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ACUTE TOXICITY TO FISH (FATHEAD MINNOW) TEST SUBSTANCE Identity:Mixturecontainingperfluorooctanesulfonatweh,ich may also be referredto as PFOS, FC-95, or as a component of FC-203CF-X or FC-203CF. (1-Octanesulfoniaccid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 101. The testsample isFC203CF. Currentinformationindicatesitisa mixtureof 1% PFOS, 20% diethyleneglycolmonobutyl ether,70.2% water,2.75% Acrylicfoamer,3% sodium octylsulfate1,% triethanolmaine, 2% sulfonatedalkylethersalts,and 0.05% tolyltdazole. The followingsummary applies to a mixture with incompletely characterized concentrations of impurities.Data may not accurately reflecttoxicityof the fluorochemkal component of the testsample. METHOD: Method: OECD Guideline203. Type: Acute static GLP: No Year completed: 1991 Species: Pimephalespromelas Supplier: AquaticResearch Organisms,DivisionofResource Analysts,Inc., Hampton, NH. Analyticalmonitoring: Temperature, pH, conductivitya,nd dissolved oxygen. Exposure period: 96-hours Statisticamlethods: No calculationpserformed,based on study results. Test fish age: Juveniles Length and weight: Average length 34 mm Average weight 0.23g (wet) Loading: 0.15g/L Pretreatment:None Test conditions: Dilutionwater: Filterendaturawlellwater Dilutionwater chemistry: Hardness: 80 mg/L as CaC03 Conductivity: 830 pmhos/cm pH: 7.7 Lighting: Cool-whitefluorescentlights.A dailyphotoperiod of 16 hours light and 8 hours dark was maintainedthroughoutthetestingperiod. Stock and testsolutionpreparation: The testmaterialwas added directly tothe dilutiownater on a weight/volume basis. Concentratiodnossingrate:Once. StabiliotfythetestchemicaslolutionsN:otnoted. Exposurevessels:20L glasasquari(a40x2Ox2c5m)containi1n5gL oftest solutionand an approximatewater depth of 18 cm. Number of replicates:2 Number of fishper replicate:10. Number of concentrations:fiveplusa blankcontrol Water chemistry during the study: pH range (0-96hours): 7.1- 7.7(controelxposure) 7.5- 7.7(1000mg/L exposure) Temperature range (0-96hours): 21.8- 22.7OC Conductivityrange (0-96hours): 830 - 850 pmhos/cm (controelxposure) 850 - 910 pmhos/cm (1000 mg/L exposure) Dissolved oxygen range (0-96hours): 6.8- 8.1mg/L (controelxposure) 2.5- 8.5mg/L (1000mg/L exposure) RESULTS Nominal concentrations: Blankcontrol,150,250, 400, 600, and 1000 mg/L Elementvalues: 96-hourLC50=>1000mg/L Element valuesbased on nominalconcentrations Remarks: Testingwas conductedon themixtureas describedinthe Test Substance Remarks field.The valuesreportedapplytothatmixtureand not the fiuorochemicaplroportionalone. CONCLUSIONS The testsample 96-hourLC50 forfatheadminnow was determinedto be >1000 mg/L. Submitter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2. Testingmeets thecriterifaorquality testing.However, thesample puritwyas notpropedycharacterizeadnd the study lacksanalyticaclonfirmatioonfthe amount offluorochemicaplroportion inthe solution. REFERENCES Thistestwas conducted by EnviroSystemsDivisionR,esource Analysts, Incorporatedo,f Hampton, NH attherequestofthe 3M Company, St.Paul, MN, Lab Request number J1884-3,1991. OTHER Last changed: 6/28/00 Study Title Static AcutO Toziciti7,of,lt-@203Crto the fatheadNinnowfp$jwpbalosprowlas Autiors Timthi J.