Document NemajaQzO7azdDmMoq8v4k8MR

ARUG-0^3 BIOACCUMULATION (ANALYTICAL ONLY) TEST SUBSTANCE Identity: Perfluorooctanesulfonylamido(ethyl)acetate; may also be referred to as PFOSAA, FC-128 or as the major component of FC-127, FC-129, FC-109, or FC-109-X. (Glycine, N-ethyl-N[(heptadecafluorooctyl)sulfonyl]-, potassium salt, CAS # 299151-7) Remarks: The test sample is FC-128. Current information indicates it is a mixture of 86% test substance and 14% C 4 - C 7 fluoroalkyl carboxylate compounds. 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. 0003^4 GC/MS analysis of ethyl acetate extracted fish exposed to plant effluent did not detect the presence of FC-128. 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., N H 3-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 Fluorochemicals 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 OTHER Last changed: 5/18/00 000.1*5 S^47, a TECHNICAL REPORT SUMMARY TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN Date 5/1/79 (Im p o rtan t - I f rep o rt sp rin ted on both sides o f paper, send tw o copies to TC C .i D iv isio n Dept. Number Project Environmental Laboratory (EE & PC) Req. No. 48765 0535 Protect Num ber Fate of Fluorochemicals Report Title 9970612600 Report Number Analysis for Fluorochemicals in Bluegill Fish To 014 A u th o r(i) D. L. Bacon Em ployee Number!) G. A. Vrasplr/Arthur Mendel Notebook Reference klo7. 6of4P1ao6es/ln4cl3ud9tn3g9 48838-18 S E C U R IT Y ^ Open (Com pany C onfidential) 0 Closed (Special A uthorization) 3M C H E M IC A L w R E G IS TR Y W New Chem ical* Reported Yea (g No KEYW ORDS: (Select term * from 3M Thesaurus. Suggest other applicable term s.) C U R R E N T O B JEC TIVE: Status Report REPO R T AB STR A C T: (200-250 words) This abstract inform ation is distributed by the Technical Com m unications Center to alert 3M 'ers to Company R&O . It is Company confidential m aterial. ' -S c:R.A.Prokop-236-33 V.Pothapragada-2 36-3A A .N.Welter-21-2W R.L.Bohon-21-2W Inform ation Liaison.. r 1Initials: ; V 0003^.6 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 (yg/g) of Fli 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. 0003^.7 -3FLOW 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 -R-o-u-t-e--3-----, Concentrate I * *-- ----------- GC/MS TLC For FC-95 FC-128 FC-143 (2) Acidify (3) Extract (Toluene) (4) Centrifuge (5) Isolate Toluene Layer (6 ) Methylate 11 GC/MS GC For FC-143 (Ester) 0003*8 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.V 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. 000349 FC (Req. 4876S)/GAV/AM -5- 5/1/79 PE/EtAc extract showed approximately 10 yg/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, "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, Mlr-roohemlr>a1 Journal r 23, 121 (1978). GAV/AM/cen Attachment 0003-0 ;y> 1* ?M3-< '*'> CENTRAL ANALYTICAL LABORATORY Report No. ...1 1 ^ ............................ Oat.. .1 .J 52?......... Subject: Oecatur Fish Bioassy. Requestor:G_ ^. r2 ! P L D e p t . Name AiJ.fS............... R e q u e st N o . . 5.50687.............. Report: D a te d .J^yember J 6 _ . J 9 7 8 _ INTRODUCTION Proj. No.[H it!#*0* Samples were received from the re q u e sto rs from a f i s h b io a ssy taken a t the Decatur P la n t. F is h were exposed to two d i f f e r e n t w ater sy ste m s: the Tennessee R ive r which flow s Ju st o u tsid e the p la n t and the p la n t e fflu e n t running into the riv e r. Ethyl ace tate e x tra c ts o f the f i s h were analyzed u sin g the GC/HS system . Toluene e x t r a c ts o f the f i s h were m ethylated w ith diazomethane and analyzed. DISUCSSION The sam ples were la b e le d R.W. #1 and P .E . #1 (m ethylated to lu e n e e x t r a c t s ) and R.W. #2 and P .E . #2 (e th y l a c e ta te e x t r a c t s ) . Samples R.W. #1 and P .E . #1 were analyzed u sin g c o n d it io n s f o r column A below and sam ples R.W. #2 and P .E . #2 were analyzed u sin g c o n d it io n s f o r column B below. instrum ent: V a ria n 2740 g a s chrom atograph Column A: 12* S . S . 20* D C -200 + 10* Ben tone 3<i on Anakrom PA Oven: 110"C isoth erm al f o r 11 m inutes then II0 -1 7 0 *C a t !0 *C /m ln. In je cto r: I65*C . v1. Column B: 6 1 S . S . CW 20H 15* on Chromosorb W Oven: 50 - 225C a t 10*C/mln. In je c to r : 220*C D etecto r: V a ria n flam e io n iz a t io n a t 290#C C a r r ie r Gas: Helium a t 25 cc/m ln. Instrum ent: duPont 2 1-4 91B mass spectrom eter Source: Chemical Io n iz a tio n Mode: a l l sam ples were run in E le c tr o n Io n iz a t io n , was a ls o run in Chemical Io n iz a t io n w ith NH. Hass Range: 18 - 617 m/e * Helium Separator: S in g le Stage J e t M u ltip lie r : Bendix C h a n n e ltro n sam ple gas P.E. #2 000351 Rc*i>orl No. m * k tWil'UI i u , i . ) / j RESULTS Samples R.W. #1 and P. E. #1 showed by GC/HS o n ly the s o lv e n t s used in e x tra c tin g and m ethylating: eth ano l, e th y l e th e r, ethoxy eth yl ecetate toluene and xylenes. I t was necessary to a i r concentrate the e th y l ace ta te e x t r a c t s In o rd e r to ob tain s u f f ic ie n t data to in te rp re t the mass spectra. The Tennessee R iv e r exposed f i s h e th y l a c e ta te e x t r a c t co n tain e d numerous G.C. peaks none o f which appeared to c o n ta in flu o r in e . The components appear to correspond to references o f 2*methy1*3*octanone, a c e tic a c id , a p h th ala te , and h igh m olecular alkanes from about C | j to a t le a s t The p la n t e fflu e n t exposed f is h e th yl ace te te e x tra c t co n tain e d cqiiapunds which appear to correspond to references o f 2>fnethyl~3**OCtenons, d p l t l c a c id , flu o ro ch e m tca ls whose h ig h e st mass fragm ents seen umire i/ e and 582 m/e, n itro g e n c o n ta in in g ftu o ro c e h m ica ls (whose h ig h e s t mass fragm ents seen were kk8 m/e and 58k m/e)^ FH 3923 (FOSE amide) and FM Jh22 (FOSE a lc o h o l) . SUMMARY T h is f is h b io a ssy showed that e x tra c ts from the f is h exposed to the Tennessee R iv e r w ater contained such compounds a s e th a n o l, e th y l e th e r , ethoxy e th y l ace ta te , toluene and xylene. The e x tra c ts from the f i s h exposed to the Decatur p la n t e fflu e n t contained FOSE amide, a lc o h o l, o th e r re la te d flu o r o c h e m lc a ls , 2-m ethyl-3-octanone, and a c e tic a c id .