Document QXm5Ledq9dwJrL5pxgeVDNVj6

AR S S 6 - 07&9 ACUTE TOXICITY TO AQUATIC INVERTEBRATES (DAPHNIA MAGNA) TEST SUBSTANCE___________________________________________ Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-203. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was not noted. The test sample is FC-203. Current information indicates it is a mixture of 1.34% PFOS, 35% diethylene glycol butyl ether, 37.85% water, 20% ethylene glycol, 2.66 % Sultone foamer, 3% sodium octyl sulfate, 0.1% sodium lauryl sulfate, and 0.05% tolyltriazole. The following summary applies to a mixture with incompletely characterized concentrations of impurities. Data may not accurately reflect toxicity of the fluorochemical component of the test sample. METHOD:____________________________________________________ Method: OECD Guideline 202 Type: Acute static GLP: No Year completed: 1991 Species: Daphnia magna Supplier: In-house cultures maintained at EnviroSystems Division, Hampton, NH. Analytical monitoring: Temperature, pH, conductivity, and dissolved oxygen. Exposure period: 48-hours Test organism age: <24 hrs. old Statistical method: LC5o values calculated using a computer program from Stephan, 1983. Test conditions: Dilution water: Filtered natural well water. Dilution water chemistry: Conductivity: 700 pmhos/cm pH: 7.9 Lighting: Cool-white fluorescent lights at 12 pEin/sec/m2. A daily photoperiod of 16 hours light and 8 hours dark was maintained throughout the testing period. Stock and test solution preparation: A 1000 mg/L primary stock solution was prepared by dissolving 2.0 g of test substance in 2000 mL of dilution water. Exposure concentrations were prepared by addition of the appropriate volume of stock solution. Exposure vessels: 250 mL beakers with 200 mL exposure solution and a water depth of approximately 6 cm. 000664 Number of replicates: 4 Number of daphnids per replicate: 5 Number of concentrations: five plus a blank control Water chemistry during the study: pH range (0-48 hours): 7.9 - 8.3 (control exposure) 8.0 - 8.3 (1000 mg/L exposure) Temperature range (0-48 hours): 19.7 - 20.6 C (control exposure) 19.4 - 20.6 C (1000 mg/L exposure) Conductivity range: (0-48 hours) 640 - 700 pmhos/cm (control exposure) 640 - 660 pmhos/cm (1000 mg/L exposure) Dissolved oxygen range (0-48 hours): 8.3 - 8.8 mg/L (control exposure) _ 8.0 - 9.9 mg/L (1000 mg/L exposure) RESULTS___________________________________________________ Nominal concentrations: Blank control, 150, 250, 400, 600, and 1000 mg/L. Element values: 48-hour EC50 = >1000 mg/L Element values based on nominal concentrations Remarks: Testing was conducted on the mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone. CONCLUSIONS ____________________________________________ The test substance 48-hour EC50was determined to be >1000 mg/L, the highest concentration tested. No mortality or sublethal effects were observed. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY_______________________________________________ Reliability: Klimisch ranking = 2. This study meets the criteria for quality testing. However, the sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in the solution. REFERENCES_______________________________________________ 00066 This test was conducted by Envirosystems Division, Resource Analysts, Inc., of Hampton, NH at the request of the 3M Company, Lab Request number H2864-9, 1991. OTHER______________________________________________________ Last