@Ward Robert t@ fteri plot junq Per a On t' lo'_eO,rpmrated 1. SORMT The &cut* toxicity of FC-203CT to the fathead MiUnOW, PiAlOphal" proseles, is described in this final report. The test was conducted for 3H Company for 96 hours during June 3 to 7, 1991, at the govirosystans Division of Resource Analysts, inc. in Rampton, Now Hampshire. It was conducted by Jeanne Magazu, Peter Kowalski, Ellen Stanford, Robert boeri, and Timothy Ward The test was performed under static conditions with five concentrations of test substance and a dilution water control at a mean temperature of 22 IOC. The dilution water was filtered natural well water collected at ffampton,Now Rampshire. Aeration was not employed to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of FC-203CY were: 0 mg/L (control), 150 ag/L, 250 mg/L, 400 mg/L, 600 mg/L. and 1,000 ag/L. Nominal concentrations were used for all calculations. Fish used in the test were purchased from a commercial supplier (Aquatic Research Organism, Rampton, Now Sampshire) and acclimated under test conditions for more than IS days. After 96 hours of exposure the control fish had an average total length of 34 m and an average wet weight (blotted dry) of 0.23 g, resulting in a loading rate of approximately 0.15 g/L. Ill fish were in good condition at the beginning of the study. Exposure of fath*ad-mi,nnom.to the test substance resulted in a 96 hour LCSO greater than @l',000'@.-mg/L-rC-203CF. Two of the twenty fish uposed to 600 mg/L at 72 bours.and all of the fish exposed to 1.000 ng/L from 24 to 96 hours'.2were lethargic and exhibited a loss of ,equilibrium. No other imbletbal effects were observed in any test .vessel during the 96 hour exposure. IZ. TULT OF CONTENTS SECTION: P&OZ Summary 2 Table of Contents Index of Tables 4 Introduction @V. Nethods and Naterials vr. Results nl References !7, -Appendix A. Water:,Qua ty Data.frft ToidcitrTest --of Ill. mgx OF TULZS Table 1. Chemical characterizationof a representative sample of natural well water used as dilution water for toxicity test PAGE 6 Table 2. Survival data from toxicity test 9 Table 3. Median I tbal concentrations (bC50s) from 10 toxicitIyletest Table R.I. Conductivity,pl, tomperatur*,and dissolved 13 oxygen concentrationmeasured during toxicity test ..so. a rv. rmoDUCTION This study was sponsored by 3m company, St. Paul, Minnesota. The objective of the studywas to determinethe acute toxicityof FC-203cr to the fathe4d minnow, a freshwater fish. The report contains sections that describe the methods and materials employed in the study, and the results of the investigation. The report also contains an appendix that presents the water quality data collected during the test. TEST SUBSTANCE-. V. METHODS An MATERIALS FC-203CY (EnviroSysteas Smple Number 2937E) was delivered to EnviroSysteas on May 23, 2991. It was contained in a 4 oz plastic bottle that was labelled with the following information: OJIS84-3, FC206CF". FC-203CT (an orange-brova liquid) was supplied by 3H Company, 935 Bush Avenue, St. Paul, Minnesota. Prior to use the test material was stored in the dark at room temperature. A reserve sample (approximately 1.0 grams) will be archived at EnviroSystess for a minimum of 10 years. DILUTION WATZR: Water used for acclimationof test organisms and for all toxicity testing was collected from wells at tavirosystems in Hampton, Now.Hampshire. water was stored in 500-gallon polyethylene tanks, where It was aerated. Results of chemical analysis of a representative sample of,@@@ water are presented in Table 1. TEST ORGANISM: Juvenile fathead minnows employed as' test organisms were fr single source and were identified using an appropriate taxonomic,,ki They were purchased- from a commercial ..supplier (the Aquatic Resiai Organism Division 'ofResource Analysts, Inc , Hampton, Now Hampshire acclimated.at the 'ZnviroSysteasfacility f;r'sore than 15 days. :Contr@ fish were - weighed.