I T HNtCAL REPORT SUMMARY. For:n .3747-1 ’t-A D ate 5/22/79 ,..., TO: TECHNICAL COMMUNICATIONS CENTER - 201-2CN (Important - If report is printed on both sides of paper, send two copies to TCC.) ~ Division ’..~’,.:’ j’Oept. Number Environmental Laboratory (EE & PC) Project Decatur, Alabama - Tennessee River Fish Report Title Bioaccumulation of Fluorochemicals in Tenn. River Fish I ~O01 ,~.,,~ To D. L. Bacon Author(s) Employee Number{s) James E. Gagnon 213531 NO. of Png~s Incl~=dingCovorshoet 51568 SECURITY ~, ’~ Open (Company Confidential) KEYWOROS; (Select terrr~ from 3M Thesaurus. Suggest other applicable terms.] EE & PC Decatur ~Sp Closed ecialAuthorization) I 10 3M CHEMICAL REGISTRY New Chemicals Reported [] Yes [] No CURRENT OBJECTIVE; Qualitative and quantitative determination of , FM-3925, and FM-3422 in fish taken from the Tennessee River above and below Wheeler Dam at 3M’s Decatur plant. Analyze for organic and inorganic £1uoride in the same samples. REPORT ABSTRACT: (200-250 words) Thi.~ abstract information is distributed by the Technical Communications Center to alert 3M’ers to Cnmpnny R&D. It is Comport\ confidential material. Channel catfish (I~talur~ punctat~) had the largest ombined total, 2.74 @pm and 1.13 ppm, of F~-3923, M-3925, and FM-3422, as determined by gas chromatography. t was shown that the three fluorochemicals of interest bioaccumulated more readily in the gastrointestinal tract, fat and reproductive system of the channel catfish, while no fluorochemicals were observed in the muscle layer. A ~ white bass (Roce~+n ol~inol,~), taken ~rom below Wheeler Dam, had a combined FM-3923, FM-3925, and F~-3422 concentration of 0.40 ppm. A white crappie (Po~o~¢s ~Z~¢s), from above Wheeler Dam, was found to contain only FM-3923, 0.004 pDm. Total organic fluoride ranged ~rom 9.7 to 16.2 ppm, white crappie. Inorganic from 6.2 ppm, white bass, to 24.6 ppm, Future Studies: cc: D.Ricker-236-2 A.Welter A,Mendel ppm, channel catfish, fluoride ranged channel catfish. TLC on above fish samples. GC/MS on channel catfish samples. Background fluorochemical analysis on a crappie from a ~innesota Lake. Exhibit 1208 I r~ f o r m at ion L~.~. ~J State of Minnesota v. 3M Co., Initials: 3M CON FIDENTIAL Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0001 Court File No. 27-CV-10-28862 3MA01409559 ’~enn. River Fish/JEG 5/22/79 INTRODUCTION It is known that 3M’s Decatur, Alabama plant effluent has high organic fluoride levels, 10.9 ppm (1)(2). It has also been shown that fluorochemicals can bioaecumulate in fish in a laboratory environment (3)(4). With these combined factors, the next step was to see if fish caught in the Tennessee River near the Decatur plant had detectable levels of fluorochemicals, RESULTS AND DISCUSSION Table 1 lists the concentration, in ppm, in fish of compounds which have the same retention time as the three fluorochemicals of interest (FM-3923, FM-3925, and FM-3422). Analysis of the results for the dissected channel catfish, Sample 3A, shows that the fluorochemicals bioconcentrate to a greater extent in the gastrointestinal tract, reproductive system, and fat. It can also be seen that the muscle layer was found not to bioaccumulate the three fluorochemicals of interest. These results agree with earlier reports (3)(4). When comparing the total fluorochemical content (TFC) for the two whole fish sampl-~s, the iarge~ e~annel catfish contained more than twice the fl~orQchemical content, 2.74 ppm vs. 1.13 ppm. Since both fish were caught in the same area, a reasonable explanation for this may be related to the high partition coefficients for channel catfish. Fluorochemicals bioaccumulate in fatty tissue, and since more fatty tissue is.present in the larger fish, more fluoroehemicals would be expected. FM-3923 is present at higher concentrations in the dissected channel catfish, sample 3A, than other samples. Since bioaccumulation rates have not been determined for FM-3923, no explanations for the higher concentrations Can be offered