Document O3w4bNRw37BeaG8jmMpyxVorM

ARZ%-ortH TOXICITY TO AQUATIC PLANTS (E.G., ALGAE) TEST SUBSTANCE Identity: Perfluorooctanesulfonate; may also be referred to as PFOS or FC-95. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt, CAS # 2795-39-3) Remarks: The test substance is a white powder. Sample was taken from 3M lot number 217. Sample was stored under ambient conditions prior to testing. Purity determined to be 90.49% by LC/MS, 1H-HMR, 19F-NMR and elemental analyses techniques. METHOD Method: OECD 201, OPPTS 850.5400, ASTM 1218-90E Test: Static acute GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Selenastrum capricornutum Source: Originally from The Culture Collection of Algae at the University of Texas at Austin, maintained in culture medium at Wildlife International Ltd., Easton, MD Analytical monitoring: PFOS measured at 0, 72, 96-hours Element basis: Reported three ways: number of cells/ml, area under the growth curve and growth rate Exposure period: 96-hours Start date: 4/12/99 End date: 4/16/99 Analytical monitoring: Test concentrations measured at 0, 72, and 96hours. Test organisms laboratory culture: Algae cultures had been actively growing in freshwater algal culture medium for at least two weeks prior to test initiation. Stock nutrient solutions were prepared by adding reagentgrade chemicals to reverse osmosis-purified well water. Test Conditions: Test temperature range: 23.6 - 25.8C 000601 Growth medium: ASTM Standard Guide 1218-90E, 1990 C om pound M g C I2 6 H 20 C a C I2 2 H 20 H3BO 3 M n C I24H 20 Zn C I2 F e C I2 6 H 20 C o C I2 6 H 20 N a M o O jZ H O C u C I2 2 H 20 N a 2E D T A 2 H 20 N aN 03 M g S O 4 7 H 20 k 2h p o 4 N aH C 03 Nom inal C oncentration 1 2 .1 6 4 .4 0 0 .1 8 5 6 0 .4 1 6 3 .2 8 0 .1 5 9 8 1 .4 2 8 7 .2 6 0 .0 1 2 0 .3 0 0 2 5 .5 0 1 4 .7 0 1 .0 4 4 1 5 .0 U n its m g/L m g/L m g/L m g/L ug/L m g/L ug/L ug/L ug/L m g/L m g/L m g/L m g/L m g/L Dilution water source: Wildlife International Ltd. well water purified by reverse osmosis. The test medium was prepared by adding the appropriate volumes of stock nutrient solutions to purified well water. The pH of the medium was adjusted to 7.5 + 0.1 using 10% HCI and the medium was sterilized by filtration (0.22pm) prior to use. Stock and test solution preparation: A primary stock solution was prepared in algal medium at a concentration of 183 mg/L. The primary stock solution was stirred with a magnetic stir plate for approximately 24 hours. After mixing, the primary stock solution was proportionally diluted with algal medium to prepare the five additional test concentrations. All final test solutions appeared clear and colorless. Exposure vessels: Sterile 250 mL polycarbonate Erlenmeyer flasks plugged with foam stoppers containing 100 mL of test solution. Agitation: Shaken continuously at 100 rpm Number of replicates: three Initial algal cell loading: 1.0 X 104cells/mL Number of concentrations: seven plus a negative control plus an abiotic control at the highest concentration tested 000602 Water chemistry: pH range (0 - 96 hours) 7.5 - 8.1 (control exposure) 7 .4 -7 .5 (179 mg/L exposure) Test temperature range (0 - 96 hours) 23.6 - 25.8C Light levels: (0 - 96 hours) 3870 - 4610 lux from continuous cool-white fluorescent lighting Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Bk control, 5.7, 11,23, 46, 91, 183 mg/L plus 183 mg/L abiotic control. Measured concentrations: <LOQ, 5.5, 11, 21,44, 86, 179, 169 mg/L Element value: 24-hour EC5o(cell density) = 163 (74-191) mg/L 24-hour EbCso (area under curve) = 122 (19-176) mg/L 24-hour ErCso (growth rate) = 136 (30 - 204) mg/L 48-hour EC50 (cell density) = 81 (72 - 90) mg/L 48-hour EbCso (area under curve) = 84 (67 - 146) mg/L 48-hour ErC 5o(growth rate) = 142 (107 - 185) mg/L 72-hour EC10 (cell density) = 37 (<0 - 64) mg/L 72-hour EbC-io (area under curve) = 46 (<0 - 56) mg/L 72-hour ErC-io (growth rate) = 53 (23 - 64) mg/L 72-hour EC50 (cell density) = 70 (44 - 78) mg/L 72-hour EbCso (area under curve) = 74 (55 - 82) mg/L 72-hour ErCso (growth rate) = 120 (103 - 132) mg/L 72-hour ECg0 (cell density) = 153 (130 - 165) mg/L 72-hour EbCgo (area under curve) = 165 (145 - 176) mg/L 72-hour ErCgo (growth rate) = >179 mg/L (C.l. not calculable) 96-hour EC10 (cell density) = 49 (43 - 50) mg/L 96-hour EbC-io (area under curve) = 49 (40 - 50) mg/L 96-hour ErCio (growth rate) = 59 (54 - 63) mg/L 96-hour EC50 (cell density) = 71 (66 - 73) mg/L 96-hour EbC5o (area under curve) = 71 (67 - 74) mg/L 96-hour ErCso (growth rate) = 126 (115 - 138) mg/L 96-hour ECgo (cell