Document RaJRK3LvLmNkZNvkEmdvYrOBX

BIOACCUMULATION (ANALYTICAL ONLY) TEST SUBSTANCE Identity:N-ethylperfluorooctanseulfonamidoethanol;may alsobe referredtoas N-ETFOSE Alcoholor FM-3422. (1-Octanesulfonamide,N-ethyl1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafluoro-N-(2-hydroxyethyl)-, CAS # 1691-99-2) Remarks: Materialisan off-whitew,axy solidofuncharactedzed purity. METHOD: Method: Devised by 3M Company todetermine ifbluegilslunfish bioaccumulatefluorochemicalfsrom 3M's Decatur,AL planteffluent (See Remarks) GLP (YIN): No .Yearcompleted: 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 collectedead (diedintest) and the remainingeighteenfishwere sacrificeidnDecatur. The river water exposed fishsurvivedthroughthe exposure periodand allwere sacrificeidnDecatur. 4. Effluent-exposedand riverwater-exposed fishwere allpreserved and shipped indry icetothe 3M EnvironmentalLaboratoryinSt.Paul,MN for analysis. 5. Fishwere homogenized usinga Tekmar Model SDT tissuemizer. 6. Two 2 g. portionswere weighed and extractedO.ne portionusing ethyl acetateand tolueneon the other. 7. Samples were analyzed by GC, TLC, and GC/MS. RESULTS AnalyticalObservations: The effluent-exposefdishweighed 25.7 grams. The riverwater-exposed fish weighed 33.0 grams. GC/MS analysisofethylacetateand tolueneextractedfishexposed to plant effluentcontained7 ppm of N-ETFOSE alcohol. No fluorochemicalswere detectedinthe riverwater-exposed fish. Remarks: Study was conducted because 3M's Decatur plantmanufactures fluorochemicals.Water used inmanufacturingislatertreatedand thetreated effluendtischargeseventuallyintothe Tennessee River. CONCLUSIONS No reliablceonclusionscan be derivedfrom thisstudy. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St.Paul, Minnesota,55133 DATA QUALITY ReliabilityK:limischranking3. Study was conducted atthe manufacturing facilitryatherthan at3M's EnvironmentalLaboratory.Effluentconcentrationosf subjectfluorochemicalswere notcharacterizedand the specificprotocolfor exposure ofthe fishwas not found. There was alsono informatioonn analysisof the Tennessee Riverwater or effluenutsed inthe study(e.g.,NH3-N ineffluenits known tocause death duringNPDES testingo,r low DO, highTDS). Additionallyi,tisnot known iftherewas any opportunityfordepurationofthefish priorto sacrificeP.resence ofthetestsubstancecould have been a normal functionofthefishconcentrationshomogenizing withthe effluenctoncentrations. No explanationisattemptedas towhat was thecause ofthe 12 dead fishinthe effluent-exposegdroup. No differentiatiwoans givento whetherthe subject chemical was actuallya'ccumulated inthe fishorsimplysorbed on the surfaceof the fishbody. Sampling locationswere not described. REFERENCES 3M TechnicalReport "AnalysisforFluuorochemicalsinBluegilFlish"G. A. Vraspir/ArthurMendel, Project9970612600, Fate of Fluorochemicals,Report Number 14,May 1, 1979 3M requested expertoverview,"BioaccumulationStudies",Dr.James Gilleft, Comell UniversityM,arch 8, 1993 OTHER Last changed: 5/17/00 3M REPORT: BIOACCUWTIATION STUDEES March 9,1"3 (v4. ReportNo. 14 (511179")Analysiosf Fluorochemicalisn BluegiUFish- Althoughthisreportofferssome detaileedxplanatioonf analyticaplrocedurest,hesamplesourcesand data are presentedina verymarghmlized manner. Itgivestheimpressionthatdes-@hydroxyethyl)atioofnN-Et FOSE alcoholoccursin the environmenl "3m Forp 6747.11 -A TO: TECHNICAL TECHNICAL REPORT COMMUNICATIONS CENTER - 201-2CN SUMMARY Do* 5/l/79 timportmt - Ifreportisprintedon bdih siM ofpqw, und tm copiesto TCC.) Division Project Report TWo Environmental Laboratory (EE & PC) Reg. No. 48765 Fate of Fluorochemicals Analysis for Fluorochemicals in Bluegill Fish DepL Numbw 0535 rrobet Number 99t;O!IF600 report mu 014 Author(s) D. L. Bacon G. A. Vraspir/Arthur Note-Wck Rofw*nce 48838-18 Mendel SECUIQITY 10, 0 Open (Company Confidential) Ll Closed (Special Authorization) 3PACHEMICAL REGISTRY KEYWORDS: (Select terms from 3M Thomrus. t other applicable ltwm7l CURRENT OBJECTIVE: Status Report EnWVoyoo Numlniiel 76416Z43939 mo. ow ragn inciwaing 7 Now ChwniesisReported Yes m No REPORT ABSTRACT: (200-M words) This d3svaa information is dotrbuted by the Technical Comtnunimtions aiwt 3M'*rs to Con"ny R&D. ItIsCompany cor.IW*ntW nmwisi. Center to. c:R.A.Prokop-236-3B V.Pothapragada-236-3A A.N.Welter-21-2W R.L'.Bohon-21-2W Information Liajso?,, Initials: 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 FC-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 by an involved three-route procedure 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. -3- FLOW 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 Route 3 GC For FM 3923 FM 3925 FM 3422 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 w 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 Fish - ethylacetate extract (R.W.