Document zQv3maeg7yrwKa3Kox7dwX58g

BIOCONCENTRATION TEST SUBSTANCE Identity:N-ethylperfluorooctanseulfonamidoethanolm;ay alsobe referredto as N-ETFOSE Alcoholor FM-3422. (1-OctanesulfonamideN,-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 solidof uncharactedzedpurity. METHOD: Method/guideline followed: 3M derivedmethod outlinedinthetestprocedure fordeterminingthe aquaticBioconcentrationFactorof N-ETFOSE alcohol. Type: Staticexposure withflow-throughclearancephase. GLP (Y/N): No Year: 1977 Species: Bluegil(lLepomis macrochirus)and channelcatfis(hictalurus punctatus) Supplier: Bluegilolbtainedfrom a privatehatcheryinBalticO,hio Channel catfishobtainedfrom a privatehatcheryinLonoke, Arkansas Analyticalmonitoring: ConcentrationofN-ETFOSE alcoholinwater and fishby GC withelectroncapture;temperature Length and weight: Bluegilwlere inthe range of 2.5-3cm and weighed 0.5-1.0grams Channel catfishwere 8-10 cm and weighed 5-10 grams Exposure period: 21 days (Bluegill&)14 days (Channel catfish) Clearance period: 14 days forboth species Statisticamlethods:' Not noted inreport. Test fishage: Not noted inreport. Loading: Not noted inreport. Pretreatment: None Test conditions: Dilutionwater: Carbon-filterewdellwater Dilutionwater chemistry: Not includedinreport. Stock and testsolution preparation:The primarystocksolutionwas. prepared by dissolving57 grams ofN-ETFOSE alcoholin50 mi Of acetone,applyingthe solutionto 10 pounds of3.5 mm glassbeads, evaporatingoffthe acetone,and circulatincgarbon filterewdellwater throughthe glass beads. The system was designed toresultina final loadingratioof0.5 grams per literA.erobicconditionwsere maintained throughoutand the system was assumed togo to saturatioanfter circulatinfgor3 weeks. Exposure vessels: 30-gallonglasstanks Number of replicates:One Number of fishper replicate:Not noted inthe report. Number of concentrations:One plusa negativecontrol Water chemistry during the study: Dissolved oxygen: >5 mg/L Test temperature:21+1 OC pH: Not noted inthe report. Photoperiod: 16 hours lighatnd 8 hours darkwitha 30 minute transition period. Remarks field:Priorto exposure, and atthe time oftheirtransfertotesttanks, the fishdidnotexhibitany symptoms ofdisease or abnormalitieosf behaviorand appearance. The fishwere fed dailyata rateof2 percentoftheirtotalbody weight witha commerciallyavailablebasicdiet(TetraMin.). RESULTS Analysisofthewhole fishshowed fluorochemicalconcentrationratiosinfishto water ofapproximately400:1. The study indicatedthatthe N-ETFOSE alcohol concentrationinthefishstabilizeadfter7 days. Fishhavingan intiial concentrationof0.213 mg N-ETFOSE alcohol/g offishplaced inwaterfreeof the testsubstance showed a decrease of0.012 mg testsubstance /g offishin 14 days (wetweight). CONCLUSIONS No reliablceonclusionscan be derivedfrom thisstudy.Althoughthe objectiveon the cover page indicatetshisreportwas todeterminetheextentoffluorochemical uptake and bioconcentrationi,tappears from the abstracton the cover page that the primarypurpose was todevelop a method fortestingbloaccumulationwith the investigatioonfbi6accumulationoffluorochemicalsinfishbeingsecondary. The study'sobjectiveisnot detailedinthe reportbody. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331,St.Paul, Minnesota,55133 DATA QUALITY ReliabilityK:limischranking3. There isa lackofdescriptiofnorthe study's methodology. The vigorousmethod used tosolubiliztehetestsubstance may have resultedina supersaturatedsolution.The uptake and clearanceofthe test substance by bothtestorganisms was quiterapid,leadingone toquestionif perhaps testsubstance was sorbed tothefishratherthan beingtaken up from solution.Test substancepuritywas notcharacterized.The initiwaalter qualityof the dilutiownater ismissing.The testsubstanceconcentratiounsed inthetestis greaterthan thesolubilitTyh.e recoverytank used appears to be the same for both speciestested. Thirdpartyreviewby Dr.James W. GilletotfComell Universit(yobservations includedwithreport)also indicatea lackof reliability. REFERENCES 3M TechnicalReport "Bioconcentratioonf FM 3422 inBluegilSlunfishand in Channel Catfish"M. T. Einabarawy,Project9970612600, Fate of Fluorochemicals,Report Number 01, May 17, 1977 3M TechnicalReport "AquaticFate ofA Fluorochemical:FM 3422."A. N. Welter, Project9970512623, Fate of Fluorochemicals,Report Number 002, October 14, 1977 3M requested expertoverview,"BioaccumulationStudies",Dr.James Gillett, Comell UniversityM,arch 8,1993 OTHER Last changed: 5/18/00 30m~ Foeff. 