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AR R26 - 0726 ACUTE TOXICITY TO AQUATIC INVERTEBRATES (EASTERN OYSTER) IEsTSuBSTANCE 00000000 Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-203. aOctanesulfonic 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% ethyleneglycol, 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. M0 ETHOO:0000000 Method: Standard Practice for Conducting Static Basic AcuteToxicity 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 adultoysters 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. ECso 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: 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: 1L 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: Sedgwick-Rafter celNumber of normally developed larvae, counted with Water Tcehmempiesrtartyurdeurriannggteh(e0s-4t8udhyo:urs): 21 1 C (temperature controlled Salinity range (0-48 hours): 20 ppt environmental chamber). pDiHssroalnvgeed(o0x4y8gehonurrsa)n:ge7.(7074.88hours): >74% saturation R0 Eswrs 00000 Nominal concentrations: Bk control, 6, 10, 32, 56, and 100 mg/L. Element values: 48-hour ECs, = 47 (10 - 234) mg/L Element values based on nominal concentrations SRuebmsatraknsc:e TReesmtianrgkwsafsielcdo. nTduhcetveadlouenstrheepmoirxtteudraepapslydetosctrhiabtemdixintutrheeaTnedstnot the fluorochemical proportion alone. c0 onclusio0 ns 0000 The test substance Confidence Interval 48-hour of 10 to ECs; was 234 mg/L. determined to be 47 mg/L with a 95% Submitter: 3M Paul, Minnesota, C55o1m3p3any, Environmental Laboratory, P.O. Box 33331, St. p0 araQu0 aury 000000 Rteesltiianbgi.litHyo:weKvieirm,istchhersaankmipnlge=pu2r.itTyhiwsasstunodty pmreoeptesrltyhcehcarriatcertiearfiozredquaalnidtythe study in the lsoalcuktsioann.alytical confirmation of the amount of fluorochemical proportion - R0 EFER0 ENCE0 S 000000000 TPeesntsawcaoslac,onFdLuactttehdebryeqEuGe&stG,ofBtihoen3oMmicCsomMpaarniyne, RLeasbeaRrecqhueLsatbonruamtobreyr, 4o9f71, `Sample 5729, 1980. o0 wer 00000 Last changed: 6/27/00 000628 3 . ! Acute toxicity of Sample #5729 to embryos-larvae of eastern oysters (Crassostrea virginica) FcC-223 & FS9/9-/ ce xos-zr Toxicity Test Report Submitted to 3M Company st. Paul, MN 55133 Project Number LSS Report Number BP-80-7-117 MEaGr&iGneBioRneosmeiacrsch Laboratory P1e0n3s0a7coGlual,f BFleoarcihdaHig3h2w50a7y July 1980 000629 1 \ A marine toxicity test was conducted at EGSG 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 ofthe.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 (1) 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 % 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 3 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 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 and distributed into the spawning or test chambers. The chemical composition of BURL 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 mi embryo suspension:9 mi of seawater) from the spawning chamber. All concentrations and the control containers vere 1-1glassbeakers, cach were triplicated. of WHICH contained Test ~ 300 ni - 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. Bach test container was inoculated with an estimated 25,600 embryos within 1 hour after fertilization and then maintained at 21410C 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 mt of filtered seawater, and preserved with 1 ms of neutralized formalin. The number of normilly developed 48-hous larvae was determined by a Sedgwick-Rafter count from each triplicate test and control container. Percentage reduction of normal larvae was determined as follows: ) Percentage . mNiinunmubesearchtoheftensnuotmrbmceaorlncoe4fn8t-nhrooarutmriaolncon4t8r-ohlourlarlvaarevae reduction ~Numberofnormal 48-hourcontrollarvae X 100 The test vas 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 - linits 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 J 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 y aid occur in concentrations 332 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 AST StandardE 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, Iondon. 333 pp. ) 000634 6 TABLE 1. Toyosxtiecristy (oCfraSsasmopslterea$5v7i2r9gitnoiceam)bryeoxsp-olsaerdvafeorof48eahsoturesrn wianssttahteicr,eduuncateiroantedof stehaewantuemrb.er Tofhenocrrmiatleriloanrvafeorienffect ctoensttrocloncleanrvtarea.tionTsheasincitoimaplareidnoctouluthmewansumb2e5r,60o0f neomrbmrayols. Salinity was 20 /oo and temperature was 219C. concNeonmtirnaatlion (mg/%;ppm) norNmuamlberlaorfvae THean sb noRremdaulcti4o8n-hoofur larvae (3) Control 21,612 2,478 = 6 22,657 2,151" --- 10 22,373 2,010 -- 32 16,530 2,707 24 s6 11,258 1,054 8 100 380 142 58 ) _ _ standard deviation. ) 000635 PREPARED BY: AUDITED BY: ) REVIEWED BY: APPROVED BY: J) 7 Terry A. Hollister "To Study (Pirec Tom Heitmuller : deSom utancee on Peter J. Shuba, Ph.D. v Technical Clordinator Rod Parrish A ila 000636 | fr-- Results ofComCthievmeiecnatla"AnaInlysBelsonsfeoirosRoHuatsinieneChPaorascetesriTsoartairoantoofrySeSlaescsteend:Chenical | c--o ---------------------------------------- Chemical constituent: Immctonceentran tion C(ea/atipg--e) foafeisoenete Conger pr0%S.eo0oot1Lt0ea.ad0:0]088 $oo0.tan018 fSfhiaseaskerl <oS<S(B86ioo06rrkboe:b00sh807 0088::::6008880 | eM$NoirroSae1thteshoMSMisetaprshoeaetcenennasaassey oG0obtntt0.2 & op5:ni63 TSeoitHaarsPotioters iybcbcarbons $83 Gi9n PeFseltyiecriidoerstnated iphenyie NNoonnee ddeetteecrtreeddb NNoonnee ddeetteercrteedd Wfatrerresa:mprlensnvpezeicolt lectetd fron sionoaics Harine Research Laboratory seawater SRange of concentrations are based on tuo sampling periods. og 8 cpT hFEeonaslartueicCE hchhiaildmooek,rdsia:nDonaOsEc',emBdHaCl`yT,tBoizsl<h0i(enB0nd00ya80in)se:,oD/Oh3EAe,1pctpoaB0lce:ohCrlhoeorR,y1C0R1h6Le,Eprt:a12c3hc2lh,odsrs1e2eL4p6ot,xainsd1e2,6H0e,asls1di2ra2in0n,,, dCoimealcdrriinns,, eAndrEinD, , 1242, ana 1254 x .: Scr wn sic own seat AepENDIX B onsGteonpnasnaassTmoceioTmEST sHeeckien5 cs"h of 10m sceSr movecrro: LSS= west won: Seagle 5709 oe wou wor JOYS wevest wees ose: __//00 HOUR VEASUREIENTS In CONTROL (REPLICATE A) samme 30 : worse, ~_97 J adhopus - (se Tbetoo er pt 7) E ae - 2 : beN | . : . 5 uy 2 oe [22 Lasfac Tor | 8 were[BRoLt p01soo| shuts po sz wow | Cex Ta z sown |os|t wm |o oswom oemn y | ow | O wh {7rok0[| 2w0er [20 | 22 se vscl Joae | o Bo tog nape Lae ferLos Los ne pele loo|20 sce asLos [20 | I og 20 [oo 50 |20 | 59|50|gu [oo| TT J moe n btr n EHi E "wcnarton = 2a2 vid $0 ge, a iy e = 10.36c0antonc Th TE = 85,60 cade 000638