Document 1gp19gxG86jr437db63OvkqDo

AM%-0S3& TOXICITY TO AQUATIC PLANTS (E.G., ALGAE) TEST SUBSTANCE Identity: Perfluorooctylsulfonate, didecyldimethylammonium salt; may also be referred to as Fluoroalkyl ammonium derivative. [1Decaminium, N-decyl-N,N-dimethyl-, salt with 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafluoro-1 octanesulfonic acid (1:1), CAS # 251099-16-8] Remarks: The 3M production lot number was Lot 1. The test sample is L-14394 referred to by the test laboratory as P3025. The sample was labeled F-11615, Lot 1. The test sample is a mixture of the test substance in water (approximately 30-40% test substance, 60-70% water, and 0-5% of residual perfluorochemicals). All values reported relate to this mixture. The test sample appears to be a 2-phase dispersion (clear liquid with opaque solid) which rapidly separates after agitation. No calculations were made to adjust for the actual concentration of the test substance in the test sample. METHOD Method: OECD 201 Test: Static acute GLP: No Date exposure completed: 1/17/97 (Repeat of earlier study completed 1/11/97) Species: Selenastrum capricornutum Source: Originally from The American Type Culture Collection (ATCC), Strain 22662. Element basis: Growth rate and number of cells/mL. Analytical monitoring: Nominal concentrations only. pH, conductivity, temperature were monitored. Exposure period: 96-hours Start date: 1/13/97 End date: 1/17/97 Test organisms laboratory culture. Algae cultures were growing in OECD-recommended culture medium at AScI Corporation, Duluth, MN for 4 days before test initiation. Test Conditions: Test temperature range: 24.6 - 25.6C Dilution water source: The algal medium was prepared to OECD recommended concentrations by spiking deionized water with nutrient stocks (including Na2EDTA). The medium was sterilized 004254 via 0.22 pm filtration prior to use in the test. The pH of the synthetic algal medium at test initiation was 7.95. Stock and test solution preparation: Water accommodated fractions. Test solutions were prepared for each test concentration by mass addition of test substance in 500 ml_ of algal medium. The solutions were vigorously stirred for 21 -hours (vortex 1/2 solution depth). The aqueous phase was pipetted from the vessels at mid depth after the solutions had settled for 1-hour. Exposure vessels: Sterile 250 mL borosilicate Erlenmeyer flasks plugged with foam stoppers containing 100 mL of test solution. Agitation: Shaken continuously at 100 rpm Number of replicates: Six for control, three per test substance loading Initial algal cell loading: 1.2 X 104cells/mL Number of concentrations: Five plus a negative control Water chemistry: pH range (0 - 96 hours) 7.95 - 9.43 (control exposure) 7.91 - 7.78 (400 mg/L exposure) Conductivity range (0 - 96 hours) 134- 150 pmhos/cm (control exposure) 1 5 4 -1 6 6 pmhos/cm (400 mg/L exposure) Test temperature range (0 - 96 hours) 24.6 - 25.6C Light levels: (0 - 96 hours) 730- 750 ft-c from continuous cool-white fluorescent lighting Remarks: Inconsistent growth inhibition at increasing exposure concentrations was seen when the study was first run using exposure concentrations from 5.2 to 40 mg/L. In this first study, the 48, 72, and 96hour ErLso (growth rate) could not be calculated, because less than 50% growth inhibition was seen at all exposure concentrations. The study was repeated at higher test substance concentrations. In the second study, complete inhibition was seen at every exposure concentration loading except the lowest two concentrations, 86.4 (growth rate) and 51.8 mg/L (cell density, growth rate). As a result, the 48, 72, and 96-hour EL5o(cell density) could not be calculated and the ErLso at each time period was slightly higher than the lowest exposure