6747 11 A TECHNICAL REPORT SUJWMARY c"';/ 17/77 : TECHNICAL COMMUNICATIONS CENTER - 201-2CN (important -- If report is printed on both sides of paloor, ond r" copies to TCC.) Division P,Oloct we-p-o-,T-ift10 Environmental Laboratory Fate of Fluorochemicals (EE 4 PC) oiot N-u-mi6@- 0222-78 rrol*ct Number 9970612600 Report Number Bioconcentration of FM 3422 In Bluegill Sunfish and In Channel Catfish 01 To Author(s) A. N. Welter - 21-2W (58) EnV@Nse Numberts) Notebook M. T. Elnabarawy - 2-3E Fleforence 46981 Including Cowef*Wot 42669, Pages 17-26 rI Open SECURITY 10,- 11 Closed lr;onvanCyonfidential)(Special Authorization) CHEMICAL REGISTRY 10. Now ChemkolsReported 0 Yes M No KEYWORDS: '(Seletcetrmsfrom3M Thesaurus. SuW@st other applicabletorna.) CURRENT OBJECTIVE: The purpose of this pilot study was to Oetermine the extent of fluorochemical (FM 3422) uptake and/or bioconcentration by Bluegill sunfish (Lepomis macrochirus) and Channel catfish (Ictalurus punctatus). Fluorochemical Biology/Activity -).,Pscreening t,aioaccumulation) (Fish) L-E & PC Div. REPORT ABSTRACT:(200-2w5o0rds) This abstractinformation is distributedbV the Technical Comrnunications alert3M'ers to Company R&D. It isCompany confidentialrmteris1. . Centetro A modified technique is proposed for monitoring fluorochemicals in suspect aquatic environments by whole f'ish or tissue analysis. Bluegiil sunfish (Lepomis macrochirus) and Cliannel catfish (Ictal'urus gunctatus) contained concentrations of FM 3422 greater than those found in their water environment, achieving ratios of approximately 400:1. Concent'rations of FNI 3422 by these fish species had also plateaued within seven days of exposure. When whole fish (Lepomis macrochirus) were analyzed for FM 3422 uptake following a two-minute exposure in the test tank, an insignificant uptake was noted (.0006 mg/g),. Bioconcentration s,,udiesare useful to qualitatively monitor fluorochemicals in water. Whole fish or tissue analysis from fish exposed to fluorochemicals in their environment may prove to be a useful tool in evaluating the mobility of these chemicals in an aquatic envirorment. @nfL,riyiat,Losna,son n tials -2- Experimental: A sample of FM 3422 (57 grams dissolved in 50 ml acetone) was applied to 10 pounds of 3h mm glass beads. The glass beads were then placed under the hood and the acetone was allowed to evaporate over a 48-hour period. The FM 3422 coated beads were spread evenly in an all-glass 30-gallon tank (114 liters), forming a layer of approximately 2 cm above the undergravel filter. The test tank was filled to its capacity with carbon-filtered well water. Chemical composition of the water is available upon request. The amount of test compound applied was designed to give a final loading ratio of 0.5 grams per liter. For 3 weeks and under continuous aerobic conditions (aeration was maintained at all times),this dynamic system generated saturated water solution of FM 3422 without organic solvents. Under identical test conditions, a control tank was set up in a similar manner containing no FM 3422. After the 3-week period of aerobic aging, test fish were introduced into the tanks (an assumption was made that equilibrium in the system had been reached). Fish used in this experiment were obtained from private hatcheries: Source: Size: Weight: Bluegill sunfish (Lepomis macrochirus) Baltic, Ohio 2.5-3 cm .5-1.0 grams Channel catfish Clctalurus punctatus) Lonoke, Arkansas 8-10 cm 5-10 grams The fish were held and cared for in adequately aerated water (dissolved oxygen was greater than S mg/1). The fish were acclimated to test sonditions; water temperature was maintained at room temperature 70+20 F.