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AR.A2.6 _ 0SS6 TO XICITY TO AQUATIC PLANTS TEST SUBSTANCE Identity: Mixture containing perfluorooctanesulfonate which may also be referred to as PFOS, FC-95, or as a component of FC-3003. (1Octanesulfonic acid) (CAS # 2795-39-3). Remarks: The 3M production lot number was not given. The test sample is FC-3003, identified by the laboratory as R1331. Current information indicates it is a mixture of 1% PFOS, 15% diethylene glycol butyl ether, 79.39% water, 2% Sultone foamer, 1.5% sodium octyl sulfate, 1% sodium decyl sulfate, 0.06% sodium lauryl sulfate, and 0.05% benzotriazole. The following summary applies to a mixture with incompletely characterized concentrations o f impurities. Data may not accurately reflect toxicity o f the fiuorochemicai component o f the test sample. METHOD:________________________________________________________ Method: OECD Guideline 201 Type: Acute Static GLP: No Y ear study performed: 1997 Species: Selenastrum capricomutum Source: Originally obtained from ATCC, Rockville, MD, maintained in culture medium at AScl-DETD, Duluth, MN Elem ent basis: Algal cell counts (cells/ml), specific growth rates. Exposure period: 96 hours Statistical Methods: Statistical analyses were performed using the Trimmed Spearman-Karber method. Analytical m onitoring: pH, conductivity, and temperature Test Conditions: Algal Nutrient Medium: Nutrient medium per OECD. This nutrient medium provided all mineral nutrients essential for algal growth and also served as the diluent for all algal operations. The pH of this synthetic algal medium was 8.0. Stock and test solution preparation: A 2000 mg/L primary stock solution was prepared by dissolving 700 mg in 350 mL algal medium, followed by mechanical stirring. Each test concentration was made by volume addition of the stock to algal medium. Exposure vessels: 250 ml Erlenmeyer flasks containing 100 ml test solution, sealed with foam stoppers. Agitation: 100 rpm. Num ber o f replicates: 3 for exposure concentrations, 6 for blank control. Initial cell loading: 1.0 x 104 cells/mL (1.3 x 104 cells/mL actual). 002721 Num ber o f concentrations: five plus a blank control Lighting: Continuous lighting at 740 ft-candles using cool-white fluorescent lamps. W ater Chemistry: pH range (0-96 hours): 8.00 - 9.90 (control exposure) 4.72 - 7.90 (1000 mg/L exposure) Conductivity range (0-96 hours): 1 5 1 -1 8 6 pmhos/cm (control exposure) 1 5 9 -1 7 0 pmhos/cm (1000 mg/L exposure) Test tem perature range (0-96 hours): 24.8 - 25.8 C (incubator) RESULTS Nom inal concentrations: Blank control, 130, 2 f6 , 360, 600, and 1000 mg/L A lgal Growth ECso Values Exposure (C o n ta c t) Hours 24 48 72 96 C ell-C ount m g/L (95% C .l.) 23 3(1 59-342 ) <130 (Not Calculable) <130 (Not Calculable) <130 (Not Calculable) A verage Specific Growth Rate m g/L (95% C .l.) 1 6 0(1 50-170 ) 251 (204 - 309) 572 (434 - 756) 644 (528 - 834) Elem ent values: 96-hour EC50 = <130 mg/L (cell count) 96-hour ErCso = 664 (528 - 834) mg/L (growth rate) 96-hour NOEC = 23 mg/L (cell count) 96-hour NOEC = 96 mg/L (growth rate) Element values are based on nominal concentrations. Reversibility o f Growth Inhibition: Aliquots of the 1000 mg/L test solution were diluted with algal medium and cultured for an additional period. Based on growth observed in the recovery phase, the effect on algal growth was found to be algistatic. 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 002722 FC-3003 exhibits a 96-hour EC50 cell count value of <130 mg/L and a 96-hour E1C 50growth rate value of 664 (528 - 834) mg/L. The 96-hour No Observed Effect Concentration (NOEC) is 23 mg/L for cell count and 96 mg/L for growth rate. This test substance was determined to be algistatic. Subm itter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY___________________________________________________ Reliability: Klimisch ranking = 2. Testing 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____________________________________________________ This test was conducted by AScI Corporation, Duluth Environmental Testing Division, Duluth, MN at the request of the 3M Company, Lab Request number R 1 3 3 1 ,1997. OTHER_____________________________________________________ _ _ Last changed: 6/26/00 002723 AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 STUDY TITLE GROWTH INHIBITION OF R 1331 FOR GREEN ALGA (Selenastrum capncomunm) BATAjSTANPARP OECD GUIDELINE 201 STUDY COMPLETED April 3, 1997 TESTING FACILITY AScI Corporation/AScI-Duluth Environmental Testing Division 4444 Airpark Boulevard Duluth, MN 55811 Tel. No. (218) 722-4040 Fax No. (218) 722-2592 STUDY IDENTIFICATION NUMBERS AScI Study ID# 5010-068 3M Company Lab Request# R1331 Sponsor: 3M Company Sponsor Study ID# R1331 y 002724 AScI Corporation/AScl-Dululh Environmental Testing Division AScI Study ID# 5010-068 1.0 GENERAL INFORMATION Study Title Data Standard Sponsor Sponsor's Representative Sponsor's Lab Request# Testing Facility Principal Investigator Project Director Testing Facility Director Definitive Test Dates Data Validation and Report Review Growth Inhibition of R1331 for Green Alga (Selenastrum capricomuium). OECD Guideline 201. 3M Environmental Technology and Safety Services, Building 2-3E-09, 935 Bush Avenue, St. Paul, MN 55133; Tel No. (612) 778-7452. Susan Beach, 3M Environmental Technology and Safety Services, Building 2-3E-09, 935 Bush Avenue, St. Paul, MN 55133; Tel No. (612) 778-7452. r' R1331. AScI Corporation/AScI-Duluth Environmental Testing Division, 4444 Airpark Boulevard, Duluth, MN 55811; Tel. No. (218) 722-4040. Todd McGuire Joe Amato Donald Mount March 25-29, 1997. :j Alan Mozol Report Signature Retention of Raw Data and Final Report Location of Raw Data and Final Report Two years from the date study is completed. AScI Corporation/AScI-Duluth Environmental Testing Division, 4444 Airpark Boulevard, . Duluth, MN 55811; Tel. No. (218) 722-4o4o. Sponsor: 3M Company Sponsor Study ID# R1331 2 . 3 002725 2.0 OBJECTIVE(S) AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 To determine the 4-d median effect concentration (ECS0) and the 4-d no observable effect concentration (NOEC) of the test substance, R1331, for green alga (Selenastrum capricomutum) under static test conditions. This study was conducted according to OECD Guideline 201 (OECD 1993). Test Substance Test Organisms fi Algal Medium Exposure Chambers 3.0 TEST METHODS ;> Properties'. R1331, clear amber-colored liquid, specific gravity =1.02 (water=1), stable, and completely soluble in water. Test concentrations: 0 (algal medium control), 130, 216, 360, 600, and 1000 mg/L. A stock solution was prepared by mass addition to algal medium followed by mechanical stirring. Each test concentration was made by volume addition of the stock to algal medium. Source: ATCC 22662 Inoculum preparation: Algal stock was subcultured in OECD recommended medium for 4 days before test initiation. Use in test: Stock culture was diluted and spiked into test solutions to obtain an initial cell density of 1.0 X 104 cells per ml (1.3 X 104 actual concentration). 4 Preparation: Deionized water was spiked with nutrient stocks (including; Na2EDTA) to OECD recommended concentrations. The most recent chemical analysis for deionized water is in Raw Data Package. Characteristics: At test time, the medium had a pH of 8.00. Sterilized via 0.22 pm filtration prior to use in the test. Type: Sterilized via autoclave 250-ml borosilicate-glass Erlenmeyer flasks sealed with foam stoppers. Use in test: 100 ml of algal medium or test solution/chamber. Six control replicates and three replicates per test substance concentration were __ _ employed. During the test, kept stoppered, except when experimental observations were made. Sponsor: 3M Company Sponsor Study ID# R1331 3 002726 Incubation Observations Endpoint Calculations AScI Corporation/AScI-Dululh Environmental Testing Division AScI Study ID# 5010-068 Duration: 4 days. Daily photoperiod: Continuous at 740 ft-candles using cool-white fluorescent lamps. Test chambers held on shaker table set at 100 ipm. Temperature: 25.0 2C. Biological: Daily cell counts using direct microscopic enumeration via Improved Neubauer hemacytometer. Water chemistry: (1) At test initiation and termination- pH, conductivity, and temperature, (2) daily- temperature. All determinations were made according to APHA methods (1995). u H' 1-, 2-, 3-, and 4-d EC50's, and 4-d NOEC based on both cell counts and', average specific growth rates (jx). Statistical analyses performed using Trimmed Spearman-Karber Method (EC50's) and ICp analysis. 