concentration. RESULTS Nominal concentrations: Bk control, 51.8, 86.4, 144, 240, 400 mg/L. Element value: 24-hour EL50 (cell density) = 55.8 (50.2 - 62.1) mg/L 24-hour Erl_5o (growth rate) = 57.7 (54.2-61.5) mg/L 48-hour EL50 (cell density) = <51.8 mg/L (Cl not calculable) 48-hour ErLso (growth rate) =54.8 (50.4-59.6) mg/L 72-hour EL50 (cell density) = <51.8 mg/L (Cl not calculable) 72-hour ErLso (growth rate) = 61.4 (58.2-64.9) mg/L 96-hour EL50 (cell density) = <51.8 mg/L (Cl not calculable) 96-hour ErLso (growth rate) = 64.6 (62.5-66.9) mg/L 96-hour NOEL (cell density): 7.7 (7.5-8.1) mg/L* 96-hour NOEL (growth rate): 29.3 (27.7-31.6) mg/L* *NOEL values are approximations using the IC10 as the endpoint All element values based on nominal concentrations. Statistical methods: Cell densities, growth rates and percent inhibition values were calculated using an Excel spreadsheet. The Trimmed Spearman-Karber method and the Inhibition Concentration (ICp) approach (Version 2.01, USEPA-ERL, Duluth, MN) were used for EL and NOEL determinations. Control response: satisfactory Biological observations after 96-hours: N om inal Loading Test C oncentration, m g/L Mean Number o f C ells per mL Percent Inhibition via D ensity Percent Inhibition via Growth R ate B lank Control 5 1 .8 8 6 .4 144 240 400 6 ,1 3 0 ,0 0 0 2 ,0 4 0 ,0 0 0 2 0 ,0 0 0 1 0 ,0 0 0 1 7 ,0 0 0 7 ,0 0 0 . -67 -100 -1 0 0 -100 -100 . -1 8 -9 2 -100 -94 -100 Observations: Algal cell counts in each test vessel were determined by means of direct microscopic counts with an Improved Newbauer hemacytometer. CONCLUSIONS The test sample 96-hour EL50 interval for Selenastrum capricornutum was determined using two calculation methods. By cell density, it was <51.8 004256 mg/L and by growth rate 64.6 (62.5 - 66.9) mg/L. The 96-hour NOEL was determined to be 7.7 mg/L using cell density and 29.3 mg/L using growth rate. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY Reliability: Klimisch ranking 3. The study lacks analytical measurement of test substance concentrations in the test solutions and sample purity is not sufficiently characterized. Accurate EL5o's could not be calculated at each observation period due to the almost complete inhibition of cell growth at all exposure concentrations. Additionally, data is for a mixture and toxicity cannot be positively attributed to didecyldimethylammonium Perfluorooctylsulfonate salt alone. Report quality: Two definitive studies were conducted on this test substance at the same laboratory, in the same time period. A careful examination of the raw data revealed several errors during report preparation. These are: 1. The "study completed" dates printed on the report covers (January 29, 1997 and February 21,1997) are switched with each other. The sections under "1.0 General Information" for "Definitive Test Dates" are correct for each report, however. 2. The discussion under section "3.0 Test Methods" for "Test loadings" are switched. However, the test concentrations listed in this section are correct in each report. 3. The algicidal/algistatic determination test was only conducted after the second study. The discussion under "4.0 Results" in the report with the cover reading January 29, 1997 should not have been included. No evidence was seen in the raw data that the algicidal/algistatic determination was conducted during the first study. The raw data from the second study revealed that the test substance was, in fact, algicidal. REFERENCES This study was conducted at AScI Corporation, Environmental Testing Division, Duluth, MN at the request of the 3M Company. OTHER Last changed: 5/24/00 G04257 * AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 STUDY TITLE GROWTH INHIBITION OF P 'fo 'r f GREEN ALGA (Selenastrum capricomutum) DATA STANDARD OECD GUIDELINE 201 STUDY COMPLETED JANUARY 29, 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# P3025 Sponsor: 3M Company Sponsor Study ID# P3025 1 004258 AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 l.o 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 p3c^S" Selenastrum capricomutum). 