(21+1 C). A i)@otoperiod of 16-hour light and 8-houi-dark was provided with a 30-minute transition period. The fish were fed,daily at a rate of 2 percent of their total body weight with a commercially available basic diet (Tetra Min). Prior to exposure, and at the time of their transfer to test tanks, the fish did not exhibit any symptoms of disease or abnormalities of behavior and appearance. A criterion for sampling was set up On various days of increasing periods of exposure, representative water samples at various depths were collected and fish were-sacrificed at random. At the end of the exposure period, remaining fish were transferred to a clean aquarium, which was continuously filled with fresh water (a flow-throughsystem) to determine the clearance (recovery) rate. At sacrifice, the total fresh body weight was recorded. Some channel catfish were dissected, and various parts were removed and retained for analysis. Results: Extracts of water samples and of sacrificed fish were analyzed for FM 3422 concentrations by the GC teciiniquewith an electron capture detector. Extractions and GC analytical techniques have been perfomed in-house under the direct supervision of A. Mendel. Fish-to-water concentration ratios of FM 3422 were also calculated. Detailed description of this work is attached. Additional information on this project can be found in M. T. Elnabarawyls technical notebook, No. 42669, pp. 17-26. -3- Discussion: Throughout the experiment, all fish appeared generally healthy and active in both test and control tanks. During periods of exposure, S bluegill sunfish died (2 from test tank and 3 from control tank). The mortality was most likely due to physical injury and was not considered to be test compound related. Water was added periodically to supplement the loss due to sampling and evaporation. Concentrations of FM 3422 in both fish species had reached a plateau by 7 days of exposure. Measured concentrations of FM 3422 in the viscera illustrated the importance of fish size on bioaccumulation (the test compound tends to bioconcentrate in higher ratios in larger fish). After transferring the remaining bluegill sunfish to the recovery tank, and before introducing channel catfish, 5 pounds of sea-sand (washed and ignited) were evenly dispersed on the bottom covering the FM 3422 coated beads. Analysis of water samples taken before and after adding the sand showed no change in FM 3422 concentration. Whole fish analysis of 3 bluegill sunfish after a two-ainute dip in the test tank showed insignificant uptake (.0006 mg/g). If you have any questions, please contact me on 3-9186. MTE/cel Attachments -4- Bluegill Sunfish (Lepomis macrochin!a Treatment FM 3422 Concentrations Fish-to-Water Ratio L)ptake(euosure dayl: In Fish (mg/&)* 0 No background 8 .118 14 .117 21 .213+ .005 Clearance irecovery days): 0 .213 7 .007 .012 In Water (ppm)** .329+ .003 (air-off) .290!..005(air-off) 407:1 .428 (air-off) 273:1 .580 (air-off) 367:1 Values are means and statidarddeviations from the analysis of 3 (whole) fish. Values are means and standard deviations from the analysis of 3 water samples representing depths of--36, 20 and 6 cm from-bottom. Channel Catfish (Ictalurusgunctatus) Treatment FM 3422 Coricentrations Fish-to-Water Ratio L)ptake(exposure d4ys): In Fish (!S/g)* 0 No background 7 .128+.Ol 14 Clearance (Recovery days): 0 7 14 .100 .100 .023 .003 In Water (ppi!)** .340 (air-off) .3'&1(5,-ir-off) 1.075 (air-on) 394:1 119:1 .546+.02 (air-off) 183:1 .62C+.05 (air-on) 161:1 Values are means from the analysis of 2 (whole) fish. Values are means and st.Lndarddeviations from the analysis of 3 water samples representing depthtof 36, 20 and 6 cm from bottom. Forni b 147 11 A TECHNICAL REPORT SUMMARY 14/77 Tt):TECHNICAL COMMUNICATIONS CENTER - 201-2CN (Important Ifreportisprintedon both sideosfpaper,sendt" copiesto TCC.) Division Project R*P-O-rt-Titio Environmental Laboratory (EE & PC) Fate of Flunrochamicals Dept Number 'Fr-oTue-cMtN bar 9970612623 Report Number Aguatic Fate of A Fluorochemical: FM 3422 To Author(s) D. L. Bacon lEmPlOV@-Number(s) Notebook Reference A. N. Welter SECURITY 10 0 Open 0 Closed (CompanyConfidential) fspeciaAluthorization) KEYVIORDS: (Selecttermsfrom 3M