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ACUTE TOXICITY TO AQUATIC INVERTEBRATES (DAPHNIA MAGNA) TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredtoas PFOS, FC-95, or as a component ofFC-206CE. (IOctanesulfonicacid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was not given.The testsample isFC-206CE (5/23/91).It'psu(itywas not sufficientclhyaracterizedt,hough currentinformationindicatesitisa mixtureof 0.92% PFOS, 15% diethylene glycolbutylether,76.09% water,4% urea,1.28% Sultonefoamer,1.55% sodium octylsulfate,1.05% polyoxyethylenemonooctylphenylether,0.06% sodium laurylsulfatea,nd 0.05% tolytriazole. The followingsummary applies to a mixture with incompletely charecteilzedconcentrations of impuilties.Data may not accurately reflectthe toxicityof the nuoruchemical component of the testsample. METHOD: Method: OECD Method 202 Type: Acute static GLP: No Year completed: 1991 Species: Daphnia magna Supplier:In-housecultures. Analyticalmonitoring: Temperature,pH, conducbvitya,nd DO Exposure period:48-hours Test organism age: <24 hoursold Statisticamlethod: Not applicableS.urvivalwas greaterthan 50% atall testconcentrabons. Test conditions: Dilutionwater: Filterednaturalwellwater Dilutionwater chemistry: Conductivity: 860 pmhos/cm pH: 7.6 Lighting: A 16 hour lightand 8 hour dark photoperiodwas automatically maintainedwithcool-whitefluorescentlighttshatprovideda lightintensitoyf 20 uEs-1M-2. Stock and testsolutionpreparation:A 1000 mg/L primarystocksolution was prepared indilutiownater. Aliquotswere then added directltyo dilution water intestvesselsto createtestsolutions. Exposure vessels: 250 mL Pyrex glass beakers containing200 mL oftest soluton and a water depth of-6 cm. Number of replicates:4 Number of daphnids per replicate:5 Number of concentrations: Fiveplusa blankcontrol Water chemistryduringthestudy: Conductivitryange(0-48hours): 860- 880 pmhos/cm(contreoxlposure) 860 - 880 pmhos/cm (1000 mg/L exposure) pH range (0-48hours): 7.6 - 8.3 (controelxposure) 7.7- 8.3(1000 mg/L exposure) Temperature range (0-48hours): 20 - 21 OC 19.0- 20.2 OC (controelxposure) 19.0- 20.1 -C (1000 mg/L exposure) Dissolved oxygen range (0-48hours): 8.5 - 9.2 mg/L (controelxposure) 8.7- 9.3 mg/L (1000 mg/L exposure) RESULTS Nominal concentrations:Bk control1,50,250,400, 600, and 1000 mg/L Element values: 48-hour EC50 = >1,000 mg/L Element values based on nominalconcentrations Remarks: Tesbng was conducted on a mixtureas describedinthe Test Substance Remarks field.The values reportedapply tothatmixtureand not the fluorochemicalproporfionalone. CONCLUSIONS The FC-206CE 48-hour EC50 was determined tobe >1000 mg/L, the highest concentrationtested.No mortalitoyr sublethaleffectswere observed atthis concentration. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY Reliability:Klimischranking2. Testingmeets the criterifaorqualitytesting. However, sample puritywas not properlycharacterizedand the study lacks analyticaclonfirmationofthe amount offluorochemicalproporton inthe solution. REFERENCES Test was conducted by EnviroSystems Divisionor Resource Analyst, IncorporatedH,ampton, NH atthe requestofthe 3M Company, St.Paul,MN, Lab Request number J1884-2, 1991. OTHER Last changed: 6/26/00 Study Title Static Acute Toxicity of I-C-206CE to the Dapbnid, Daphnja gaiyna Authors Timothy J. Vard Robert L. Boeri Study completed June, 199l.. Performina Laborit6' ftvirosystens'Divisi'o:n@'l. escurce Analystg;-.ICiOrporated On o Lafayett'eload i@.@ Upton, @4,54t ..t, r. SURMT The acute toxicity of FC-206CE to the daphnid, Dapbais Aragna, is described in this final report. The test was conducted for 3H Company for 48 hours during Juno 10 to 12, 1991, at the EnviroSysteas Division of Resource Analyst*, Inc. in Hampton, New Hampshire. it was conducted by Jeanne Nagazu, Peter Kowalski, Ellen Stanford, Robert goeri, and Timothy Ward. The test was performed under static conditions with five concentrations of test substance and a dilutionwater control at a temperature of 20 IOC. The dilution water was filtered natural well water collected at Hampton, New Hampshire. Aeration was not employed to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of rc-206CE were: 0 mg/L (control),150 mg/L, 250 no/L, 400 mg/L, 600 mg/L, and 1,000 mg/L. Nominal concentrations were used for all calculations. Daphnids used in the test were produced from an in-house culture and were less than 24 hours old at the start of the test. After 48 hours of exposure the control dapbnids bad an average wet weight (blotted dry) of 0.0006 g, resultingin a loading rate of 0.015 g/L. All dapbuids were if.good condition at the beginning of the study. The test substance was not toxic to daphnids at tested concentrations. Exposure of daphnids to the test substlnce resulted in a 48 hour EC50 greater than 1,000 ag/L FC-206CE. 