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ACUTE TOXICITY TO AQUATIC INVERTEBRATES (GRASS SHRIMP) TEST SUBSTANCE Identity:A mixturecontainingperfluorooctanesulfonatweh,ich may alsobe referredto as PFOS, FC-95, or as a component of FC-203. (1Octanesulfonicacid)(CAS # 2795-39-3). Remarks: The 3M productionlotnumber was 1. The testsample isFC203. Currentinformationindicatesitisa mixtureof 1.34% PFOS, 35% diethyleneglycolbutylether,37.85% water,20% ethyleneglycol,2.66 % Sultone foamer, 3% sodium octylsulfate0,.1% sodium laurylsulfate,and 0.05% tolyltdazole. The followingsummary applies to a mixture with incompletely characterized concentrations of impurities.Data may not accurately reflecttoxicityof the fluorochemical component of the testsample. METHOD: Method: Methods forAcute ToxicityTests withFish,Macroinvertebrates, and Amphibians (U.S.EPA 1975). Type: Acute static GLP: No Year completed: 1979 Species: Palaemonetes pugio Supplier: Fieldcollectefdrom BigLagoon,a GulfofMexico estuaryadjacent to the laboratoryP,ensacola,FL. Analyticalmonitoring: Temperature, pH, salinitya,nd DO Exposure period: 96-hours Test organism age: 16-22 mm rostrum-telsonlength Statisticamlethod: EC50 values calculatedusingthe Stephan computer program, 1977 and 1978. Test conditions: Dilutionwater: Filterendaturalseawater,from Big Lagoon, FL Dilutionwater chemistry: Salinity: 25 -26 ppt Temperature: 22 - 25 OC Lighting: Not given. Stock and testsolutionpreparation: Exposure concentrationsprepared by directadditionoftestsubstance. Exposure vessels: 19 L glassjarscontaining15 L oftestsolution. Number of replicates:2 Number of organisms per replicate:5 Number of concentrations: fiveplusa blankcontrol Water chemistry during the study: Salinityrange: (0-96hours): 26 ppt Temperature range (0-96hours): 22 *C pH range(0-9h6ours): 7.8- 8.0(contreoxlposure) 7.5- 8.1 (1000mg/L exposure)* Dissolvedoxygen range (0-96hours): 4.8- 6.4mg/L (controelxposure) 1.2- 6.3mg/L (1000 mg/L exposure)* 1000 mg/L (secondhighestconcentratione)xposurevaluesused because the highestconcentratiornesultedintotalmortalitbyy 72 hours. RESULTS Nominal concentrations:Bk control2,20, 360, 600, 1000, and 1800 mg/L. Element values: 96-hour EC50 = 510 (360 - 710) mg/L Element valuesbased on nominalconcentrations Remarks: Testingwas conductedon themixtureas describedinthe Test Substance Remarks field.The valuesreportedapplytothatmixtureand not the fluorochemicaplroportionalone. CONCLUSIONS The testsubstance96-hourEC5o was determinedto be 510 mg/L witha 95% confidenceIntervaolf 360 to710 mg/L. Submifter: 3M Company, EnvironmentalLaboratory,P.O. Box 33331, St. Paul,Minnesota,55133 DATA QUALITY ReliabilityK:limischranking= 2.This studymeets thecriterifaorquality testing.However, sample puritywas notpropedy characterizeadnd the study lacksanalyticaclonfirmatioonf theamount offiuorochemicaplroportioninthe solution. REFERENCES Testwas conducted by EG&G, BionomicsMarine Research Laboratoryof Pensacola,FL atthe requestofthe3M Company, Lab Request number 4971 S, Sample 7902, 1979. OTHER Last changed: 6/27/00 Acute toxicity of 3M Company's sample 7902 to grass shrimp (Palaemonetes pugio) F 2,-30 LR '49 7/ CAX 792,clS Toxicity Test Report Submitted to 3M Company St. Paul, Minnesota. Pioject Number H90-500 Report Number BP-79-8-121. EG&G, Bionomics Marine Research Laboratory Route 6, Box 1002 Pensacola, Florida 32507 August 1979 A marine toxicity test was conducted at 'Bionomics Marine Research Laboratory (BMRL),'Pensacola, Florida, to determine the acute effect of 3M Company's sample 7902 on grass shrimp (Palaemonetes pugio). The criterion.for effect was death. Results of the test are expressed as 24-, 48-, 72-, and<,9-6-hourLC50's (the concentrations of the test material estimated to be lethal to 50% of the test animals.at the specified times). All data related to this test are stored