Olsen, GW, Schmickler, M, Tierens, JM, Logan, PW, Burris, JM, Burlew, MM, Lundberg, JK, Mandel, JH. 2001e. Descriptive summary of serum fluorochemical levels among employee participants of the year 2000 Antwerp fluorochemical medical surveillance program. Final Report. March 19, 2001.

Olsen, GW, Madsen, DC, Burris, JM, Mandel, JH. 2001f. Descriptive summary of serum fluorochemical levels among 236 building employees. Final Report. March 19, 2001. ENV/ JM/ RD( 2002) 17/ FINAL 88 Olsen, GW, Hansen, Clemen, LA, Burris, JM, Mandel, JH. 2001g. Identification of Fluorochemicals in Human Tissue. Final Report. Epidemiology, 220 3W 05, Medical Department, 3M Company, St. Paul, MN 55144.

Olsen, GW, Burris, JM, Lundberg, JK, Hansen, KJ, Mandel, JH, Zobel, LR. 2002a. Identification of fluorochemicals in human sera. I. American Red Cross adult blood donors. Final report. 3M Medical Department.

Olsen, GW, Burris, JM, Lundberg, JK, Hansen, KJ, Mandel, JH, Zobel, LR. 2002b. Identification of fluorochemicals in human sera. II. Elderly participants of the Adult Changes in Thought Study, Seattle, Washington. Final Report. 3M Medical Department.

Olsen, GW, Burris, JM, Lundberg, JK, Hansen, KJ, Mandel, JH, Zobel, LR. 2002c. Identification of fluorochemicals in human sera. III. Pediatric participants in a Group A Streptococci clinical trial investigation. Final Report. 3M Medical Department.

Rusch, G. M., Rinehart, W. E. and Bozak, C. A. 1979. An Acute Inhalation Toxicity Study of T 2306 CoC in the Rat. Project No. 78 7185, Bio/ dynamics Inc.

Seacat, AM. Analytical Laboratory Report from the 26 Week Capsule Toxicity Study with Perfluoroctanesulfonic Acid Potassium Salt ( T 6295) in Cynomolgus Monkeys on the Determination of the Presence and Concentration of Perfluorooctanesulfonate ( PFOS) in Liver and Serum Samples. 3M Medical Department Study: T 6295.7; Covance In Life Study:# 6329 223. Analytical Study; FACT TOX 030; 3M Laboratory Request No. U2279.

Simmon, V. F. 1978. In Vitro Microbiological Mutagenicity Assays of 3M company Compounds T 2247 CoC and T 2248 CoC. Final Report. SRI Project: LSC 4442 016. SRI International Menlo Park, California 94025.

Sohlenius, A. K., Eriksson, A. M., Högström, C., Kimland, M. and De Pierre, J. W. 1993. Perfluorooctane sulfonic acid is a potent inducer of peroxisomal fatty acid B oxidation and other activities known to be affected by peroxisome proliferators in mouse liver. Pharmacol. Toxicol. 72, 90 93 Thomford, P. J. 1998. 4 Week Capsule Toxicity Study with Perfluoroctane Sulfonic Acid Potassium Salt ( PFOS; T 6295) in Cynomolgus Monkeys. Unaudited Draft. Study No. 6329 222 for 3M, St. Paul, MN, by Covance Laboratories Inc., Madison, WI. FYI 0500 1378.

Thomford, P. J. 2000. 26 Week Capsule Toxicity Study with Perfluorooctane Sulfonic Acid Potassium Salt ( PFOS; T6295) in Cynomolgus Monkeys. Unaudited Draft Final Report Prepared for 3M, St Paul, Minnesota by Covance Laboratories, Inc., Madison Wisconsin 53704 2595. Study Number 6329 223. April 12, 2000. FYI 0500 1378 USEPA 1999. New pesticide fact sheet lithium perfluorooctane sulfonate ( LPOS). EPA 730 F99 009, August 1999.

Wallace, K. B. and Starkov, A. 1998. The effect of perfluorinated arylakylsulfonamides on bioenergetics of rat liver mitochondria. Department of Biochemistry and Molecular Biology, University of MN School of Medicine, Duluth, MN 55812, USA. Supported by a grant from 3M Company.

Wetzel, L. T. 1983. Rat Teratology Study, T 3351, Final Report. Hazelton Laboratories America, Inc. Project Number: 154 160, December 19, 1983. ( 8EHQ 0399 374). ENV/ JM/ RD( 2002) 17/ FINAL 89 Annex 1. Ecological Studies The following lists of study reports were contained on a 3M Fluorochemical EPA Submissions CDs dated 12th July 2000, 28th June 2001, and February 2002 12th July 2000 1. 96 Hour Static Acute Toxicity Test with the Fathead Minnow ( Pimephales promelas) 2. 96 Hour Toxicity Test with the Freshwater Alga ( Selenastrum capricornutum) 3. 48 Hour Static Acute Toxicity Test with the Cladoceran ( Daphnia magna) 4. 96 Hour Shell Deposition Test with the Eastern Oyster ( Crassostrea virginica) 5. 96 Hour Static Acute Toxicity Test with the Freshwater Mussel ( Unio complamatus) 6. Activated Sludge, Respiration Inhibition Test 7. 96 Hour Static Acute Toxicity Test with the Saltwater Mysid ( Mysidopsis bahia) 8. Early Life Stage Toxicity Test with the Fathead Minnow ( Pimephales promelas) 9. Semi Static Life Cycle Toxicity Test with the Cladoceran ( Daphnia magna) 10. Flow through Life Cycle Toxicity Test with the Saltwater Mysid ( Mysidopsis bahia) 11. Dietary LC50 Study with the Mallard 12. Dietary LC50 Study with the Northern Bobwhite 13. Multi Phase Exposure / Recovery Algal Assay Test 14. The Effects of Continuous Aqueous Exposure to 14C 78.02 on Hatchability of Eggs and Growth and Survival of Fry of Fathead Minnow ( Pimephales promelas / Summary of histopathological examinations of Fathead Minnow ( Pimephales promelas) exposed to 78.02 for 30 Days 15. Effect of Potassium Perfluorooctanesulfonate on Survival, etc. ( Daphnid reproduction) 16. Pimephales promelas 96 hour Toxicity Test Data Summary. Sample FC 94 X ( Li salt of PFOS) 17. 48 HR Acute Toxicity to Daphnia, Daphnia magna. FC 94 X ( Li salt of PFOS) 18. Microbics Microtox Toxicity Test. Sample : FC 94 X ( Li salt of PFOS) 19. Evaluation of FC 94 X by OECD Activated Sludge Respiration Inhibition Test # 209 Review of OECD 209 and BOD/ COD Test Results for FC 94 X, test data sheets ( Li salt of PFOS) 20. 96 Hour Acute Toxicity Test on Bluegill Sunfish ( FC 99, DEA salt of PFOS) 21. Acute Toxicity to Activated Sludge ( FC 99, DEA salt of PFOS) 22. Microtox data for FM 3820 ( 28% PFOS) 23. Acute Toxicity to Daphnia magna for FM 3820 ( 28% PFOS) 24. Toxicity to Algae ( Selenastrum capricornutum) for FC 3820 ( 28% PFOS) 25. Final Comprehensive Report : FC 95 26. Data from Fathead Minnow Study on FC 93 ( 25% NH4 salt of PFOS in IPA and water), 3M Environmental Lab, Aug. 2, 1974. 27. Data from Fathead Minnow Study on FC 93 ( 25% NH4 salt of PFOS in IPA and water), 3M Environmental Lab, Oct. 19, 1974. 28. Acute toxicity of P3025 developmental material to Fathead minnow ( Pimephales promelas). 29. Acute toxicity of P3025 developmental material to Daphnia magna.

28th June 2001 30. Acute toxicity of PFOS to Rainbow trout in saltwater 31. Acute toxicity of PFOS to Rainbow trout in freshwater 32. Acute toxicity of PFOS to Artemia sp. 33. Acute toxicity of PFOS to Daphnia magna 34. Perfluorooctanesulfonate, Potassium salt ( PFOS): An acute oral toxicity study with the Honey bee 35. Perfluorooctanesulfonate, Potassium salt ( PFOS): An acute contact toxicity study with the Honey bee 36. PFOS: A 96 hour toxicity test with the freshwater alga ( Anabaena flos aquae) 37. PFOS: A 7 day toxicity test with Duckweed ( Lemna gibba G3) 38. PFOS: A 96 hour toxicity test with freshwater diatom ( Navicula pelliculosa) ENV/ JM/ RD( 2002) 17/ FINAL 90 39. PFOS: A 96 hour toxicity test with the marine diatom ( Skeletonema costatum) 40. PFOS: A frog embryo teratogenesis assay   Xenopus ( FETAX) 41. Perfluorooctanesulfonate, Potassium salt ( PFOS): A flow through bioconcentration test with the Bluegill ( Lepomis macrochirus)

February 2002 42. Perfluorooctanesulfonate, Potassium salt ( PFOS): 96 Hour Static Acute Toxicity Test with the Rainbow Trout ( Oncorhynchus mykiss) in freshwater 43. Perfluorooctanesulfonate, Potassium salt ( PFOS): 96 Hour Semi Static Acute Toxicity Test with the Sheepshead Minnow ( Cyprinodon variegatus) in saltwater ENV/ JM/ RD( 2002) 17/ FINAL 91 Annex 2. Robust Summaries of Key Ecotoxicology Studies Study reference number Title 1 96 Hour Static Acute Toxicity Test with the Fathead Minnow ( Pimephales promelas) 2 96 Hour Toxicity Test with the Freshwater Alga ( Selenastrum capricornutum) 3 48 Hour Static Acute Toxicity Test with the Cladoceran ( Daphnia magna) 4 96 Hour Shell Deposition Test with the Eastern Oyster ( Crassostrea virginica) 5 96 Hour Static Acute Toxicity Test with the Freshwater Mussel ( Unio complamatus) 6 Activated Sludge, Respiration Inhibition Test 7 96 Hour Static Acute Toxicity Test with the Saltwater Mysid ( Mysidopsis bahia) 8 Early Life Stage Toxicity Test with the Fathead Minnow ( Pimephales promelas) 9 Semi Static Life Cycle Toxicity Test with the Cladoceran ( Daphnia magna) 10 Flow through Life Cycle Toxicity Test with the Saltwater Mysid ( Mysidopsis bahia) 11 Dietary LC50 Study with the Mallard 12 Dietary LC50 Study with the Northern Bobwhite 13 Multi Phase Exposure / Recovery Algal Assay Test 14 The Effects of Continuous Aqueous Exposure to 14C 78.02 on Hatchability of Eggs and Growth and Survival of Fry of Fathead Minnow ( Pimephales promelas / Summary of histopathological examinations of Fathead Minnow ( Pimephales promelas) exposed to 78.02 for 30 Days 15 Effect of Potassium Perfluorooctanesulfonate on Survival, etc. ( Daphnid reproduction) 16 Pimephales promelas 96 hour Toxicity Test Data Summary. Sample FC 94 X ( Li salt of PFOS) 17 48 HR Acute Toxicity to Daphnia, Daphnia magna. FC 94 X ( Li salt of PFOS) 20 96 Hour Acute Toxicity Test on Bluegill Sunfish ( FC 99, DEA salt of PFOS) 23 Acute Toxicity to Daphnia magna for FM 3820 ( 28% PFOS) 26 Data from Fathead Minnow Study on FC 93 ( 25% NH4 salt of PFOS in IPA and water), 3M Environmental Lab, Aug. 2, 1974. 27 Data from Fathead Minnow Study on FC 93 ( 25% NH4 salt of PFOS in IPA and water), 3M Environmental Lab, Oct. 19, 1974. 28 Acute toxicity of P3025 developmental material to Fathead minnow ( Pimephales promelas). 29 Acute toxicity of P3025 developmental material to Daphnia magna. 30 Acute toxicity of PFOS to Rainbow trout in saltwater 31 Acute toxicity of PFOS to Rainbow trout in freshwater 32 Acute toxicity of PFOS to Artemia sp. 33 Acute toxicity of PFOS to Daphnia magna 34 Perfluorooctanesulfonate, Potassium salt ( PFOS): An acute oral toxicity study with the Honey bee 35 Perfluorooctanesulfonate, Potassium salt ( PFOS): An acute contact toxicity study with the Honey bee 36 PFOS: A 96 hour toxicity test with the freshwater alga ( Anabaena flos aquae) 37 PFOS: A 7 day toxicity test with Duckweed ( Lemna gibba G3) 38 PFOS: A 96 hour toxicity test with freshwater diatom ( Navicula pelliculosa) 39 PFOS: A 96 hour toxicity test with the marine diatom ( Skeletonema costatum) 40 PFOS: A frog embryo teratogenesis assay   Xenopus ( FETAX) 41 Perfluorooctanesulfonate, Potassium salt ( PFOS): A flow through bioconcentration test with the Bluegill ( Lepomis macrochirus) 42 Perfluorooctanesulfonate, Potassium salt ( PFOS): 96 Hour Static Acute Toxicity Test with the Rainbow Trout ( Oncorhynchus mykiss) in freshwater 43 Perfluorooctanesulfonate, Potassium salt ( PFOS): 96 Hour Semi Static Acute Toxicity Test with the Sheepshead Minnow ( Cyprinodon variegatus) in saltwater 44 Bioconcentration test of Salt ( Na, K, Li) of perfluoroalkyl ( C= 4 12) sulfonic acid [ This test was performed using Perfluorooctane sulfonic acid, potassium salt ( Test substance number K 1520)] in carp ENV/ JM/ RD( 2002) 17/ FINAL 92 Robust Study Report Reference No. 1   96 Hour Static Acute Toxicity Test with the Fathead Minnow (

