.iocRheneaml. eTxocxriectoilo.n of 7(1):31-36. VpearnfdlueonriHneautveedlf,atJ.tPy.acKiudsslitkoipsr,oBt.eiLn,sainndtPheetpelrassomna,,R.lEiv.er19a9n1da.tesCtoevsoaflernatts.biCnhdeimn-gBoifol. Interact. 82317328, `diVsatnrdiebnutiHoenu,vmeelt,a1b.Po.,lisKmu,slaiknids,clBi.mLi,naVtainonoRaffpeelrgfhleumo,roMo.cLt.anaonidc aPceitderisnonm,alR.eEa.nd19f91ebm.alTeirsassu.eJ Biochem. Toxicol. 6(2):83-92. 9V9ra7s0p6i1r2,6G0.0A:.,FMateendoeflF,luAorrtohcuhre.mic1a9l7s9.. 3AMnaTleycshinsicfoarl FRleupoorrotcRheepmoirctalNsuimnbeBlru1g4.ilMaFiysh.| Project = gl PWiamredp,hTa.l,eNsepvrioumeslJ,a.sa.ndTRR.. BWoielrib.ury199L6aab.oraAtcoruitees,toIxnicc.itLyaobf FReCq-u1e0s1t5ntuomtbheerfaPt1h6e2a4d. mi3nMnow, Company, St. Paul, MN. Ward, T., Nevius, J. and and the freshwater alga, SRe.leBonearsi.iru1m9c96abp.ricGorrnouwttuhm.andT.rR.eprWoidlubcutriyonLtaobxoircaittoyriteesst, with Inc. FC-1015 Lab Request number P1624. 3M Company, St. Paul, MN. mWaagrnda,.T., T.NRe.vWiiulsbJ,u,ryanLdabRo.raBtooerriie.s,1I9nc9.6c.LaAbcruetqeuetsotxinciutmyboefrFPC1-612041.53toMDCaopmhnpiadn,yD,aSpt.hnPiaaul, MN EWenlvsihr,onSmKe.ntal19L7a8b.orTaetcohrnyi.ca3l MReCpoormtpaSnuymmParorjyec-tA9d9s7o0r6p1t2i6o3n3o:fFFaCt-e9o5faFnlduoFrCo-c1h4e3miocnalsso,l. Report Number 1. St. Paul, MN. February 27. hYeapgaetorc,ar1.cD.inJro.geannedsiYsagienrf,eRm.al1e98S0p.raOgruale-cDoanwtrlaecyeprtatisv,eCsatnecroeirdsRaess.pr4o0m:o36t8e0r-s36o8f5. uVriiinneanr,y eMx.c,reHtainohniojafrpveir,flH.u, oJraoaokcotnanaohioc,aIc,idanind tPheeurmaa,leP.ra1t9.89P.haSrtmiamcuolla.tiToonxibcyole.st6r5ad:i2o7l4o-f27t7h.e tYhieinreant,afM.e,rKsoijnogl,eAa.n,dHsaunbhcijhdrrovnii,cHa.dmainndisPteruatriao,nP.. B1u9l9l0..EnDviisrpoons.itCioonnotafmp,erTfolxuiocroolo.c4t4a:n4o6i-c5a3c.id in 76 8