at BMRL. MATERIALS AND METHODS Test Material The sample was received at BMRL on 3 July 1979,.aiid was contained in a*500-milliliter (mt) NALGENEO bottle labeled "3M SAMPLE 7902, BIOASSAYS: 96ht LCSO-GRASS SHRIMP (PALAEMONETES VULGARIS); 48hr LC50-ATLANTIC OYigTER LARVAE (CRASSOSTREA VIRGINICA)." The sample was a medium orange liquid. Concentrations are reported here as milligrams (mg) of.,*.'whole test material per I of seawater or as parts per million (ppm). Test animals Grass shrimp, 16-22 millimeters rostrum-telson length, were collected from Big Lagoon, a Gulf of Mexico estuary adjacent to BMRL. The shrimp were held in the laboratory in flowing, natural seawater. During the 48-hour period prior to testing, salinity was 25-26 parts per thousand (0/oo) and temperature, 22-25 degrees Celsius (OC). No mortality was observed during the holding period. It should be noted that Palaemonetes pugio were tested instead. of P. vulgaris based on approval of Dr. Eric Reiner, 3M Company. 2 Test water Water used for holding and testing was natural seawater which was pumped from Big Lagoon. The pump intake was 85 meters (m) of fshore at a depth of approximately 3 m. Test water was pumped by a #316 stainless steel pump through hard polyvinylchloride (PVC) pipess, through sandfilled fiberglass filters, and through 10-micrometers (um) pore size polypropylene core filters into an elevated fiberglass reservoir. Water was continously and vigorously 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 test chambers. The chemical composition of BMRL seawater is characterized in Appendix A. Test conditions Methods for the 96-hour static test were those described in Methods for Acute Toxicity Tests with Fish,, Macro invertebrates, and Amphibians (U.S. EPA, 1975). The test was conducted in 19-t uncovered glass jars which contained 15 1 of filtered natural seawater. Salinity was 26 O/oo and temperature was maintained at 220C. Five shrimp were tested per jar and all treatments were duplicated. There was no aeration. Dissolved oxygen (DO) concentrations were near 90% of saturation at initiation of the test. Based on a range-finding test, shrimp were definitively tested at nominal concentrations of 220-1,800 ppm. Test concentrations were .prepared by adding appropriate weighed amounts of sample 7902 directly into seawater in the test containers. No solvent/carrier was used. The seawater control received no test material. 3 Test animals were not fed during the definitive test which was conducted 8-12 August 1979. Statistical analyses Based on the results of the test, 48-, 72-, and 96-hour LCSO's and their 95% confidence limits were calculated. The computer program (Stephan, 1977 and 1978 personal communication) estimated LC50 values by using one of three statistical methods in the following order: moving average angle analysis, probit-analysis, or binomial probability. The method selected was determined by the characteristics of the data, that is, the presence or absence of 0% and 100% mortality and the number of concentrations in tqhich >0%<100% mortality occurred. The computer scanned the data, identified the most suited method, and performed the analyses. For this test, the moving average angle method was used-to analyze the 72- and 96-hour data and the probit analysis method was used for the 48-hour data. RESULTS AND DISCUSSION 3M Company's sample 7902 was "practically nontoxic" to grass shrimp, based on EPA Hazardous Substances Guidelines, FEDERAL REGISTER, ------------------ 30 December 1975-..