Document 0gR3DEMnm876mYNV48oGGQ3Xx

BIOACCUMULATION (ANALYTICAL ONLY) AR23&0354 TEST SUBSTANCE_______________ Identity: N-ethylperfluorooctane sulfonamidoethanol; may also be referred to as N-EtFOSE Alcohol or 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: Material is an off-white, waxy solid of uncharacterized purity. METHOD:_____________________________________ ________________________ Method: Devised by 3M Company to determine if bluegill sunfish bioaccumulate fluorochemicals from 3M's Decatur, AL plant effluent (See Remarks) GLP (Y/N): No Year completed: 1978 Remarks: The following method was devised and used: 1. Two lots of 30 fish each were used. 2. One lot was exposed to Decatur plant effluent from Oct. 5 to Oct. 25. The other lot was exposed to Tennessee River water only from Oct. 3 to Oct. 25. 3. Twelve fish from the effluent exposed lot were collected dead (died in test) and the remaining eighteen fish were sacrificed in Decatur. The river water exposed fish survived through the exposure period and all were sacrificed in Decatur. 4. Effluent-exposed and river water-exposed fish were all preserved and shipped in dry ice to the 3M Environmental Laboratory in St. Paul, MN for analysis. 5. Fish were homogenized using a Tekmar Model SDT tissuemizer. 6. Two 2 g. portions were weighed and extracted. One portion using ethyl acetate and toluene on the other. 7. Samples were analyzed by GC, TLC, and GC/MS. RESULTS________________________________________________________ Analytical Observations: The effluent-exposed fish weighed 25.7 grams. The river water-exposed fish weighed 33.0 grams. 006276 GC/MS analysis of ethyl acetate and toluene extracted fish exposed to plant effluent contained 7 ppm of N-EtFOSE alcohol. No fluorochemicals were detected in the river water-exposed fish. Remarks: Study was conducted because 3M's Decatur plant manufactures fluorochemicals. Water used in manufacturing is later treated and the treated effluent discharges eventually into the Tennessee River. CONCLUSIONS__________________________________________ No reliable conclusions can be derived from this study. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY_________________________________________________________ Reliability: Klimisch ranking 3. Study was conducted at the manufacturing facility rather than at 3M's Environmental Laboratory. Effluent concentrations of subject fluorochemicals were not characterized and the specific protocol for exposure of the fish was not found. There was also no information on analysis of the Tennessee River water or effluent used in the study (e.g., NH3-N in effluent is known to cause death during NPDES testing, or low DO, high TDS). Additionally, it is not known if there was any opportunity for depuration of the fish prior to sacrifice. Presence of the test substance could have been a normal function of the fish concentrations homogenizing with the effluent concentrations. No explanation is attempted as to what was the cause of the 12 dead fish in the effluent-exposed group. No differentiation was given to whether the subject chemical was actually accumulated in the fish or simply sorbed on the surface of the fish body. Sampling locations were not described. REFERENCES__________________________________________________________ 3M Technical Report "Analysis for Fluuorochemicals in Bluegill Fish" G. A. Vraspir / Arthur Mendel, Project 9970612600, Fate of Fluorochemicals, Report Number 14, May 1,1979 3M requested expert overview, "Bioaccumulation Studies", Dr. James Gillett, Cornell University, March 8, 1993 O T H E R ______________________________________________________________ Last changed: 5/17/00 006277 3M REPORT: BIOACCUMULATION STUDIES March 8, 1993 4. Report No. 14 (5/1/79) 'Analysis of Fluorochemicals in BiuegiU Fish * Although this report offers som e detailed explanation o f analytical procedures, the sample sources and data are presented in a very marginalized manner. It gives the impression that des-(hydroxyethyl)ation o f N-Et FOSE alcohol occurs in the environment. O 006278 3E3 ` TECHNICAL REPORT SUMMARY O ra 5/1/79 TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN (Im p o rtan t - f f re p o rt sp rin ted on both sides o f paper, send two copies to TCCJ Division Project Environmental Laboratory (EE Fate of Fluorochemicals Report Title Analysis for Fluorochemicals To A u th o rial D. L. Bacon G. A. Vraspir/Arthur Mendel Notebook Reference 48838-18 & PC) Req. in Bluegill No. 48765 Fish Dept. Number 0535 Prelect Number ,,9970612600 Report Number 014 Employee Number!t) 76416/43939 N a oDPagee Including Coversheet 7 S E C U R IT Y ^ G Open (Com pany Confidential) G D o tad (Special A uthorization) 3M C H E M IC A L w R E G IS T R Y " Now Cham icali Reported Yee IS No KEYW ORDS: (Select terms from 3M Thesaurus. Suggest other applicable terms.) C U R R E N T O B JEC TIVE: Status Report REPO RT A B STRACT: (200*250 words) This abstract inform ation is dtetributed by tha Technical Com m unications Cantar to . alert 3M 'ers to Company R & D . It is Company confidential m aterial. v. ? c :R .A .Prokop-236-313 V .Pothapragada-2 36-3A A.N.Welter-21-2W R .L .Bohon-21-2W Information Uaiforp,. Initials: r( 006279 FC (Req. 4876S)/GAV/AM -2- 5/1/79 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 chroma tography, 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 Rive* 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. 