Document 4JOkYwv6R47yZYebRpZNVjnbe

BIOCONCENTRATION - REVISED 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: Sample from 3M production lot number217. The test substance is a white powder. Puritydeterminedto be 86.9% by LC/MS, 1H-HMR, 19F-NMRand elemental analyses techniques. METHOD: Method/guideline followed: US EPA OPPTS 850.1730 and OECD 305 Type: Flow-throughexposurewith flow-throughdepurationphase. GLP (Y/N): Yes Year: 2001, Report amended 2002 Species: Bluegill(Lepomis macrochirus) Supplier: Osage Catfisheries, Inc., Osage Beach, Missouri Length and weight at test termination: Mean length= 62 mm, range 56-66 mm Mean weight - 2.70 g, range 2.03 - 3.32 g Loading: 0.48 g fish/L/day(based on initialloadingof 90 fish per tank, using mean fish weight at the end of the studyand volume of water that passed through test chamber in 24-hours). Fish age: Approximately7 monthsat test initiation Analytical monitoring: Concentrationof PFOS in water and fish. Pretreatment: None Number of concentrations: Two plus a negative control Test concentrations (mean measured): Negative control,0.086 and 0.87 mg/L Uptake period: 62-days (0.086 mg/L exposure) 35-days (0.87 mg/L exposure- this exposureended after 35-days due to fish mortality) Depuration period: 56-days (0.086 mg/L exposure) None (0.087 mg/L exposure) Test conditions: Dilution water: Moderately-hard well water Dilution water chemistry: Specific conductance: 313 (310 - 315 umhos/cm) Hardness: 130 (128- 132 mg/L) Alkalinity: 178 (176 - 178) pH: 8.1 (8.0 - 8.2) Measured duringthe 4-week period immediatelyprecedingthe test. 4" Test conditions (cont.): Stock and test solution preparation: Two stock solutionswere prepared at 10 and 100 mg a.i./L. Stock solutionsstirredwith an electrictop-downmixer to aid in the solubilizationof the test substance. After mixing,the stocksappeared clear and colorless. Stockswere prepared at approximatelyweekly intervalsduringthe uptake phase. Stocksinjected into the dilutermixingchambersat a rate of 3.5 mL/minutewhere they were mixed with dilutionwater at a rate of 350 mL/minuteto achieve the desiredtest concentrations. All final test solutionsappeared clear and colorless. Diluter flow rate: Approx. 6.3 volume additionsper 24-hours Exposure vessels: 104 L stainlesssteel aquada filled with approximately80 L solution. Number of replicates: None - one vessel per concentration Number of fish per vessel: 90 Diet: Flake food, Ziegler Brothers, Inc., Gardners, PA Water chemistry ranges during the study: Dissolved oxygen, mglL: 6.8 - 8.6 6.8 - 8.6 6.4 - 8.2 Temperature r C: 21.8 - 22,0 21.7 - 22.0 21.7 - 21.9 I pH: N7e.g9.-C8o.n2trol 07.098-6 8m.2g/L 07.897- m8.g2/L Photoperiod: 16 hours lightand 8 hoursdark with a 30 minute transitionperiod. Light intensity: 278 lux at surface of the negative control vessel at test initiation Collection of tissue samples: Fish were collected from test chambers by randomselection at 12 time pointsduringthe 62-day uptake phase. They were euthanized, blotteddry, weighed and measured. Fish then rinsedwith dilutionwater, blotted dry again and dissectedintoedible and nonedibletissuefractions. The fractionswere individuallyweighed. The head, fins and viscerawere consideredto be nonedibletissue. The remainingtissue, includingskinwas consideredto be edible tissue. Statistical methods: Whole fish concentrationswere calculated based on the sum of the edible and nonedibleparts. Steady-state bioconcentrationBCF values were originallycalculatedfrom the tissue concentrationsat apparent steady-stateusingthe BIOFAC model. Upon further investigation,this model was deemed inappropriatefor use with this data set. The tissue concentrationshad reached a point where there were not statisticallysignificantdifferencesbetween the lastthree sample days. However,a plot of the data clearly shows a trend of increasing concentrationsin the tissues. In addition,the BIOFAC programis not accepted as appropriatefor surfactants. An amended report was issued with BCFK values calculated as outlined in the draft OPPTS 850.1730 Guidance Document. The kinetic bioconcentretionfactor (BCFK), uptake rate (kl) and depurationrate (k2) were calculatedfor the edible, nonedible and whole fish exposed to 0.086 mg/L. These rate constantswere then used to calculate a BCFK (BCFK = KI/K2) and half-lives for clearance for each tissue type. The resultsfrom this data reanalysisare presented below. RESULTS Nominal concentrations: Negative control,0.1 and 1.0 mg/L Mean measured concentrations: < 0.05, 0.086 and 0.87 mg/L Kinetic Bioconcentration factors (BCFK): 