@at the conclusion of,.. toxicityt*xt.. Pr ior*to-4 tooting,. fish a were-,maintained in.100% diltitionwater under flow:thi@6igh' T" :.,Condition in" an@'@ill glass aquarius.,@During acclimation fish.wero not4 treated for disease and,they wore fro* of apparent sickness.injuriii;i,i" abnormalities at the beginning of the test. During the acclimation peribd' 14" @days prior- to tbi@test initiation the temperature ranged fran 21-.81114'tii- 23'60C and the dissolved -oxygen 'concentration was, aaintained-.'aio@v ,;7.6 fish were fed a commercial fish,,food-(Znvirotrateaslot@ TNOI) once or twice daily before the test."--. e, 5@ @T Cbesical ebaracter"zation@oi@ 4 r resentativo sagple of natural.wellwater used as dilutionwaterfor toxicitytest Parameter ps Conductivity Vardness Organoeblorine pesticides Orgah6pbospborus asticides Unit of Neasuroment Reporting Linit PR units Unhas/en mg/L as CaCot ug/L 0.5 ug/L 0.S.@. ug/L 6!'5 Measured Value 7.7 830 so ND ND 'j es@ -1"; ND apt detected at'-Ota,bov*L@hi ng.. ssi:@@.and.cooductivi,!,:wrere;aeasured t@,i3atiit'@.vos@sieixo@r@t-o r octioli@-ofj'i6., m - ,and TOXICITY TESTING: A screening test with the test substancewas conducted during May 24 to 23, 1991. The test was conducted at 1, 10, 100, and 1,000 ag/16 and after 96 hours there was 100% survival at all concentrations. The definitive toxicity test was performed during June 3 to 7, 1991. it to based on procedures of the OECD (1984). The test was conducted at a target temperature of 22 t IOC with five concentrations of test substanceand a dilution water control. No stock solution was prepared as test material was added directly to dilution water contained in the test vessels without the use of a solvent. Nominal concentrations of the test material were: 0 mg/L (control),150 ag/L, 250 mg/L, 400 ag/L, 600 mg/L, and 1,000 mg/L. Twenty fish were randomly and equally distributed among two replicates of each treatment. The test was performed in 20 liter glass aquaria (approximately 20 co in width, 40 co in length, and 25 ca in height) that contained 15 liters of test solution (waterdepth was approximately 19 ca). Test vessels were randomly arranged in an incubator during the 96 hour test (a random numbers table was used to select the location of each vessel). A 16 hour light and 9 hour dark photoperiod was automatically maintained with cool-mbite fluorescent lights. Aeration was not employed to maintain dissolved oxygen concentrations above acceptable levels. Irishwere not fed during the test. The number . of surviving organisms and the occurrence of sublethal effects (loss of equilibrium,I erratic swimming, loss of reflex, excitability, discoloration, or, change in behavior) were determined visually and recorded initiallyand after 24, 48, 72, and 96 hours. . Dissolved oxygen (YSI Model 57 setii; instrument number P]tL-3). pa (Backman model pEl 12 aster; instrument number M-51), conductivity (Cole -,,Parmormodel number 1481-60,, instrument nuamr PRL-50)',and tempira tiir4 (ASTN mercury thei@someter,@,t.h.e-r,m.o,meter nuum r 1686) were, measured and recorded daily,'..iinach-test.,chamberthat containod.live aniaals'- STATISTICAL MMODS: y Results of the t@xicit test could not bi 'interpreted.'b standard statistical techniques (Stophan,;@@,1983) because