density) = 137 (105 - 153) mg/L 96-hour EbCgo (area under curve) = 145 (125 - 155) mg/L 96-hour ErCgo (growth rate) = >179 mg/L (C.l. not calculable) 72-hour NOEC (growth rate, cell density, area under the curve): 44 mg/L 96-hour NOEC (growth rate, cell density, area under the curve): 44 mg/L 000603 All element values based on mean measured concentrations Statistical methods: Cell densities, area under the growth curve values, growth rates and percent inhibition values were calculated using "The SAS System for Windows", Release 6.12. These values were then analyzed by linear interpolation using TOXSTAT Version 3.5 to estimate the EC10, EC50, and EC9o values and 95% confidence limits at 72 and 96. Cell densities, areas under the growth curve and growth rates at 72 and 96 hours were also evaluated for normality and homogeneity of variances using the Shapiro-Wilks's test and Bartlett's test, respectively. The treatment groups were then compared to the control using Dunnett's test. Results of the statistical analyses were used to determine the NOEC values. Analytical Methodology: Analyses of test solutions were performed at Wildlife International Ltd. using high performance liquid chromatography with mass spectrometric detection (HPLC/MS). When determining the concentration of the test substance in the test solutions, the same and most prominent peak response for perfluorooctanesulfonate was used. No attempt was made to quantify on the basis of individual isomeric components. The LOQ (limit of quantitation) was 0.115 mg/L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 99.1. Samples collected at test initiation had measured values from 82.0 to 98.1% of nominal. Measured values for samples taken at 72-hours ranged from 91.7 to 105% of nominal. Measured values for samples taken at 96-hours ranged from 90.3 to 102% of nominal. For the abiotic controls, measured values for samples taken at 72-hours ranged from 81.9 - 105% of nominal and for samples taken at 96-hours, 90.3 - 103% of nominal. Summary of analytical chemistry data: N om inal T est C o n c e n tra tio n , m g/L N e g a tiv e Control M easured V alues at 0, 72, and 96hours, Respectively, m g/L All < L O Q M ean M easu red C o n c e n tra tio n , m g/L <LOQ Percent of Nom inal - 5 .7 4 .7 3 , 6.04, 5.84 5.5 96 11 10 .7, 11 .2, 12.1 11 1 0 0 23 19.8, 2 3 .1 ,2 0 .7 21 91 46 4 2 .7 ,4 1 .9 , 46.3 44 96 91 83.3, 86.0, 88 .3 86 95 183 179, 186, 172 179 98 18 3 (abiotic) not analyzed, 150, 188 169 92 000604 Control response: satisfactory Biological observations after 96-hours: M ean M easured C oncentration, m g/L Mean Number o f Cells per mL Percent Inhibition via D e n s ity Percent Inhibition via Area Under the Curve Percent Inhibition via Growth R ate N eg ativ e Control 5 .5 11 21 44 86 179 2 ,7 4 0 ,0 0 0 3 ,0 4 0 ,0 0 0 2 ,8 8 0 ,0 0 0 3 ,2 4 0 ,0 0 0 3 ,0 8 0 ,0 0 0 6 2 6 ,6 6 7 3 3 ,6 6 7 . -11 -5.1 -1 8 -1 2 77 99 -8 .5 -3 .3 -1 3 -5 .3 75 98 -1 .9 -0 .8 4 -3 .0 -2 .0 27 79 Observations: After 96 hours of exposure, there were no signs of aggregation, flocculation or adherence of the algae to the flasks in the negative control or any test treatment group. In addition, there were no noticeable changes in cell color or morphology when compared to the negative control, although a few cells appeared enlarged in the 86 and 179 mg/L treatment groups. Reversibility of Growth Inhibition: The 179 mg/L treatment group was maximally inhibited after 96-hours. Aliquots of the test solution were diluted with algal medium and cultured for five days. Based on the growth observed in the recovery phase, the effect on algal growth was found to be algistatic. CONCLUSIONS The potassium perfluorooctanesulfonate 96-hour ECso and 95% confidence interval for Selenastrum capricomutum was determined using three calculation methods. By cell density, it was 71 (66-73) mg/L, by area under the growth curve it was 71 (67-74) mg/L and by growth rate 126 (115-138) mg/L. The 96-hour NOEC was determined by Dunnett's procedure (p < 0.05) to be 44 mg/L using all three methods. No signs of aggregation, flocculation, or adherence were noted in any of the test solutions or the controls. This test substance was determined to be algistatic. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 000S05 DATA QUALITY Reliability: Klimisch ranking = 1 REFERENCES______________________________________________ This study was conducted at Wildlife International Ltd., Easton, MD at the request of the 3M Company. OTHER____________________________________________________ Last changed: 5/3/00 000806