 )

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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: OECD 203 and OPPTS 850.1075 Type: Static acute GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Pimephales promelas Supplier: In house cultures, Wildlife International, Ltd., Easton, MD, USA Analytical monitoring: PFOS measured at 0, 48, 96 hours Exposure period: 96 hours Statistical methods: LC50 values calculated, when possible, by probit analysis, moving average method or binomial probability with non linear interpolation using the computer software of C. E. Stephan. Test fish age: Approximately 126 days old Length and weight: 35 ( 30 38) mm, 0.36 ( 0.21 0.49) g Loading: 0.24 g fish/ L Pretreatment: None Test Conditions Dilution water: 0.45 µ m filtered well water Dilution water chemistry ( during the 4 week period immediately preceding the test): Hardness: 131 ( 128 136) mg/ L as CaCO3 Alkalinity: 177 ( 176 178) mg/ L as CaCO3 pH: 8.3 TOC: < 1.0 mg/ L Conductivity: 311 ( 310 315) µ mhos/ cm Stock and test solution preparation: Primary stock prepared in dilution water at 27 mg/ L and mixed for ~ 22 hours prior to use. After mixing, primary stock solution was proportionally diluted with dilution water to prepare the four additional test concentrations. Concentrations dosing rate: Once Stability of the test chemical solutions: Extremely stable Exposure vessels: 25L polyethylene aquaria containing approximately 15L of test solution; water depth approximately 17.6 cm. Number of replicates: two Number of fish per replicate: ten Number of concentrations: five plus a negative control Water chemistry during the study: Dissolved oxygen range ( 0   96 hours): 7.8   8.8 mg/ L ( control exposure) ENV/ JM/ RD( 2002) 17/ FINAL 93 7.7   9.0 mg/ L ( 28 mg/ L exposure) pH range ( 0   96 hours) 8.3   8.6 ( control exposure) 8.4   8.5 ( 28 mg/ L exposure) Test temperature range ( 0   96 hours) 20.4   22.1oC ( control exposure) 21.3   22.3 oC ( 28 mg/ L exposure) Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative control, 3.6, 5.9, 9.9, 16, 27 mg/ L Measured concentrations: < LOQ, 3.3, 5.6, 9.5, 17, 28 mg/ L Element value: ( 95% confidence interval is given in brackets) 24 hour LC50 = > 28 mg/ L ( C. I. not calculable) 48 hour LC50 = > 28 mg/ L ( C. I. not calculable) 72 hour LC50 = 27 ( 22   41) mg/ L 96 hour LC50 = 9.5 ( 8.0   11) mg/ L All element values based on mean measured concentrations Statistical evaluation of mortality: Confidence limits for 24 and 48 hours could not be calculated due to lack of mortality. The 72 hour LC50 value is questionable because a concentration effect relationship was not demonstrated over a reasonable range of percent dead. The 24 and 48 hour LC50 values were determined by visual interpretation. Probit was used to calculate the 72 hour LC50 and Moving Average for the 96 hour LC50.

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 0.458 mg/ L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 97.9. Samples collected at test initiation had measured values from 85.3 to 117% of nominal. Measured values for samples taken at 48 hours ranged from 86.3 to 101% of nominal. Measured values for samples taken at 96 hours ranged from 87.6 to 98.3% of nominal.

Summary of analytical chemistry data:

Nominal Test Concentration, mg/ L Measured Duplicate Values at 0, 48, and 96 hours, Respectively, mg/ L Mean Measured Concentration mg/ L Percent of Nominal Negative Control All < LOQ < LOQ  3.6 3.16, 3.53, 3.08, 3.22, 3.46, 3.13 3.3 92 5.9 6.05, 5.07, 5.48, 5.89, 5.70, 5.55 5.6 95 9.9 8.99, 9.47, 9.88, 9.33, 9.70, 9.52 9.5 96 16 18.2, 19.3, 15.0, 15.6, 14.8, 16.2 17 106 27 28.5, 28.5, 27.0, 27.8, 26.8, 26.6 28 104 Biological observations after 96 hours: Fish in the negative control and the 3.3 mg/ L exposure concentration appeared normal. Some or all of the surviving fish were observed to be swimming erratically ( 4/ 16 in 5.6 mg/ L exposure, 10/ 10 in 9.5 mg/ L, 4/ 4 in 17 mg/ L) at test termination. ENV/ JM/ RD( 2002) 17/ FINAL 94 Cumulative percent mortality:

Mean Measured Test Concentration mg/ L 24 hours 48 hours 72 hours 96 hours Neg. Control 0 0 0 0 3.3 0 0 0 0 5.6 0 0 0 20 9.5 0 0 0 50 17 0 0 15 80 28 0 0 50 100 Lowest concentration causing 100% mortality: 28 mg/ L Mortality of controls: None CONCLUSIONS The potassium perfluorooctanesulfonate 96 hour LC50 for fathead minnow was determined to be 9.5 mg/ L with a 95% confidence interval of 8.0   11 mg/ L. The 96 hour no mortality and no effects concentration was 3.3 mg/ L.

Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA 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 ENV/ JM/ RD( 2002) 17/ FINAL 95 Robust Study Report Reference No. 2   96 Hour Toxicity Test with the Freshwater Alga (

           )

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonicacid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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, ' H 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, USA 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 96 hours. 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 reagent grade chemicals to reverse osmosis purified well water. Test Conditions: Test temperature range: 23.6 25.8oC Growth medium: ASTM Standard Guide1218 90E, 1990 Compound Nominal concentration Units MgCl26H20 12.16 mg/ l CaCl22H2O 4.40 mg/ l H3BO3 0.1856 mg/ l MnCl24H2O 0.416 mg/ l ZnCl2 3.28 µ g/ l FeCl26H2O 0.1598 mg/ l CoCl26H2O) 1.428 µ g/ l Na2MoO42H2O 7.26 µ g/ l CuCl22H2O 0.012 µ g/ l Na2EDTA2H2O 0.300 mg/ l NaNO3 25.50 mg/ l MgSO47H2O 14.70 mg/ l K2HPO4 1.044 mg/ l ENV/ JM/ RD( 2002) 17/ FINAL 96 NaHCO3 15.0 mg/ 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 ( 0.22 µ m) 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 104 cells/ mL Number of concentrations: six plus a negative control plus an abiotic control at the highest concentration tested 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  25.8oC 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: Negative 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: ( 95% confidence interval is given in brackets) 24 hour EC50 ( cell density) = 163 ( 74  191) mg/ L 24 hour EbC50 ( area under curve) = 122 ( 19  176) mg/ L 24 hour EC50 ( growth rate) = 136 ( 30  204) mg/ L 48 hour EC50 ( cell density) = 81 ( 72  90) mg/ L 48 hour EbC50 ( area under curve) = 84 ( 67  146) mg/ L 48 hour ErC50 ( growth rate) = 142 ( 107  185) mg/ L 72 hourEC10 ( cell density) = 37 (< O  64) mg/ L 72 hourEbC10 ( area under curve) = 46 (< O  56) mg/ L 72 hour ErC10 ( growth rate) = 53 ( 23  64) mg/ L 72 hourEC50 ( cell density) = 70 ( 44  78) mg/ L 72 hourEbC50 ( area under curve) = 74 ( 55  82) mg/ L 72 hour ErC50 ( growth rate) = 120 ( 103  132) mg/ L 72 hourEC90 ( cell density) = 153 ( 130  165) mg/ L 72 hourEbC90 ( area under curve) = 165 ( 145  176) mg/ L 72 hour ErC90 ( growth rate) = > 179 mg/ L ( C. I. not calculable) 96 hourEC10 ( cell density) = 49 ( 43  50) mg/ L 96 hourEBC10 ( area under curve) = 49 ( 40  50) mg/ L 96 hourErC10 ( growth rate) = 59 ( 54  63) mg/ L ENV/ JM/ RD( 2002) 17/ FINAL 97 96 hourEC50 ( cell density) = 71 ( 66  73) mg/ L 96 hourEbC50 ( area under curve) = 71 ( 67 74) mg/ L 96 hour ErC50 ( growth rate) = 126 ( 115  138) mg/ L 96 hour EC90 ( cell density) = 137 ( 105  153) mg/ L 96 hour EbC90 ( area under curve) = 145 ( 125  155) mg/ L 96 hour ErC90 ( growth rate) = > 179 mg/ L ( C. I. 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 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 ECIO, EC50, and EC90 values and 95% confidence limits at 72 and 96 hours. 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 at72 hours ranged from 81.9 to 105% of nominal and for samples taken at 96 hours, 90.3 to 103% of nominal.

Summary of analytical chemistry data:

Nominal test concentration, mg/ l Measured values at 0, 72, and 96 hours respectively, mg/ l Mean measured concentration, mg/ l Percent nominal Negative control All < LOQ < LOQ  5.7 4.73, 6.04, 5.84 5.5 96 11 10.7, 11.2, 12.1 11 100 23 19.8, 23.1, 20.7 21 91 46 42.7, 41.9, 46.3 44 96 91 83.3, 86.0, 88.3 86 95 183 179, 186, 172 179 98 183 ( abiotic) Not analyzed, 150, 188 169 92 Control response: satisfactory ENV/ JM/ RD( 2002) 17/ FINAL 98 Biological observations after 96 hours:

Mean measured concentration, mg/ l Mean number of cells per ml Percent Inhibition via Density Percent Inhibition via Area Under the Curve Percent Inhibition via Growth Rate Negative control 2,740,00    5.5 3,040,000  11  8.5  1.9 11 2,880,000  5.1  3.3  0.84 21 3,240,000  18  13  3.0 44 3,080,000  12  5.3  2.0 86 626,667 77 75 27 179 33,667 99 98 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 EC50 and 95% confidence interval for Selenastrum capficornutumwas 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, USA 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 ENV/ JM/ RD( 2002) 17/ FINAL 99 Robust Study Report Reference No. 3  48 Hour Static Acute Toxicity Test with the Cladoceran (

)

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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: OECD 202 and OPPTS 850.1010 Test type: Static acute GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Daphnia magna Analytical monitoring: PFOS measured at 0, 24, 48 hours Statistical methods: EC50 values calculated, when possible, by probit analysis, moving average method or binomial probability with non linear interpolation using the computer software of C. E. Stephan. Test daphnid source: Obtained from cultures maintained by Wildlife International Ltd., Easton, MD. Identification of the original brood stock was verified by the Academy of Natural Sciences, Philadelphia, PA. , USA Test daphnid age at study initiation: < 24 hours Test conditions Dilution water: 0.45 µ m filtered well water Dilution water chemistry ( during the 4 week period immediately preceding the test): Hardness: 132 ( 128 136) mg/ L as CaCO3 Alkalinity: 178 ( 176 178) mg/ L as CaCO3 pH: 8.3 ( 8.2 8.3) TOC: < 1.0 mg/ L Conductivity: 313 ( 310 315) µ mhos/ cm Ca/ Mg ratio: 35/ 13.5 Na/ K ratio: 21.3/ 6.62 Lighting: Colortone ® 50 fluorescent lights, intensity approximately 359 lux. Photoperiod of 16 hours light, 8 hours dark with a 30 minute transition period. Stock and test solutions preparation: A primary stock solution was prepared in dilution water at 91 mg/ L. It was mixed for ~ 19.5 hours prior to use. After mixing, the primary stock was proportionally diluted with dilution water to prepare the four additional test concentrations. All test solutions appeared clear and colorless. Exposure vessels: 250 mL plastic beakers containing 240 mL of test solution. The approximate depth of test solution was 6.4 cm. Number of replicates: two Number of daphnids per replicate: ten Number of concentrations: five plus a negative control Water chemistry during the study: Dissolved oxygen range ( 0   48 hours): 8.6   8.9 mg/ L ( control exposure) ENV/ JM/ RD( 2002) 17/ FINAL 100 8.6   9.1 mg/ L ( 91 mg/ L exposure) pH range ( 0   48 hours) 8.2   8.5 ( control exposure) 8.5   8.6 ( 91 mg/ L exposure) Test temperature range ( 0   48 hours) 19.5   20.2oC ( control exposure) 19.3   20.1oC ( 91 mg/ L exposure) Element basis: mortality and immobilization Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative control, 12, 20, 33, 55, 91 mg/ L Measured concentrations: < LOQ, 11, 20, 33, 56, 91 mg/ L Element value: ( 95% confidence interval is given in brackets) 24 hour EC10 = 82 ( 81 83) mg/ L 24 hour EC50 = > 91 mg/ L ( C. I. not calculable) 24 hour EC90 = > 91 mg/ L ( C. I. not calculable) 48 hour EC10 = 53 (< 11> 91) mg/ L 48 hour EC50 = 61 ( 33 91) mg/ L 48 hour EC90 = 63 (< 11> 91) mg/ L All element values based on mean measured concentrations Statistical evaluation: The EC50 values and 95% confidence intervals were calculated when possible by probit analysis, the moving average method or binomial probability with non linear interpolation using the computer software of C. E. Stephan. The EC10 and EC90 values were calculated when possible using the Bruce Versteeg method because there were less than two concentrations with partial mortality or immobility. 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 4.58 mg/ L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 96.2. Samples collected at test initiation had measured values from 85.5 to 112% of nominal. Measured values for samples taken at 24 hours ranged from 92.2 to 115% of nominal. Measured values for samples taken at 48 hours ranged from 91.6 to 106% of nominal.