ihosaurusS.uggestother applicablwems.) CURRENT OBJECTIVE: 3M CHEMICAL REGISTRY 0 362 Nm of-Pap-n Including Covershost 9 Now ChemicalsReported Yes Ea No LE & PC - Div. Fluorochemicals (Aquatic) Toxicity @Bioconcentration) Progress Report REPORT ABSTRACT:(200-2w5o0rdsT)hiasbstracitnformatioinsdistributbeyd theTechnicalCommunicationsCenterto aler3Mt'ersto ConvarvyR&O. ItisCompany confidentimaalterial. :n,fa,mat 'In L 1,NTRODUCTION ,I'hesubject compound (FM 3422) was selected for testing based on its importance as an intermediate in the synthesis of other commercially important fluorochemicals. These compounds represent-a major commitment by the Commercial Chemicals Division. With increasing governmental regulations pertaining to the influence of chemicals on the environment, consideration of the environmental impact of this chemical class was mandated. Furthermore, since FM 3422 might qualify as a 3M "critical chemical," extensive laboratory investigations were performed to assess its possible environmental impact. Physicochemical data have been utilized to predict the behavior of chemicals in the environment in the absence of experimental data. Available physicochemical data for FM 3422 include the following (1):s water solubility - 0.05 ppm, partition coefficient in n-octanol/water system - >10 . These data would suggest that FM 3422 would be persistent, relatively insoluble in water and possess lipophilic properties. FM 3422 was nontoxic within its solubility limits when submitted for aquatic toxicity determinations. It is the purpose of this report to present data relative to the bioconcentration potential, uptake and clearance rates of FM 3422 in either the bluegill (Lepomis macrochirus) and/or the channel catfish (Ictaluruz punctatus). ME-'niODS. M. T. Elnabarawy has recently described the standard methods utilized in the Environmental Laboratory relative to the acclimation period, care of aquatic organisms and method of chemical exposure of these organisms (2). Specific protocols for the determination of bioconcentration factors (BCF), uptake and clearance rates used in this study follow: Bluegill BCF were determined at the 8, 14 and 21 days of exposure to FM 3422. Clearance values were evaluated on the 7 and 14 days of depuration. Channel catfish BCF were ot)tainedat the 7th and 14th day of exposure with clearance values being determined after identical periods of depuration. Bioconcentration factors for specific organs of the channel catfish were determined after either one or four weeks of exposure to the test fluorochemical. Water samples were obtained at three different levels within the experimental tank on those days when fish sanq)leswere obtained. The analytical techniques used-for the determination of FM 3422 levels were those routinely used by the Environmental Laboratory and will be the subject of a report (1). Ri:suurs Biucoticentrationfactors, uptake and clearance rates were monitored in both the bluegill (Lepourisnxzcrochii@@,a-n)d channel catfish (Ictaturuspunctatus) at varying time intervals during exposure to 0.5 g/i FM 3422 in the aquatic environment as well as during depuratioii (Tables 1, 2, Figures 1, 2). Graphic representation of these data indicates that a rapid uptake of FM 3422 by both organisms had occurred (Figs. 1, 2). The initial sampling period (Days 7 and 8) values indicate that a steady state has been attained at some earlier time period. Of greater importance was the rapid clearance of the test material from both organisms which was quite apparent after seven days of depuration. The channel catfish cleared FM 3422 somewhat more completely than did the bluegills. Bioconcent ration factors and clearance values determined for whole organs were quite similar and seemingly independent of exposure periods (Table 1). The elevated fluorochemical C values observed in the channel catfish experiments were due to a single seriwesof outlier values. When these values were ignored, fluorochemical concentrations achieved in both experimental tanks were identical, 0.4 ppm. In the channel catfish, the more lipophilic organs bioconcentrate FM 3422 to a