Document 1gk2VBV6mrNEa550w0n8Y7Gma

AH 8.2.6 - 072j> ACUTE TOXICITY TO AQUATIC INVERTEBRATES (EASTERN OYSTER) 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 (F5919-1). 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: Standard Practice for Conducting Static Basic Acute Toxicity Tests with Larvae of Four Species of Bivalve Molluscs (ASTM 1980). Type: Acute static GLP: No Year completed: 1980 Species: Crassostrea virginica Supplier: Induced spawning in laboratory-maintained mature adult oysters Analytical monitoring: Temperature, pH, salinity, and DO. Exposure period: 48-hours Test organism age: Embryos, within 1 hour after fertilization. Statistical method: Test concentrations converted to logarithm and corresponding percentage reduction of normal larvae was converted to a probit. EC50values then calculated using linear regression. Test conditions: Dilution water: Filtered natural seawater pumped from Big Lagoon, a Gulf of Mexico estuary adjacent to the laboratory, Pensacola, FL. Dilution water chemistry: Salinity: 20 ppt Lighting: Not given. Stock and test solution preparation: A primary stock solution was prepared by adding a weighed amount of test substance to filtered seawater. Exposure concentrations were then prepared by addition of the appropriate volume of stock solution. Exposure vessels: 1 L glass beakers containing 900 mL of test solution. Number of replicates: 3 Number of organisms per replicate: Approx. 25,600 embryos Number of concentrations: five plus a blank control ~ 000627 Element basis: Number of normally developed larvae, counted with Sedgwick-Rafter cell Water chemistry during the study: Temperature range (0-48 hours): 21 1 C (temperature controlled environmental chamber). Salinity range (0-48 hours): 20 ppt pH range (0-48 hours): 7.7 -7.8 Dissolved oxygen range (0-48 hours): >74% saturation RESULTS____________________________________________________ Nominal concentrations: Bk control, 6, 10, 32, 56, and 100 mg/L. Element values: 48-hour EC50 = 47 (10 - 234) 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 EC50 was determined to be 47 mg/L with a 95% Confidence Interval of 10 to 234 mg/L. 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________________________________________________ Test was conducted by EG&G, Bionomics Marine Research Laboratory, of Pensacola, FL at the request of the 3M Company, Lab Request number 4971, Sample 5729, 1980. OTHER______________________________________________________ Last changed: 6/27/00 000628 Acute toxicity of Sample #5729 to embryos-larvae of eastern oysters (Crassostrea virginica) -- ----- J SrO S - z Jr Toxicity Test Report Submitted to 3M Company St. Paul, MN 55133 Project Number L55 Report Number BP-80-7-117 EG&G Bionomics Marine Research Laboratory 10307 Gulf Beach Highway Pensacola, Florida 32507 July 1980 000629 1 A marine toxicity test was conducted at EG&G Bionomics Marine Research Laboratory (BMRL), Pensacola, Florida, to determine the effect of Sample #5729 on embryos-larvae of eastern oysters (Crassostrea virginica). The criterion for effect was reduction of the number of normal larvae (those which developed to the fully-shelled, straight-hinged veliger stage within 48 hours) in test concentrations as compared to the number of normal control larvae. Results of the.test are expressed as a 48-hour EC50 (the concentration of test material estimated to be effective in preventing normal development of 50% of the exposed embryos-larvae). Data from the test are maintained at BMRL. MATERIALS AND METHODS Test material The sample, received at BMRL on 12 June 1980, was contained in a 0.9-liter () clear glass bottle labeled "3M Company, Sample #5729." The sample was an orange liquid. Test concentrations