--...calculated 9 - our LC50 was with 95% Li! P-PM confidence limits of 3@60-07-710 ppm (Table 10) After 96 hours of ex'posure, i1n0tphpemco(nTtarbole 1) mortality ranged from 0% in the con ro 100% in 1,BOO ppm (Table 2). Low DO concentrations could have contributed to the toxicitv of sample 7902. At 24 hours of testing and later, the@DO concentrations generally decreased as the test concentrations increased and after 96 hours ranged from 16% of saturation in the 1,000 ppm test concen- tration to 66% of saturation in the control (Table 3). Loading of 4 test animals (0.013 g/t) was not sufficient to cause the low DO concentrations as evidenced by the 66% of saturation measurement in the control after 96 hours of testing. Apparently two factors, the inherent chemical nature.of the test material and the increased oxygen demand of shrimp stressed by exposure to sample 7902, resulted in the low DO concentrations. The pH in all treatments was within an acceptable range. After 96 hours, pH was from 7.5-7.8 (Table 3). 5 REFERENCES Stephan, C.E. 1977. Methods for.calculating an LC50. ASTM, Aquatic Toxicology and Hazard Evaluation, ASTM STP 634. P.L..Mayer and J.L. Hamelink, eds. pp. 65-84. U.S. Environmental Protection Agency. 1975. Methods for acute toxicity test@ with fish, macroinvertebrates, and amphibians. EPA-600/3-75-009. 61 p. 6 TABLE 1. Calculated LC50's for grass shrimp (Palaemonetes pugio) exposed to 3M Company's sample 7902 in static: unaerated seawater. Salinity was 26 O/oo and temperature, 220C. Values were calculated based on nominal concentrations of whole material. Hour 24 48 72 96 LC50 95% confidence limits ... (mg/..t-;Ppm) (mg/t) >1,800 --- 2,400 --- 570 .410-790 sio 360-710 TABLE 2. Mortality of grass shrimp (Palaemonetes pugio) exposed to 3M Company's sample 7902 in stat3@-C,-unaerated seawater. Salinity was 26 O/oo and temperature, 220C. Test concentrations are given as whole material. Nominal concentration (mg/t,;Ppm),Control 220 360 600 1,000 11800 Mortality 24 h 48 h 72 h 0 0 @o 0 10 10 0 0 40 0 0 30 0 20 80 0 so 100 96 h 0 10 40 40 90 100 7 TABLE 3. Measured concentrations of dissolved oxygen and pH during a 96-hour exposure of grass shrimp (Palaemonetes pugio) to 3M Company's sample 7902. The test was conducted in static, unaerated seawater. Salinity was 26 0/00 and temperature, 220C. Test concentration s are given as whole material. Dissolved oxygen concentr'ations are the average of measurements in Replicates A and B. The pH was measured- in Replicate A only. Ncminal ccncentration (mg/.E;PPM) Oh Dissolved oxygen Of saturaticn) pH 24 h 48 h 72 h 96 h 0 h 96H Control 6.4 (88) '6.0(82) 6.0 (82) 5.8 (79) 4.8 (66) 8.0 7.8 220 6.2 (85) 5.6 (77) 5.0 (68) 3.8 (52) 2.8 (38) 8.1 7.7 360 6.3 (86) 5.2 (71) 3.8 (52) 2.2 (30) 1.6 (22) 8.0 7.6 600 6.4 (88) 5.4 (73) 3.6 -(50) 2.@2 (30) 1.6 (22) 8.1 7.6 isooo 1,800 6.3 (86) 5.0 (68) 2.4 (33) 1.4 (19) 1.2 (16) 8.1 7.5 6.4 (88) 4.8 (66) 1.8 (25) 1.0 (14) -a 8.1 -a aNo measurement because 100% mortality had occurred. APPENDIX A Results of Chemical Analyses for'Routine Characterization of Selected Chemical Constituents in Bionomics Marine Research Laboratory Seawater Constituent ..Concentratiork (mg7Y.; ppm) 30 January 1979 14 June 1979 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 ; - Pesticidei Polychlorinated Biphenyls <0.001 0.002 0.0075 0.023 0.0007 0.02 0.05 <0.001 <0.02 0.42 <0.01 <0.01 <5.0 <1.0 None detecteda None detectedb 0.006 <0.01 <0.01 <0.01 <0.0005 <0.01 <0.02 <0.02 <0.02 0.14 <0.01 <0.01 <5.Oq <1.0 None detected None detected Water samples were collected from Bionomics marine Research Laboratory seawater system after the mixing-station in the wet lab. apesticides: BHC, lindane, heptacl@lor, heptachlor epq.-@.ide, aldrin,dieldrin, endrin, perthane, DDE, TDE (DDD), DDT, methoxychlor, endosulfan, strobane, toxaphene, kelthane, and chlordane all <0.005 bPolychlorinated Biphenyls: Aroclore 1016, 1232, 1248, 1260, 1221, 1242, and 1254 all-<0.05 ug/.E. cpetroleum hydrocarbon sample collected 10 July 1979. PREPARED BY: AUDITED BY: REVIEWED BY: APPROVED BY: 8 Tom Heitmuller Study Dikd-ctor G. Scott Ward Quality Assurance:Unit Peter Shuba, Ph.DProject Coordinator Rod Parrish Di@:e-ctor