changed: 6/27/00 000666 Amended 8/9/91 BEST COPY AVAILABLE Stndy Title Static Acute Toxicity of FC 203 to the Daphnid, Daphnia magna Authors Timothy J. Yard Robert L. Boeri BEST COPY AVAILABLE I. GOOD LRBOK1TOKT PRACTICE S T A T U R This study was conducted according to OECD Good Laboratory Practice Regulations. Neither the Study Director nor the sponsor are aware of any circuastances that would affect the integrity of this study. l/Hfrt ____ and Study Director Robert L. Coauthor 7 -$><*1 000668 t BEST COPY AVAILABLE n. gmiorr ismucx n m n n Submitted by: EnviroSystems Division Resource Analysts, Incorporated One Lafayette toad Hampton, Hew Hanpsbire 03842 Certification: Data presented in this report were derived by nethods and with naterials identified in the section of the report entitled "Methods and Materials." The test was performed in accordance with EnviroSystems Protocol 90174-3 and the Product Registration Agnatic Toxicology Laboratory Standard Operating Procedures Manual. The toxicity test was performed by Ellen Stanford, Jeanne Magazu, Peter Kowalski, Robert Boeri, and Timothy Hard. The original raw data and final report will be archived at Resource Analysts, Inc. for at .least 10 years. All data transcribed from the raw data to the report were checked for accuracy, and all data were verified by Quality Assurance Auditors. Quality assurance audits were performed on Reported to Study Director Reported to Management 11/14/90 , , 11/15/90.' m i fcrttt " , 1/10/91 1/10/91; . 2/ 7/91 : ,2/ 7/91 : ; ' 7/30/91 % s 7/30/91 arti* $ 7/30/92 7/30/91 . 7/30/91 - 4: 000669 BEST COPY AVAILABLE III. TABLE OF CORTOTS SECTION: I. Good Laboratory Practice Statement II. Quality Assurance Statement III. Table of Contents Index of Tables V. Summary . VI. Introduction^ r ::s VII. 'Methods;and Materials - Page 2 3 4 5 6 7 7 > H 000670 1 BEST COPY AVAILABLt IT. HSSZ OF T1BLKS Table 1. Table 2. Chemical characterization o a representative saaple of natural veil water used as dilution water for toxicity test Survival data fro toxicity test Table 3. Median effective concentrations (EC50s) fro toxicity test Table A.l. Conductivity, pH, teeperatore, and dissolved oxygen concentration neasured during toxicity test . Page 8 12 13 16 000671 Amended 8/9/91 BEST COPY AVAILABLE Y. SUMMARY The acute toxicity of FC 203 to the daphnid, Daphnia magma, is described in this final report. The test was conducted for 3M Company for 48 hours during March 19 to 21, 1991, at the EnviroSystems Division of Resource Analysts, Inc. in Hampton, New Hampshire. It was conducted by Ellen Stanford, Jeanne Magazu, Peter Kowalski, Robert Boeri, and Timothy Hard according to the protocol developed for EnviroSystems Study Number 90174-3. The test was performed under static conditions with five concentrations of test substance and a dilution water control at a temperature of 20 1C. The dilution water was filtered natural well water collected from wells at Hampton, New Hampshire. Aeration was not employed to maintain dissolved oxygen concentrations above an acceptable level. Nominal concentrations of test substance were: 0 mg/L (control), 150 mg/L, 250 mg/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 daphnids had an average wet weight (blotted dry) of 0.0017 g, resulting in a loading rate of 0.004 g/L. All daphnids were in good condition at the beginning of the study. Exposure of daphnids to the test substance resulted in a 48 hour median effective concentration (EC50) greater than 1,000 mg/L FC 203, the highest tested concentration. 