0.086 mglL exposure BCFK: Edible 1124 Nonedible 4013 Time to reach 50% clearance: 86 days 116 days Whole Fish 2796 112 days 0.87 mg/L exposure AlthoughBCF valueswere calculated usingthe BIOFAC software, the resultsare not reported here. All of the fish had died or been sampled priorto achieving steady-state. As a result,the BCF valueswere underestimatedand are notrelevant. PFOS Concentrations in Tissues of Bluegill Exposed to 0.086 mglL Values are from 4 individualfish at each sample period. Uptake Day Edible Tissue, mg/kg Nonedible Tissue, mglkg 0 (4-hours) 1 3 7 14 21 0.167, 0.155, 0.144, 0.182 0.734, 0.726, 0.631, 0.806 1.73, 2.07, 2.03, 2.11 3.73, 4.25, 4.73, 62.5 11.4, 9.07, 13.7, 12.6 11.7, 12.0, 12.9, 10.6 0.415, 0.519, 0.417, 0.497 1.68, 1.85, 1.72, 2.07 4.59, 5.50, 5.47, 5.97 10.2, 10.6, 11.9, 15.2 27.3, 23.2, 35.3, 32.6 33.3, 22.7, 24.6, 24.4 28 18.3, 13.7, 23.9, 23.1 49.4, 40.7, 65.3, 57.9 35 22.6, 27.7, 23.8, 20.6 67.1,73.3, 62.0, 59.1 42 27.6, 25.3, 21.2, 27.6 64.0, 68.1,54.4, 79.6 49 33.3, 36.2, 39.0, 30.6 85.0, 95.1,93.1, 77.7 56 48.3, 38.9, 44.1, 38.3 122, 94.2, 73.2, 106 62 42,46, 6.2, 42.2, 39.2 101, 112, 105, 96.4 Depuration Day 14 I 48.5, 31.8, 31.6, 42.0 124, 79.4, 81.8, 113 28 26.0, 33.3, 38.7, 55.8 85.7, 95.1, 85.7, 94.8 42 24.13, 1.2, 30.0, 33.0 71.7, 80.6, 78.3, 82.1 56 21.1,37.6, 32.9, 31.2 57.7, 80.3, 85.4, 84.4 Whole Fish Conc., mglkg 0.293, 0.351, 0.286, 0.363 1.26, 1.34, 1.29, 1.53 32-1,4.04, 4.18, 4.38 7.33, 7.66, 8.73, 11.4 20.2, 16.9, 26.0, 24.6 23.3, 18.4, 19.8, 18.5 35.3, 29.2, 45.4, 44.1 46.3, 53.8, 46.6, 40.9 50.1,49.4, 40.9, 56.3 62.8, 69.6, 70.8, 57.4 90.6, 71.6, 63.3, 74.8 77.0, 92.7, 79.6, 73.1 90.3, 60.4, 61.6, 85.3 58.2, 70.1, 68.1, 81.1 51.4, 61.4, 61.0, 62.2 41.6, 66.5, 65.8, 62.1 PFOS Concentrations in Tissues of Bluegill Exposed to 0.87 mg/L Values are from 4 individualfish at each sample period. Uptake Edible Tissue, Nonedible Tissue, Day mg/kg mglkg 0 (4-hours) 1.46, 1.48, 1.19, 1.39 3.52, 4.37, 4.22, 4.06 1 4.68, 6.59, 5.56, 5.64 11.1, 14.2, 13.3, 12.1 Whole Fish Conc., mglkg 2.71,3.08, 2.64, 2.89 8.00, 10.9, 10.2, 9.47 3 17.3, 15.8, 19.0, 20.8 39.3, 42.0, 43.8, 51.8 30.5, 30.7, 34.5, 39.1 7 42.0, 44.0, 57.7, 46.8 100, 102, 102, 120 74.9, 77.0, 85.3, 89.8 14 87.1, 81.6, 90.7, 73.3 177, 207, 245, 214 141, 157, 180, 158 21 28(1) 79.4, 117, 104, 102 102, 131,107, 133 201,278, 246, 229 289, 372, 320, 361 146, 210, 185, 172 205,267, 232, 263 Sampling of fish stopped after Uptake Day 28 due to mortality. Test organism mortality: Negative control: None duringthe uptake phase (62 days) or depuration phase (35 days) 0.086 mg/L exposure: One fish died after 49 days and one after 59 days exposure in the uptake phase, none duringthe depurationphase (total 2.2% mortality duringthe study). 0.87 mg/L exposure: Mortality first noted on Day 9 and continued through Day 35 of the uptake phase at whichtime all of the fish had either died or had been sampled Analytical methodology: Analyses of test solutionsand fish tissues were performed at Wildlife International,Ltd. Water sampleswere diluted and analyzed by HPLC withsingle quadrupole mass spectrometric detection. Tissue sampleswere homogenized,extracted,dilutedand analyzed by HPLC with triple quadrupolemass spectrometricdetection. When determiningthe concentrationof the test substancein the samples, the same and most prominentpeak responsefor perfluorooctanesulfonate was used. No attemptwas made to quantifyon the basis of individual isomeric components. The LOQ was 0.05 mg/L for water in this study. For tissue samples, the LOQ was calculated on an individualbasisfor each sample sinceeach entire submittedsample,of differingweight, was extracted withoutan adjustmentto constantweight. Recovery was excellent in both water and fish tissues, rangingfrom 84.9 to 122% of fortificationlevels. Analyticalresultswere notcorrectedfor proceduralrecovery. CONCLUSIONS PFOS bioconcentratedin the tissuesof bluegillsunfishduringthis study. The BCFK values calculatedfor the edible, nonedibleand whole fish tissuesfrom the 0.086 mg/L exposurewere calculatedto be 1124, 4013, and 2796, respectively. PFOS depurated slowly. The BIOFAC estimates for the time to reach 50% clearance for edible, nonedibleand whole fish tissuesfrom the 0.086 mg/L exposurewere 86, 116 and 112 days, respectively. DATA QUALITY , Reliability: Klimischranking = 1 REFERENCES This study was conducted at Wildlife International,Ltd., Easton, MD at the request of the 3M Company, Lab Request number U2723. The reportwas amended and reissuedJune 6, 2002. OTHER Last changed: 6/11/02