Document wggxnKLxKKybgwLa3KDLjpv14

AfU ^-c^ CHRONIC TOXICITY TO EARLY LIFE STAGE OF FISH TEST SUBSTANCE Identity: Perfluorooctanesulfonate; may also be referred to as PFOS or FC-95. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,77,8,8,8heptadecafluoro-, potassium salt, CAS #2795-39-3) Remarks field: 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, 1HHMR, 19F-NMR and elemental analyses techniques. METHOD Method: OECD 210, OPPTS 850.1400 Type: Flow-through chronic GLP. Yes Year completed: Study completed 1999. Report completed 2000 Species: Pimephales promelas Supplier: In-house cultures, Wildlife International, Ltd., Easton, MD Analytical monitoring: PFOS measured on days 0, 4, 7, 14, 21,28, 35, 42, and 47 Exposure period: 47 days Statistical methods: Discrete-variable data were analyzed 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 were evaluated for normality using ShapiroWilk'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. Test fish age: eggs < 24-hours old at test initiation Pretreatment: None Test conditions: Dilution water: 0.45 urn filtered well water Dilution water chemistry (during the 4-week period immediately preceding the test): hardness: 126 (124-128) mg/L as CaC03 alkalinity: 172 (170-172) mg/L as CaC03 pH: 8.2 (8.2-8.3) TOC: <1.0 mg/L Conductivity: 321 (315-330) ^mhos/cm GC0 3 3 2 Stock and test solution preparation: Primary stock prepared in dilution water at 88.4 mg/L and mixed until all test substance dissolved prior to use. After mixing, the primary stock solution was proportionally diluted with dilution water to prepare five additional stock solutions at concentrations of 44.2, 22.1, 11.0, 5.52, and 2.76 mg/L. Stock solutions were prepared every three to four days during the test. The six stocks were injected into the diluter mixing chambers (at a rate of 6.0 mL/minute) where they were mixed with dilution water (at a rate of 116 mL/minute) to achieve the desired test concentrations. Flow through rate: Approximately six volume additions of test water every 24-hours Stability of the test chemical solutions: Extremely stable Exposure vessels: 9L glass aquaria filled with approximately 7 L of test solution with a depth of approximately 17 cm. Embryo incubation cups were constructed from glass cylinders approximately 50 mm in diameter with 425 pm nylon screen mesh attached to the bottom with silicone sealant. The cups were suspended in the water column of each 9L glass aquarium and attached to a rocker arm with a reciprocating motion of approximately 2 rpm. Number of replicates: four Number offish per replicate, twenty Number of concentrations: six plus a negative control Feeding: Live brine shrimp nauplii. Fed 3 times per day during the first 7 days post-hatch. On days 8 through 40 post-hatch, fed 3 times daily on weekdays and 2 times daily on weekends. Not fed for at least 48 hours prior to the termination of test to allow for gut clearance prior to weight measurements. Water chemistry during the study: Dissolved oxygen range (0 - 47 days). 7.6 - 8.2 mg/L (control exposure) 7.6 - 8.2 mg/L (1.2 mg/L exposure) pH range (0 - 47 days) 8.0 - 8.4 (control exposure) 8.0 - 8.4 (1.2 mg/L exposure) Test temperature range (0 - 4 7 days) 24.4 - 24.7C (control exposure) 24.3 - 24.7C (1.2 mg/L exposure) Method of calculating mean measured concentrations, arithmetic mean RESULTS Nominal concentrations: Bk control, 0.14, 0.29, 0.57, 1.1,2.3, 4.6 mg/L Measured concentrations: <LOQ, 0.15, 0.30, 0.60, 1.2, 2.4, 4.6 mg/L 'r>o( '? *...> Element value: 5-day hatchability NOEC = 4.6 mg/L 42-day post-hatch survival NOEC = 0.30 mg/L 42-day post-hatch growth NOEC = 0.30 mg/L 42-day post-hatch survival LOEC = 0.60 mg/L All element values based on mean measured concentrations Statistical Evaluation of Mortality: The statistical difference for growth at concentrations equal to and higher than 0.60 mg/L was not evaluated due to a significant effect on survival. No statistically significant difference between the negative control and the highest concentration tested was seen for hatchability. 