000672 Amended 8/9/91 BEST COPY AVAILABLE vi. introduction This report was amended to include data from the final definitive toxicity test that was conducted during March 19 to 21, 1991. The study was sponsored by 3M Company, St. Paul, Minnesota. The objective of the study was to determine the acute toxicity of FC 203 to the daphnid, 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. VII. METHODS AMD MATERIALS TEST SUBSTANCE: FC 203 (EnviroSystems Sample Humber 2699E) was delivered to EnviroSystems on November 15, 1990. It was contained in a 250 ml glass bottle that was labelled with the following information: "H 2864-9, FC 203**. The test substance was supplied by 3M Company, St. Paul, Minnesota. Prior to use it was stored in the dark at room temperature. A reserve sample will be archived at EnviroSystems for a minimum of 10 years. DILUTION WATER: . ,, ; ; Water used for acclimation of test organisms and for all toxicity testing vwas collected from wells at EnviroSystems in Hampton, Mew 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: M-*1 , S Ut. V 1;^:" JiAif Juvenile'daphnids employed as test organisms were from a single source and; |';were identified using an appropriate^taxonomic key. Daphnids used in the test ' T- produced from an in-house culture,and were less than 24;hours old at. the of the test.' Control daphnids!.were weighed'at the,conclusion 'of"the toxicity test. Prior to testing/ daphnids were maintained in 100% dilution water* conditions in 4 liter/glass^axsVf;During acclimationdaphnids were to not treated for disease and they werefree'.of apparent sickness,Unjuries,'and, abnomalitiesat -the beginning of^'the'Atestt$?During the acclimation period 24;':t hours prior,to the test initiation the*temperature was 19.6C'.!|Daphnid cultures '; were'fed yeast/trout chow and/or-the'freshwater alga Selenastrumicapricornutum .... . .... . turner?w rri **5?2. .............. ,, . ItoviroSystems Study Humber ,90X74- t. th.t K*'9 > 4&**v*, emr.aKf '< VA*-* * m- W M - ; BesoacAnatyAlnc.SubsldtavofMujc Csi' i f i t V-&& . < 000673 Amended 8/9/91 BEST COPY AVAILABLE Table 1: Chemical characterization of a representative sample of natural veil water used as dilution water for toxicity test Parameter Unit of Heasurement Reporting Limit Measured Value pH pH units -- 7.9 Conductivity umhos/cm -- 700 Organochlorine pesticides ug/L 2 HD Organophosphorus pesticides ug/L 5.0 HD Polychlorinated biphenyls *-/ , ....._ 'Ms`-' ug/L 0.6 .if-\ - HD Notes:.1. HD Hot detected above the reporting limit. and conductivity values were detemined in a sample of dilution water collected from a control test vessel at the beginning of .the toxicity test.', Additional chemical parameters were measured in^dilution mater during October, 1990 as part of routine biannual y ^ter^quUi^vanalysi^v^lJ^i:5^-| * j .TSr ! .. / v t r ;1 S ... v 1 >'**1\ \ 4. $4? ,, J r, V ~ %, 5 *V s , V-v f W EnviroSystems'Study Humber 90174-3 ` iySHK'i 4 MtifStK< .Wg,,f-'if'?'