Summary of analytical chemistry data:

Nominal Test Concentration mg/ L Measured duplicated values at 0, 24, and 48 hours respectively, mg/ L Mean Measured Concentration mg/ L Percent Of Nominal Negative Control All < LOQ < LOQ  12 10.5, 10.6, 11.5, 12.5, 10.9, 12.0 11 92 20 17.2, 18.1, 22.8, 21.6, 21.4, 18.8 20 100 33 30.2, 34.1, 34.0, 36.1, 31.3, 34.0 33 100 55 50.5, 49.9, 57.0, 63.0, 56.8, 56.4 56 102 91 87.6, 102, 90.1, 84.4, 88.7, 92.4 91 100 Biological observations after 48 hours: Daphnids in the negative control, the 11 and the 20 mg/ L ENV/ JM/ RD( 2002) 17/ FINAL 101 treatments appeared healthy and normal throughout the test with no mortality, immobility or overt clinical signs of toxicity. Five percent mortality was observed at 48 hours in the negative control. The effects noted in this study were mortality; no immobilization was noted at any test concentration.

Cumulative percent mortality:

Mean Measured Test Concentration mg/ L 24 hours 48 hours Negative Control 0 5 11 0 0 20 0 0 33 0 0 56 0 35 91 35 100 Control response: satisfactory CONCLUSIONS The potassium perfluorooctanesulfonate 48 hour EC50 for Daphnia magna was determined to be 61 mg/ L with a 95% confidence interval of 33 91 mg/ L. The 48 hour no immobilization and no observed effect concentration was 33 mg/ L.

Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA 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 ENV/ JM/ RD( 2002) 17/ FINAL 102 Robust Study Report Reference No. 4  96 Hour Shell Deposition Test with the Eastern Oyster (

)

TEST SUBSTANCE Identity: Potassium pertluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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 850.1025 Type: Static acute GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Crassostrea virginica Supplier: P. Cummins Oyster Company, Inc., Baltimore, MD, USA Shell grinding: Prior to test initiation, recently deposited shell at the rounded ( ventral) end was removed using a small electric grinder. Care was taken to remove the shell rim uniformly to produce a smooth, rounded, blunt profile. Analytical monitoring: PFOS measured at 0,48,96 hours Exposure period: 96 hours Statistical methods: Shell growth inhibition was calculated for each treatment group as the percent reduction in shell growth relative to mean shell growth in the negative control. The EC50 value was estimated by visual inspection of shell growth inhibition data. The shell growth data was evaluated for normality and homogeneity of variances using the Chi Square test and Bartlett's test, respectively. Dunnett's test was used to identify treatment groups that had a statistically significant (< 0.05) reduction in shell growth as compared to the control. Test oyster age: unknown Length: 33.8 ( 27.8 41.5) mm, Pretreatment: None Test conditions: Dilution water: Natural seawater diluted to a salinity of 20 O/ oo with well water Dilution water chemistry ( during the 4 week period immediately preceding the test): salinity: 21 ( 20 21) O/ oo pH: 8.1( 8.0 8.2) TOC: < 1.0 mg/ L Stock and test solution preparation: Primary stock prepared in dilution water at 9.1mg/ L and mixed for ~ 24 hours prior to use. After mixing, primary stock solution appeared clear and colorless with some white particulate material suspended throughout the solution. It was proportionally diluted with dilution water to prepare the four additional test concentrations. All test solutions appeared clear and colorless. Due to the relatively low solubility of PFOS in natural seawater, the highest concentration attainable with this matrix is approximately 3.3mg/ L. Concentrations dosing rate: Once Exposure vessels: 52L polyethylene aquaria containing approximately 40L of test solution; water depth approximately 21 cm. Each chamber was continuously stirred to circulate the supplemental ENV/ JM/ RD( 2002) 17/ FINAL 103 algae diet using an electric paddle mixer. Feeding: Algal cells ( Thalassiosira pseudonana, Skeletonema sp., Chaetoceros sp., and lsochrysis sp.) were provided to supplement naturally occurring algae and to maximize oyster growth rates during the test. Number of replicates: one Number of oysters per replicate: twenty Number of concentrations: five plus a negative control Water chemistry during the study: Dissolved oxygen range ( 0 96 hours): 6.6  7.6mg/ L ( control exposure) 6.1  7.7mg/ L ( 3.0 mg/ L exposure). pH range ( 0 96 hours): 7.6  8.1 ( control exposure) 7.6  8.1 ( 3.0 mg/ L exposure) Test temperature range ( 0 96 hours): 22.2  22.3 oC ( control exposure) 21.8  22.7 oC ( 3.0 mg/ L exposure) Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative control, 1.2, 2.0, 3.3, 5.5, 9.1 mg/ L Measured concentrations: < LOQ, 0.36, 0.40, 1.3, 1.9, 3.0 mg/ L Element value: ( 95% confidence interval is given in brackets) 96 hour EC50 = > 3.0 mg/ L ( C. I. not calculable) 96 hour NOEC = 1.9 mg/ L All element values based on mean measured concentrations Statistical evaluation of shell growth: EC50 values could not be calculated due to insufficient shell growth inhibition at the highest attainable concentration.

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. Samples collected at test initiation had measured values from 28 to 46% of nominal. Measured values for samples taken at 48 hours ranged from 15 to 41 % of nominal. Measured values for samples taken at 96 hours ranged from < LOQ to 52% of nominal.

Summary of analytical chemistry data:

Nominal Test Concentration, mgll Measured Duplicate Values at 0, 48 and 96 hours, Respectively, mg/ L Mean Measured Concentration, mg/ l Percent of Nominal Negative Control All < LOQ < LOQ  1.2 0.331, 0.353, 0.341, 0.429, < LOQ, < LOQ 0.36 30 2.0 0/ 622, 0.633, 0.299, 0.40 20 ENV/ JM/ RD( 2002) 17/ FINAL 104 0.313, 0.249, 0.257 3.3 1.36, 1.15, 0.924, 0.878, 1.58, 1.72 1.3 39 5.5 2.42, 2.53, 2.02, 2.24, 1.45, 0.970 1.9 35 9.1 3.39, 3.44, 3.01*, 3.74, 3.57, 1.99, 2.19 3.0 33 * 3 replicates analyzed at time 0 Biological observations after 96 hours: Oysters in the negative control and all PFOS treatment groups appeared normal and healthy throughout the exposure period.

Shell deposition and shell growth Inhibition at test termination:

Mean Measured Concentration, mg/ l Shell Deposition Mean + SD, mm Percent Inhibition in Shell Growth Negative Control 2.67 + 0.824  0.36 2.50 + 0.933 6.4 0.40 2.40 + 0.820 10 1.3 2.51 + 0.919 6.0 1.9 2.13 + 0.804 20 3.0 1.91 + 0.591 28 Mortality of controls: None CONCLUSIONS The potassium perfluorooctanesulfonate 96 hour EC50 for the Eastern Oyster was determined to be > 3.0 mg/ L, the highest concentration tested and the practical limit of solubility in unfiltered seawater. The 96 hour no effect concentration was 1.9 mg/ L.

Submitter: 3M Company, Environmental Laboratory, P. O. Box 33331, St. Paul, Minnesota, 55133, USA 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/ 3100 ENV/ JM/ RD( 2002) 17/ FINAL 105 Robust Study Report Reference No. 5  96 Hour Static Acute Toxicity Test with the Freshwater Mussel (

   )

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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: The study was conducted using a protocol based on procedures outlined in U. S. Environmental Protection Agency Series 850   Ecological Effects Guidelines, OPPTS Number 850.1075 ; OECD 203: Fish, Acute Toxicity Test; and ASTM Standard E729 88a, Standard Guide for Conducting Toxicity Tests with Fishes, Macroinvertebrates and Amphibians. Test type: Semi static Renewal GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Unio complamatus Analytical monitoring: Test substance concentrations measured by LCMS at 0, 48, 96 hours Statistical methods: LC50 values calculated, when possible, by probit analysis, moving average method or binomial probability with non linear interpolation using the computer software of C. E. Stephan. Test organism source: Obtained from Carolina Biological Supply Company, Burlington, North Carolina, USA. Carolina collected from the wild. Test organism age at study initiation: Unknown Test Conditions Dilution water: 0.45 µ m filtered well water Dilution water chemistry ( during the 4 week period immediately preceding the test): Hardness: 126 ( 120 132) mg/ L as CaCO3 Alkalinity: 174 ( 170 178) mg/ L as CaCO3 pH: 8.3 ( 8.1 8.5) TOC: < 1.0 mg/ L Conductivity: 321 ( 310 330) µ mhos/ cm Ca/ Mg ratio: 35/ 13.5 Na/ K ratio: 21.3/ 6.62 Lighting: Colortone ® 50 fluorescent lights, intensity approximately 369 lux. Photoperiod of 16 hours light, 8 hours dark with a 30 minute transition period. Stock and test solutions preparation: A primary stock solution was prepared in dilution water at 91 mg/ L. It was mixed for approximately 24 hours prior to use. After mixing, the primary stock was proportionally diluted with dilution water to prepare the four additional test concentrations. All test solutions appeared clear and colorless. Exposure vessels: 25 liter polyethylene aquaria containing approximately 20 L of test solution. The approximate depth of test solution was 23.2 cm. Number of replicates: two Number of test organisms per replicate: ten Number of concentrations: five plus a negative control Water chemistry during the study: ENV/ JM/ RD( 2002) 17/ FINAL 106 Dissolved oxygen range ( 0   96 hours): 5.8   8.5 mg/ L ( control exposure) 5.0   8.6 mg/ L ( 79 mg/ L exposure) pH range ( 0   96 hours): 8.0   8.4 ( control exposure) 7.9   8.5 ( 79 mg/ L exposure) Test temperature range ( 0   96 hours): 21.4   21.8oC ( control exposure) 21.8   23.7 oC ( 79 mg/ L exposure) Element Basis: Mortality. Mussels with open shells and not responding to gentle prodding were considered dead. The number of individuals exhibiting clinical signs of toxicity or abnormal behavior also were evaluated. Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: < LOQ, 5.7, 11, 23, 46, 91 mg/ L Measured concentrations: < LOQ, 5.3, 12, 20, 41, 79 mg/ L Element value: ( 95% confidence interval is given in brackets) 96 hour LC50 = 59 mg/ L ( 51 68 mg/ L)

Statistical evaluation: The LC50 values and 95% confidence intervals were calculated when possible by probit analysis, the moving average method or binomial probability with non linear interpolation using the computer software of C. E. Stephan.

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 94.7%. Samples collected at test initiation had measured values from 73.7% to 96.0% of nominal. Measured values for samples taken at 48 hours ranged from 81.2 to 98.9% of nominal. Measured values for samples taken at 96 hours ranged from 88.5 to 130% of nominal.

Summary of analytical chemistry data:

Nominal Test Concentration , mg/ L Measured duplicated values at 0, 48, and 96 hours respectively, mg/ L Mean Measured Concentration, mg/ L Percent of Nominal Negative Control All < LOQ < LOQ  5.7 5.47, 4.93, 5.18, 5.70, 5.24, 5.26 5.3 93 11 11.4, 10.1, 11.2, 10.5, 10.9, 15.4 12 109 23 19.0, 16.8, 18.7, 18.7, 22.9, 22.4 20 87 46 37.2, 40.6, 37.1, 39.5, 48.2, 40.5 41 89 91 69.0, 74.7, 81.3, 77.6, 88.2, 85.7 79 87 Biological observations after 96 hours: Mussels in the negative control, the 5.3, 12 and the 20 mg/ L treatments appeared healthy and normal throughout the test with no mortality or overt clinical signs of toxicity. Five percent mortality was observed at 96 hours in the 41 mg/ L treatment and 90% mortality was observed in the 79 mg/ L treatment. No abnormal behavior was noted in these concentrations. ENV/ JM/ RD( 2002) 17/ FINAL 107 Cumulative percent mortality:

Mean Measured Test Concentration mg/ L 24 Hours 48 Hours 72 Hours 96 Hours Negative Control 0 0 0 0 5.3 0 0 0 0 12 0 0 0 0 20 0 0 0 0 41 0 0 0 5 79 30 40 50 90 Control response: Satisfactory CONCLUSIONS The potassium perfluorooctanesulfonate 96 hour LC50 for the Freshwater Mussel, Unio complamatus was determined to be 59 mg/ L with a 95% confidence interval of 51 68 mg/ L. The 96 hour no mortality concentration was 20 mg/ L.

Submitter: 3M Corporation, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA 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 ENV/ JM/ RD( 2002) 17/ FINAL 108 Robust Study Report Reference No. 6   Activated Sludge, Respiration Inhibition Test TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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 209 Test type: Static acute GLP: Yes Year Completed: Study completed 1999. Report completed 2000 Analytical monitoring: Dissolved oxygen concentrations. Statistical methods: Probit analysis using the computer software of C. E. Stephan. Test organism source. Activated sludge collected from the Prospect Bay Wastewater Treatment Facility, Grasonville, Maryland, USA.

Test conditions Dilution water: NANOpure ® Synthetic Sewage: 1 liter municipal water 16.0 g peptone 11.0 g meat extract 3.0 g urea 0.7 g NaCl 0.4 g CaCI2 2H20 0.2 g MgSO4 7H20 2.8 g K2HPO4 Reference and test solution preparation: A stock solution of the reference substance, 3,5 dichlorophenol, was prepared by dissolving 500 mg in 10 mL of 1N NaOH, diluted to 30 mL with NANOpure   water, then brought to the point of incipient precipitation with 1 NH2S04, and diluted to 1 L with NANOpure4   water. The pH of the reference solution was measured to be 7.18. PFOS was added directly to test vessels rather than volumetric addition of a stock solution. This method was deemed appropriate based on the observed solubility of the test substance in water. Test vessels: Mixtures were prepared and aerated in 500 mL Erlenmeyer flasks and then transferred into 300 mL Biochemical Oxygen Demand ( BOD) bottles Number of concentrations: 7 plus 3 reference controls and 2 Blank controls Temperature: 19 21oC Total Suspended Solids and pH for sludge on day of testing: 4380 mg/ L and 7.87 respectively. Element Basis: Respiration inhibition as determined by dissolved oxygen concentration.

