Document YrOQz3br1nemQa0ZLZVovDgxE

p. 1 Ae- zZ~-37"7 ACUTE TOXICITY TO THE OLIGOCHAETE WORM (Lumbriculus variegatus) TEST SUBSTANC E 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 86 .9% by LC/MS, 'H-HMR, 19F-NM R and elemental analyses techniques . The impurities were identified as : metals (1 .45%) ; lower chain-length (C4-C7) perfluorinated sulfonates (8.41 %) ; inorganic fluoride (0 .59%) ; NMR impurities (1 .905%); perfluorooctanoate anion (0 .33%); and the total of tri-, penta-, hepta-, and nonapentafluoropropanoic acid (0.38%) . METHOD Method: ASTM E729-96 Type : Static renewal GLP: Yes Date completed : Study completed 2007 Species: Lumbriculus variegatu s Supplier : Wildlife International, Ltd . Cultures (originally from Environmental Consulting and Testing (ECT), Superior, WI) Analytical monitoring : PFOS measured at initiation, prior to and after renewal at 48 hours and at test termination (96-hours ) Exposure period : 96-hours Statistical methods : Bionomial probability method used to calculate all LC50 values . The NOEC determined by visual interpretation of mortality and observation data . Test organism age : Adult Average wet weight at test initiation : Not provided Loading : One worm per chamber 7P- retreatment : None Acclimation : Prior to the test, the test organisms were acclimated to test conditions for 2 weeks by holding them in a container with paper toweling and water from the same source and at approximately the sam e temperature that would be used in the test . Temperatures ranged from 23.3 to 24.4 C . The pH ranged from 8 .0 to 8 .4. Dissolved oxygen ranged from 7 .2 to 7 .9 mg/L . Organisms were fed a mixture of yeast, cereal grass media, and trout chow during acclimation . Test conditions : CONTAINS NO CBI ~3 ef G -12 p. 2 Dilution water : Filtered, aerated, well wate r Dilution water chemist ry (during the 4-week period immediately preceding the test) : Specific Conductance : 291 (285 - 300) pmhos/cm Hardness : 136 (132 - 140) mg/L as CaCO3 Alkalinity : 182 (180 - 184) mg/L as CaCO3 pH : 8 .1 (7.9 - 8.1 ) Stock and test solution preparation : A primary stock solution was prepared at a nominal concentration of 12 mg active ingredient per liter using a calculated amount of PFOS, and mixed by sonication for 15 minutes followed by inversion . After mixing, the primary stock solution was proportionally diluted with dilution water to prepare each test concentration . Each test solution was mixed by inversion . All test solutions were adjusted to 100% active ingredient during preparation, based on test substance purity . All test solutions appeared clear and colorless at test initiation, renewal and termination . Solvent : non e Concentrations dosing rate : Static renewal after 48-hours Stability of the test chemical solutions under test conditions : Stable as demonstrated by analytical monitoring . Exposure vessels : 30-mL Nalgene polypropylene plastic beakers filled with approximately 20 mL of water ; no substrate provided . Beakers covered with plastic wrap . Water depth was approximately 2 .2 cm . Number of replicates : Ten Number of worms per replicate : one Number of concentrations : Five plus a negative contro l Photo period : 16 hours light and 8 hours dark with a 30-minute transition in simulated natural sunlight from fluorescent bulbs . Feeding schedule : Test organisms were not fed during the test Water chemistry during the study : Dissolved oxygen range (0 - 96 hours) : 8 .0 - 8 .3 mg/L (negative control exposure) 7 .4 - 8 .4 mg/L (all test substance exposures) pH range (0 - 96 hours ) 8.2 - 8 .6 (negative control exposure) 8.2 - 8 .6 (all test substance exposures) Test temperature range (0 - 96 hours ) 22 .4 - 23.8C (negative control exposure) 22 .5 - 24.0C (all test substance exposures) RESULT S Nominal concentrations : Bk control, 0 .80, 1 .5, 3 .0, 6.0, and 12 mg/L q4- p. 3 Measured concentrations : <LOQ, 0.71, 1 .4, 2 .8, 5 .6, 11 mg/L Element value: 24-hour LC50 = 11 (C .I . = >5.6) mg/L 48-hour LC50 = 7 .9 (C .I . = >2 .8) mg/L 72-hour LC50 = 6 .5 (2 .8 - 11) mg/L 96-hour LC50 = 5 .6 (27 - 51) mg/L 96-hour NOEC = 2 .8 mg/L All element values based on mean measured concentrations Statistical Evaluation of Mo rtality: Binomial probability with nonlinear interpolatio n Analytical Methodology : Analyses of test solutions were performed at Wildlife International Ltd . The method consisted of dilution of the samples 1 :1, v/v with acetonitrile, followed by secondary dilution using acetonitrile : HPLC-grade bottled water (50 :50, v/v) and analysis by direct injection high performance liquid chromatography with mass spectrometric (LC/MS/MS) detection . The LOQ (limit of quantitation) was 0 .200 mg/L in this study . The measured concentrations of matrix fortifications analyzed concurrently during sample analysis were 96 .4-108% of nominal . Samples collected in exposure vessels at test initiation had measured values from 92 to 97% of nominal . New samples collected at 48 hours had measured concentrations from 85 to 94% of nominal . Old samples collected at 48 hours had measured concentrations ranging from 88 to 93% of nominal . Measured concentrations of PFOS in samples collected at test termination were 88 to 97% of nominal . Summary of analytical chemist ry data : Nominal Test Measured Values of New and Old Mea n Concentration, Test Solutions, mg/L(l) Measured Percent of mg/L Concentration, Nominal mg/L Negative Control All < LOQ( 2 ) <LOQ - 0 .80 0.749, 0 .707, 0 .683, 0 .700 0 .71 88.8 1 .5 1 .37, 1 .35, 1 .39, 1 .36 1 .4 93.3 3 .0 2 .87, 2 .80, 2 .73, 2 .88 2 .8 93.3 6 .0 5 .80, 5 .33, 5 .64, 5 .80 5 .6 93.3 12 11 .4,11 .0,11 .3,11 .6 11 91 .7 (1) Samples are listed in this order : Day 0 (new), 48-hours (old), 48-hours (new), 96-hours (old ) (2) LOQ = Limit of Quantitation = 0 .200 mg/L active ingredien t Biological observations after 96-hours : All oligochaetes in the negative control and the 0 .71, 1 .4, and 2.8 mg/L treatments appeared healthy and normal throughout the exposure period with no mortalities or overt signs of toxicity observed . At test termination, mortality in the 5 .6 and 11 mg/L treatment group was 50 and 100%, respectively . Signs of toxicity observed in the affected oligochaetes from the 5 .6 and 11 mg/L treatment groups included pale appearance and occurrence of fragmentation . 75 p. 4 Cumulative percent mo rtality: Mean Measured 24-hours 48-hours 72-hours 96-hours Test Conc. , mg/L Negative 0 0 0 0 Control 0 .71 0 0 0 0 1 .4 0 0 0 0 2 .8 0 0 0 0 5 .6 0 30 30 50 11 50 70 100 100 Lowest concentration causing 100% mo rtality: 11 mg/ L Mortality of controls : None CONCLUSIONS The potassium perfluorooctanesulfonate 96-hour LC50 for the oligochaete worm Lumbriculus variegatus was determined to be 5.6 mg/L . The 96hour no mortality concentration was 2 .8 mg/L and no observed effects concentration was 2.8 mg/L . Submitter : 3M Environmental Laboratory, Maplewood, Minnesota, 55144 DATA QUALITY Reliability: Klimisch ranking = 1 REFERENCE S This study was conducted at Wildlife International Ltd ., Easton, MD at the request of 3M . OTHER Last changed : 07/10/07