006280 -3- FLOW DIAGRAM 1 ANALYSIS OF BLUEGILL FISH 30 Fish (1) Weigh (2) "Tissueinize" (1) Weigh 2 g. (2) Extract (ETOAc) (3) Centrifuge (4) Isolate ETOAc Layer GC For FM 3923 FM 3925 FM 3422 Route 3 "1 Concentrate l GC/MS "\k 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 ________ I____ GC/MS % GC For FC-143 (Ester) 006281 FC (Req. 4876S)/GAV/AM -4- 5/1/79 EXPERIMENTAL Two lots of thirty fish each, wrapped in aluminum foil, respec tively, 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 Water Holding Tank Exposed Fish - ethylacetate extract (R.W.-EtAc) River Water Holding Tank Exposed Fish - toluene extract (R.W.-Tol.) Plant Effluent S-l Exposed Fish - ethylacetate extract (P.E.-EtAc) Plant Effluent S-l 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 cone. 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 examined using the following GC conditions: Instr. HP 5713 with Ni electron capture detector. Column - 6''.X 1/8" S.S.* 10% Carbowax 20 M on 60/80 mesh Chromosorb W-AW, Ini. Port Temp - 200 C., Def. Temp - 300 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 conditions 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 developed 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 examined using the following GC conditions: Instr. HP 5713 with Ni electron capture detector. Column - 12'x 1/8" S.S. 20% DC-200 and 10% Bentone 34 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. 006282 FC (Req. 4876S)/GAV/AM -5- 5/1/79 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 FC-95. REFERENCES (1) a) M. T. Elnabarawy, ''Biocon cent ration 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," 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, Miorochemlnal Journal. 23, 121 (1978). GAV/AM/cen Attachment 006283 CENTRAL ANALYTICAL LABORATORY Report No. ...1L2A ... Date . . _2*.J-9.7? Subject: Decatur Fish Bioassy. R e q u e s to r A '. .? ? '!*1 Ai.ifSDept. Name .................. P r o j . Mo. ! 5 5 i ` !2 ? * r t R e q u e s t No. ....... ' Dated C .'A'- R e p o rt: INTRODUCTION Samples were received from the requestors from a fish bioessy taken at the Decatur Plant. Fish were exposed to two different water system: the Tennessee River which flows Just outside the plant and the plant affluent running into the river. Ethyl acetate extracts of the fish were analysed using the GC/HS system. Toluene extracts of the fish were methylated with diazomethane and analyzed. D1SUCSSI0N The samples were labeled R.W. #1 and P.E. #1 (methylated toluene extracts) and R.W. #2 and P.E. #2 (ethyl acetate extracts). Samples R.W. #1 and P.E. #1 were analyzed using conditions for column A below and samples R.W. #2 and P.E. H Z were analyzed using conditions for column B below. Instrument: Varian 2740 gas chromatograph Column A: 12* S.S. 202 DC-200 + 102 Bentone 3k on Anakrom PA Oven: 110*C isothermal for II minutes then 110-170*C at 10*C/m(n. Injector: 165*C y,i Column 6: 6* S.S. CW 20H 152 on Chromosorb V Oven: 50 - 225'C at 10*C/mln. Injector: 220#C v. < Detector: Varian flame ionization at 290*C Carrier Gas: Helium at 25 cc/mln. Instrument: duPont 21-491B mass spectrometer Source: Chemical ionization Mode: all samples were run in Electron Ionization, sample P.E. #2 was also run In Chemical Ionization with NH. gas Mass Range: 18 - 617 m/e Helium Separator: Single Stage Jet Multiplier: Bendlx Channeltron( 006284 Report No. y 24 k lcilt.ll Ci, i RESULTS Samples R.W. #1 and P. E. #1 showed by GC/MS only the solvents used in extracting and methylating: ethanol, ethyl ether, ethoxy ethyl acetate toluene and xylenes. It was necessary to air concentrate the ethyl acetate extracts tn order to obtain sufficient data to Interpret the mass spectra. The Tennessee River exposed fish ethyl acetate extract contained numerous G.C. peaks none of which appeared to contain fluorine. Th components appear to correspond to references of 2'methyl-3~octanone, acetic acid, a phthalate, and high molecular alkanes from about to ijt-least The plant effluent exposed fish ethyl acetate extract contained c(pounds which appear to correspond to references of 2-methyl-3~OCtphone> Ctlc acid, fluorochemicals whose highest mass fragments seen ''ifrir and, 582 m/e, nitrogen containing fluorocehmicals (whose highest mass fragments seen were 448 m/e and 584 m/e), FH 3923 (FOSE amide) and FH 3422 (POSE alcohol). SUMMARY This fish bioassy showed that extracts from the fish exposed to the Tennessee River water contained such compounds as ethanol, ethyl ether, ethoxy ethyl acetate, toluene and xylene. The extracts from the fish exposed to the Decatur plant effluent contained FOSE amide, alcohol, other related fluorochemicals, 2-methyI-3-octanone, and acetic acid. s 'h . , ]C 'j. N. SchroepTer /ch 006285