The two fish samples which had cores taken from them will not be rigorously compared to whole fish samples. The reason for this is that the core samples may not have representative concentrations of fluorochemicals (whole fish,values may be higher or lower). Since core samples were taken from the approximate same location, the results can be rigorously qompared. The white bass from below Whee%eq. Dam,~ sample IB, had a whole fish TFC of 0.40 ppm, while the white crappie from above Wheeler Dam, sample 2A, had a whole fish TFC of 0.004 ppm. ~N~~~~C~-~ small statistical samples, it would be difficult to say that the larger TFC is due only to the White bass living in the presence of higher fluorochemical concentration, downstream from the plant. Other possible explanations for the higher TFC could be the following: Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0002 3MA01409560 Tenn. River Fish/JEG 5/22/79 TABLE i FLUOROCHEMICAL CONCENTRATION (ppm) IN TENNESSEE RIVER FISH FM-3925 & FM-3422 (i) Total Combined FC i~ Fish (ppm) (2) Sample FM-3923 IA - Whole fish 0.40 0 73 1.13 1B - Core (3) 0.82 3 31 0.40 (4) 2A 0.06 N D. 3A - Gills 1.48 0.80 3A - Liver 2.17 0 38 3A - Parts (7) 1.33 0 43 3A - Muscle N.D. N D. 3A - Fat (8) 13.85 6 12 3A - Gall bladder 1.57 0 74 Water blank N.D. N D. Ethyl acetate blank N.D. N D. - Core Footnotes (i) (2) (3) (4) (5) (6) (7) (8) (9) (5) to Table (6) o.oo4 (4) 2.74 (9) i: FM-3925 and FM-3422 cannot be resolved with GC parameters used; therefore, a combinec~ value is reported. Based on frozen weight of 1~he fish. Sample core, 3.61 cm, id contained skin, filet, reproductive organs, and parts of kidney, rectum, and backbone. Assumes that the concentralions obtained in the core are representative of the rest ~of the fish. Sample core, 3.61 cm id contained filet, vertebrae, skin, and bile. N.D. = Not detected. Consisted of muscle, skin, hi.cod, bone, and cartilage. Consisted of gastrointestinal tract, reproductive system, and fat. Based on the actual weight of sample used, 18.8% less than frozen weight, and weight percent o~ each part. Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0003 3MA01409561 Tenn. River Fish/JEG -4- 5/22/79 Longer river residence time, older fish. Longer location residence time. Different species a) Different ~eeding and life styles b) Contains larger weight percent of organs which tend to bioaccumulate fluorochemicals o) Larger fluorochemical partition coefficients If the core samples are representative of whole fish concentrations, then it can be postulated that channel catfish bioaccumulate fluorochemicals to a greater extent than either white bass or crappie. Reasons for this are the same as listed above. Table 2 gives the results of the organic (RF) and inorganic fluoride (Fv) concentration, in ppm, in the fish samples. TABLE 2 (5) ORGANIC (RF) AND INORGANIC (F0) FLUORIDE CONCENTRATIONS (ppm) Sample RF IA 9.7 24.6 2A 16.2 13.3 IB I0.5 6.2 Water N.L. 0.01 F@ Jon Belisle points out that the high inorganic fluoride values seem rather surprising. His only explanation was that fish flour previously analyzed, for a different requestor, was shown to have inorganic fluoride values higher than organic fluoride. Jon also states that high inorganic fluoride values would make it difficult to calculate low levels of organic fluoride. Comparison of the organic and inorganic fluoride content shows that samples from above Wheeler Dam have just as high, if not higher, values than for the sample from below the dam. There are no clear cut explanations for this observation. An earlier analysis of Tennessee River water showed high organic fluoride concentrations upstream from the plant. At that time, it was thought the samples may have been mislabeled. With these results, Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0004 3MA01409562 Tenn. River Fish/JEG -b- 5/22/79 it would seem to indicate that the concentration of fluorochemicals may actually be less below Wheeler Dam. This may be caused by volatilization