are reported here as milligrams (mg) of whole material per Z of seawater or as parts per million (ppm). Test animals ' Oyster embryos were obtained by induced spawning of sexually mature adult oysters which were maintained in flowing, unfiltered seawater at BMRL until testing began. Test water Water used for spawning and testing was natural seawater which was pumped from Big Lagoon, a Gulf of Mexico estuary adjacent to BMRL. The pump intake was about 80 meters (m) offshore at a depth 000630 2 of approximately 3 m. Seawater was pumped by a #316 stainless steel pump through hard polyvinylchloride (PVC) pipes, through sandfilled fiberglass filters, and through 10-micrometers (pm) pore size polypropylene core filters into an elevated fiberglass reservoir. Water was aerated in the reservoir and flowed by gravity through PVC pipes into the laboratory. There it was pumped through a 5-pm pore size polypropylene core filter and distributed into the spawning or test chambers. The chemical composition of BMRL seawater is characterized in Appendix A. Test conditions Methods for the 48-hour oyster embryo-larvae test were based on Standard Practice for Conducting Static Basic Acute Toxicity Tests with Larvae of Four Species of Bivalve Molluscs (ASTM, 1980). Individual, sexually mature female oysters . were induced to spawn by placing them in glass chambers containing 1 of filtered seawater at 25 degrees Celsius (C) and increasing the water temperature to approximately 30C in the presence of viable sperm excised from the gonad of a sexually mature male oyster. Fertilization occurred upon release of the eggs into the spawning chambers and was confirmed microscopically. Fertilization success was estimated to be >.90%. Density of the embryos was determined by a Sedgwick-Rafter count of a 1:10 dilution (1 mil embryo suspensions mi. of seawater) from the spawning chamber. All concentrations and the control were triplicated. Test containers were 1-Jl glass beakers, each of which contained 900 mi, of filtered, natural seawater. A primary stock was prepared by adding a weighed amount of test material to a known volume of 000631 3 filtered seawater and the appropriate volumes were added to each test container to obtain the desired test concentrations. Each test container was inoculated with an estimated 25,600 embryos within 1 hour after fertilization and then maintained at 211C in a temperature-controlled environmental chamber. After 48 hours of exposure, the larvae from each container were collected in a 37-ym mesh size sieve, rinsed into a plastic bottle with 24 m2, of filtered seawater, and preserved with 1 m2, of neutralized formalin. The number of normally developed 48-hour larvae was determined by a Sedgwick-Rafter count from each triplicate test and control container. .Percentage reduction of normal larvae was determined as follows: Number of normal 48-hour control larvae minus the number of normal 48-hour larvae ) Percentage _ in each test concentration_____________ reduction Number of normal 48-hour control larvae X 100 The test was conducted 1-3 July 1980. Statistical analyses Each test concentration was converted to a logarithm and the corresponding percentage reduction of normal larvae was converted to a probit (Finney, 1971). The 48-hour EC50 and 95% confidence limits were then calculated by linear regression. RESULTS AND DISCUSSION The calculated 48-hour EC50 for embryos-larvae of eastern oysters exposed to Sample #5729 in static, unaerated seawater significant reduction of embryos-larvae which developed normally to the straight hinged veliger stage after 48 hours occurred in 000632 4 6 or 10 ppm, but a significant reduction of normal larvae did occur in concentrations >32 ppm (Table 1). Measured concentrations of dissolved oxygen remained >74% saturation and the pH was from 7.7-7.8 after 48 hours of exposure (Appendix B ) . 