-EtAc) Water Holding Tank Exposed Fish - toluene extract (R.W.-Tol.) Effluent B-1 Exposed Fish - ethylacetate extract (P.E.-EtAc) Effluent S-1 Exposed Fish - toluene 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 ml's of pesticide grade ethylacetate were added to one and 2 ml conc. HC1 plus 2 ml of AR grade toluene were added to the other. Each sample was mixed approximately 1 min. on a Vortex Genie mixer. The samples were centrifuged for 15 min. at 10,000 rpm's. The ethylacetate and toluene extracts were drawn off and placed in glass vials, respectively. The ethylacetate extract was exa6ned using the following GC conditions: Instr. HP 5713 with Ni electron capture detector. Column - 6,lx 1/811 S.S.'@' 10% Carbowax 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 PC-95, FC-143, and FC-128 on E. Merck silica gel plates. They were first developeg in 10% ETOH in EtAc and dried. Then the plates were turned 90 and developed 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 exnlned using the following GC conditions: Instr. HP 5713 with Ni electron capture detector. Column - 12'x 1/81'S.S. 20% DC-200 and 10% Bentone 3 or 80/90 mesh Anakrom PA. Inj. Port 0 a 0 200 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 reference PC-95. REFERENCES (1)a) M. T. Elnabarawy, "Bioconcentration of FM 3422 in Bluegill Sunfish and In Channel Catfish," EE &PC. Report., 5/17/77. b) A. N. Welter, "Aquatic Fate of A Fluorochemical, FM 3422,11 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. Macioszczyk, 23, 121 (1978). VAa@ #,/lq GAV/AM/cen Attachment CENTRAL ANALYTICAL LABORATORY Report No. --_1124------------?4rch 28, 1-9-79----------- Subject: Decatur Fish gloassy. Requeotor. G. Vraspir, A. Mendai Request No. -------- D opt. Name _E-g--&-P-C-------D&tod November 16, 1978 no. -9-15-0-1-6-190404* Report: INTRODUCT ION Samples were rec ived from the requestors from a fish bloassy tak*n at the Decatur Plant. ;Ish were exposed to two differentwater elfattm: the Tennessee River W%ich flows just outside the plant and the plant effluent running Into the river. Ethyl acetate extracts of the fish were &Wlyzed using the GC/MS system. Toluene.extracts of the fish were mothylated with diazomethane and analyzed. DISUCSSION The samples were labeled R.W. 11 and P.E. 11 (mothylatod toluene extracts) and R.W. 02 and P.E. 12 (ethyl acetate extracts). Samples R.W. 11 and P.E. 11 were analyzed using conditions for column A below and sw*les R.W. 12 orw P.E. #2 were analyzed using conditions for column B below. Instrument: Varian 2740 gas chromatograph Column A: 121 S.S. 20% OC-200 + 10% Bentone 34 on Anakrom PA Oven: 110*C isothermal for 11 minutes then 110-170*C at 10*C/sin. Injector: 165*C Column 6: 61 S.S. CW 20M 15% on Chromosorb W Oven: 50 225*C at ]O*C/min. Injector: 220*C Detector: Varian flame Ionization at 290*C Carrier Gas: Helium at 25-cc/min. Instrument: dupont 21-491B mass spectrometer Source: Chemical Ionization Mode: all samples were run in Electron Ionization,s&Wle was also run in Chemical Ionizationwith NN gas Mass Range: 18 - 617 m/e Helium Separator: Single Stage t Multipli.or:Bandix Channeltron(9 P.E. 12 Relit)rLNo. 124 RESULTS Samples R.W. 11 and P. E. 01 showed by CC/MS only the solvents used In extracting and mthylating: ethanol, ethyl other, sthoxy ethyl watat* toluene and xylones. it was necessary to air concentrate the ethyl watate extracts In order to obtain sufficient data to Interpret the mass spectra. The Tennessee River exposed fish ethyl acetate oxtr"t,contained iol@marous -G.C. peaks none of which appeared to contain fluorine. The components appear to correspond to references of 2-mthyl-3-octanonso wetic acids 41 phthalate, and high molecular alkanes from about C to at 1,04stC24-'*' The plant effluent exposed fish ethyl watate extract cAoAtal'i*,d s which appear to correspond to references of 2-mthyl-3@. 'tic acid, fluorochomicals whose highest mass fragments seen im'fe 3. and 582 mte, nitrogen containing f)Liorocohmicals (whose highest *Ass firogmnts seen were 448 m/o and 584 m/e@ FM 3923 (FOSE &old*) and FM 3422 (.ft$galcohol)-. SUMMARY This fish bloossy showed that extracts from the fish exposed to the Tennessee River water contained such compounds as ethanol$ othyl other, othoxy ethyl acetate, toluene and xylone. The extracts from the fish exposed to the Decatur plant effluent contained FOSE amide, alcohol, other related fluorochomicalso 2-methyl-3-octanorag and acetic acid. "J. N. Schroop r /ch