Method Remarks: Stock solutions of PFOS that were prepared at nominal concentration of approximately 500 and 1 000 mg/ L in NANOpure   water contained test material that was not in solution after 20 minutes of sonication. Therefore, direct weight addition was employed to administer PFOS to the test system. ENV/ JM/ RD( 2002) 17/ FINAL 109 Test mixtures were prepared at 15 minute intervals and aerated until the contact time of the test substance with the activated sludge was three hours. After 3 hours of contact time, dissolved oxygen was measured over a period of up to 10 minutes.

RESULTS Nominal concentrations: Two blank controls, three reference substance controls, 0.90, 2.7, 9.0, 27, 90, 271. 905 mg/ L test material solutions.

Statistical Analyses: EC50 values were calculated for the reference material by probit analysis using the computer software of C. E. Stephan. An EC50 value could not be calculated for the test substance.

Analytical Methodology: Analysis of DO concentrations in all test solutions were performed at Wildlife International Ltd. using a YSI Model 50B Dissolved Oxygen Meter. Dissolved oxygen readings were recorded every 10 seconds for 10 minutes or until the dissolved oxygen dropped below 1.0 mg/ L Respiration Rates and Percent Inhibitions Treatment Respiration Rate mg 02/ L/ hour Percent Inhibition Control 1 39.6 NA Control 2 41.1 NA 3,5 dichlorophenol 3mg/ L 31.1 22.9 3,5 dichlorophenol 15 mg/ L 14.6 63.8 3,5 dichlorophenol 50 mg/ L 5.1 87.4 Test substance 0.90 mg/ L 38.9 3.6 Test substance 2.7 mg/ L 35.1 13.0 Test substance 9.0 mg/ L 33.5 17.0 Test substance 27 mg/ L 37.9 6.1 Test substance 90 mg/ L 32.7 19.0 Test substance 271 mg/ L 28.1 30.4 Test substance 905 mg/ L 24.7 38.8 Control response: satisfactory CONCLUSIONS The test substance exhibited a maximum inhibitory effect of 38% upon respiration at a nominal test substance concentration of 905 mg/ L. The EC50 ( respiration inhibition) is therefore greater than the solubility of the test substance.

Submitter: 3M Company, Environmental Laboratory, P. O. Box 33331, St. Paul, Minnesota, 55133, USA DATA QUALITY Reliability: Klimisch ranking 1.

REFERENCES This study was conducted at Wildlife International, Ltd. Easton, MD at the request of the 3M Company. ENV/ JM/ RD( 2002) 17/ FINAL 110 OTHER Last changed: 5/ 3/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 111 Robust Study Report Reference No. 7  96 Hour Static Acute Toxicity Test with the Saltwater Mysid (

)

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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: OPPTS 850.1035 Type: Static acute GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Mysidopsis bahia Supplier: In house cultures, Wildlife International, Ltd., Easton, MD, USA Analytical monitoring: PFOS measured at 0, 48, 96 hours Exposure period: 96 hours Statistical methods: LC50 values calculated, when possible, by probit analysis, moving average method or binomial probability with non linear interpolation using the computer software of C. E. Stephan. Test fish age: < 24 hours old Pretreatment: None Test Conditions: Dilution water: Natural seawater diluted to 20% o with well water, 0.45 µ m filtered. Dilution water chemistry ( during the 4 week period immediately preceding the test): Salinity: 20 ( 20 20) % o pH: 8.2 ( 8.1 8.2) TOC: < 1.0 mg/ L Stock and test solution preparation: Primary stock prepared at 8.2 mg/ L and mixed for ~ 22 hours prior to use. After mixing, primary stock solution was proportionally diluted with dilution water to prepare the four additional test concentrations. All test solutions appeared clear and colorless. Concentrations dosing rate: Once Stability of the test chemical solutions: Extremely stable Exposure vessels: 2L polyethylene aquaria containing approximately 1000mL of test solution; water depth approximately 6.6 cm. Number of replicates: two Number of mysids per replicate: ten Number of concentrations: five plus a negative control Feeding: Live brine shrimp nauplii daily Water chemistry during the study: Dissolved oxygen range ( 0   96 hours): 6.8   7.4 mg/ L ( control exposure) 6.8   7.3 mg/ L ( 5.4 mg/ L exposure) pH range ( 0   96 hours): ENV/ JM/ RD( 2002) 17/ FINAL 112 8.1   8.2 ( control exposure) 8.1   8.2 ( 5.4 mg/ L exposure) Test temperature range ( 0   96 hours): 24.2   25.4oC ( control exposure) 23.8   24.5oC ( 5.4 mg/ L exposure) Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative control, 1.1, 1.8, 3.0, 4.9, 8.2 mg/ L Measured concentrations: < LOQ, 0.57, 1.1, 1.9, 3.0, 5.4 mg/ L Element value: ( 95% confidence interval is given in brackets) 24 hour LC50 = > 5.4 mg/ L ( CI not calculable) 48 hour LC50 = > 5.4 mg/ L C. I. not calculable) 72 hour LC50 = 4.4 ( 3.6 6.2) mg/ L 96 hour LC50 = 3.6 ( 3.0 4.6) mg/ L All element values based on mean measured concentrations Statistical evaluation of mortality: LC50 values could not be calculated for 24 and 48 hours of exposure due to the lack on an adequate concentration response pattern. The probit method was used to evaluate mortality at 72 and 96 hours.

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 0.115 mg/ L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 97.4. Samples collected at test initiation had measured values from 52.4 to 70.7% of nominal. Measured values for samples taken at 48 hours ranged from 43.5 to 71.0% of nominal. Measured values for samples taken at 96 hours ranged from 35.5 to 71.1% of nominal.

Summary of analytical chemistry data:

Nominal Test Concentration, mg/ L Measured duplicate values at 0, 48, and 96 hours respectively, mg/ L Mean Measured Concentration mg/ L Percent of Nominal Negative Control All < LOQ < LOQ  1.1 0.575, 0.622, 0.605, 0.640, 0.391, 0.580 0.57 52 1.8 1.12, 1.19, 1.10, 1.09, 1.04, 1.13 1.1 61 3.0 1.92, 1.99, 1.92, 1.91, 1.79, 1.91 1.9 63 4.9 3.05, 2.66, 2.96, 3.35, 3.11, 3.11 3.0 61 8.2 5.82, 5.78, 3.58, 5.85, 5.22, 5.86 5.4 66 Biological observations after 96 hours: Mysids in the negative control, and the 0.57 and 1.1 mg/ L ( mean measured concentrations) treatment groups appeared normal and healthy during the test. ENV/ JM/ RD( 2002) 17/ FINAL 113 Cumulative percent mortality:

Mean Measured Test Concentration, mg/ L 24 hours 48 hours 72 hours 96 hours Negative Control 0 0 0 0 0.57 0 0 0 0 1.1 0 0 0 0 1.9 0 0 5 10 3.0 5 15 30 40 5.4 15 45 60 75 Mortality of controls: None CONCLUSIONS The potassium perfluorooctanesulfonate 96 hour LC50 for saltwater mysids was determined to be 3.6 mg/ L with a 95% confidence interval of 3.0   4.6 mg/ L. The 96 hour no mortality and NOEC concentration was 1.1 mg/ L.

Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA 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 ENV/ JM/ RD( 2002) 17/ FINAL 114 Robust Study Report Reference No. 8  Early Life Stage Toxicity Test with the Fathead Minnow (

 )

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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: 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, USA 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 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. Test fish age: eggs < 24 hours old at test initiation Pretreatment: None Test Conditions Dilution water: 0.45 µ m filtered well water Dilution water chemistry ( during the 4 week period immediately preceding the test): Hardness: 126 ( 124 128) mg/ L as CaCO3 Alkalinity: 172 ( 170 172) mg/ L as CaCO3 pH: 8.2 ( 8.2 8.3) TOC: < 1.0 mg/ L Conductivity: 321 ( 315 330) µ mhos/ cm 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 µ m nylon screen mesh attached to the bottom with ENV/ JM/ RD( 2002) 17/ FINAL 115 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 of fish 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   47 days): 24.4   24.7oC ( control exposure) 24.3   24.7oC ( 1.2 mg/ L exposure) Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative 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 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. ENV/ JM/ RD( 2002) 17/ FINAL 116 Summary of analytical chemistry data:

Nominal Test Concentration, mg/ L Measured duplicate values at 0, 4, 7, 14, 21, 28, 35, 42, and 47 Days respectively, mg/ L Mean Measured Concentration, mg/ L Percent of Nominal Negative Control All < LOQ   0.14 0.147, 0.160, 0.141, 0.140, 0.144, 0.148, 0.134, 0.135, 0.153, 0.143, 0.160, 0.158, 0.179, 0.173, 0.157, 0.160, 0.147, 0.155 0.15 107 0.29 0.287, 0.277, 0.270, 0.289, 0.292, 0.296, 0.269, 0.266, 0.307, 0.315, 0.343, 0.341, 0.311, 0.325, 0.319, 0.313, 0.296, 0.276 0.30 103 0.57 0.571, 0.576, 0.619, 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 0.60 105 1.1 1.14, 1.13, 1.21, 1.25, 1.13, 1.23, 1.03, 1.10, 1.19, 1.24, 1.30, 1.31, 1.30, 1.31, 1.14, 1.19, 1.13, 1.09 1.2 109 2.3 2.21, 2.27, 2.52, 2.46, 2.43, 2.38, fish all dead at Day 7 2.4 104 4.6 4.56, 4.40, 4.79, 4.79, 4.46, 4.76, fish all dead at Day 7 4.6 100 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 Mean Measured Concentration mg/ L Number of Eggs Exposed Number Hatched, Day 3 Number Hatched, Day 4 Number Hatched, Day 5 Total Number Hatched Percent Hatching Success Negative Control 80 0 20 54 74 93 0.15 80 0 18 58 76 95 0.3 80 0 14 58 72 90 0.6 80 0 28 48 76 95 1.2 80 0 25 49 74 93 2.4 80 0 16 59 75 94 4.6 80 0 14 60 74 93 ENV/ JM/ RD( 2002) 17/ FINAL 117 Larval Survival Mean Measured Concentration, mg/ L Percent Survival, Day 42 Negative Control 88 0.15 79 0.3 81 0.6 66 1.2 5.4 2.4 0 4.6 0 Growth Mean Measured Concentration, mg/ L Number of Surviving Larvae Total Length Mean + SD, mm Wet Weight Mean + SD, mg Dry Weight Mean + SD, mg Negative Control 65 26.5 + 0.721 158 + 9.10 32.5 + 1.20 0.15 60 26.6 + 0.208 160 + 3.10 33.3 + 0.900 0.30 58 26.6 + 0.813 167 + 11.9 34.2 + 2.70 0.60 50 26.5 + 0.399 166 + 11.3 33.5 + 2.70 1.2 4 26.7 + 2.02 185 + 33.8 35.4 + 6.66 2.4 0    4.6 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, MN 55133, USA.

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 ENV/ JM/ RD( 2002) 17/ FINAL 118 Robust Study Report Reference No. 9  Semi Static Life Cycle Toxicity Test with the Cladoceran (

)

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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 850.1300, OECD Guideline 211, and ASTM Standard E 1193 87. Type: Semi Static Life Cycle Toxicity GLP: Yes Year completed: Study completed 1999. Report completed 2000 Species: Daphnia magna Supplier: In house cultures, Wildlife International, Ltd., Easton, MD, USA Analytical monitoring: PFOS measured on days 0, 2, 11, 14, 18, and 21. Exposure period: 21 days Statistical methods: Survival data was evaluated on first generation daphnids, the number of live young and the length and dry weight of the surviving first generation daphnids. Survival data were analyzed using Fisher's exact test. Reproduction and growth ( length and dry weight) data 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 was used to identify treatment groups that were statistically significant in comparison to the negative control ( p < 0.05). All statistical tests were performed using a personal computer with SPSS/ PC Version 2.0 or " TOXSTAT Release 3.5" statistical software. Test organism age: < 24 hours old at test initiation Pretreatment: None Test Conditions Dilution water: 0.45 µ m filtered well water passed through a UV sterilizer to remove microorganisms and fine particles Dilution water chemistry ( during the 4 week period immediately preceding the test): Hardness: 124 ( 120 128) mg/ L as CaCO3 Alkalinity: 169 ( 164 172) mg/ L as CaCO3 pH: 8.2 ( 8.0 8.3) TOC: < 1.0 mg/ L Conductivity: 329 ( 315 340) µ mhos/ cm Ca/ Mg ratio: 35/ 13.5 Na/ K ratio: 21.3/ 6.62 Stock and test solution preparation: Primary stock solution was prepared in dilution water at 46 mg/ L. It was stirred until all test substance was dissolved prior to use. After mixing, the primary stock solution was proportionally diluted with UV sterilized dilution water to prepare five additional stock solutions at nominal concentrations of 1.4, 2.9, 5.7, 11, and 23 mg/ L. All test solutions appeared clear and colorless. Renewal rate: Every Monday, Wednesday and Friday. Exposure vessels: 250 mL plastic beakers containing approximately 200 mL test solution. The depth was approximately 5 cm. Number of replicates: 10 ENV/ JM/ RD( 2002) 17/ FINAL 119 Number of test organisms per replicate: 1 Number of concentrations: 6 plus a negative control Feeding: Each test chamber was fed 0.3 mL of YCT ( a mixture of yeast, Cerophyll ® , and trout chow at 1800 mg TSS/ L) and 0.60 mL of Selenastrum capricornutum ( 3.5 x 107 cells/ mL) once daily. Lighting: Colortone ® 50 fluorescent lights. Intensity ranged from 329  383 lux at the water surface. Photoperiod of 16 hours light, 8 hours dark with a 30 minute transition period. Water chemistry of new and old solutions during the study: Dissolved oxygen range ( 0   21 days): 8.3   8.9 mg/ L ( negative control exposure) 8.3   9.0 mg/ L ( 12 mg/ L exposure) 8.4   8.9 mg/ L* ( 48 mg/ L exposure) pH range ( 0   21 days): 8.1   8.4 ( negative control exposure) 8.2   8.5 ( 12 mg/ L exposure) 8.4   8.5* ( 48 mg/ L exposure) Test temperature range ( 0   21 days): 19.4   20.1oC ( negative control exposure) 19.4   20.1oC ( 12 mg/ L exposure) 19.4   19.5 oC* ( 48 mg/ L exposure)

* ( Measurements discontinued at Day 3 due to 100% mortality.