'Page 2 of 14 Resource 11. TileLl0jrcorruts SECTION: r. Summary Table of Contents rii. Index of Tables IV. Introduction V. Methods and Materials vr. Results vir. References Appendix A. gaterQualityData from Toxicity Test PACR 2 3 4 5 5 8 12 ps moot, tr_ MMLLS iii.rnixOFTABLIS AND ricnzs Table 1. Chemical characterization of a representative sample of natural well water used as dilution water for toxicity test Table 2. Survivaldata from toxicity test Table 3. Median effective concentrations (EC50s) from toxicity test Table A.l. Conductivity, ps, temperature, and dissolved oxygen concentration measured during toxicity test PAGE 6 9 10 13 of 14 &"Cfty d .. rT. RMODUCTION This study was sponsored by 3H Company, St. Paul, Minnesota. The objective of the study was to determine the acute toxicity of FC-206CI to the dapbuid, a freshwater invertebrate. 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. V. METHODS AND MATERIALS TEST SUBSTANCE: FC-206CE (EnviroSystess Sample Number 2936E) was delivered to EnviroSystess on May 23, 1991. It was contained in a 4 oz. plastic bottle that was labelled with the following information:-JlBS4-2, FC206CZ-. FC-206CE (a yellow 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 (approximately1.0 gram) will be archived at EnviroSystemi for a minimum of 10 years. DILUTION WATER: Water used for acclimation of test organismsand for all toxicity testing was collected from wells at EnviroSysteas in Hampton, New Hampshire. Water was stored in 500-gallon polyethylene tanks where it was aerated. Results of cbesical analysis of a representativesample of water are presented in Table 1. TEST ORGAXISM: Juvenile dapbuids employed as test organisms were from a single source and were identified using an appropriate taxonomic key. Daphnids used in the test were I psrtoadrutcoefd ftrhoemtaensti.n-Choonutsreocludlatpuhreniadnsdwweerreeweliegsshetdhaatn 2t4he hcouornscluoslidonaotfthtehe. toxicity test. Prior to testing daphnids were maintained -in 100% dilution water under static conditions inI 4 liter glass jars. During acclimation dapbnids were not treated for disease and they vere free of apparent sickness, injuries, and abnormalities at the -,K. beginning of the test During the acclimation period 24 hours prior to,. the test initiationth; temperature was 19.5*C. Daphnids were fed yeast, trout cbow, and/or the freshwater alga. Solonastruncapricornutusonce. or twice daily before the test. page, S"of 14. Rowu= MotyftW- &bO*q d, '@m Table 1. Ch-@nicalcharacterization of a representative sample of natural well water used as dilution water for toxicity test Parameter PH Conductivity organocblorine pesticides organophospborus pesticides Polychlorinated bipbonyls Unit of Measurement PH units ushos/cm ug/L Reporting Limit -0.5 ug/L 0.5 ug/L 0.5 Measured Value 7.6 860 ND ND ND Notes: 1. None detected at or.above the reporting limit. 2. pb an& conductivity were measured in dilution water collected from a control test vessel prior to introduction of organisms and pesticide and PCB data is collected during routine biannual water quality testing. IF' j(* Fag* 6 of pig_ TOXRCITY TESTING: 1991. The definitive toxicity toot was performed during June 10 to 12, It is based on procedures of the OECD (1984). The test was conducted at a target temperature of 20 t IOC with five concentrations of test substance and a dilution water control. solution was formulated without the use of a sAol1v,e0n0t0.mg/L stock concentrations of the test material war*: 0 mg/L (control),15N0ommign/abl, 250 so/L, 400 mg/L, 600 mg/L, and 1,000 mg/L. Twenty daphnids were randomly and equally distributed among four replicates of each treatment. The test was performed in 250 al glass beakers that contained 200 al of test solution (water depth was approximately 6 co). Test vessels re randomly arranged in an incubator during the 48 hour test (a r:lodonnumbers table was used to