Document xjqqo0ryLO37jZz5g4KnBeOaJ

AnalysisforfluorochemicalisnBluegilflish. BIOACCUMULATION (ANALYTICAL ONLY) TEST SUBSTANCE Identity:Perfluorooctanesulfonatmea;y alsobe referredto as PFOS or FC-95. (1-Octanesulfoniaccid,1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-p,otassium salt,CAS # 2795-39-3) Remarks: The testsubstance isa white powder. No information was recordedon the purity. METHOD Method: Devised by 3M Company to determine ifbluegilslunfish bioaccumulatefluorochemicalsfrom 3M's Decatur,AL plant effluen(tSee Remarks) GLP (Y/N): No Year completed: 1978 Remarks: The followinmgethod was devisedand used: 1 . Two lotsof30 fisheach were used. 2. One lotwas exposed to Decatur planteffluenftrom Oct.5 toOct. 25. The otherlotwas exposed toTennessee Riverwater onlyfrom Oct.3 to Oct.25. 3. Twelve fishfrom the effluenetxposed lotwere collecteddead and the remainingeighteenfishwere sacrificeidnDecatur. The river water exposed fishsurvivedthroughthe exposure periodand all were sacrificeidnDecatur. 4. Effluent-exposedand riverwater-exposed fishwere allpreserved and shipped indry icetothe 3M EnvironmentalLaboratoryinSt. Paul,MN foranalysis. 5. Fishwere homogenized usinga Tekmar Model SDT tissuemizer. 6. Two 2 g.portionswere weighed and extractedusing ethylacetate on one and tolueneon theother. 7. Samples were analyzed by GC, TLC, and GC/MS. RESULTS AnalyticalObservations: The effluent-exposedfishweighed 25.7 grams. The riverwater-exposed fishweighed 33.0 grams. TLC qualitativeilnydicatedthe presence of perfluorooctanesulfonatbeu,t was notquantified. No fluorochemicalswere detectedinthe riverwater-exposed fish. Remarks: Study was conducted because 3M's Decatur plant manufacturesfluorochemicalsW.ater used inmanufacturingislater treatedand the treatedeffluendtischargeseventuallyintotheTennessee River.. DATA QUALITY ReliabilityK:limischranking3. Effluenctoncentrationsof subject fluorochemicalswas not charactedzedand the specificprotocolfor exposure ofthe fishwas notfound. There was also no informatioonn analysisoftheTennessee Riverwater used inthe study.Additionallyi,tis not known iftherewas any opportunityfordepurationofthe fishpriorto sacdfice.Presence ofthe testsubstance could have been a normal functionofthefishconcentrationshomogenizing withthe effluent concentrations.No explanationisattemptedas towhat was the cause of the 12 dead fishintheeffluent-exposegdroup. No differentiatiwoans giventowhether the subjectchemical was actuallyaccumulated inthe fish or simplysorbed on the surfaceofthe fishbody. CONCLUSIONS No conclusionscan be made about the uptake of perfluorooctanesulfonate from Decatur effluenftorreasons given above. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St.Paul,Minnesota,55133 REFERENCES Study conducted by 3M Company. OTHER Last changed: 5/4/00 Form 6747-11-A TECHNICAL REPORT SUMMARY .,TTEoC:HNICALCOMMUNICATIONSCENTER - 201-2CN (import-anItfreporitsprintoendboM sideosfpapers,endtvmcopietsoTCC.J 5/l/79 oivision ProJect Report Title Environmental Laboratory Fat'e of Fluorochemicals (EE & PC) Reg. Analysis for Fluorochemicals To AuthorT%F D. L. Bacon G. A. Vraspir/Arthur -9'otak-wokReference Mendel 48838-18 in Bluegill SET:URITYIO, 0 - Open Vo"d 3M CHEMICAL (CompanCyonfidentiatls)mciAaulthorizna)tio REGISTRY KOYWORDS: (Seletcetrmf$rom3M ThesaurSuusg.psotther applicablteerms.) CURRENT OBJECTIVE: Status Report No. 48765 Fish Dept um 0535 PfaiectNumbw 9970612600 Report Number 014 Efnplaym Number(s) 76416143939 No. of FqM IncludingCovershoot 7 Now Chemicals Reported Yes IN No REPORT ABSTRACT:12DO-2w6o0rd4ThiasbstriancftomutiiosdnistribbuyttehdeTechnicCaolmmunicatiCoennstetro aler3tM'ortsoCompanyR&D.: ,:R.A.Prokop-236-3B V.Pothapragada-236-3A A.N.Welter-21-2W R. L'.Bohon- 21- 2W InformatioLniaisop Initials: t -. I FC (Req. 4876S)/GAV/AM -2- 5/l/79 OBJECTIVE The purpose of this investigation was to determine if bluegill fish, after exposure to our Decatur plant effluent, bioaccumulated fluorochemicals. SUMMARY Extracts from bluegill fish exposed to Decatur plant effluent (PE) contained 10 ppm (pg/g) of FM 3923 and 7 ppm