greater degree than the relatively lipid-free materials (Table 2). Thus, the oil layer obtained from the skin and the viscera (gut) possessed the highest fluorochemical bioconcentration factors following one week of exposure to FM 3422. Brain tissue, which was analyzed only following four weeks of exposure to the fluorochemical showed similar elevated BCF'S. The gills had attained high levels of FM 3422 at both exposure pe*riods, which is probably indicative of a large surface area available for binding. The remaining organs which were tested - muscle skin, skeleton - achieved similar levels of FM 3422. These values for BCF were a'pproximately one order of magnitude less than that found for the more lipophilic materials. DISCUSSION In these studies, uptake of FM 3422 by the bluegills and channel catfish probably occurred via the gills, oral and/or cutaneous routes. Transient exposure, < 2 min.,ot-fish to a fluorochemical-aquatic environment did demonstrate uptake of this fluorochemical. Uptake may result from the penetration of the lipophilic gill epithelium by this lipophilic molecule. During long-term (days) FM 3422 exposure, the fluorochemical could enter the gill circulation and thence be transported to various sites within the organism. Granmo and Kollberg (3) have discussed the uptake mechanisms of nonionic surfactants in the cod. In their studies, it was demonstrated that rapid uptake of this chemical by the gills had occurred and that blood was the principal transport medium to the various tissues/ organs where deposition occurred. Bass and Heath (4) postulated that the gills may be damaged by exposure to toxic materials resulting in tissue hypoxemia which culminated in death of the test organism. Bass et al (5), in a subsequent.paper, conclusively demonstrated that hyperplasia with Ta-n@e-Ilafrusion of gill filaments ajidedema did occur in the presence of a toxicant. The resulting hypoxic condition wzis due to impaired respiratory gas transport. These papers stress the importance of the gills in the uptake of foreign chemicals while also enumerating potential lesions which may result, nanifesting themselves as a toxic response. Oral uptake of FM 3422 (feeding) would afford a direct route for the absorption of this fluorochemical by the gastrointestinal tract (gut). Movement of the fluorochemical into the intestinal circulation would result in the transport of this chemical by the blood to otlierorgans of the body; liver, gall bladder, inesentery,etc. Circulation of the fluorochemical would then result in the selective deposition of this material in the more lipophilic tissues of the organism. It has been suggested, Chiou et al (6) that toxicity associated with e:@posureto lipophilic materials may be the result of the long-term slow release ol'the chemical or a metabolic product into the circulation of the organism. Percutaneous penetration of FM 3422 may explain the bioaccumulation of this material in the oil layer of the skin. It may be speculated that FM 3422 may remain It)calizedin this area with a subsequent leaching effect resulting in its eventual clearance. In these pilot studies, we did not quantitate uptake rates of FM 3422 either in the intact organism or specific organ systems. However, the data did indicate that in the intact organism a steady state (intake=output) had been attained in less than seven days. Uptake rates observed are a function of metabolic rate, age, weight, water temperature, feeding habits, etc. These variables must be controlled when replicating these experiments. Clearance (elimination) rates were not determined during the initial days of depuration; therefore, it can only be stated that FM 3422 was cleared in <7 days. Of interest would be the determination of clearance rates at the earlier time intervals in an effort to ascertain whether or not these values would indicate the presence of a two-phase system. The initial rapid phase indicative of the elimination of free or loosely bound fluorochemical may be followed by a slower phase which would be presumptive evidence for the release of tightly bound material. ' It should be noted