/ 9$$' AX. :'`fii S'- ifieoautCB Analysis,Inc, Subskflay at MUJPORE - - Pf. ;/,,y ;-s^ 000674 Amended 8/9/91 BEST COPY AVAILABLE TOXICITY TESTING: A screening test with the test substance was conducted during December 5 to 7, 1990. Nominal concentrations of test substance were: 0.1, 1.0, 10, 100, and 1,000 mg/L. After 48 hours of exposure at least 70% survival occurred at all tested concentrations. A definitive test was attempted during January 9 to 11, 1991. Nominal concentrations of test substance were: 150, 250, 400, 600, and 1,000 mg/L. After 48 hours of exposure sublethal effects were noted at all tested concentrations (sublethal effects were not seen at 1.000 mg/L during the screening test) and the test was repeated. The definitive toxicity test was performed during Harch 19 to 21, 1991, according to EnviroSystems Test Protocol 90174-3 (Static, Acute Toxicity Test With FC 203 And The Daphnid, Panhnia waqna). which was signed by the Study Director on November 16, 1990. It is based on procedures of the OECD (1984). The test was conducted at a target temperature of 20 1C with five concentrations of test substance and a dilution water control. A 1,000 mg/L stock solution was prepared by combining 2.0 g of test substance with dilution water and adjusting the final concentration to 2,000 ml without the use of a solvent. Nominal concentrations of the test material were: 0 mg/L (control), 150 mg/b, 250 ng/b. 400 mg/b, 00 ng/b. and 1,000 mg/L FC 203. Twenty daphnids were randomly and equally distributed among four replicates of each treatment. The test was performed in 250 ml glass beakers that contained 200 nl of test solution (water depth was approximately 6 cm). Test vessels were randomly arranged in an incubator during the 48 hour test (a random numbers table was used to select the location of each vessel). A 16 hour light and 8 hour dark photoperiodvas automatically maintained with cool-white fluorescent lights that provided a light intensity of 12 uEs'V*. Aeration was not required to maintain `X;. dissolved oxygen concentrations above acceptable levels. Daphnids vere not fed during.the^test. t 1#^" wESW'?j''-"A** : v-;` t , f-Ar" The number of surviving organisms and the,occurrence ofnblethal effects end immobilization (inability to swim within 15 seconds after gentle"agitation) orrother^sublethal effects^(loss of equilibrium.^erratic swimming,; loss of- reflexi'Xexcitability,;,discoloration',;*or ,change in*'behavior);were^determinedX ^^Xvisually and recorded initially)andafter24 and 48 hours.1- Dead test'organisms/ l^^were^removed)-:when first)observed^ Dissolved oxygen (TSIHodel:57 meter; minstrument ,nuid>eri;PRb-3), pH (Beckman model pHI .12 meter; instrument number)1 fPRL-4) conductivity,;(Labcomp)jSCT||meter,:'instrument -nunberj^?RIr-9)vShnd]) .temperature (ASTMmercury thermometer; thermometer number 2265)werymeasured and .recorded daily in each test chamber'that contained live,animals mm s.isv* -<;*4??i r, Reiouce A n & fA .V. SubSdksyoMUFORE - '*;;&-vl ,, 000675 BEST COPY AVAILABLE ended 8/9/91 STATISTICAL METHODS: Results of the toxicity test could not be interpreted ly standard statistical techniques (Stephan, 1983) because less than 50% of the daphnids exposed to the highest concentration were killed or immobilized. nominal concentrations of test substance and the number of daphnids killed or immobilized vere used to determine the EC50 value. 000676 Amended 8/9/91 BEST COPY AVAILABLE Till. RESULTS All test vessels containing FC 203 appeared clear initially and regained free of insoluble material throughout the test. Biological and water quality data generated by the acute toxicity test are presented in 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 0.0017 g at the end of the test. Loading rate during the toxicity test was approximately 0.004 g/L. Exposure of daphnids to the reference toxicant sodium dodecyl sulfate resulted in a 48 hour median lethal concentration (LC50) of 7.9 mg/L. The 24 and 48 hour EC50s for daphnids exposed to FC 203 are presented in Table 3. The 48 hour EC50 was greater than 1.000 mg/L FC 203, the highest tested concentration. 