of the fluorochemical when going over the dam (I), settling of fluorochemicals before the dam. Comparison of organic fluoride values from Tables l.and 2 show no correlation. For example, the highest organic fluoride value, 16.2 ppm for sample 2A, had the lowest TFC, 0.004 ppm, for the fluorochemicals analyzed. A possible explanation is that there are organic fluorides present in very high concentrations which were not analyzed for individually. The species which had the highest fat content, channel catfish, had the lowest organic fluoride concentrations. With limited sample population (2 fish of one species and one of each of two other species), it is difficult to draw any meaningful conclusions. The only definite conclusion is that the fluorochemicals studied do appear to bioaccumulate in river fish under natural conditions. EXPERI MENTAL I. Sample materials Fish 1,.A - Small channel catfish (Ictalurus punctatus), caught above Wheeler Dam in Tennessee River. IB - White bass (Roccus chrHsops), Dam in Tennessee River. caught below Wheeler 2A - White crappie (Po.;om£u annuZari~), Dam in Tennessee River. 3A - Large channel above Wheeler caught above Wheeler catfish (Ictalurus punctatus), Dam in q’ennessee River. caught St andards FM-3923, FM-3924, and FM-3422. Ten ppm standards of FM-3923, FM-3925, and FM-3422 were prepared by diluting 1 ml of a I00 ppm standard, in ethyl acetate, to mark with ethyl acetate in separate I0 ml volumetric flasks. Analysis Instrument~terials Blender: Waring Commercial blender, Model #91-263, available from Waring Products Division, Route 44, New Hartford, CT 06057. Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0005 3MA01409563 Tenn. River Fish/JEG 5/22/79 Tissuemizer: Mode] #SDT0 available from Tekmar Company, 37202, Cincinnati, OH 45222. P. O. Box Dinker Die: 3.61 cm id AISI-02 high carbon steel cutting die made by Jerry Guthrie in Central Research Labs, described in 3M Technical Notebook #51568-35. Mixer: "Vortex Genie" Model #K-550-G, available from Scientific Industries, Inc., Bohemia, NY i1716. Centrifuge: Damon-IEC Model #B-20A, available from Damon-IEC Corporation, Needham Heights, MA. Bottles" Four-ounce widemouthed clear glass bottle sealed with aluminum foil and aluminum foil-lined caps. 125-mi linear polyethylene (LPE) plastic bottle with polyseal caps. Gas Chromatograph: Chromatograph - Hewlet~Packard Model 5713 GC. Integrator - Hewlett-Packard Model 33SOA integratorprinter. Both o£ the above available from llcwlcLL-Packard 150 Page Mill Road, Pale Alto, CA 94304. Co., Column - Six-foot, 1/8 inch OD, stainless steel, packed with 10% CW20M on 60/80 Chromasorb W-AW. Column Temperature - Isothermal 180° C. Injector - On-column at 200° C. Detector - Electron Capture at 300° C. Flow - %40 cciminute of Argon:Methane (95/5). Ethyl Acetate" "Li Chrosolv" chromatography solvent available from MC/B Manufacturing Chemists, 2909 Highland Avenue, Norwood, OH 45212, as Catalog #6008688M. Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0006 3MA01409564 Tenn. River Fish/JEG -%- 5/22/79 Water: Deionized water. 3. Procedure (6) Procedures used below, except for minor modifications, were obtained from earlier 3M Technical Report summaries (7). Samples IA through 3A and IB were removed from the freezer and placed in large aluminum pans, in a fume hood, and allowed to thaw. A whole channel catfish, sample IA, was cut into 5 sections and homogenized in a blender with 200 ml water. Sample IB had a dinker die core sample taken just off the lateral line behind the gill plate. Contents of the 20.591 gram sample were skin, filet, small part of backbone, reproductive organs, part of kidney, and rectum. Sample 2A had a dinker die core sample taken behind the gill plate. The 16.684 grsm sample contained filet, vertebrae, skin, and bile. Samples IB and 2A were homogenized with I0 ml of water in a "tissuemizer." Sample 3A was dissected, and the various individual parts were homogenized with water. Individual parts weighing more than 