Element basis: Survival, reproduction and growth. Effect concentrations based on survival. Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative control, 1.4, 2.9, 5.7, 11, 23, 46 mg/ L Measured concentrations: < LOQ, 1.5, 2.9, 5.6, 12, 24, 48 mg/ L Element value: 21 day NOEC = 12 mg/ L 21 day LOEC = 24 mg/ L 21 day MATC = 17 mg/ L 2nd generation acute survival NOEC = 12 mg/ L 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 the test substance was used. No attempt was made to quantify on the basis of individual isomeric components. The LOQ ( limit of quantitation) was 0.458 mg/ L in this study. The mean procedural recovery of matrix fortifications analyzed concurrently during sample analysis was 104%. Measured values of new samples ranged from 94 to 121% of nominal. Measured values from the old solutions ranged from 90 to 108% of nominal values. PFOS was stable throughout the renewal periods. ENV/ JM/ RD( 2002) 17/ FINAL 120 Summary of analytical chemistry data:

Nominal Test Concentration mg/ L Measured duplicate values at 0, 2, 11, 14, 18, and 21 Days respectively, mg/ L Mean Measured Concentration mg/ L Percent of Nominal Negative Control All < LOQ   1.4 1.78, 1.72, 1.58, 1.56, 1.38, 1.47, 1.36, 1.32, 1.38, 1.43, 1.50, 1.45 1.5 107 2.9 3.20, 3.05, 3.01, 3.07, 2.75, 2.77, 2.85, 2.71, 2.79, 2.81, 2.81, 2.82 2.9 100 5.7 5.97, 5.87, 5.65, 5.72, 5.63, 5.59, 5.36, 5.39, 5.58, 5.75, 5.24, 5.37 5.6 98 11 11.5, 11.5, 11.6, 11.8, 11.3, 11.3, 11.2, 11.6, 11.8, 11.6, 11.5, 11.3 12 109 23 24.2, 23.1, 24.0, 24.6, 22.8, 22.5, 23.6, 23.1, 24.8, 25.0, all daphnids dead after 18 days exposure 24 104 46 47.3, 48.0, 49.1, 49.4, all daphnids dead after 2 days exposure 48 104 NOTE: Mean measured concentrations were determined from new ( renewal solutions) and corresponding old solutions during each week of the test. Days 0, 11, and 18 are " new" and days 2, 24, and 21 are " old".

Biological Observations Survival: All surviving first generation daphnids appeared normal at test termination. Survival in the 24 and 48 mg/ L treatments was statistically significantly different from the negative control group. Reproduction: Daphnids in the control and treatment groups < 12 mg/ L started producing neonates on Day 9. The Bonferroni t test showed that reproduction was not significantly reduced in any treatment group < 12 mg/ L ( p > 0.05). The 24 and 48 mg/ L treatment groups were not included in the statistical analysis of the reproduction data due to a statistically significant effect on survival. Growth: The Bonferroni t test showed that mean length and dry weight in the treatment groups < 12 mg/ L were not significantly reduced in comparison to the negative control ( p > 0.05). Second Generation Acute Exposure: After 48 hours of exposure, survival in the negative control was 95%. Survival in the 1.5, 2.9, 5.6, 12, and 24 mg/ L treatment groups was 100, 100, 100, 90, and 0% respectively. Survival in the 24 mg/ L treatment group was significantly different from the negative control ( p < 0.05).

Summary of Percent Mortality Mean Measured Concentration, mg/ L Day 7 Day 14 Day 21 Negative Control 0 0 0 1.5 0 0 10 2.9 0 0 10 5.6 0 0 10 12 0 10 10 24 70 90 100 48 100 100 100 ENV/ JM/ RD( 2002) 17/ FINAL 121 Second Generation Mortality Mean Measured Concentrations, mg/ L Total Number Exposed Number Alive after 48 hours Cumulative Percent Dead Negative Control 20 19 5 1.5 20 20 0 2.9 20 20 0 5.6 20 20 0 12 20 18 10 24 8 0 100 Summary of Length and Dry Weight of Surviving Individually Exposed First Generation Daphnids Mean Measured Concentration, mg/ L Number of Surviving Daphnids Total Length, Mean + SD, mm Dry Weight, Mean + SD, mg Negative Control 10 4.65 + 0.111 0.695 + 0.100 1.5 9 4.66+ 0.118 0.669 + 0.0623 2.9 9 4.62 + 0.100 0.724 + 0.110 5.6 9 4.61 + 0.124 0.727 + 0.0665 12 9 4.59 + 0.102 0.723 + 0.0661 24 0     48 0

Reproduction Mean Measured Concentration, mg/ L Number of Surviving Daphnids Mean Live Young/ Surviving Adult Daphnid (+ SD) First Day of Reproduction Total Number of Dead / Immobile Neonates Total Number of Aborted Eggs Negative Control 10 122 + 19.2 9 0 0 1.5 9 142 + 24.7 9 0 0 2.9 9 136 + 17.9 9 0 0 5.6 9 132 + 19.5 9 0 0 12 9 119 + 26.5 9 1 0 24 0   11 10 0 48 0   None

CONCLUSIONS There were no adverse effects on survival, reproduction or growth of Daphnia magna exposed to the test substance at concentration < 12 mg/ L for 21 days. Daphnia magna exposed to 24 and 48 mg/ L had significantly reduced survival.

Author and/ or submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA.

DATA QUALITY Reliability: Klimisch ranking 1. ENV/ JM/ RD( 2002) 17/ FINAL 122 REFERENCES This study was conducted at Wildlife International, Ltd. Easton, MD at the request of the 3M Company.

OTHER Last changed: 5/ 3/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 123 Robust Study Report Reference No. 10  Flow through Life Cycle Toxicity Test with the Saltwater Mysid (

)

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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, USA 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% o with well water, 0.45 µ m filtered. Dilution water chemistry ( during the 4 week period immediately preceding the test): Salinity: 20 ( 20 20) % o TOC: < 1.0 mg/ L 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 postpairing 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. ENV/ JM/ RD( 2002) 17/ FINAL 124 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 of fish 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   25.2oC ( control exposure) 24.4   25.1oC ( 1.3 mg/ L exposure)

Method of calculating mean measured concentrations: arithmetic mean RESULTS Nominal concentrations: Negative 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 0.0458 mg/ L in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 92.8. Samples collected at pretest 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. ENV/ JM/ RD( 2002) 17/ FINAL 125 Summary of analytical chemistry data:

Nominal Test Concentration, mg/ L Measured duplicate values at 0, 7, 14, 21, 28, 35, Days respectively, mg/ L Mean Measured Concentration, mg/ L Percent of Nominal Negative Control All < LOQ   0.086 0.0694, 0.0578, 0.0478, 0.0619, 0.0606, 0.0614, 0.0554, 0.0509, 0.0515, 0.0569, 0.0580, 0.0514 0.057 66 0.17 0.125, 0.114, 0.0778, 0.125, 0.124, 0.127, 0.0970, 0.112, 0.122, 0.128, 0.124, 0.119 0.12 71 0.34 0.289, 0.286, 0.231, 0.197, 0.276, 0.253, 0.227, 0.212, 0.262, 0.271, 0.278, 0.251 0.25 74 0.69 0.562, 0.659, 0.581, 0.450, 0.543, 0.542, 0.516, 0.528, 0.529, 0.544, 0.556, 0.583 0.55 80 1.4 1.23, 1.32, 1.13, 1.20, 1.35, 1.27, 1.23, 1.15, 1.39, 1.39, 1.26, 1.20 1.3 93 2.7 2.56, 2.79, 2.58, 2.30, 2.54, 2.69, all mysids dead after 14 days exposure 2.6 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.

Percent Survival Mean Measured Concentration, mg/ L Juvenile Pre Pairing Survival, Day 20 Adult Post Pairing Survival, Day35 Negative Control 78 92 0.057 92 96 0.12 75 90 0.25 82 97 0.55 83 95 1.3 32 57 2.6 0  ENV/ JM/ RD( 2002) 17/ FINAL 126 Second Generation Survival Mean Measured Concentrations, mg/ L Total Number Exposed Number Alive after 96 hours Percent Survival Negative Control 71 68 96 0.057 65 63 97 0.12 83 79 95 0.25 62 59 95 0.55 13 13 100 Adult Mysid Growth Mean Measured Concentration, mg/ L Number of Surviving Mysids/ Number Exposed Total Length, Mean + SD, mm Dry Weight, Mean + SD, mg Negative Control 36/ 39 6.43 + 0.0634 0.634 + 0.0510 0.057 44/ 46 6.43 + 0.0729 0.599 + 0.0276 0.12 36/ 40 6.56 + 0.105 0.641 + 0.0241 0.25 36/ 37 6.40 + 0.0548 0.622 + 0.0227 0.55 35/ 37 6.14 + 0.0794 0.562 + 0.00624 1.3 8/ 14 5.85 + 0.178 0.436 + 0.0441 Reproduction Mean Measured Concentration, mg/ L Replicate Number of Reproductive Days Number of Young Mean Number of Young/ Reproductive Day Overall Mean+ SD Negative control A 70 18 0.257 0.315 + 0.0925 B 53 14 0.264 C 70 20 0.286 D 42 19 0.452 0.57 A 60 17 0.283 0.261 + 0.0873 B 70 14 0.200 C 70 13 0.186 D 56 21 0.375 0.12 A 70 21 0.300 0.361 + 0.101 B 46 22 0.478 C 54 22 0.407 D 70 18 0.257 0.25 A 70 19 0.271 0.252 + 0.0723 B 56 12 0.214 C 61 21 0.344 D 56 10 0.179 0.55 A 54 3 0.0556 0.0559 + 0.0376 B 56 6 0.107 C 70 3 0.0429 D 56 1 0.0179 ENV/ JM/ RD( 2002) 17/ FINAL 127 1.3 A 22 0   B 14 0  C 0 0  D 11 0

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, MN 55133, USA 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: 7/ 9/ 01 ENV/ JM/ RD( 2002) 17/ FINAL 128 Robust Study Report Reference No. 11   Dietary LC50 Study with the Mallard duck TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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, ' H HMR, 19FNMR and elemental analyses techniques.

METHOD Method: OPPTS 850.2200, OECD 206, and FIFRA Subdivision E. Section 71 2 Type: Dietary acute GLP: Yes Year completed: 2000 Species: Anas platyrhynchos Supplier: Whistling Wings, Inc., Hanover, IL, USA Analytical monitoring: PFOS measured on Day 0 for homogeneity in feed and verification, and Day 5 for stability. Test phases: Acclimation  9 days Exposure  5 days Post exposure observation  3 or 17 days Statistical methods: LC50 values calculated by probit analysis using the computer software of C. E. Stephan. Body weight data were compared by Dunnett's test using TOXSTAT software. No statistical analyses were applied to feed consumption data. Test bird age: 10 days Pretreatment: None Test conditions: Housing and environmental conditions: Indoors In batteries of thermostatically controlled brooding pens. Floor space of each pen measured approximately 62 x 90 cm. Ceiling height was approximately 25.5 cm. External walls, ceilings and floors were constructed of galvanized steel wire and sheeting. ldentification: Each group of birds Identified by pen number and test concentration. Individuals identified by wing bands. Number of replicates: Six for controls, two for each treatment group Number of ducks per replicate: five Number of concentrations: Eight plus a negative control Feed and water: Game bird ration formulated as below, water from the town of Easton public water supply. Both provided ad libitum during acclimation and testing. ENV/ JM/ RD( 2002) 17/ FINAL 129 Game Bird Ration Ingredients Percent Fine Corn Meal 44.83 Soy Bean Meal, 48% Protein 30.65 Wheat Midds 6.50 Protein Base 6.00 Agway Special, 60% Protein 4.00 Alfalfa Meal, 20% Protein 3.00 Dried Whey 2.50 Ground Limestone 0.90 Eastman CalPhos 0.60 Methionine Premix + Liquid 0.35 Vitamin and Mineral Premix ( see below) 0.32 GL Form ( Fermatco) 1 0.25 Salt iodized 0.10 1Fermentation by products ( source of unidentified growth factors)

Vitamin and Mineral Premix Vitamin or Mineral Amount Per Ton Vitamin D3 2,000,000 I. C. U. Vitamin A 7,000,000 I. U. Riboflavin 6 g Niacin 40 g Pantothenic Acid 10 g Vitamin B12 8 mg Folic Acid 600 mg Biotin 64 mg Pyridoxine 1.2 g Thiamine 1.2 g Vitamin E 20,000 I. U. Vitamin K ( Menadione dimethylpyrimidinol bisulfite) 5.8 g Manganese 102 g Zinc 47 g Copper 6.8 g Iodine 1.5 g Iron 51 g Selenium 182 g Prophylaxis: None Brooding compartment mean temperature: 38 ± 2oC Ambient room mean temperature: 25.2 ± 0.7oC Average relative humidity: 53 ± 18% Photoperiod: Sixteen hours light per day Lighting: fluorescent lights which closely approximate noon day sunlight; average approximately 207 lux. ENV/ JM/ RD( 2002) 17/ FINAL 130 Test diet preparation: Test substance mixed directly into the ration by means of a Hobart mixer. No carrier was used.

