Document MJOy7n80RRY6Bx7oEwXOmDwLV

mi-oioD CHRONIC TOXICITY TO THE SALTWATER MYSID TEST SUBSTANCE Identity: Perfluorooctanesulfonate; may also be referred to as PFOS or FC-95. (1-Octanesulfonic add, 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, 19FNMR and elemental analyses techniques. METHOD Method: OPPTS 850.1350 Type: Flow-through chronic GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Mysidopsis bahia Supplier: In-house cultures, Wildlife International, Ltd., Easton, MD Analytical monitoring: PFOS measured on days 0, 7, 14, 21,28, and 35 Exposure period: 35 days Statistical methods: Survival data was evaluated (prior to pairing and after pairing) using 2 X 2 contingency tables to identify treatment groups that showed a statistically significant difference (p<0.05) from the negative control group. All continuous-variable data (reproduction and growth) were evaluated for normality using Shapiro-Wilk's test and for homogeneity of variance using Bartlett's test. Analysis of variance and Dunnett's test were used to evaluate differences between treatment and control means. All statistical tests were performed using a personal computer with SPSS/PC Version 2.0 or "TOXSTAT Release 3.5" statistical software. Test mysids age: < 24-hours old at test initiation Pretreatment: None Test conditions: Natural seawater diluted to 20%0with well water, 0.45pm filtered. Dilution water chemistry (during the 4-week period immediately preceding the test): Salinity: 20 (20-20) /00 TOC: < 1.0 mg/L 001184 Stock and test solution preparation: Primary stock prepared at 0.0895 mg/L and mixed for approximately 24 hours prior to use. After mixing, the primary stock solution was proportionally diluted with dilution water to prepare five additional stock solutions at concentrations of 0.0447, 0.0224, 0.0112, 0.00559, and 0.00280 mg/L. The six stocks were injected into the diluter mixing chambers (at a rate of 4.60 mL/minute) where they were mixed with dilution water (at a rate of 150 mL/minute) to achieve the desired test concentrations. Flow through rate: Approximately eleven volume additions of test water every 24- hours Stability of the test chemical solutions: Extremely stable Exposure vessels: Prior to pairing, mysids placed in glass beakers with nylon mesh screen attached to two holes on opposite sides. After reaching sexual maturity, pairs placed in glass petri dishes with sides of nylon mesh screen attached with silicone adhesive. Both pre-pairing and post-pairing exposure vessels were placed in 9L glass aquaria filled with approximately 5 L of test solution. The depth was approximately 6.2 cm prior to pairing and 5.5 cm after pairing. The test chambers for the second generation exposure were 2L beakers with 1L of test solution which was dipped out of a test chamber from the appropriate treatment group. Number of replicates: four Number of concentrations: six plus a negative control Number offish per replicate: Fifteen juveniles before pairing, 5 pairs (10 adults) when possible after pairing. Feeding: Fed live brine shrimp nauplii 3 or four times per day. Not fed the last day of the test. Water chemistry during the study: Dissolved oxygen range (0 - 35 days): 6.0 - 6.4 mg/L (control exposure) 5.9 - 6.3 mg/L (1.3 mg/L exposure) pH range (0 - 35 days) 8.2 - 8.4 (control exposure) 8.3 - 8.4 (1.3 mg/L exposure) Test temperature range (0 - 35 days) 24.5 - 25.2C (control exposure) 24.4 - 25.1 C (1.3 mg/L exposure) Method of calculating mean measured concentrations: arithmetic mean 001155 RESULTS Nominal concentrations: Bk control, 0.086, 0.17, 0.34, 0.69, 1.4, 2.7 mg/L Measured concentrations: <LOQ, 0.057, 0.12, 0.25, 0.55, 1.3, 2.6 mg/L Element value: 20-day survival (pre-pairing) NOEC = 0.55 mg/L 35-day (post-pairing) survival NOEC = 0.55 mg/L 35-day reproduction NOEC = 0.25 mg/L 35-day growth NOEC = 0.25 mg/L 35-day reprod & growth LOEC = 0.55 mg/L 2nd generation acute survival NOEC = 0.55 mg/L (highest concentration tested) All element values based on mean measured concentrations 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 0.0.0458 mg/L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 92.8. Samples collected at pre test ranged from 57.4 to 99.3% of nominal. Samples at test initiation had measured values from 67.1 to 103% of nominal. Measured values for samples taken at test termination ranged from 59.8 to 90.0% of nominal. C0HS6 Summary of analytical chemistry data: N om inal T e st C oncentration, m g/L M easu red Duplicate V alu es at 0, 7, 14, 2 1 ,2 8 , 35, D ays, Respectively, m g/L M ean M easured C o n c e n tra tio n , m g/L Percent of N om inal N eg ative Control All < L O Q 0 .0 8 6 0.0 694 , 0.0578, 0.0478, 0.0619, 0.0606, 0.0614, 0.0 554 , 0.0509, 0.0 515 , 0.0569, 0.0 580 , 0.0514 0 .0 5 7 . 