select the location of each vessel). A 16 hour light and 8 hour dark pbotoperiod was automatically maintained with cool-white fluorescent lights that provided a light intensity of 20 uE3-lo-2. Aeration was not required to maintain dissolved oxygen concentrationsabove acceptable levels. Dapbnids were not fed during the test. The number immobilization of surviving organisms and the occurrence of (inability to swim within 15 seconds after gentle agitation) or other sublethal effects swimming, lose of reflex, (loss of equilibrium,erratic excitability, discoloration,or change in behavior) were determined visually and recorded initially and after 24 and 48 hours. Dissolved oxygen (Ysr model 57 meter; instrument number cPoRnLd-u3c)t,ivpiHty (Beckman model (Labcoup pHl 12 meter; instrument number PRL-51), SCT meter, instrument number PRL-50), and temperature (ASTH mercury thermometer; thermometer number 2265) were measured and recorded daily In each test chamber that contained live animals. STATISTICAL )MTBODS: Results of the 'i@xicity test could not be interpreted by standard statistical techniques (Stephan, 1983) because 50% survival or greater occurred in all tested concentrations. Page 7 of 14 MKLFM Vi. RESULTS All test vessels containing FC-206CZ 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 A. respectively. One hundred percent survival occurred in the control exposure. Control daphnids had an average wet weight (blotted dry) of 0.0006 9 at the end of the test. Loadingrate during the toxicity test was approximately 0.015 g/L. The 24 and 48 hour EC50s for dapbnids exposed to FC-206CE are presented in Table 3. The 48 hour EC50 was greater than 1,000 mg/L FC-206CE. @e . ... iesve 3 of @24 Table 2. Survivaldata from toxicity test Nominal Concentration (mg/L) rep. 0 (control) 1 2 3 4 150 1 2 3 4 250 1 2 3 4 400 1 2 3 4 600 1 2 3 4 1,000 1 2 3 Number Alive 0 br 24 hr 48 hr 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 s 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 s 5 5 5 5 5 5 5 5 5 5 5,@: 5 Number Affected Ohr 24hr 48hr 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0Ol., 0 0 0 0 0 o .0 page 9.0t@14 Table 3. Nedien eff*ctivo concentrations (ICSOB) from toxicity test Exposure period 95 percent EC50 confidence limits 24 hours )1,000mg/L 48 hours )1,000 mg/L Calculation method 4 1., 0' 4;1 &"dkzy Vii. llrimczs OZCD. 2984. Guidelines for Testing of Chemicals. Section 2: Effects on Biotic Systems. Method 202, Daphnid sp., Acute Immobilization Test and Reproduction Test. Adopted April 4, 1984. pa4i' 11 of 14 Ilk, CIMRLFM Appendix A. VATWR OVALITY DATA nON TOXICITY TEST AA Table A.I. Conductivity, ps, temperature, and dissolved oxygen concentration meature6 during toxicity toot Conductivity(usho/cm) PH Nominal concentration 0 24 48 (mg/L) Rep. hr hr br 0 24 48 hr br hr 0 (control) 1 860 870 880 2 860 $70 Sao 3 860 870 880 4 $60 870 ago 7.6 7.7 9.3 7.6 7.7 8.3 7.6 7.7 8.3 7.6 7.7 9.3 150 1 $60 $60 900 7.5 7.7 8.3 2 860 860 900 3 860 860 900 4 860 860 900 7.5 7.7 9.3 7.5 7.7 8.3 7.5 7.7 8.3 250 1 870 870 870 7.6 7.7 8.3 2 870 870 870 7.6 7.7 8.3 3 870 870 870 4 $70 870 870 7.6 7.7 8.3 7.6 7.7 8.3 400 1 870 $70 860 7.6 7.7 8.3 2 870 870 860 7.6 7.7 8.3 3 870 870 860 7.6 7.7 8.3 4 $70 870 860 7.6 7.7 88 .3 600 1 ago 870 860 - 7.6 7.7 .3 2 880 $70 860 7.6 7.7 8.3 3 880 870 860 7.6 7.7 83 4 880 870@':; 8"" 7.6 7.7 8:3 1,000 1 ago a6 Oi 7.7 7.7 8.3 2 .890 $70 860 7.7 7.7 S.3 3 880 $70 8 60 7 7. 77 8.3 4 $80 -870:' 860' 7*#7 @7:7 .3 Page 13, Table A.I. Continued. Nominal concentration (mg/L) Rep. Temperature (OC) 0 24 48 hr hr br 0 (control) 1 2 3 4 20.0 20.2 20.1 20.2 150 1 19.8 2 19.7 3 19.9 4 19.9 250 1 20.0 2 20.0 3 20.1 4 20.1 400 1 20.2 2 19.9 3 20.1 4 20.1 600 J-@ 20.2 2 7, 20.3@ 3, 19.8 4 10000 20.0 2i 20.1; 3 20.0' 4,@ 19.0 19.0 19.1 19.1 19.2 19.3 19.1 19.2 19.2 19.3 19.1 19.1 19.2 19.1 19.1 19.0 19.1 19.0 19.2 19.1 19.2 '--19.1 19 19 2 19.1 19.3 19.4 19.3 19.3 19.3 19.2 19.3 19.4 19.2 19.2 19.3 19.2 19.1 19.4 19.3 19.4 19.5 19.3 .19.2 19-i 19.4 19.3- Dissolved oxygen (mg/L) 0 24 48 hr hr hr 8.5 8.5 8.5 8.5 8.6 8.6 8.6 8.6 8.7 8.7 8.6 9.6 8.6 8.6 8.7 8.7 8.7 8.7 8.7 8.8 8.7 8.7 8.7 3.7 9.0 9.0 9.0 9.0 9.o 9.1 9.0 9.2 9.2 9.2 9.2 9.2 9.2 9.2 9.3 9.3 9.2 9.4 9.4 9.3 9.2 9.3 9.3 9.2 9.2 9.2 9.2 9.2 8.7 9.7 8.6 8.7 8.8 8.8 8.9 8.9 9.0 9.1 9.0 9.1 9.3 9.4 9.2 9.2 9.1 9.0: 9.0,7;@:",; 9.1