Diet sampling: Homogeneity of the test substance in the diet evaluated by collecting six samples from the 9.1 ppm and six from the lowest and highest concentration. Samples collected from the top, middle, and bottom of the left and right sections of the mixing vessel. These samples also served as the verification samples for these concentrations. Two verification samples from the remaining concentrations and one from the control were collected at preparation on Day 0. Stability samples were collected at the end of the exposure period ( Day 5) from the control ( one sample) and each treatment group ( two samples each).

RESULTS Nominal concentrations: Negative control, 9.1, 18.3, 36.6, 73.2, 146, 293, 586, and 1171 ppm Measured concentrations: < LOQ, 9.8, 19.5, 40.2, 74.5, 174, 291, 537, and 1196 ppm Element value: Dietary LC50 = 628 ( 448  958) ppm No mortality concentration 146 ppm NOEC ( body weight gain) 36.6 ppm All element values based on nominal concentrations Analytical Methodology: Diet samples were extracted with methanol. 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 1.15 ppm in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 94.7. Samples collected for determination of homogeneity in diet ranged from 102  108% of nominal. Samples collected for verification in diet had measured values from 92 to 119% of nominal. Measured values for ambient stability samples taken at Day 5 ranged from 94   130% of nominal.

Summary of analytical chemistry data Homogeneity in Avian diet Nominal Test Concentration, ppm Measured Values at Day 0, ppm Mean Measured Concentration, ppm Percent of Nominal 9.1 9.52, 9.70, 9.79, 8.09, 10.9, 10.5 9.8 108 18.3 18.5, 23.4, 18.3, 17.3, 19.4, 19.9 19.6 107 1171 1239, 1221, 1118, 1301, 1163, 1133 1196 102 Verification in avian diet Nominal Test Concentration, ppm Measured Duplicate Concentrations at Day 0, ppm Mean Measured Concentration, ppm Percent of Nominal Negative Control < LOQ   36.6 46.7, 34.6 40.2 110 73.2 77.8, 71.2 74.5 102 146 176, 172 174 119 293 274, 307 291 99 586 550, 523 537 92 ENV/ JM/ RD( 2002) 17/ FINAL 131 Ambient stability in avian diet Day 0 Mean Measured Concentration, ppm Measured Duplicate Concentrations at Day 5, ppm Mean Measured Concentration, ppm Mean Percent of Day 0 Negative Control < LOQ  9.8 12.3, 11.0 11.7 119 19.5 18.2, 19.7 19.0 97 40.2 47.9, 56.8 52.4 130 74.5 77.6, 77.9 77.8 104 174 167, 160 164 94.3 291 297, 293 295 101 537 552, 530 541 101 1196 1150, 1122 1136 95 Biological observations Survival and clinical observations: No mortalities occurred in the control group, and all birds were normal in appearance and behavior throughout the test. The first deaths occurred on day 4 in the 1171 ppm treatment. Mortality occurred through Day 8 in all dose groups > 293 ppm with some of the deaths being during the post exposure period. There were no treatment related mortalities or overt signs of toxicity at concentrations < 146 ppm. Birds at all concentrations > 293 ppm displayed signs of toxicity including reduced reaction to stimuli ( sound and motion), loss of coordination, ruffled appearance, lethargy and lower limb weakness. Birds at the 1171 ppm level also displayed prostrate posture, depression and convulsions through Day 8. Recovery with normal appearance and behavior was noted from Day 9 through test termination Body weight gain: When compared to the control group, there were no apparent treatment related effects on body weight among the birds in concentrations < 36.6 ppm. During the Day 8 15 and Day 15 22 post exposure periods, body weight gain appeared comparable among all groups. There was a statistically significant ( p < 0.05) reduction in weight gain at the 9.1 ppm level for the Day 0 5 and Day 5 8 periods. However, differences from the control group at the 9.1 ppm level appear to be due to a lower mean Day 0 body weight for the 9.1 ppm level, and were not dose responsive. Therefore, these differences were not considered treatment related. Marked, treatment related, concentration responsive effects on body weight was noted in concentrations > 73.2 ppm for Days 0 5; Day 5 8 post exposure weight gain continued to be reduced at concentrations > 293 ppm. Feed Consumption: When compared to the control group, there was a marked reduction in feed consumption in the treatment groups > 293 ppm throughout the study. Gross Necropsy: All birds that died during the study, half of those surviving at Day 8 and the rest at test termination were subjected to a gross necropsy. Necropsy results for birds found dead were similar, including thin condition, loss of muscle mass, altered spleen color, empty crops, and empty gastrointestinal tracts. These necropsy findings were considered to be treatment related.

Percent Cumulative Mortality Nominal Concentration, ppm Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8*

Negative Control 0 0 0 0 0 0 0 0 9.1 0 0 0 0 0 0 0 0 18.3 0 0 0 0 0 0 0 0 36.6 o 0 0 0 0 0 0 0 ENV/ JM/ RD( 2002) 17/ FINAL 132 73.2 0 0 0 0 0 0 0 0 146 0 0 0 0 0 0 0 0 293 0 0 0 0 0 0 10 20 586 0 0 0 0 10 20 30 30 1171 0 0 0 20 60 80 90 90 * No mortalities occurred in any of the treatment levels from Day 8 to Day 22 Bodyweight ( grams)

Exposure Period Recovery Period Nominal Concentration, ppm Mean Body Weight, Day 0 Mean Body Weight Change Day 0 5 Mean Body Weight Change Day 5 8 Mean Body Weight Change Day 8 15 Mean Body Weight Change Day 15 22 Mean Total Body Weight Change Day 8 22 Mean Body Weight Day 22 Negative Control 135 144 101 230 183 413 823 9.1 119 108 91 230 198 427 773 18.3 146 131 100 241 186 427 811 36.6 147 128 100 243 208 451 828 73.2 143 117 82 216 203 418 782 146 143 100 89 232 124 356 688 293 129 32 57 256 234 490 701 586 144  6 36 221 219 439 613 1171 147  37 1531 198 251 449 634 Mean average feed consumption Nominal Concentration, ppm Grams food/ bird/ day Days 0 5 Grams food/ bird/ day Days 6 8 Grams food/ bird/ day Days 8 15 Grams food/ bird/ day Days 15 22 Negative Control 92 125 171 180 9.1 73 117 172 198 18.3 91 132 186 204 36.6 94 125 165 179 73.2 77 101 148 173 146 105 159 159 164 293 44 63 109 132 586 36 55 114 143 1171 22 25 106 154 Gross pathological observations from Birds that died in study Finding 293ppm N = 2 586ppm N = 3 1171ppm N = 9 Crop empty 0 2 7 Emaciated 1 1 4 G. I. Tract, primarily empty 1 1 1 Gizzard contents bile stained 0 2 4 Gizzard, empty 0 0 1 Intestinal contents, black and tar like 0 0 1 Keel prominent 0 0 2 ENV/ JM/ RD( 2002) 17/ FINAL 133 Kidneys, pale 0 0 1 Loss of muscle mass 2 2 5 Spleen, grey 0 0 1 Spleen, small and pale 1 1 2 Spleen, pale 0 2 3 Thin 1 1 4 CONCLUSIONS The dietary LC50 value for Mallard Duck exposed to PFOS was determined to be 628 ppm with a 95% confidence interval of 448 to 958 ppm. The slope of the concentration response curve was 3.67 and the chi square value was 2.13. The no mortality concentration was 146 ppm. Based upon reductions in body weight gain at the 73.2 ppm test concentration, the no observed effect concentration was 36.6 ppm.

Submitter: 3M Company, Environmental Laboratory, P. O. Box 33331, St. Paul, Minnesota, 55133, USA 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/ 1/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 134 Robust Study Report Reference No. 12   Dietary LC50 Study with the Northern Bobwhite quail TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, 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, 1H HMR, 19F NMR and elemental analyses techniques.

METHOD Method: OPPTS 850.2200, OECD 205, and FIFRA Subdivision E. Section 71 2 Type: Dietary acute GLP: Yes Year completed: 1999 ( Test), 2000 ( Report) Species: Colinus virginianus Supplier: Wildlife International Ltd. Production Flock, Easton, Maryland, USA Analytical monitoring: Test substance concentration in standards and samples were determined by reversed phase HPLC and mass spectroscopy. PFOS measured on Day 0 for homogeneity in feed and verification, and Day 5 for stability. Test phases: Acclimation   10 days Exposure   5 days Post exposure observation   3 or 17 days Statistical methods: LC50 values calculated by probit analysis using the computer software of C. E. Stephan. Body weight data were compared by Dunnett's test using TOXSTAT software. No statistical analyses were applied to feed consumption data. Test bird age: 10 days Pretreatment: None Test conditions: Housing and environmental conditions: Indoors in batteries of thermostatically controlled brooding pens. Each pen's floor space measured approximately 72 x 90 cm. Ceiling height was approximately 23 cm. External walls, ceilings and floors were constructed of galvanized steel wire and sheeting. Identification: Each group of birds Identified by pen number and test concentration. Individuals Identified by leg bands. Number of replicates: Six for controls, two for each treatment group Number of bobwhite per replicate: five Number of concentrations: seven plus a negative control Feed and water: Game bird ration formulated as below, water from the town of Easton public water supply. Both provided ad libitum during acclimation and testing. ENV/ JM/ RD( 2002) 17/ FINAL 135 Game Bird Ration Ingredients Percent Fine Corn Meal 44.83 Soy Bean Meal, 48% Protein 30.65 Wheat Midds 6.50 Protein Base 6.00 Agway Special, 60% Protein 4.00 Alfalfa Meal, 20% Protein 3.00 Dried Whey 2.50 Ground Limestone 0.90 Eastman CalPhos 0.60 Methionine Premix + Liquid 0.35 Vitamin and Mineral Premix ( see below) 0.32 GL Form ( Fermatco) 1 0.25 Salt iodized 0.10 1Fermentation by products ( source of unidentified growth factors)

Vitamin and Mineral Premix Vitamin or Mineral Amount Per Ton Vitamin D3 2,000,000 I. C. U. Vitamin A 7,000,000 I. U. Riboflavin 6 g Niacin 40 g Pantothenic Acid 10 g Vitamin B12 8 mg Folic Acid 600 mg Biotin 64 mg Pyridoxine 1.2 g Thiamine 1.2 g Vitamin E 20,000 I. U. Vitamin K ( Menadione dimethylpyrimidinol bisulfite) 5.8 g Manganese 102 g Zinc 47 g Copper 6.8 g Iodine 1.5 g Iron 51 g Selenium 182 g Prophylaxis: None Brooding compartment mean temperature: 38 ± 2oC Ambient room mean temperature: 27.3 ± 1.2oCAverage relative humidity: 31 ± 14% Photoperiod: Sixteen hours light per day Lighting: fluorescent lights which closely approximate noon day sunlight; average of approximately 139 lux of illumination Test diet preparation: Test substance mixed directly into the ration by means of a Hobart mixer. No ENV/ JM/ RD( 2002) 17/ FINAL 136 carrier was used.

Diet sampling: Homogeneity of the test substance in the diet evaluated by collecting six samples from the, 18.3 pprn concentration and six from the 1171 ppm concentration. Samples collected from the top, middle, and bottom of the left and right sections of the mixing vessel. These samples also served as the verification samples for these concentrations. Verification samples of the other treatment groups ( two samples from each) and the control ( one sample) were collected at preparation on Day 0. Stability samples were collected at the end of the exposure period ( Day 5) from the control ( one sample) and each treatment group ( two samples each).

RESULTS Nominal concentrations: Negative control, 18.3, 36.6, 73.2, 146, 293, 586, and 1171 ppm Measured concentrations: < LOQ, 19.5. 40.2, 74.5, 174, 291, 537, and 1196 ppm Element value: Dietary LC50 = 220 ( 164  289) ppm No mortality concentration = 73.2 ppm NOEC ( body weight gain) = 73.2 ppm All element values based on nominal concentrations Analytical Methodology: Diet samples were extracted with methanol. 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 1.15 ppm in this study. The mean percent recovery of matrix fortifications analyzed concurrently during sample analysis was 94.7. Samples collected for determination of homogeneity in diet ranged from 102 107% of nominal. Samples collected for verification in diet had measured values from 92 to 119% of nominal. Measured values for ambient stability samples taken at Day 5 ranged from 101   122% of nominal.