that in this study uptake and clearance values were only determined in intact organisms rather than utilizing the specific organ technique. It would appear that this latter method should be utilized in an effort to determine whether or not FM 3422 is also rapidly cleared from the more lipophilic organs. One may speculate that bioconcenti-ation,per se, may be due to a multiplicity of cliemical-receptorinteractions. In receptor theory several types of receptor attachments have been identified. For example, the material may bind to the receptor in stable or labile fashion, or the material may be found in the free state. Of importance would be the determination of whether or not binding associated with FM 3422 may be reversible. This latter hypothesis may approximate the true situation.,inasmuch as I-M3422 was rapidly cleared. If clearance were prolonged, this would be indicative of an irreversible binding. This pharmacological tool may well have application in assessing applicable mechanisms and modes of chemical uptake and clearance by aquatic organisms. In these pilot studies, we have demonstrated that Ft434i2 does: 1) Bioconcentrate in lipopliilicorgans achieving levels of fluorochemical approximately one order of magnitude greater than those found in relatively less lipophilic organs. 2) Attain a steady state within 7 days. 3) Clear rapidly, channel catfish >bluegill. In tiicabsence of toxic signs and considering the relatively rapid clearance of the fluorochemical by the test organisms one may assume that this material was nontoxic under the conditions employed in the foregoing experiments. BIBLIOGRAPHY (1) f4endel,A. Personal communication. Report in progress. (2)Technical Report, May 17, 1977, M. T. Elnabarawy to A. N. Welter entitled: "Bioconcentration of FM 3422 in Bluegill Sunfish and in Channel Catfish." (3)Granmo, A. and S. Kollberg: Uptake Pathways and Elimination of a Nonionic Surfactant in Cod (Gadua Morrhea L.1 Water Research 10:189-194, 1976. (4)Bass, Michael L. and Alan G. Heath: Cardiovascular and Respiratory Changes in Rainbow Trout, Salmo gairdneri, Exposed Intermittently to Chlorine, Water Research 11: 497-502, 1977. (5)Bass, Michael L. Charles T. Berry,.Jr., and Allan G. Heath: Histopathological Effects of Intermittent Chlorine Expose on Bluegill (Lepomis macrochirus) and Rainbow trout (SaZmo gaiz-dneri),Water Research 11: 731-735, 1977. (6)Chiou, Cary T., Virgil H. Freed, David W. Schmedding and Rodger L. Kohnert: Partition Coefficients and Bioaccumulation of Selected Organic Chemicals, Environmental Sci. and Tech. 11: 475-478, 1977. LEGEND Figure 1 Abscissa in days, Bluegill Data Uptake indicates exposure period to FM 3422 Clearance indicates days of depuration C value indicates concentration of@fluorochemical w in water indicates absence of intermediate values, hence the line segment is arbitrary. Figure 2 Legend as for Figure 1, Channel Catfish Data. 77-'7 - - - - -- -77, I 'I L 71 1 I --7-.-7T7. . .. . . . . . 7 L-L . . . . . . .I t I.. . . . . . . . . . . .... . .. L- t- 17:Vl t it 7-1 t. A7 t. It t :I t I @l Tit. .-I- L i. f lb4 i@ 4- L-i Zr I-1'4 If r-4 1. -1" 29"Y4E4 I -444+++ T7 ..... ...... ....... -7 'L -7 77 =tat t LL .. .... .. .... .... . 7-1 @-H7!4-:lltEL it @77 ...... .4-+4. -7 7 7---'7 .. .. .... ... "7 7 7- ILI,- 1 -4-t i f 4 rt t t I t7 i 1.4 1 L- I II -FT77 ... . . . . . . . . . . f iI i 17Ti tn ow JIP.' re TABLE 1 FM 3422: BIOCONCENTRATION FACTORS AT VARYING EXPOSURE AND CLEAaP.NCE TIMES: CHANNEL CATFISH BLUEGILLS AND Organism BIOCONCENTRATION 8 14 FACTOR DAYS 21 Bluegillsb 4xlO2 7 Channel catfish c 4xlO2 3xlo2 4xlO2 DA Y S 14 2.102 CLEARANCE 7 2xlO-2 14 4xlO-2 7 2 7xlO - 14 9xlo-3 a Values are rounded to nearest hundredth. b Cw .4 .1 ppm, Mean SD Concentration of FM 3422 in water. c c .6 .2 ppm, Mean SD w Concentrationof FM 3422 in water. TABLE 2 BIOCONCENTRATION FACTOR OF CHANNEL CATFISH ORGANS AT VARYING EXPOSURE PERIODS OF FM 3422 Organ Muscle Viscera (Gut) Gills Skin and Skeleton Skin. Skeleton Oil Layer (Skin) Brain BIOCONCENTRATION FACTOR a 1-Week Exposure 4-Week @Msure c. 2 3xlo 2 3xlo 3 2xlo 2 7xlO 3 3xlo 2 7xlO 3 lxlo 