66 0 .1 7 0.125, 0.114, 0.0778, 0.125, 0.124, 0.127, 0.0970, 0 .1 1 2 , 0.122, 0 .1 28, 0.1 24, 0.119 0 .1 2 71 0 .3 4 0 .2 89, 0.286, 0 .2 3 1 ,0 .1 9 7 , 0.276, 0.253, 0.227, 0 .2 1 2 , 0 .2 6 2 , 0 .2 7 1 ,0 .2 7 8 , 0.251 0 .2 5 74 0 .6 9 0.562, 0.659, 0 .5 8 1 ,0 .4 5 0 , 0.543, 0.542, 0.516, 0 .5 2 8 , 0.529, 0 .5 44, 0.556, 0.583 0 .5 5 80 1.4 1 .2 3, 1 .3 2, 1 .1 3, 1.20, 1.35, 1 .2 7, 1.23, 1.15, 1.39, 1.39, 1 .2 6 ,1 .2 0 2 .7 2 .5 6 , 2 .7 9 , 2 .5 8 , 2 .3 0 , 2 .5 4 , 2 .6 9 , all m ysid s dead after 14-days exposure 1.3 2 .6 93 96 Biological observations Survival: All surviving mysids appeared normal. Survival in the 1.3 and 2.6 mg/L treatments were statistically significantly different from the negative control group. Reproduction: The day of first brood release in this study was Day 22. Dunnett's test showed that reproduction was significantly reduced in the 0.55 mg/L treatment group when compared to the negative control (p <_0.05). The 1.3 and 2.6 mg/L treatment groups were not included in the statistical analysis of the reproduction data due to a statistically significant difference in survival. Growth: Mysids exposed to PFOS at concentrations < 0.25 mg/L showed no statistically significant reductions in length or dry weight (p < 0.05). Second Generation Acute Exposure: Survival in all PFOS treatment groups was > 95% and was not statistically different from the controls. All surviving mysids in the second generation exposure appeared normal with no overt signs of toxicity. C0I37 Percent Survival M ean M easured C o n c e n tra tio n , m g/L J u v e n ile P re-Pairing S u rv iv a l, Day 20 A d u lt P o s t-P a irin g S urvival, Day 35 N eg ative Control 78 92 0 .0 5 7 0 .1 2 0 .2 5 0 .5 5 1 .3 2 .6 92 96 75 90 82 97 83 95 32 57 0_ Second Generation Survival M ean M easured Concentrations, m g/L Total N um ber Exposed N eg ative Control 71 0 .0 5 7 65 N um ber Alive after 9 6 -h o u rs 68 63 0.1 2 0 .2 5 0 .5 5 83 79 62 59 13 13 Percent S u rv iv a l 96 97 95 95 100 Adult Mysid Growth M ean M easured C oncentration, m g/L N eg ativ e Control 0 .0 5 7 0.1 2 0.2 5 0.5 5 1 .3 Number of S u rv iv in g M ysids/N um ber Exposed Total Length, Mean + SD, mm Dry W eight, Mean + SD, mg 3 6 /3 9 6.4 3 + 0 .0 6 3 4 0.634 + 0.0510 4 4 /4 6 6.43 + 0 .0 7 2 9 0.599 + 0.0276 3 6 /4 0 6 .5 6 + 0.105 0.641 + 0.0241 3 6 /3 7 6.40 + 0 .0 5 4 8 0.622 + 0.0227 3 5 /3 7 6.14 + 0 .0 7 9 4 0.562 + 0 .0 0 6 2 4 8 /1 4 5 .8 5 + 0.178 0 .4 3 6 + 0.0441 C011S3 Reproduction M ean M easured C oncentration, m g/L R eplicate N eg ativ e Control A 0 .0 5 7 B C D A 0.1 2 B C D A 0 .2 5 B C D A 0 .5 5 B C D A B C D 1.3 A B C D Number of R eproductive D ays 70 53 70 42 60 70 70 56 70 46 54 70 70 56 61 56 54 56 70 56 22 14 0 11 Number of Young Mean Number of Y oung/R eproductive Day O v e ra ll Mean + SD 18 0 .2 5 7 0.315 + 0 .0 9 2 5 14 0.264 20 0.286 19 0.452 17 0 .2 8 3 0.261 + 0.0 873 14 0.200 13 0.186 21 0 .3 7 5 21 0 .3 0 0 0.361 + 0 .1 0 1 22 0.478 22 0.407 18 0.257 19 0 .2 7 1 0.2 52 + 0 .0 7 2 3 12 0.214 21 0.3 44 10 0.179 3 0 .0 5 5 6 0.0 559 + 0.0 376 6 0.107 3 0.0429 1 0.0179 0- 000- 001189 CONCLUSIONS There were no statistically significant effects on survival, reproduction or growth of mysid shrimp exposed to potassium perfluorooctanesulfonate at concentrations < 0.25 mg/L for 35 days. Reproduction, length and dry weight were the most sensitive biological endpoints in this study. Second generation mysids exposed to PFOS during a static 96-hour exposure showed no adverse effects. Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 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 001130