Summary of analytical chemistry data Homogeneity in avian diet Nominal Test Concentration, ppm Measured Values at Day 0, ppm Mean Measured Concentration, ppm Percent of Nominal 18.3 18.5, 23.4, 18.3, 17.3, 19.4, 19.9 19.5 107 1171 1239, 1221, 1118, 1301, 1163, 1133 1196 102 Verification in avian diet Nominal Test Concentration, ppm Measured Duplicate Concentrations at Day 0, ppm Mean Measured Concentration, ppm Percent of Nominal Negative Control < LOQ   36.6 46.7, 34.6 40.2 110 73.2 77.8, 71.2 74.5 102 146 176, 172 174 119 293 274, 307 291 99 586 550, 523 537 92 ENV/ JM/ RD( 2002) 17/ FINAL 137 Ambient stability in avian diet Day 0 Mean Measured Concentration, ppm Measured Duplicate Concentrations at Day 5, ppm Mean Measured Concentration, ppm Mean Percent of Day 0 Negative Control < LOQ  19.5 19.2, 19.9 19.6 101 40.2 44.4, 53.8 49.1 122 74.5 76.4, 77.9 77.2 104 174 177, 174 176 101 291 318, 315 317 109 537 560, 665 613 114 1196 1260, 1187 1224 102 Biological observations Mortalities and clinical observations: One incidental mortality occurred in the control group as a result of a, broken leg on the morning of Day 5. It was subsequently euthanized on Day 6. Two other birds in the control group were intermittently noted with foot lesions associated with cage mate aggression. Otherwise, all control birds were observed to be normal in appearance and behavior throughout the test.

The first treatment related mortalities occurred on Day 3 in the 586 and 1171 ppm treatment groups. Mortality occurred through Day 8 in all dose groups > 146 ppm with some of the deaths being during the post exposure period. There were no treatment related mortalities or overt signs of toxicity at concentrations < 73.2 ppm.

There was 11 % mortality in the 146 ppm treatment group, and two additional birds displayed clinical signs of toxicity ( wing droop). All other birds in this test group displayed normal appearance and behavior for the duration of the test. Recovery with normal appearance and behavior occurred on Day 9 to test termination.

There was 80% mortality ( occurring on Days 5, 6, and 7) for birds in the 293 ppm treatment group. Signs of toxicity observed prior to death included a ruffled appearance, reduced reaction to stimuli ( sound and motion), lethargy, wing droop, loss of coordination, lower limb weakness and convulsions. Recovery with normal appearance and behavior occurred on Day 9 to test termination.

There was 100% mortality ( occurring from Day 3 through Day 7) for birds in the 586 ppm treatment group. Signs of toxicity observed prior to death included a ruffled appearance, reduced reaction to stimuli ( sound and motion), lethargy, depression, wing droop, loss of coordination, lower limb weakness, lower limb rigidity, prostrate posture, and convulsions.

There was 100% mortality ( occurring from Day 2 ( noted on Day 3 for Day 2 afternoon) through Day 4) for birds In the 1171 ppm treatment group. Signs of toxicity observed prior to death included a ruffled appearance, reduced reaction to stimuli ( sound and motion), lethargy, depression, wing droop, loss of coordination, lower limb weakness, and lower limb rigidity.

Body weight gain: When compared to the control group, there were no apparent treatment related effects on body weight among the birds in concentrations < 73.2 ppm. During Days 0 5 statistically significant reductions in body weight gain or body weight loss occurred in the 146, 293, and 586 ppm treatment groups. Body weight effects could not be determined for test ENV/ JM/ RD( 2002) 17/ FINAL 138 organisms in the 1171 ppm group due to total mortality.

Feed Consumption: No apparent treatment related effects were noted for feed consumption for birds in concentrations < 146 ppm. Reduced feed consumption was noted for birds In treatment groups > 293 ppm from Days 0 5. No treatment related effects on feed consumption in any of the surviving treatment groups during the Day 6 8 post exposure period were observed.

Gross Necropsy: All birds that died during the study, half of those surviving at Day 8 and the rest at test termination were subjected to a gross necropsy. Necropsy results for birds found dead were similar, including thin condition, loss of muscle mass, altered spleen color, autolysis of tissues and pale organs. These necropsy findings were considered to be treatment related. The single bird euthanized from the 293 ppm treatment was found to have treatment related' necropsy findings. Necropsy results for all other birds euthanized on Day 8 and Day 22 were unremarkable.

% Cumulative Mortality Nominal Concentration, ppm Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8*

Negative Control 0 0 0 0 0 0 0 0 18.3 0 0 0 0 0 0 0 0 36.6 o 0 0 0 0 0 0 0 73.2 0 0 0 0 0 0 0 0 146 0 0 10** 0 0 0 10 10 293 0 0 0 0 20 40 80 80 586 0 0 10 20 50 80 100 100 1171 0 0 30 100 100 100 100 100 * No mortalities occurred in any of the treatment levels from Day 8 to Day 22 ** Bird euthanized on day 3 after sustaining a broken log.

Body Weight ( grams)

Exposure Period Recovery Period Nominal Concentration ( ppm) Mean Body Weight, Day 0 Mean Body Weight Change Day 0 5 Mean Body Weight Change Day 5 8 Mean Body Weight Change Day 8 15 Mean Body Weight Change Day 15 22 Mean Total Body Weight Change, Day 8 22 Mean Body Weight, Day 22 Negative Control 20 + 10 + 8 + 23 + 22 45 82 18.3 21 + 11 + 9 + 24 + 23 47 87 36.6 20 + 11 + 8 + 26 + 24 50 89 73.2 20 + 9 + 7 + 24 + 20 44 79 146 20 + 7* + 6** + 24 + 21 45 79 293 20  2**  1** + 14 + 20 34 55 586 20  4**      1171 20

Note: numbers may not add manually due to rounding. Values for 293 ppm treatment group are impacted by the fact that only one bird remained in that group after day 8. ENV/ JM/ RD( 2002) 17/ FINAL 139 * Statistically different from the control group at p< 0.05 ( Dunnett's t test) ** Statistically different from the control group at p=< 0.01 ( Dunnett's t test) () = no data available due to mortality Mean average feed consumption Nominal Concentration, ppm Grams Feed/ Bird/ Day Days 0 5 Grams Feed/ Bird/ Day Days 6 8 Grams Feed/ Bird/ Day Days 8 15 Grams Feed/ Bird/ Day Days 15 22 Negative Control 9 10 9 13 18.3 9 11 10 12 36.6 8 12 14 15 73.2 10 13 13 15 146 9 10 11 14 293 5 9 8  586 6 19   1171 4    () = No data available due to mortality.

Gross Pathological Observations from Birds that Died In Study Male, Female, and Undetermined ( ppm) Finding Control N = l 146 N = 2 293 N = 8 586 N = 10 1171 N = 10 Abdominal cavity, some autolysis 0 0 2 2 4 Abdominal cavity, autolysis throughout 0 0 0 1 1 Crop, empty 0 0 2 5 2 Emaciated 0 0 2 5 8 Fractured leg 1 1 0 0 0 G. I. Tract, empty 0 0 1 1 0 Gizzard contents bile stained 0 0 2 5 1 Heart, anterior portion mottled white color 0 0 1 0 0 Heart, pale . 0 0 0 2 1 Intestinal contents tar like 0 0 0 2 0 Keel, prominent 0 0 1 3 10 Kidneys, pale 0 0 0 2 0 Liver, pale and mottled 0 1 0 0 0 Loss of muscle mass 0 0 4 7 9 Muscular skeletal, pale 0 1 0 0 0 Small in stature 0 0 3 0 0 Spleen, black 0 0 0 1 0 Spleen, dark 0 0 0 0 2 Spleen, grey 0 0 0 1 0 Spleen, grey brown 0 0 0 0 1 Spleen, pale 0 0 1 0 1 Spleen, small 0 0 0 0 1 Spleen, small and pale 0 0 0 3 0 Thin 0 0 0 4 2 Not remarkable 0 0 1 0 0 ENV/ JM/ RD( 2002) 17/ FINAL 140 CONCLUSIONS The dietary LC50 value for Northern Bobwhite exposed to perfluorooctanesulfonate was determined to be 220 ppm with a 95% confidence Interval of 164 to 289 ppm. The slope of the concentration response curve was 7.005 and the chi square value was 0.023. The no mortality concentration was 73.2 ppm. Based upon treatment related mortality, signs of toxicity and effects upon body weight gain at the 146 ppm test concentration, the no observed effect concentration was 73.2 ppm.

Author and/ or submitter: 3M Corporation, Environmental Laboratory, P. O. Box 33331, St. Paul, Minnesota, 55133, USA 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 ENV/ JM/ RD( 2002) 17/ FINAL 141 Robust Report Reference No. 13  Multi Phase Exposure / Recovery Algal Assay Test TEST SUBSTANCE Identity: Potassium perfluorooetanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfbnic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, potassium salt, CAS # 2795 39 3)

Remarks field: The test substance is a white powder ( 3M Lot 583) of uncharacterized purity. The following summary is abbreviated due to the fact that this study has been superceded by a more recent test.

METHOD Method: Test protocol utilized was modified after those described by USEPA  600/ 9 78 018, 1978; ASTM E 35.23, 1981; OECD 201,1979. ASTM STP # 667 Type: Semi chronic GLP: No Year completed: 1981 Species: Selenastrum capricornutum RESULTS EC50 Values, mg/ L ( 95% confidence interval is given in brackets)

Exposure ( contact) Days Cell Dry Weight Cell Count 4 115 ( 18  65) 82 ( No Conf. Limits) 7 122 ( 41 366) 99 ( 19  398) 10 128 ( 46  372) 98 ( 16  431) 14 146 ( 33  350) 95 ( 12  455)

Remarks: The statistical program used was questionable.

DATA QUALITY Reliability: Klimisch ranking = 2. This study satisfied criteria for quality testing at the time performed, but actual concentrations were not measured. Results were based on nominal concentrations. Additionally, sample purity was not properly characterized.

REFERENCES This study was conducted by the 3M Company, Environmental Laboratory, 1981.

OTHER Submitter: 3M Company, Environmental Laboratory, P. O. Box 33331, St. Paul, Minnesota, 55133, USA Last changed: 5/ 3/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 142 Robust Study Report Reference No. 14  The Effects of Continuous Aqueous Exposure to 14C 78.02 on Hatchability of Eggs and Growth and Survival of Fry of Fathead Minnow (

 / Summary of histopathological examinations of Fathead Minnow (

 ) exposed to 78.02 for 30 Days TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as 14C 78.02, PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, potassium salt, CAS # 2795 39 3)

Remarks: The test substance is a white powder. Sample was radiolabeled. Sample purity was not characterized.

The following summary is abbreviated due to the fact that this study has been superceded by a more recent test.

METHOD Method: Method was developed by E G & G, Bionomic and closely followed those presented in the " Proposed recommended bioassay procedure for egg and fry stages of freshwater fish", U. S. EPA, 1972. Type: Flow through chronic GLP: No Year completed: 1978 Species: Pimephales promelas RESULTS 30 Day NOEC: 1 mg/ L 30 Day LOEC: 1.9 mg/ L 30 Day MATC: > 1 mg/ L and < 1.9 mg/ L DATA QUALITY Reliability: Klimisch ranking = 2. This study satisfied criteria for quality testing at the time performed, but the analytical methodology was questionable.

REFERENCES This study was conducted at E G & G, Bionomics, Aquatic Toxicology Laboratory in Wareham, Massachusetts at the request of the 3M Company.

OTHER Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA.

Last changed: 5/ 3/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 143 Robust Study Report Reference No. 15  Effect of Potassium Perfluorooctanesulfonate on Survival, etc. ( Daphnid reproduction)

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, potassium salt, CAS # 2795 39 3)

Remarks field: The test substance is a white powder of uncharacterized purity.

The following summary is abbreviated due to the fact that this study has been superceded by a more recent test.

METHOD Methods: ASTM, 1981, Proposed Standard Practice for Conducting Renewal Life Cycle Toxicity Tests with Daphnia magna; OECD, 1981, Daphnia sp., 14 day Reproduction Test. Type: Acute static and Chronic renewal GLP: No Year completed: 1984 Species: Daphnia magna RESULTS ( 95% confidence interval is given in brackets)

48 hour EC50 ( immobilization): 27 ( 25 28) mg/ L 14 day EC50 ( reproduction)*: 14.7( 12 18) mg/ L 21 day EC50 ( reproduction)*: 12.4( 11 14) mg/ L 28 day EC50 ( reproduction)*: 11.4( 10 13) mg/ L 28 day NOEC**: 7 mg/ L 14, 2 1 and 28 day MATC: 11.2 mg/ L * Cumulative ( Young/ Adult) ** This concentration applies to no observed effects on Cumulative results of Young/ Adult and Broods/ Adult and on the total number of Young/ Adult.

DATA QUALITY Reliability: Klimisch ranking = 2. This study satisfied all criteria for quality testing at the time performed, but actual concentrations were not measured. Results were based on nominal concentrations. Additionally, sample purity was not adequately characterized.

REFERENCES This study was conducted by the 3M Company, Environmental Laboratory, 1984.

OTHER Submitter: 3M Company, Environmental Laboratory, P. O. Box 33331, St. Paul, Minnesota, 55133, USA Last changed: 5/ 3/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 144 Robust Study Report Reference No. 16

 96 hour Toxicity Test Data Summary. Sample FC 94 X ( Li salt of PFOS)

TEST SUBSTANCE Identity: Perfluorooctanesulfonate, Lithium salt; may also be referred to as PFOS Li salt, FC 94, or FC 94 X. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, lithium salt, CAS # 29457 72 5)

Remarks: Test sample was taken from 3M production lot # 1. The test sample is a mixture of the test substance in water ( approximately 24.5% test substance and 75.5% water). No calculations were made to adjust for the actual concentration of the test substance in the test sample.