000677 Amended 8/9/91 Table 2. Survival data from toxicity test BEST COPY AVAILABLE Nominal Concentration (mg/L) rep. Number Alive 0 hr 24 hr 48 hr Number Affected Ohr 24hr 48hr 0 (control) 1 2 3 4 555 555 555 555 0 00 000 0 00 0 00 150 1 5 5 4 0 0 0 2 5 5 5 000 3 55 4 000 4 5 5 5 000 250 1 554 000 2 554 000 3 55 5 0 00 4 5 5 4 000 400 1 5 4 3 000 - -- ,. 2 ..- 5 5 * 4 ... 0 0 0 3 .; 5 --;, . - . 4 ;; . 5 ..4 -1 ' 0 5 -S 0 0 0 0 600 1 y 5 :5 5 000 '' r^v!vtc':-- 1 .V < VfP f; ' ; 5 ' .5 - '''%2Af:. ; 42?-->\.t;!''.-... 0 0 5a 0 0 <0, 0 .'0 0 .i iv. : ? :> - * ^ rvJdfcxi ^-.ooqjfe^.;^. i j y-, 5 t:-'V > ^ : v*5 ? -S 0 0 * -i' &*V-$*. 5t S r i s m ^ * 0 $ 0 o v-vv (*^KPiHPlNMp^r:J&sa u&<~I-.VS'k''.5 0 4`~ o T U., ' 4 ?\ ''o ; .* ,0V:-; o;;,v, .o ''<-'> '-If*. w . . ::- rwSLr f*.KV" *-'9x&1%$ ,r!, t'?-5' A jtfcvr.^- V,.&t^- ^ f_ r r f< .>"5f#*ct.*,>,lNrl.'v'XVs|its,&^''^'fs *r , 4 ?->*.' 'jcct .vwr.- -EnviroSystems;Study"Number;90174-3 vPafi .. ...... ^taaf--*rt i'YW &%?:>4M . M \ - ,"** -5 ;.''a -.* ' - Peexjce A re***.Inc, Subsktay oMUJPORE &' '" '_V, :.> ' f> lf ^ % 'r' tF- 000678 BEST C O P Y A V A IU W Amended l/J/91 Table 3. Median effective concentrations (EC50s) from toxicity test Exposure period 95 percent Calculation EC50 confidence limits method 24 hours 48 hours >1,000 mq/L >1,000 mg/L -- -- -- -- 000679 BEST COPY AVAILABLE IX. KSmXRCES OECD. 1981. Decision of the Council Concerning the Mutual Acceptance of Data in the Assessaent of Chemicals. Annex 2. OECD Principles of Good Laboratory Practice. C (81) 30 (Final). OECD. 1984. 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. Stephan, C.E. 1983. Computer program for calculation of LC50 values. Personal communication. 000680 * S T e 0 'A W lM 8 lt Appendix A. TATEA QUALITY DATA FiOM TOXICITY TEST 000681 -tVW Amended 8/9/91 BEST c o p y a v a il a b l e Table A.l. Conductivity, pH, temperature, and dissolved oxygen concentration measured during toxicity test Nominal concentration (mg/L) Rep. Conductivity (umhos/cm) ------------------ 0 24 48 hr hr hr pH -------------- _ 0 24 48 hr hr hr 0 (control) 150 250 400 1 2 3 4 1 2 3 4 1 2 3 4 1 2 3 .4 700 660 640 7.9 8.3 8.0 700 660 650 7.9 8.3 8.0 700 660 650 7.9 8.3 8.0 700 660 660 7.9 8.3 8.0 660 630 650 8.0 8.2 8.0 660 630 650 8.0 8.2 8.0 660 630 650 8.0 8.2 3.0 660 640 660 8.0 8.2 8.0 660 640 660 8.0 8.2 8.0 660 640 660 8.0 8.2 8.0 660 640 660 8.0 8.2 8.0 660 640 660 8.0 8.2 8.0 660 i 640 660 8.0 8.2 8.0 660 640 660 8.0 8.2 8.0 660 640 660 8.0 8.2 8.0 660 640 660 8.0 '8.2 8.0 0006S2 Amended 8/9/91 Table A.l. Continued. BEST COPY AVAILABLE nuniuai concentration (mg/L) Rep. Temperature (C) 0 24 48 hr hr hr Dissolved Oxygen (mg/L) 0 24 48 hr hr hr 0 (control) 1 20.3 19.8 19.7 8.5 8.8 8.3 2 20.3 19.9 20.1 8.4 8.8 8.3 3 20.2 19.9 20.5 8.5 8.8 8.5 4 20.4 19.8 20.6 8.5 8.8 8.3 150 1 20.4 19.7 19.4 8.8 8.7 8.1 2 20.3 19.8 19.3 8.7 9.0 8.2 3 20.3 19.7 19.2 8.6 9.2 8.0 4 20.3 19.3 19.3 8.7 9.2 8.1 250 1 20.3 19.3 19.3 8.6 9.4 8.0 2 20.2 19.3 19.3 8.6 9.4 8.0 3 -20.1 19.4 19.3 8.5 9.5 8.1 4 20.3 19.7 19.3 8.6 9.6 8.1 400 \ ' .20.4 19.8 19.4 8.7 9.6 8.1 .2 -20.3 19.7 19.6 8.5 9.7 8.1 jt.120.2 19.9 19.4 8.5 9.7 8.1 '*a^.i."J.A'*vy-*-.i*-. -.4>O'.J-;20.3 19.8 19.5 8.5 ^ 3 $ 9.6 8.2 000683