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. P3025. AScI Corporation/AScI-Duluth Environmental Testing Division, 4444 Airpark Boulevard, Duluth, MN 55811; Tel. No. (218) 722-4040. Todd McGuire Joe Amato Donald Mount January 13-17, 1997. 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 5581 1; Tel. No. (218) 722-4040. Sponsor: 3M Company Sponsor Study ID# P3025 2 004259 AScI Corporaticm/AScI-Duluih Environmental Testing Division AScI Study ID# 5010-068 2.0 OBJECTIVE(S) To determine the 4-d median effect loading (EL50) and the 4-d no observable effect loading (NOEL) of the test substance, " 'fc 3-<, for green alga (Selenastrum capricomutum) under static test conditions. This study was conducted according to OECD Guideline 201 (OECD 1993). Test solution preparation was water-accommodated fraction (WAF) per Girling & Whale 1994. 3.0 TEST METHODS Test Substance Properties: , two phase liquid, specific gravity ca. 1.2 (water = 1), stable, and insoluble in water. Test Organisms Test loadings: 0 (algal medium control), 51.8, 86.4, 144, 240, and 400 mg/L. Test loading range was determined from range-finding test results. Each test loading was prepared by mass addition of test substance to 500-ml aliquots of algal medium. Test substance was thoroughly homogenized before each mass was determined. Each solution was vigorously stirred (vortex 1/2 of flasks' depth) for 21 hours to obtain WAF's. Test solution obtained from flasks by pipetting from mid-depth. Source: ATCC 22662 Inoculum preparation: Algal stock was subcultured in OECD recommended medium for 4 days before test initiation. Algal Medium 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 (1.2 X 104 actual concentration). 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 7.95. Sterilized via 0.22 filtration prior to use in the test. Sponsor. 3M Company Sponsor Study ID# P3025 3 0*04260 Exposure Chambers Incubation Observations Endpoint Calculations AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 Type: Sterilized 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 loading were employed. During the test, kept stoppered, except when experimental observations were made. Duration: 4 days. Daily photoperiod: Continuous at 740 ft-candles using cool-white fluorescent lamps. Test chambers held on shaker table set at 100 rpm. Temperature: 25.0 2 C. Biological: Daily cell counts using direct microscopic enumeration via Improved Neubauer hemocytometer. Water chemistry: (1) At test initiation and termination- pH, conductivity, and temperature, (2) daily- temperature. All determinations were made according to APHA methods (1995). 1-, 2-, 3-, and 4-d EL50's, and 4-d NOEL based on both cell counts and average specific growth rates (ji). Sponsor: 3M Company Sponsor Study ID# P3025 4 004261 AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 4.0 RESULTS Test Substance Toxicity (Pertinent data are in Tables 1, 2, and 3) Test Conditions Cell Count 1-d EL5Q: 55.8 mg/L (50.2-62.1). H 1-d EL5Q: 57.7 mg/L (54.2-61.5). Cell Count 2-d EL50: < 51.8 mg/L. fi 2-d ELS0: 54.8 mg/L (50.4-59.6). Cell Count 3-d EL50: < 51.8 mg/L. H 3-d ELS0: 61.4 mg/L (58.2-64.9). Cell Count 4-d EL50: < 51.8 mg/L. H 4-d EL5Q: 64.6 mg/L (62.5-66.9). Cell Count 4-d NOEL: 7.7 mg/L (7.5-8.1). ft 4-d NOEL: 29.3 mg/L (27.7-31.6). NOEL values are approximations using the IC 10 as the endpoint. Effect of test substance was determined to be algistatic based on the results of a post definitive test exposure. Temperature (C): 24.6-25.6. Light intensity ft-cdls: 730-750. pH: 7.78-9.43. Specific conductivity (imhos/cm): 134-166. Sponsor: 3M Company Sponsor Study ID# P3025 5 094262 5.0 TEST QA/QC AScl Corporation/AScI-Duluth Environmental Testing Division AScl Study ID# 5010-068 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 191. 