3 1.6xlO 2 3xlO 2 5xlo 2 9xlo 2 SX10 3b 3xlO (1) 3 10 a Specimens obtained from 2 channel catfish Single analysis c Analysis of (1) channel catfish 3M Report:BioaccumulationStudies J.W.Gillett ReporNto.1 (5/17/'7B7i)oconcmiroafFtMi3o4n22inBhiegSiulnlfiasnhdinChanneClalfiasnhd' (3)1. ProgreRsesporNto.2 (101141*7F7a)toefFluorochemicals, A modifi(efdrowmhat?e)xposu(r?e)techniwqauseusetdoestimaBtCeFtobluegawndchanncealtfish (whole body) and particulatrissuesfollowingexposureto a supersaturatesdolution(suspensiono)f FM 3422 (N-Et FOSE alcohol).Experimentaldetailare lackingin many areasforwhich thereare textstatementsand conclusions.Even for itstime thisset of experimentswas only marginallyuseful,at best. Problems include the following: a. 'Me characterand sourceof the FM 3422 isnever stated,particularwliyth regardto purity,similarlyt,he assaytechniqueisundocumented as to precisionand sensitiviftoyr the purposesto which itwas appliecl b. 7le materialtaken up and bioaccumulatedwas never identifieads the nominal toxicanto which the fish were exposed,as faras readersare concerned. Could itbe a metaboliteand/orbound residue? c. Geo. Chapman and Chuck Warren at Oregon StateUniv. had demonstratedthe utilitoyf saturatedglass bead deliveryin a flow-throughsystem in the early1970's.In the course of thoseexperimentstheywere ableto demonstratethepresenceof a verymuch more highlytoxiccomponent of tech.dieldrinwhich was not the nominal compound (HEOD). Ileirmethod was to saturatetheglassbeadswith tech.dieldrinby remoxil of acetonesolventin the flashevaporator,then placethe beads ina largecolumn over glasswool and a @ortionof cleanbeads. Ile water-jacketecdolumn was brought to experimentaltemperatureand portionsof thewater stream assayedfordieldri(nasHEOD) by extractioannd GC4EC A compound with lower EC responsethan HEOD foritstoxicit(yor much more tojdcfor itsEC response)came offthe column duringthe equilibratiopnhase (1-2weeks),but was not detectableby 34 weeks when the HEOD concentratiownas stabilize(ditremained constantforca.10 weeks). Use of theBrungs-Mount diluterwith thissaturatedsolutionproduced consistentresultsifthe diluterapparatuswas equilibratewdith the test solution.The point isthatthe colilmncan generatesuspensionsand solutionsof changingcomposition and toxicitya,s well as concentration."Me staticsystem employed in thesepapers cannot equilibratien the same sense as does a flow-throughsystem. d. 'Me authorsused the 'air-offcuoncentrationass'thatforthe BCF calculationa,lthough theyclearlyhad a problem withvaluesof FM 3422 15-300% (!?)higherwith "air-on.0Ile latterwas probablythe *well aerated*water to which testfishwere cqxrea. This underlinesthe extensiveproblem supersaturation* creates. e. Itislikelythatthe valuesreportedhave littlteo do with bioaccumulationor even uptake per se,at least asexpositedwithinthereports.Whole body residuesmight includeentr,apped material(colloidaplarticles in the gills)t;issueresiduesdepend upon@not only correctdissectionb,ut alsoprotectionfrom surficial contamination. t 'Me number of fishconstitutinagsample,the number of fishper tank,the size(lengthw,eight),ofeach fish,and whether measured fishwere liveor among thedead removed allare alludedto but never detailed by theauthors.Neverthelesst,heyproceed to note thatlargerfishaccumulatemore FM 3422 than smaller fishand thatsome (3)of the controlsdied. Heavierfishmight have a higherpercentageof fatthan younger,leanerfish.Itwould be usefulto know ifthe testsubjectshad about the normal lipidcontenl g. It is not clear why fish were dipped for 2 min, then analyzed. Tte experiment is without much meaning with respect to protection, toxicityanalysis,etc. I know of no database including such a measurement for comparison with othertoxicants. h. The citedvaluefor K,. appearsin error,itmay involventis-measuremenl 3M REPORT: BIOACCUMUIATION STUDEES March 9,1"3 i. Some of the tissueshow no increasein BCF between I and 4 weeks; others(gillasnd gut)increasedramaticary.The data in tissuesdo not support the assumption thata true eqaflibriumisreached between the fishand itsenviromment or within the fishbetween tissues.