25.0 grams were homogenized in a blender, while those of lesser weight were homogenized in a "tissuemizer." Table 3 lists the sample, sample weight, and amount of water added for homogenizing each sample. All of the above samples, after homogenization, were divided into five aliquots and placed in precleaned bottles, (dichromate/acid, water rinse, dry, toluene, dry). Three aliquots were placed In LPE Do~Ules, while the other two were placed inoglass bottles. Samples were in a refrigerator at 4.5 C. until needed. stored Samples analyzed for FM-3923, FM-3925, and FM-3422 were prepared according to the following procedure. See Table 4 for weight of sample and milliliters of ethyl acetate used ior extractions. A previously homogenized sample, stored in a glass bottle, was weighed (no larger than 4.00 g) and added to & 30-ml precleaned glass centrifuge tube. A volume of ethyl acetate was added at the rate of 1.0 ml ethyl acetate per gram of homogenate. The ethyl acetate/fish homogenate were mixed for 1.5 minutes in a mixer at a speed setting of 3. The samples were removed and centrifuged at 1500 rpm at Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0007 3MA01409565 Tenn. River Fish/JEO 5/22/79 21° C. for i0 minutes. After centrifuging, the ethyl acetate layer was separated, by use of a pipet, and placed in a vial. Five ~i of sample (standard) was injected for gas chromatographic analysis. Samples IA, 2A, and IB homogenates, plus a water blank, in LPE bottles, were sen~ to Jon Belisle of the Central Research Laboratory for organic and inorganic fluoride analysis. REFERENCES (i) 3M Technical Report Summary, August 30, 1978, Arthur Mendel to R. L. Bohon, "Fate of Fluorochemicals Project - Progress Report." (2) Centra! Research Laboratory Report Number 6902, 1978, Jon Belisle. (3) "Bioaoncentration Channel Catfish," May 17, 1977. of FM-3422 in Bluegill M. T. Elnabarawy to A. April 20, Sunfish and N. Welter, in (4) 3M TRS, August 16, 1978, A. N. Welter I:o D. L. l~aaon, "Evaluation of the Bioconc~ntration Potential of FM-3422." (5) Central Research Laboratory Report by Jon Belisle, May 7, 197@. on Request #A72199 (6) Experimental work done in cooperation with A. N. Welter of the Environmental Laboratory (EE & PC), who per£ormed the dissections and homogenizations. (7) 3M Technical Report Summary, November 15, 1977, A. Mendel to D. L. Bacon, "Analytical Methodology on ~M-3422." ~cen Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0008 3MA01409566 Tenn. River Fish/JEG -9- 5/22/79 TABLE 3 £ISH WEIGHTS AND WATER VOLUMES USED FOR IIOMOGENIZATION Sample .D,e_scription Initial Whole Frozen Weight Actual Sample Weight Used ml Water Used IA 146.0 g 2A 266.5 g 16 684 g (3) I0 IB 210.0 g 20 591 g (3) I0 3A - Muscle 752.0 g 3A - Gall bladder 752.0 g 1 37S g I0 3A - Liver 752.0 g 5 949 g I0 3A - l~at 752.0 g 52 230 g i00 3A - Parts 752.0 g 321 57 g 300 3A - Gills 752.0 g 19 38 g I00 Whole £ish (1)(2) 209 93 g 200 200 Footnotes" (I) A :Fish hook, with no apparent rust or line, gish and was removed be£ore homogenization. (2) The fish appeared to be sl%ghtly to constant air flow over surface weight of fish used may h&ve been (3) Sample core 3.61 am dehydrated was ~ound (possibly i.n due of fish) so the actual less than frozen weight. id. Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0009 3MA01409567 Tenn. River Fish/JEG -I0- 5/22/79 TABLE 4 FISH WEIGHTS AND ETI~L ACETATE USED FOR EXTRACTIONS Sample Description VOLUMES Weight of Fish % Water Homogen at e in ( grams ) Homogen at e Actual Fish Wt. Extracted (rag) ml EtOAc 3A - Gall Bladder 1.20 87.9 145.2 1.2 3A - Liver 2.2o 62.7 820.6 2.2 3A - Muscle 2.40 48.8 1228.8 2.4 3A - Fat 2.40 65.7 823.2 2.4 3A - Parts 3. O0 48 3 1551.0 3.0 3A - Gills 3.00 83 8 486.0 3.0 Water Blank 2.40 i00 0 IA 2.40 57 8 1012.8 2.4 IB 2.40 32 7 1615.2 2.4 2A 2,40 37 5 1500.0 2.4 Made Available by 3M for Inspection and Copying as Confidential Information: Subject to Protective Order In Palmer v. 3M, No. C2-04-6309 1208.0010 2.4 3MA01409568