greater tb;;,-,@ 50% survival occurred in all tested concentrations. of ?a I Ti. USMTS All test vessels containing FC-203CY were initially clear and remained clear throughout the test. Biological and water quality data generated by the acute toxicity test are presentedin Table 2 and Appendix h, respectively. one hundred percent survivaloccurred in the control exposure. Control fish had an average total length of 34 M, and an average met weight (blotted dry) of 0.23 9 at the end of the test. Loading rate during the toxicity test was approximately 0.15 9/10. The 24, 48, 72, and 96 hour LCSOs for fish exposed to rC-203CY are presented In Table 3. The 96 hour LCSO value is greater than 1,000 mg/L FC-203CY. Two of the twenty fish exposed to 600 mg/L at 72 bourn and all of the fish exposed to 1,000 mg/L from 24 to 96 hours were lethargic and exhibited a loss of equilibrium. No other sublethal effects were observed in any text vessel during the 96 hour exposure. Tabl* 2. Survival data froe toxicity@t*st Nominal Concentration rep. Number Alive Obr 24hr 4$hr 72hr 96br Number iffected Ohr 24hr 48br 72hr 96br 0 (control) 1 2 iso 1 2 250 1 2 400 1 2 600 1 2 11000 1 2 10 10 10 10 10 10 10 10 to 10. 10 10 10 10 10 10 10 10 10 10 10 20 10 to 20 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 to 10 10 9 9 9 10 10 9 9 9 00 0 0 0 0 00 0 0 0 0 00 0 00000 00000 0 00 0 0 0 00 0 0 00000 00010 00 0 1 0 0 10 9 9 9 0 10 9 9 9 Note:'-.Affected fish were lethargic and,w&ibited a loss of equilibrium. 'at !FT Table".3. N*dian lethal:"" tr ons (,LCSOSf)rom toxicitytest ltxposure period LCSO 24 hours )1,000 mg/L .49 hours >1,000 mg/L,. 95 percent confidence limit 72 hours )1,000 ag/L 96 hours >1,000 mg/L loc5o calculation notbod. viz. Rznmcllgs OECD. l9S4. Guidelinesfor Testing Of Cbmic&ls- Section2: EffectsOn Biotic systems. Notbod 203,-FishAcute ToxicityTest. Adopted April 4, 1994. its OMNI 'I lippenix A. VATN O@ATM DATA FROM TOX-.CITY TICST paa"* Table I.I.Conductivity,pN, tompolature, and dissolved oxygen concentrationmeasuredduring toxicity test Nominal concentration (mg/L) Rep. conductivity(usho/cm) 0 24 48 72 96 br br hr br br PH 0 24 48 72 96 br br br br hr 0 (control) 1 2 150 1 2 250 1 2 400 1 2 600 1 2 1,000 1 2 830 830 $40 $40 @850 830 $40 $40 850 850 $40 $40 850 850 860 840 840 850 850 860 840 840 850 850 $70 840 $40 $50 OSO $60 840 840 850 $60 860 $40 $40 850 850 860 $40 840 850 860 860 $40 $40 850 860 870 860 850 900 860 870 880 $50 900 910, 910 7.7 7.3 7.1 7.1 7.4 7.7 7.4 7.1 7.1 7.4 7.7 7.5 7.3 7.2 7.4 7.7 7.5 7.3 7.2 7.4 7.7 7.5 7.3 7.3 7.5 7.7 7.5 7.3 7.3 7.6 7.7 7.5 7.3 7.4 7.6 7.7 7.5 7.4 7.4 7.5 7.7 7.6 7.4 7.5 7.5 7.7 7.6 7.5 7.5 7.6 7.7 7.7 7.7 7.S 7.6 7.7 7.7 7.7 7.5 7.5 li, 4@! i@i .2 Table A.I. Continued. Nominal concentration (mg/L) Rep. Temperature (OC) 0 24 48 12 96 hr br br hr hr 0 (control) 1 2 ISO 1 2 250 1 2 400 1 2 600 1 2 1,000 1 2 22.2 22.2 22.1 22.2 22.4 21.8 22.0 22.0 22.1 22.4 22.4 22.0 21.9 22.0 22.4 22.4 22.1 22.1 22.1 22.5 22.0 22.7 21.8 22.2 22.3 22.4 22.2 22.1 22.4 22.5 22.3 22.0 22.0 22.1 22.5 22.3 22.1. 22.2 22.3 22.5 22.0 22.0 22.1.22.2 22.5 21.9 22.2.22.3 22.4 22.6 22.1 21.9@'21.9 22.0 22.4 22.3 22.4 22@3 22.5 22.7 Dissolved Oxygen (mg/L) 0 24 48 72 96 br br br br br 8.1 3.1 7.2 7.0 6.8 8.1 8.0 7.3 7.2 6.8 8.4 8.0 7.0 6.8 6.3 8.4 9.0 7.0 6.8 5.2 8.7 8.0 7.0 6.8 6.2 8.7 8.0 7.0 6.8 6.2 8.5 8.0 7.0 6.4 5.6 9.5 8.0 7.0 6.5 5.6 8.5 8.0 7.0 6.2 4.4 9.5 8.0 7.0 6.3 5.5 8.5 8.0 7.0 6.0 2.5, 8.5 8.0 7.0 6.2 2.6 'All 41i@ age,