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ARR2G - 0725 ACUTE TOXICITY TO AQUATIC INVERTEBRATES (EASTERN OYSTER) JEsTsu 0000B 0000s 0000T 0 ANCE 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. METHOO: 000000 Method: Standard Practice for Conducting Static Basic Acute Toxicity Tests with Larvae of Four Species of Bivalve Mollusks (ASTM, Draft No. 5). Type: Acute static GLP: No Year completed: 1979 Species: Crassostrea virginica Supplier: Induced spawning in the laboratory of field-collected mature adults from an estuary adjacent to Biloxi, Mississippi. Maintained at BMRL until testing. Analytical monitoring: Temperature, pH, salinity, and DO. Exposure period: 48-hours Test organism age: Empryos, within 1 hour after fertilization. Statistical method: Test concentrations converted to logarithm and corresponding percentage reduction of normal larvae was converted to a probit. ECs values 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: 22 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. 23,400 embryos 000615 Number of concentrations: five plus a blank control Element basis: Number of normally developed larvae, counted with `Sedgwick-Ratter cell Water chemistry during the study: Temperature range (0-48 hours): 20 1 C (Temperature controlled Salinity range (048 hours): 22 ppt water bath) PH range (048 hours): 8.08.1 Dissolved oxygen range (048 hours): > 67% saturation Reswvs 0000 Nominal concentrations: Bk control, 0.6, 1.0, 3.2, 5.6, and 10 mg/L. Element values: 48-hour ECso = 3.5 (0.5 - 22) 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 the fluorochemical proportion alone. reported apply to that mixture and not conclusions 00000 The test substance 48-hour ECso was determined to be 3.5 mg/L with a 95% Confidence Interval of 0.5 to 22 mg/L. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 paTaQuALTY 0000000 Reliability: Klimisch ranking = 2. This study meets the criteria for quality testing. However, sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in the ~ solution. REFERENCES 000 Test was conducted by EG&G Bionomics Marine Research Laboratory, Pensacola, FL at the request 49718, Sample 7902, 1979. of the 3M Company, Lab Request number orWER Last changed: 6/27/00 OOOOO00 000616 ) Acute toxicity of 34 Company's Sample 7902 to embryos-larvae of eastern oysters (Crassostrea virginica) Fe zon 7 | FAT 79245 LR 497s Toxicity Test Report Submitted to : 34 Company st. Paul, Minnesota ) Project Number HS0-500 ; Report Number BP-79-8-123 EMGa&rGi,neBiRoensoemairccsh Laboratory PReonustaeco6l,a,BoxFlo1r0i02da 32507 August 1979 000617 . 1 A marine toxicity test was conducted at Bionomics Marine ) Research Laboratory (BMRL), Pensacola, Florida, to determine the effect of Sample 7902 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(fi-hourECSD)(the concentration of Sample 7902 estimated to be effective in preventing normal development of 50% of the exposed embryos-larvae). Data from the test are maintained at BMRL. MATERTALS AND METHODS Test material ) The sample was received at BURL on 3 July 1979, and was con- tained in a 500-milliliter (mf) NALGENE bottle labeled "3M SAMPLE 48hr LCS0-ATLANTIC OYSTER LARVAE (CRASSOSTREA VIRGINICA)." The 3 sample was a medium orange liquid. Concentrations are reported here as milligrams (ng) of whole test material per % of seawater or as parts per million (ppm). Test animals Oyster embryos were obtained by induced spawning of sexually mature adult oysters which had been collected from an estuary adjacent to Biloxi, Mississippi, on 20 July 1979 and maintained in flowing, unfiltered seawater at BMRL until testing began. J 000618 . 2 Test water 5 Water used for spawning and testing was natural seawater which was pumped from Big Lagoon, a Gulf of Mexico estuary adjacent to BURL. The punp intake was about 85 meters (m) offshore at a depth . of approximately 3 m. Seavater vas pumped by a #316 stainless steel pump through hard polyvinylchloride (PVC) pipes, through sandfilled fiberglass filters, and through l0-micrometers (um) pore size polypropylene core filters into an elevated fiberglass reservoir. Water was continuously and vigorously aerated .in the reservoir and flowed by gravity through PVC pipes into the laboratory. There it was pumped through a 5-um pore size polypropylene core filter ana distributed into test chambers. - : The chemical composition of BMRI 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, Draft No. 5). Individual, sexually mature female oysters were induced to spawn by placing them in glass chambers containing 1 & of filtered (S-ym), 26 degrees Celsius (C) seawater and increasing the water temperature to 32C 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 390%. Density of the embryos was determined by a Sedgwick-Rafter count of a 1:10 dilution (1 me J embryo suspension:9 mi seawater) from the spawning chamber. 