Document kmn36Kna1BrVyZO97XkggwEwB

Attain-ooGI BIOACCUMULATION (ANALYTICAL ONLY) TEST SUBSTANCE______________ Identity: Perfluorooctanesulfonate; may also be referred to as PFOS or FC-95. (1-Octanesulfonicadd, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt, CAS # 2795-39-3) Remarks: The test substance is a white powder. No information was recorded on the 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 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 using ethylacetate on one 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. 000273 TLC qualitatively indicated the presence of perfluorooctanesulfonate, but was not quantified. 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. DATA QUALITY____________________________________________ Reliability: Klimisch ranking 3. Effluent concentrations of subject fluorochemicals was 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 used in the study. 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. CONCLUSIONS______________________________________________ No conclusions can be made about the uptake of perfluorooctanesulfonate from Decatur effluent for reasons given above. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 REFERENCES________________________________________________ Study conducted by 3M Company. OTHER_____________________ Last changed: 5/4/00 000279 3SSr r747tiA TECHNICAL REPORT SUM M ARY TO; TECHNICAL COMMUNICATIONS CENTER - 201-2CN -(Im p o rta n t I f re p o rt is p rin te d on both sides o f paper, send tw o copies to TCC.) Date 5/1/79 D ivisio n Dept. Num ber Project Environmental Laboratory (EE & PC) Req. No. 48765 0535 Project N um ber Fate of Fluorochemicals Report Title 9970612600 Report Num ber Analysis for Fluorochemicals in Bluegill Fish To 014 A uthoril D. L. Bacon " " Em ployee Num ber(i) ________ G. A. Vraspir/Arthur Mendel N otebook Reference 48838-18 76416/43939 TTo. o f Pa ge t In c lu d in g C o ve rth e e t 7 i ^QbPirUilKR ii T V y hk Q(ComOppeanny Confidential) dosed ispecial Authorization) 3M CHEMICAL w REGISTRY ^ New Chemicals Reported Yes H No KEYWORDS: (Select terms from 3M Thesaurus. Suggest other applicable terms.) CURRENT OBJECTIVE: Status Report ) R EP O R T ABSTRACT: (200-250 words) This abstract information is distributed by the Technical Communications Center to alert 3M'ers to Company R&O. : :R.A.Prokop-236-33 V .Pothapragada-2 36-3A A.N.Welter-21-2W R.L.Bohon-21-2W J Information Liaison C002S0 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 (ng/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 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. C002S1 -3FLOW DIAGRAM 1 ANALYSIS OF BLUEGILL FISH 30 Fish (1) Weigh (2) "Tissuemize" (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 I * -- ------- ^ 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) C002S2 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 No 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 Ni0 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. C0G233 FC (Req. 4876S)/GAV/AM -5- 5/1/79 PE/EtAc extract showed approximately 10 y g /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. Elhabarawy, "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," 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, Ml oroohftmi r.al Journal. 23, 121 (1978). GAV/AM/cen Attachment 000284 ' _ , wn CENTRAL ANALYTICAL LABORATORY Report N o . ___7I_24_-- Date . . -Mf .r5t. 2 8 J 979 Subject: Decatur Fish Bioassy. R equestor:GJ.yr?s.Pj.0.A/.5"!!-*1 Request No. ................... * Dept. N a m e .!E.A.P5........ D ated .A.v5^r.JA.i3Z5-- Proj. No. R e p o rt: INTRODUCTION Samples were received from the requestors from a fish bloassy taken at the Decatur Plant. Fish were exposed to two different water systems: the Tennessee River which flows just outside the plant and the plant effluent running into the river. Ethyl acetate extracts of the fish were analyzed using the GC/HS system. Toluene extracts of the fish were methylated with diazomethane and analyzed. DISUCSSION 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. ^ #2 were analyzed using conditions for column B below. Instrument: Varian 2740 gas chromatograph Column A: 121 S.S. 20$ DC-200 + 10% Ben tone 34 on Anakrom PA Oven: 110C isothermal for 11 minutes then 1I0-170*C at IO*C/mIn. Injector: 165"C Column 6: 6* S.S. CW 20M 15% on Chromosorb W Oven: 50 - 225*C at 10C/m!n. Injector: 220C Detector: Varian flame ionization at 290C Carrier Gas: Helium at 25 cc/mln. Instrument: duPont 21-4918 mass spectrometer Source: Chemical Ionization Mode: all samples were run in Electron Ionization* sample P.E. #2 was also run in Chemical Ionization with MH. gas Mass Range: 18 - 617 m/e . * Helium Separator: Single Stage Jet Multiplier: Bendix Channeltron J C002S5 R e p o rt No. '/ I 2*1 rv.n t v , . ,i; RESULTS Samples R.W. #1 and P. E. #1 showed by GC/HS 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 In order to obtain sufficient data tt> interpret the mass spectra. The Tennessee River exposed fish ethyl acetate extract contained numerous G .C ; peaks none of which appeared to contain fluorine. The components appear to correspond to references of 2-methyl-3~octanonet acetic acid, a phthalate, and high molecular alkanes from about Cjg to at least Cjj,* The plant effluent exposed fish ethyl acetate extract contained compounds which appear to correspond to references of 2-methyl-3~octanonek acetic acid, fluorochemicals whose highest mass fragments seen were 483 m/e and. 582 m/e, nitrogen containing fluorocehmicais (whose highest mass fragments seen were 448 m/e and 584 m/e), FM 3923 (FOSE amide) and FH 3422 (FOSE alcohol) SUMMARY T h i s fish bioassy showed that extracts from the fish exposed to the Tennessee R i v e r water contained such compounds as ethanol, ethyl ether, ethoxy ethyl a c e t a t e , toluene and xylene. The extracts from the fish exposed to the Decatur p l a n t effluent contained FOSE amide, alcohol, other related fluorochemicals, 2-methyl-3_octanone, and acetic acid. /ch 0002SG