of FM 3422. TLC indicated the presence of KC-95, while FC-143 and FC-128 were -not detected. Extracts from bluegill fish exposed to river water (RW) contained ethanol, ethyl ether, ethoxyethyl acetate, toluene, and xylene. No fluorochemicals were detected. A copy is attached of CRL report 7124 which detailed GC/MS experiments. INTRODUCTION The uptake *and clearance of specific radiolabeled fluorochemicals by fish were reported (1). The current report details the analyses for fluorochemicals in fish previously exposed to our Decatur plant effluent and in fish exposed only to Tennessee River water (2). Thin-layer chromatography (TLC), gas chromatography, and gas chromatography/mass spectroscopy (GC/MS) were used. .DISCUSSION Our Decatur plant manufactures fluorochemicals. Water used in manufacturing is later treated and the treated effluent discharges eventually into the Tennessee River. Bluegill fish, chosen for these bioassay experiments, were exposed to P.E. Another lot of fish were exposed only to the Tennessee River.water (2), The fish were then. analyzed for fluorochemicals b:V"an involved three-route proc' edure as outlined in Flow Diagram 1 and detailed in the experimental section. Briefly, fish were homogenized, and some of the homogenate was extracted with toluene.(Route 1) while some of the homogenate was extracted with ethyl acetate (Route 2). A portion of the latter extract was concentrated Route 3) for GC/MS and TLC. Results are given in the summary section. -3FLOW DIAGRAM 1 ANALYSIS OF BLUEGILL FISH 30 Fish (1) Weigh (2) "Tissuemizell e 0 (1) Weigh 2 g. (2)Extract (ETOAC) (3) Centrifuge (4)Isolate ETOAC Layer F- GC For FM 3923 Fli 3925 FM 3422 Route 3 Concentrate GC/MS TLC For FC-95 FC-128 FC-143 (1).Weigh 2 g.. (2).Acidify (3) Extract (Toluene) (4)Centrifuge (5) Isolate Toluene Layer (6) Methylate GC/MS GC For FC-143 (Ester) FC (Req. 4876S)/GAV/AM -4- 5/l/79 EXPERIMENTAL Two lots of thirty fish each, wrapped in aluminum foil, respectively, and shipped in dry ice, were received on November 3, 1978, by the Environmental Laboratory. The fish were kept frozen until analyzed (Nov. 3, 1978). The effluent-exposed lot (Oct. 5 to Oct. 25) weighed 25.7 g. Twelve fish were collected dead and eighteen fish were sacrificed in Decatur. The river-exposed lot (Oct. 3 to Oct. 25) weighed 33.0 grams and were all sacrificed at Decatur. To simplify,the experimental one lot is discussed. The other was analyzed in exactly the same manner. The following codes were used: River River Plant Plant Water Holding Tank Exposed Water Holding Tank Exposed Effluent S-1 Exposed Fish Effluent S-1 Exposed Fish Fish - ethylacetate extract (R.W.-EtAc) Fish - toluene extract (R.W.-Tol.) ethylacetate extract (P.E.-EtAc) toluehe extract (P.E.-Tol.) The fish were homogenized using a Tekmar Model SDT tissuemizer. Two 2 g. portions were weighed into separate polyethylene centrifuge tubes. Two mlls of pesticide grade ethylacetate were added to one and 2 ml cone. HC1 plus 2 ml of AR grade toluene were'added to the other. Each sample was mixed approximately 1 min. on a VortQx Genie mixer. The samples were centrifuged for 15 min. at 10,000 rpmls. The ethylacetate and toluene extracts were drawn off and placed in.glass vials, respectively. The ethylacetate extract was exawined using the following GC conditions: Instr. HP 5713 with Ni , electron capture detector. Column - 6'x 1/811S.S. 10% CarbowBx 20 M on 60/80 mesh Chromosorb W-AW.' Ing. Port-Temp - 200 C., Def. Temp - 3000 C., Oven Temp - 150 C isothermal, Flow - 40 cc/min. Argon/Methane (95/5). After GC analysis of EtAc extract, a portion was concentrated under N, purge for two dimensional TLC analyses. The following TLC con itions,were used. The concentrated EtAc extracts were spotted alongside standards of FC-95, FC-143, and FC-128 on E. Merck silica gel plates. They were first developeg in 10% ETOH in ttac and dried. Then the plates were turned 90 and develope' d in 50% ETOH in EtAc. The dried plates were visualized by spraying with 0.01% New Fuchsin in water (3). The toluene extract was methylated with diazomethane