4.0 RESULTS Test Substance Toxicity (Pertinent data are in Tables 1, 2, and 3) Cell Count 1-d ECS0: 233 mg/L (159-342). ix 1-d EC50: 160 mg/L (150-170). Cell Count 2-d ECS0: < 130 mg/L. ix 2-d EC5Q: 251 mg/L (204-309). Cell Count 3-d ECS0: < 130 mg/L. fx 3-d ECS0: 572 mg/L (434-756). ;f (; Cell Count 4-d ECS0: <130 mg/L. ix 4-d EC5Q: 664 mg/L (528-834). - Cell Count 4-d NOEC (IC10): 23 mg/L (22-24). ix 4-d NOEC (1C10): 96 mg/L (88-105). Effect was determined to be algistatic based on production of cells exposed during the definitive test to the 1000 mg/L test substance concentration. Sponsor: 3M Company Sponsor Study ID# R1331 4 C; 002727 1 Test Conditions AScl Corporation/AScl-Duluth Environmental Testing Division AScl Study ID# 5010-06 Temperature (C): 24.8-25.8. Light intensity ft-cdls: 740. . pH: 4.72-9.90. Specific conductivity (umhos/cm): 129-186. 5.0 TEST QA/QC QA/QC Criteria Algal counts in control must increase by at least a factor of 16 during the first 3-d of the exposure. Test duration must be 4-d. Data - During the first 3-d of the test control algal counts increased by a factor of approximately 157. The test duration was 4-d. 6.0 REFERENCES American Public Health Association (APHA). 1995. Standard Methods for the Examination of Water and Wastewater. APHA, Washington, D.C. Organization for Economic Cooperation and Development (OECD). 1993. OECD Guidelines fpr Testing of Chemicals. OECD Publication Information Center, Washington, D.C. .. Hamilton, M.A., R.C. Russo, and R.V. Thurston, 1977. Trimmed Spearman-Karber Method for Estimating Median Lethal Concentrations in Toxicity Bioassays. Environ. Sci. Technol. t-h 11(7): 714-719; Correction 12 (4): 417 (1978). US EPA. 1993. A Linear Interpolation Method for Sublethal Toxicity: The Inhibition Concentration (ICp) Approach (Version 2.0). Environmental Research Laboratory, Duluth, Minnesota. ; Sponsor: 3M Company Sponsor Study ID# R1331 5 002728 AScI Corpontion/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 TABLE 1. EC5q'S, 95% Confidence Intervals, and 4-d NOEC'S from Algal Growth Inhibition Test with R1331 * Values Based on Nominal Test Concentrations Sponsor: 3M Company Sponsor Study ID# R1331 6 JSV 002729 AScl Corporation/AScI-Duluth Environmental Testing Division AScl Study ID# 5010-068 TABLE 2. Cells per ml and Percentage Difference from Mean Control Value for Selenastrum capricomutum Exposed to R1331 1 Teat Cone, 1 (oeg/L) Control 130 216 360 600 1000 Replicate A B C D E F Mean A B C Mean A B C Mean A B C Mean A B C Mean A B C Mean Id 4 2 3 5 3 5 3.7 2 4 2 2.7 (-28) 2 1 l 1.3 (-64) 3 1 2 2.0 (-46) 1 1 2 1.3 (-64) 2 1 2 1.7 (-55) Cells/ml (104) 2d 3 d 35 200 35 215 35 200 40 195 31 205 30 210 34.3 204 13 17 14 14.7 (-57) 69 61 64 64.7 (-68) 5 6 5 5.3 (-84) 39 49 40 42.7 (-79) 4 6 5 5.0 (-85) 3 4 5 4.0 (-88) 23 25 32 26.7 (-87) 18 14 14 15.3 (-92) 4 4 2 3.3 (-90) 10 6 10 8.7 (-96) 4d 726 660 627 616 704 638 662 : 310 270 275 285 (-57) 195 : 171 165 177 (-73) 75 ,, 78 * 93 82.0 (-88) 41 39 33 37.7 (-94) i2 13 14 13.0 (-$8) Parenthetic Values = % Difference from Control Mean Value * 0-d Mean Algal Count = 1.3 X 104 Sponsor: 3M Company Sponsor Study ID# R1331 7 00E730 AScl Corporation/AScI-Duiuth Environmental Testing Division AScl Study ID# 5010-068 TABLE 3. Average Specific Growth Rates (p) and Percentage Difference from Mean Control Value for Selenastrum capricomutum Exposed to R1331 I Teat Cono. Id Average Specific Growth Rat (/<)* 2d 3 d 4d Control 1.046 1.636 1.686 1.558 130 0.731 (-30) 1.213 (-26) 1.302 (-23) 1.348 (-13) 216 0 (-100) 0.703 (-57) - 1.164 (-31) 1.228 360 0.431 (-59) 0.674 (-59) 1.007 (-40) 1.036 (-34) :, 600 0 (-100) 0.562 (-66) 0.822 (-51) 0.842 (-46) ' 1000 0.268 (-74) 0.466 (-72) 0.634 (-62) 0.576 (-63) - * /t = In Cells per ml t,, - In Cella/ml U * -*0 b Parenthetic Values -- % Difference from Control Mean Value * O-d Mean Algal Count = 1.3 X 104 V; Sponsor: 3M Company Sponsor Study ID# R1331 8 002331
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