000633 5 REFERENCES ASTM Standard E 724-80, Practice for Conducting Static Acute Toxicity Tests with Larvae of Four Species of Bivalve Molluscs. American Society for Testing and Materials, 1916 Race Street, Philadelphia, PA 19103. Finney, D.J. 1971. Probit Analysis. Cambridge University Press, London. 333 pp. ) 000634 6 TABLE 1. Toxicity of Sample #5729 to embryos-larvae of eastern oysters (Crassostrea virginica) exposed for 48 hours in static, unaerated seawater. The criterion for effect was the reduction of the number of normal larvae in test concentrations as compared to the number of normal control larvae. The initial inoculum was 25,600 embryos. Salinity was 20 /oo and temperature was 21C. Nominal concentration (mg/;ppm) Control 6 10 32 56 100 Number of normal larvae Mean SD3 21,612 2,478 22,657 2,151 " 22,373 2,010 16,530 2,707 11,258 1,054 380 142 Reduction of normal 48-hour larvae (%) --_ -- -- 24 48 98 aStandard deviation. `v 000635 PREPARED BY AUDITED BY: REVIEWED BY: APPROVED BY: J Terry A. Hollister 7 Tom Heitmuller Quality Peter J. Shuba, Ph.D. v, Technical Coordinator Rod Parrish 000636 APPENDIX A Results of Chemical Analyses for Routine Characterization of Selected Chemical Constituents in Bionomics Marine Research Laboratory Seawater Chemical Constituent Arsenic Cadmium Chromium Copper Mercury Nickel Zinc Lead Total Phosphate as P Ammonia Nitrogen as N Nitrate Nitrogen as N Nitrite Nitrogen as N Total Petroleum Hydrocarbons Sulfides Pesticides Polychlorinated Biphenyls Concentration (mq/Jl;ppm)________ 1979 Range3- 1 April 1980 <0.001-0.006 <0 .01-0.002 <0.01-0.008 <0 .01-0.02 <0.0005-0.0007 <0 .01-0.02 <0.02-0.05 <0 .02 <0.02 0.14-0.42 <0.01 <0.01 <5.0 <1.0 None detected13 None detected0 0.013 < 0.01 < 0.01 < 0.001 0.002 0.02 0.02 0 .05 0.02 < 0.01 0.10 < 0.01 <5.0 < 0.01 None detected None detected Water samples were collected from Bionomics Marine Research Laboratory seawater system after the mixing station in the wet lab. aRange of concentrations are based on two sampling periods. ^Pesticides: BHC, lindane, heptachlor, heptachlor epoxide, aldrin, dieldrin, endrin, perthane, DDE, TDE (DDD), DDT, methoxychlor, endosulfan, strobane, toxaphene, kelthane, and chlordane all <0.005 yg/A;Ppb. cPolychlorinated Biphenyls: Aroclor 1016, 1232, 1248, 1260, 1221, 1242, and 1254 all <0.05 yg/;ppb. i > i 000637 3JECT: 48-HOUR STATIC DATA SHEET APPENDIX B QUALITY ASSURANCE FORM OYSTER-EMBRYO TOXICITY TEST C g y r ^ ^CLIENT: 3 / r ? ____________________ ________ PROJECT NO: NO: BP-QAF-OE-OOl Effective: April 1980 Revision: #1 Page: TEST MATERIAL: . ^ ^ 7 s - y ____ _______________ ______________________ TIME INOCULUM ADDED: /O V S __________ ^____________________________________________ TIME TEST MATERIAL ADDED: fHO 0 HOUR MEASUREMENTS IN CONTROL (REPLICATE A) SALINITY: .0 O TEMPERATURE: _ (See Table Below for Data by) . -- / &Y*pjArJfc/ -- atwJL gj<L <Zk c g C TEST HOUR 0 HOUR DATE/TIME DATA BY n tn is o \ .. / ^ u ASURF.MENT 3NCENTRATI0N DISSOLVED OXYGEN (mg/1) REP A REP B REP c PH REP A 48 HOUR li Inj# VV Bo 1 HoO DISSOLVED OXYGEN (mg/1) REP A REP B REP C pH REP A 7 ,f 6?i * vfo !Offh~ 0,0 O.X 5(a /Pph a 0.0 /o 0 1pqW' 0.0 n.o o.i i.x o.> 7,0 1,1 7,0 0,0 on U >t(o M b X I X 7 f 6,r b.H 6.1 7 .f L .H te.~b 6,Ip 1 J 7? . 3 ____ 0>.5 7i t * 6.0 6, f 1J 1.0 S .9 5,1 6 ./ o n r I BEST COPY AVAILABLE V- . TIAL INOCULUM: Number of normal embryos found in 1 ml number of fields X Sedgwick Rafter x Factor CALCULATION: *//,1 7 / _______ ___________ Dilution Factor Total number of nor mally developed embryos per ml / ,<^60 S .,suL(ubP to -A ^ ^ as.o/co^x 000638