and then exglined using the following GC conditions: Instr. RP 5713 with Ni electron capture detector. Column - 12'x 1/8" S.S. 20-m ,t DC-900 and 10% Bentone 38 or 80/90 mesh Anakrom PA. Inj. Port 200 C. Det. temp - 300 C., Oven - 110 C Isothermal, Flow 40 cc/min. Argon/Methane (95/5). The ethylacetate and methylated toluene extracts were submitted to Central Research Analytical for GC/MS analysis (Req. No. C-50687). See attached CR Report No. 7124 for GC/MS conditions. FC (Req. 4876S)/GAV/AM -5- 5/l/79 PE/EtAc extract showed approximately 10 ug/g of FM 3422. RW & PE-Tol. showed no difference. No FC-143 or FC-128 was detected by TLC of the PE EtAc, but a spot was noticed whose Rf Was that of refere'nee FC-95. REFERENCES (l)-a) M, T. Elhabarawy, "Bioconcentration of FM 3422 in Bluegill Sunfish and In Channel Catfish," EE & PC Report, 5/17/77. A. N. Welter, "Aquatic Fate of A Fluorochemical, FM@34@2,11 EE & PC Report, 10/14/77. (2) Decatur Effluent Fish Toxicity Test Protocols, dated Oct. 18, 1978 and Feb. 5, 1979;-M. T. Elnabarawy. (3) J. Sliwiok.and A. Macioszezyk, Minrnahpminal Journal, 23, 121 (1978). VAa@ glzfllq GAV/AM/cen Attachment. Pon CENTRAL ANALYTICAL LABORATORY Report No. __ 7124 March-2-8,- 1-9-79--------------------- Subject: Decatur Fish Bloassy. Requ e stor:@.-Y-raspir-, A.-@-M.-a-nd-el Requejst No.'-@@0687 Report: INTRODUCTION Dept. Name -t@-16--P---C----Dated November 16, 1978 ]Proj.No. 5-0-)-6-19-0-404 Samples. were received from.the requestors from.a fish bloossy. taken'st the Decatur Plant. Fish were exposed to two different water syst@: the TennesseeRiverwhich flows just outside the plant and the planteffluent running Into the river. Ethyl acetate extracts of the fish were analyzed using the GC/MS system. Toluene extracts of the fish were mai@ylated with diazomethane and analyzed. OISUCSSION The samples were labeled R.W. #I and P.E. #I (mothylatedtoluene extracts) and R.W. 12 and P.E. 12 (ethyl acetate extracts). S&Mles R.W. 01 and P.E. #I were analyzed using conditions for column A below and s&Wies R.W. 12 and P.E. #2 were analyzed using conditions for column 8 below. 'Instrument: Varian 2740 gas chromatograph Column A: 121 S.S. 20% DC-200 + 10% Denton* 34 on Anakrom PA Oven: 110*C isothermal for 11 minutes then 110-170*C at )O*C/min. Injector: 165*C Column 6: 61 S.S. CW 20M 15% on Chromosorb W Oven: 50 2*25'*C at 10*C/min. Injector: 220*C Detector: Varian flame ionization at 290'C Carrier Gas: Helium at 25 cc/min. Instrument: dupdnt 21-4915 mass spectrometer "Source: Chemical Ionization Mode: all samples were run In Electron Ionization,sample P.E. 12 . was also run In Chemical Ionizationwith KH gas Mass Range: 18 - 617 m/e - 3 Helium Separator: Single StageALat Multiplier: gendix ChannaltrontU RESULTS Samples R.W. 11 and P. E. 11 showed by GC/MS only the solvents used in extracting and mothylati,ng:ethanol, ethyl other, othoxy ethyl acetate toluene and xylones. It was necessary to air concentrate the ethyl acetate extracts In order to obtain sufficient data th Interpret the mass spectra. The Tennessee River exposed fish ethyl acetate extract cmtalned numerous G.Ci peaks none of which appeared to contain fluorine.- The components appear to correspond to references of 2-mothyl-3-octenons. acetlc acid. a phthalate, and high,molocular alkanes from about C to at least C 24* The plant effluent exposed fish 6thyl.acetat6 extract contained Compounds. which appear to correspond to references of 2-mothyl-3-octanonat acetic acid, fluorochomicals whose highest mass fragments seen tiara 483 M/o and, 582 m/e, nitrogen containing fluorocchmicals (whose highest mass fragments seen were 448 m/a and 584 m/e)l FM 3923 (FOSE amid*) and FM 3422 (FOSE alcohol). SUAMARY .This fish bioassy showed that extracts from the fish exposed to the.Tennessee River water contained such compounds as ethanol, ethyl other, ethoxy ethyl acetate, toluene and xylene. The extracts from the fish exposed to the Decatur plant effluent contained FOSE amide, alcohol, other related fluotochomica)s, 2-met.hyl-3-octanone, and acetic acid. J. N. Scbroep@er /ch