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. Girling, A .E., G.F. Whale, and D.M.M. Adema. 1994. A Guideline Supplement for Determining the Aquatic Toxicity of Poorly Water-Soluble Complex Mixtures Using WaterAccommodated Fractions. Chemosphere 29(12):2645-2649. Organization for Economic Cooperation and Development (OECD). 1993. OECD Guidelines for 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. 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# P3025 6 AScI Corpo ration/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 TABLE 1. ELso'S, 95% Confidence Intervals, and 4-d NOEL'S from Algal Growth Inhibition Test with iJ503-5 Endpoint Cell Count Time: " 1-d EL50 2-d EL50 3-d ELS0 4-d ELS0 4-d NOEL Data* (mg/L) 55.8 < 5 1 .8 < 5 1 .8 <51.8 7.7 95% Confidence Interval (mg/L) 50.2-62.1 Not Calculable Not Calculable Not Calculable 7.5-8.1 Endpoint Average Specific Growth Rate (p.) 1-d EL50 2-d EL50 3-d ELj0 4-d EL50 4-d NOEL" (mg/L) 57.7 54.8 61.4 64.6 29.3 * NOEL values are approximations using the IC10 as the endpoint. 95% Confidence Interval (mg/L) 54.2-61.5 50.4-59.6 58.2-64.9 62.5-66.9 27.7-31.6 Sponsor: 3M Company Sponsor Study ID# P3025 7 * 004264 ASc! Corporation/AScI-Duluth Environmental Testing Division AScI Study ID// 5010-068 TABLE 2. Cells per ml and Percentage Difference from Mean Control Value for Selenastrum capricomutum Exposed to A Test Loading (tng/L) Control 51.8 86.4 144 240 400 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 1d 8 5 7 5 5 6 6.0 (-) 3 5 2 3.3 (-44)* 1 0 0 0.3 (-94) 2 0 1 1.0 (-83) 1 2 1 1.3 (-78) 0 0 0 0 (-100) Cells/ml (104) 2d 3 d 38 36 37 34 33 36 35.7 (-) 200 232 256 232 200 256 229 (-) 10 7 7 8.0 (-78) 59 42 41 47.3 (-79) 1 1 2 1.3 (-96) 2 1 3 2.0 (-99) 2 1 0 1.0 (-97) 1 1 3 1.7 (-99) 1 1 0 0.7 (-98) 1 2 0 1.0 (-100) 1 0 0 0.3 (-99) 1 0 0 0.3 (-100) 4d 585 615 675 570 600 630 613 (-) 198 222 192 204 (-67) 3 1 2 2.0 (-100) 0 1 2 1.0 (-100) 1 2 2 1.7 (-100) 1 0 1 0.7 (-100) * Parenthetic Values = % Difference from Control Mean Value * 0-d Mean Algal Count = 1.2 X 104 Sponsor: 3M Company Sponsor Study ID# P3025 8 AScI Corporation/AScI-Duluth Environmental Testing Division AScI Study ID# 5010-068 TABLE 3. Average Specific Growth Rates ip.) and Percentage Difference from Mean Control Value for Selenastrum capricomutum Exposed to f jc ^ (mg/L) Control 51.8 86.4 144 240 400 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 1d 1.897 1.427 1.764 1.427 1.427 1.609 1.609 (-) 0.916 1.427 0.511 1.012 (-37)b -0.182 0 0 -1.386 (-100) 0.511 0 -0.182 -0.182 (-100) -0.182 0.511 -0.182 0.080 (-95) 0 0 0 0.000 (-100) Average Specific Growth Rate O')* 2d 3 d 1.728 1.701 1.714 1.672 1.657 1.701 1.696 (-) 1.705 1.755 1.788 1.755 1.705 1.788 1.751 (-) 1.060 0.882 0.882 0.949 (-44) 1.298 1.185 1.177 1.225 (-30) -0.091 -0.091 0.255 0.040 (-98) 0.170 -0.061 0.305 0.170 (-90) 0.255 -0.091 0 -0.091 (-100) -0.061 -0.061 0.305 0.116 (-93) -0.091 -0.091 0 -0.269 (-100) -0.061 0.170 0 -0.061 (-100) -0.091 0 0 -0.693 (-100) -0.061 0 0 -0.462 (-100) 4d 1.547 1.560 1.583 1.541 1.554 1.566 1.559 (-) 1.276 1.305 1.269 1.284 (-18) 0.229 -0.046 0.128 0.128 (-92) 0 -0.046 0.128 -0.046 (-100) -0.046 0.128 0.128 0.087 (-94) -0.046 0 -0.046 -0.135 (-100) * n = In Cells per ml - In Cells/ml U -o b Parenthetic Values = % Difference from Control Mean Value * 0-d Mean Algal Count = 1.2 X IO4 Sponsor: 3M Company Sponsor Study ID# P3025 9 004266