000619 . 3 All concentrations and the control were triplicated. Test ) containers were 1-1 glass beakers, each of which contained 900 mi of filtered (5-um), natural seawater. A primary stock solution was prepared by adding a weighed amount of Sample 7902 to a known volume of 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 23,400 embryos within 1 hour after fertilization and then maintained at 201C in a temperature-controlled water bath. 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 mi of filtered seawater, and preserved with 1 me, : of neutralized formalin. The number of normally developed 48-hour larvae was determined by a Sedgwick-Rafter count from each tripli| cate test and control container. Percentage reduction of normal embryos was determined as follows: mNiunmubsertohfe nnourmbmearl o4f8-hnoourrmalcon4t8r-ohlourlalravarevae : v Percentage _ in each test concentration 100 reduction Numberofnormal 48-hourcontrollarvae The test was conducted 1-3 August 1979. 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 ECS50 and 95% confidence limits were then calculated by linear regression. J 000620 4 3) RESULTS AND DISCUSSION The calculated 48-hour EC50 for embryos-larvae of eastern oysters exposed to Sample 7902 in static, unaerated seawater was 3.5 ppm with 95% confidence limits of 0.5-22 ppm) Reduction of embryos-larvae which developed normally to the straight-hinged veliger stage after 48 hours was from 12% in 0.6 ppm to 74% in 10 ppm (Tables 1 and 2). Measured concentrations of dissolved oxygen remained 367% of saturation and the pH was from 8.0-8.1 after 48 hours of exposure. ) J 000621 . 5 N REFERENCES American Society for Testing and Materials Committee E-35 on Pesticides. August 1978. Standard Practice for Conducting Static Basic Acute Toxicity Tests with Larvae of Four Species of Bivalve Molluscs. Draft No. 5. Finney, D.J. 1971. Probit Analysis. Cambridge University Press, London. 333 p. ) 000622 3) TABLE 1. Toxicity of 3M Company's sample 7902 to embryos larvae of eastern oysters (Crassostrea virginica) exposed for 48 hours in static, unaerate seawater. Tnchouemmpbacerrreidtofertoinoonrtmhaeflonrulmaebrfevfraeectofiwnansotremstathlecorcnoecndeturcnottliroantliaoognfvsaet.haes Salinity was 22 /o0 and temperature, 20t1C. seers i--S---- concNeonmtirnaatlion (mg/2; ppm) Control Pneorrcmeanlta4g8e-hroeudrucltairovnaeodf --- 0.6 12 1.0 Et 3.2 44 5.6 61 10 74 ) mNiunmeubseorto ohfe nnourmmbaelr o4e f8-nhoorumralcon4t8r-p ohlour-- larlva-- arevae preerdcuecnttiaogne _= Ninumbeeacrhoftesntormcaolncent48r-ahotuironcontrol Tarvae -- x 1iL00 6 : > J 000623 . 7 dD TABLE 2. Calculated virginica) number larvae of normal following eastern oysters (Crassostrea 48 hours of exposutroe3M nCuonmbpearnsywserSeampblaesed79o0n2 Sinedgstwaitcikc-,Rafutnearcccaotuendtoso.mmaItneist.ialThe 2i0n1o1cuCl.um was 23,400. Salinity was 22 9/oo and temperature, "cWonecmenmtraaltion (mg/2;ppm) Control 0.6 1.0 3.2 5.6 10 Rep A 19,380 15,39 13,822 12,112 8,408 5,771 NRuempberB_ 22,515 18,098 11,970 8,978 6,484 3,919 ofRneoprmCa_l 18,382 19,238 15,818 12,825 8,761 6,270 Mlaeravane 20,092 17,575 13,870 11,305 7,885 5,320 _SD3_ 2,157 1,976 1,924 2,046 1,226 1,239 standard deviation. ) ) 0C0Z4 APPENDIX A hl RCeshualrtasctoefrizCahteimiocnalofAnSaellyescetsed fCohremRiocuatline ConstituentsLianborBaitoonroymicSseaMwaatreirne Research Constituent CJoanncueanrtyration (mJgu/nEye 15 ACrasdenniiucn 00..000012 <00l.00106 CChorpopmeirum 00..0020375 <<00..0011 MNeircckeulry 00..002007 0<0..000105 ZLienacd 000.0051 <<00..0022 TAomtmaolniaPhNoispthraotgeenasas P N <00..0422 <00.10a2 NNiittrraittee NNiittrrooggeenn aass NN <0<0l.0011 <00.0011 TsoutlaflidePsetroleum Hydrocarbons a<l5.o0 <51008 . PPoelsytcihcliodreisnated Biphenyls NNoonnee ddeetteecctteeddb? NNoonnee ddeetteecctteedd --e ----e--s ---------------------------------------- ` LWaatbeorratsoarmyplesseawwearteercoslylsetecmtedaftferromtBhieonmoimxiicnsg MsatraitnieonReisneathrech wet lab. pesticides: aBlHdC,rinl,idnidealnder,inh,epteandcrhilno,r, pheertphtaanceh,lorDDEe,poxDiEde, . t(oDxDaD)p,hen`DeD,T, kmeletthhaonxey,chlaonxd, chelnodorsdualnfeana,ilst<r0o.b0a0n5e, va/e. polychlorinate*d Biphenyls: A1r26o0c,lo1r2e211,0161,2421,232a,nd1214285,4 . all 0.05 ng/2. - Cpetroleun hydrocarbon sample collected 10 July 1979. J 000625 . : ) PREPARED BY: AupIIED BY: ) REVIEWED BY: APPROVED BY: J 8 Terry A. Hollister AegaPneake 6. Scott wara GrTey aAesdmancePGatae s Peter Shuba, Ph.D. --C [Ade rR, Rod Parrish RA Benen 000626