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 perfluorooctane sulfonate was used. No attempt was made to quantify on the basis of individual isomeric components. The LOQ (limit of quantitation) was 0.0458 mg/L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 102. Samples collected at pre test ranged from 91.4 to 105% of nominal. Samples at test initiation had measured values from 95.5 to 114% of nominal. Measured values for samples taken at test termination ranged from 95.2 to 111% of nominal. Summary of analytical chemistry data: N o m in a l T e st C on ce n tra tio n , m g/L M ean M easured M e a su re d D u p lic a te V a lu e s a t 0, 4 , 7, 1 4 ,2 1 , 28, 35, C o n c e n tra tio n , 42, and 4 7 D ays, R espe ctively, m g/L m g/L P ercent o f N om inal N e g a tive C ontrol A ll < LO Q 0 .1 4 0.147, 0.160, 0 .1 4 1 ,0 .1 4 0 , 0.144, 0.148, 0.134, 0.135, 0.153, 0 .1 43, 0.160, 0.158, 0.179, 0.173, 0.1 57, 0.160, 0.1 47, 0.155 0 .2 9 0.287, 0.277, 0.270, 0.289, 0.292, 0.296, 0.269, 0.266, 0.307, 0.315, 0.343, 0 .3 4 1 ,0 .3 1 1 , 0.325, 0.319, 0.313, 0.296, 0.276 0 .5 7 0.5 71, 0.576, 0.6 19, 0.659, 0.597, 0.642, 0.539, 0.535, 0.608, 0.580, 0.639, 0.617, 0.646, 0.644, 0.575, 0.576, 0.545, 0.543 1.1 1 .1 4 , 1 .1 3 , 1 .2 1 , 1 .2 5 , 1 .1 3 , 1 .2 3 , 1 .0 3 , 1 .1 0 , 1 .1 9 , 1 .2 4, 1,30, 1,31, 1.30, 1.31, 1.14, 1.19, 1.13, 1.09 . 0 .1 5 0 .3 0 0 .6 0 1 .2 107 103 105 109 2 .3 2 .2 1 ,2 .2 7 , 2 .5 2 , 2 .4 6 , 2 .4 3, 2 .3 8 , fish all d e a d at Day 7 4 .6 4 .5 6 , 4 .4 0 , 4 .7 9 , 4 .7 9 , 4 .4 6 , 4 .7 6 , fis h all d e a d at Day 7 2 .4 4 .6 104 100 00030- Biological observations Hatching success and time to hatch: All viable fathead minnow embryos hatched on Day 4 or 5. There were no apparent differences between the time to hatch in the negative control and the PFOS treatment groups. Survival: All fish surviving to test termination appeared normal with no overt signs of sublethal toxicity. Fish which did not survive generally appeared to be swimming erratically prior to death. Growth: Fish exposed to PFOS at concentrations of 0.15 or 0.30 mg/L for 42 days post-hatch showed no statistically significant reduction in total length, wet weight or dry weight in comparison to the negative control. Hatchability M ean M easured C o n ce n tra tio n , m g/L Number of Eggs Exposed N um ber H atched, Day 3 N um ber H atched, Day 4 N ega tive C ontrol 80 0 20 0 .1 5 80 0 18 0.3 80 0 14 0.6 80 0 28 1.2 80 0 25 2.4 80 0 16 4 .6 80 0 14 N um ber H atched, Day 5 54 58 58 48 49 59 60 T o ta l N um ber H atched 74 76 72 76 74 75 74 P ercent H a tch in g Success 93 95 90 95 93 94 93 Larval Survival M ean M easured C o n ce n tra tio n , m g/L P ercent S urvival, D ay 42 N ega tive C ontrol 88 0 .1 5 0 .3 0 .6 1 .2 2 .4 4 .6 79 81 66 5 .4 0 0 Growth M ean M easured C o n ce n tra tio n , m g/L Number of S urviving L a rv a e T otal Length Mean + SD, mm W e t W e ig h t Mean + SD, mg Dry W e ig h t Mean + SD, mg N ega tive C ontrol 0 .1 5 0 .3 0 0 .6 0 1.2 2 .4 4 .6 65 2 6 .5 + 0.721 158 + 9.10 3 2 .5 + 1.20 60 26 .6 + 0.208 160 + 3.10 33 .3 + 0.900 58 26.6 + 0.813 167 + 1 1 .9 3 4 .2 + 2.70 50 2 6 .5 + 0.399 166 + 11.3 33.5 + 2.70 4 26.7 + 2.02 185 + 33.8 35,4 + 6.66 _0 . . 0__ CONCLUSIONS Fathead minnows exposed to potassium perfluorooctanesulfonate at concentrations < 0.30 mg/L for 42 days post-hatch showed no statistically significant reductions in time to hatch, hatching success, survival or growth. The most sensitive endpoint in this study was post-hatch survival. 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