METHOD Method: Not noted. Type: Static acute GLP: No Year completed: 1994 Species: Pimephales promelas Supplier: Aquatic Biosystems Inc., Fort Collins, CO, USA Analytical monitoring: pH and DO content Exposure period: 96 hours Statistical methods: LC50 values calculated by Trimmed Spearman  Karber. Test fish age: 79 days. Length and weight: Average length = 2.1 + 0.3cm Average weight = 0.069 + 0.03 g Loading: 0.69 g fish / L Pretreatment: None Test Conditions Dilution water: Carbon filtered well water Dilution water chemistry: pH: 8.4 DO: 8.1 mg/ L Stock and test solution preparation: A primary stock solution was prepared in dilution water to yield a test sample concentration of 400 mg/ L. All test solutions were made by diluting the appropriate amount of stock solution with dilution water to make 1 L of solution per concentration. Stability of the test chemical solutions: Not noted. Exposure vessels: 2 L glass beakers Number of replicates: two. Number of fish per replicate: ten Number of concentrations: six plus a negative control Water chemistry during the study: Dissolved oxygen range ( 0   96 hours): 6.0 7.2 mg/ L ( control exposure) 4.8 7.9 mg/ L ( 56.0 mg/ L exposure) pH range ( 0   96 hours): 8.0 8.4 ( control exposure) 8.0 8.4 ( 56.0 mg/ L exposure) Test temperature range ( 0   96 hours): 19.2 19.5oC ENV/ JM/ RD( 2002) 17/ FINAL 145 RESULTS Nominal concentrations: Negative control, 3.2, 5.6, 10.0, 18.0, 32.0, 56.0 mg/ L Element value ( 95% confidence interval is given in brackets): 96 hour LC50 = 19 mg/ L ( 16 24) Mortality of controls: None Remarks: Values reported are for the test sample. No calculations were made to adjust for the concentration of the test substance in the test sample.

CONCLUSIONS The test sample containing 24.5% Perfluorooctanesulfonate, Lithium salt exhibited a 96 hour LC50 for fathead minnow of 19 mg/ L. This value must be divided by 4 in order to express the result in terms of the concentration of PFOS.

Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA.

DATA QUALITY Reliability: Klimisch ranking 2. This study, while well conducted, lacks analytical data for: determination of the test substance concentration in the test solutions; and determination of the sample purity.

REFERENCES This study was conducted by the 3M Company, Environmental Laboratory, Lab Request number M1018, 3/ 25/ 94.

OTHER Last changed: 7/ 9/ 01 ENV/ JM/ RD( 2002) 17/ FINAL 146 Robust Study Report Reference No. 17  48 hour Acute Toxicity to Daphnia,

. FC 94 X ( Li salt of PFOS)

TEST SUBSTANCE Identity: Perfluorooctanesulfonate, Lithium salt; may also be referred to as PFOS Li salt, FC 94, or FC 94 X. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, lithium salt, CAS # 29457 72 5)

Remarks: The test sample is a mixture of the test substance in water ( approximately 24.5% test substance and 75.5% water). No calculations were made to adjust for the actual concentration of the test substance in the test sample.

METHOD Method: Not noted. Test type: Static acute GLP: No Year completed: 1994 Species: Daphnia magna Analytical monitoring: pH and DO content Statistical methods: EC50 values calculated using Trimmed Spearman Karber method Test daphnid source: Obtained from U. S. EPA NETAC, Duluth, Minnesota, USA. Test daphnid age at study initiation: < 24 hours Test Conditions Dilution water: Carbon filtered well water Dilution water chemistry: pH: 8.4 DO: 8.6 mg/ L Stock and test solutions preparation: A primary stock solution was prepared in dilution water to yield a test sample concentration of 1000 mg/ L. All test solutions were made by diluting the appropriate amount of stock solution with dilution water to make 50 mL of solution per concentration. Exposure vessels: 100 mL glass beakers containing 50 mL of test solution. Number of replicates: 4 Number of daphnids per replicate: 5 Number of concentrations: five plus a negative control Water chemistry during the study: Dissolved oxygen at test termination: 7.0 mg/ L ( control exposure) 7.8 mg/ L ( 1000 mg/ L exposure) pH at test termination: 8.6 ( control exposure) 8.6 ( 1000 mg/ L exposure) Test temperature range ( 0   48 hours) 20.1 21.0 C Element basis: mortality and immobilization RESULTS Nominal concentrations: Negative control, 100, 180, 320, 560, 1000 mg/ L ENV/ JM/ RD( 2002) 17/ FINAL 147 Element value: ( 95% confidence interval is given in brackets) 24 hour EC50 = 330 ( 290 370) mg/ L 48 hour EC50 = 210 ( 190 230) mg/ L 48 hour NOEC = 100 mg/ L Statistical Evaluation: The EC50 values and 95% confidence intervals were calculated using the Trimmed Spearman Karber method with trim set to 0%. Mortality of controls: None Remarks: Values reported are for the test sample. No calculations were made to adjust for the concentration of the test substance in the test sample.

CONCLUSIONS The test sample containing 24.5% Perfluorooctanesulfonate, Lithium salt exhibited a 48 hour EC50 for Daphnia magna of 210 mg/ L. This value must be divided by 4 in order to express the result in terms of the concentration of PFOS.

Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA.

DATA QUALITY Reliability: Klimisch ranking 2. This study, while well conducted, lacks analytical data for: determination of the test substance concentration in the test solutions; and determination of the sample purity.

REFERENCES This study was conducted by the 3M Company, Environmental Laboratory, Lab Request number M1018, 2/ 10/ 94.

OTHER Last changed: 7/ 9/ 01 ENV/ JM/ RD( 2002) 17/ FINAL 148 Robust Study Report Reference No. 20  96 hour Acute Toxicity Test on Bluegill Sunfish ( FC 99, DEA salt of PFOS)

TEST SUBSTANCE Identity: Perfluorooctanesulfonate, DEA salt; may also be referred to as PFOS DEA salt, FC 99, or 3M Sample No. 2. ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, compd. with 2,2' iminobis[ ethanol] ( 1: 1), CAS # 70225 14 8)

Remarks: Test sample is a mixture of the test substance in water ( approximately 25% test substance and 75% water). All values reported relate to this mixture. No calculations were made to adjust for the actual concentration of the test substance in the test sample.

METHOD Method: Environmental Protection Agency, Ecological Research Series EPA 660/ 3 75 009, April, 1975. Standard Methods. Type: Static acute GLP: Yes Year completed: 1979 Species: Lepomis macrochirus Supplier: Osage Catfisheries, Inc. in Osage Beach, Missouri, USA. Analytical monitoring: pH and DO / ammonia content Exposure period: 96 hours Statistical methods: Probit analysis. Test fish age: Not noted. Length and weight: Average length = 28.6 + 2.17 mm. Average weight = 0.60 + 0.15 g Loading: 0.2 g fish / L Pretreatment: None Test Conditions Dilution water: Laboratory well water Dilution water chemistry: Dissolved oxygen: 9.3 mg/ L Hardness: 255 mg/ L as CaCO3 Alkalinity: 368 mg/ L as CaCO3 pH: 7.8 Conductivity: 50 µ mhos/ cm Stock and test solution preparation: Primary stock prepared in deionized water at a concentration of 150 mg/ mL. The test concentrations were prepared by transferring appropriate aliquots of the stock standard directly to the test chambers. The test solutions were noted to foam when stirring in toxicant aliquots. Test concentrations were prepared based on total sample, not on percent concentration of the test substance in the test sample. Concentrations dosing rate: Once Stability of the test chemical solutions: Not noted Exposure vessels: 40 liter glass aquaria containing 30L of test solution. Number of replicates: one Number of fish per replicate: ten Number of concentrations: six plus a negative control ENV/ JM/ RD( 2002) 17/ FINAL 149 Water chemistry during the study: Dissolved oxygen range ( 0   96 hours): 6.0  8.4 mg/ L ( control exposure) 5.8  8.3 mg/ L ( 18 mg/ L exposure) pH range ( 0   96 hours): 8.2  8.3 ( control exposure) 8.3  8.3 ( 18 mg/ L exposure) Test temperature: Temperature held constant at 22oC through use of a water bath for test vessels.

RESULTS Nominal concentrations: Negative control, 18, 37, 75, 160, 320, 650 mg/ L Element value: ( 95% confidence interval is given in brackets) 24 hour LC50 = 460 ( 370 580) mg/ L 48 hour LC50 = 370 ( 290 470) mg/ L 96 hour LC50 = 31 ( 22 43) mg/ L 96 hour NOEC = 18 mg/ L ( C. I. not calculated)

All element values based on nominal concentrations Statistical evaluation of mortality: Probit analysis was used to calculate LC50 values and the corresponding confidence limits.

Quality Check for Test Organism Health: The bluegill sunfish were challenged with a reference compound, Antimycin A. The observed 96 hour LC50 and 95% confidence limits ( C. I.) were within the 95% confidence limits reported in the literature, indicating that the fish were in good condition.

Cumulative percent mortality:

Nominal Test Concentration mg/ L 24 hours 48 hours 72 hours 96 hours Neg. Control 0 0 0 0 18 0 0 0 0 37 0 0 10 80 75 0 0 30 90 160 0 0 70 100 320 0 20 100 100 650 100 100 100 100 CONCLUSIONS The test sample 96 hour LC50 for bluegill sunfish was determined to be 31 mg/ L with a 95% confidence interval of 22 43 mg/ L. The 96 hour no observed effect concentration was 18 mg/ L. These values must be divided by 4 in order to express the results in terms of the concentration of PFOS. Submitter: 3M Company, Environmental Laboratory P. O. Box 33331 St. Paul, MN 55133, USA.

DATA QUALITY Reliability: Klimisch ranking 2. This study, while well conducted, lacks analytical data for determination of the test substance concentration in the test solutions and determination of the sample purity. There were ENV/ JM/ RD( 2002) 17/ FINAL 150 also gaps in the measurement of water quality criteria for a number of the concentrations at given time intervals.

REFERENCES This study was conducted by Analytical BioChemistry Laboratories, Inc. of Columbia, Missouri on behalf of the 3M Company.

OTHER Last changed: 5/ 2/ 00 ENV/ JM/ RD( 2002) 17/ FINAL 151 Robust Study Report Reference No. 23   Acute toxicity to Aquatic Invertebrates ( e. g. Daphnia)

TEST SUBSTANCE Identity: Potassium perfluorooctanesulfonate; may also be referred to as PFOS or FC 95 or as part of the mixed product FM 3820 ( see Remarks). ( 1 Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8 heptadecafluoro, potassium salt, CAS # 2795 39 3)

Remarks: The test sample is FM 3820, a mixture of the test substance in diethylene glycol butyl ether and water ( approximately 24 28% test substance in diethylene glycol butyl ether and water). Calculations were made to adjust test values using the upper limit concentration of the test substance ( 28%) in the test sample and no adjustment was made for the presence of the diethylene glycol butyl ether or water when noted below. These calculations assumed that all toxicity was due to the presence of the Perfluorooctanesulfonate substance.

METHOD Method: OECD 202 Test type: Static acute GLP: Yes Year completed: 1991 Species: Daphnia magna Analytical monitoring: DO, pH, Conductivity, and temperature were monitored daily. Statistical methods: EC50 values calculated, when possible by standard statistical techniques ( Stephan, 1983) Test daphnid source: Obtained from cultures maintained by EnviroSystems Division, Resource Analysts, Inc., Hampton, NH, USA. Test daphnid age at study initiation: < 24 hours Test Conditions Dilution water: Well water from wells at EnviroSystems in Hampton, New Hampshire. Dilution water chemistry: pH: 7.8* Conductivity: 1200 µ mhos/ cm* TOC: < 2.0 mg/ L * Values measured at time of test.

Lighting: Cool white fluorescent lights, intensity 23 µ E/ s/ m2. Photoperiod of 16 hours light, 8 hours dark. No transition period noted. Stock and test solutions preparation: A primary stock solution was prepared in dilution water at 1000 mg/ L. The primary stock was proportionally diluted with dilution water to prepare the five test concentrations. Exposure vessels: 250 mL plastic beakers containing 200 mL of test solution. The approximate depth of test solution was 6 cm. Number of replicates: Four Number of daphnids per replicate: Five Number of concentrations: Five plus a negative control Water chemistry during the study: Dissolved oxygen range ( 0   48 hours): 8.2   8.5 mg/ L ( control exposure) 8.1   8.5 mg/ L ( 150 mg/ L exposure) ENV/ JM/ RD( 2002) 17/ FINAL 152 pH range ( 0   48 hours): 7.8   8.6 ( control exposure) 7.8   8.6 ( 150 mg/ L exposure) Test temperature range ( 0   48 hours): 20.8   21.0oC ( control exposure) 20.7   20.9oC ( 150 mg/ L exposure)

Conductivity range ( 0   48 hours): 1200   1300 µ mhos/ cm ( control exposure) 1200   1300 µ mhos/ cm ( 150 mg/ L exposure)

Element basis: mortality RESULTS Nominal concentrations: Negative control, 25, 40, 60, 100, 150 mg/ L Element values: 24 hour EC50 = > 150 mg/ L ( C. I. not calculable) 48 hour EC50 = 49 ( 43 56) mg/ L Perfluorooctanesulfonate concentration adjusted element value: 24 hour EC50 = > 42 mg/ L 48 hour EC50 = 14 mg/ L All element values based on nominal concentrations Biological observations: Ninety five percent survival occurred in the control exposure. The number of surviving organisms and the occurrence of sub lethal effects and immobilization or other sublethal effects were determined visually and recorded initially and after 24 and 48 hours.

Cumulative percent mortality:

Nominal Test Concentration mg/ L 24 hours 48 hours Neg. Control 0 5 25 0 0 40 5 25 60 0 25 100 0 100 150 20 100 Control response: Satisfactory CONCLUSIONS The test substance 48 hour EC50 for Daphnia magna was determined to be 49 mg/ L with a 95% confidence interval of 43 56 mg/ L. If you assume all toxicity of the mixture is due to the Perfluorooctanesulfonate, the adjusted 48 hour EC50 value is 14 mg/ L ( 49 mg/ L x 0.28). Submitter: 3M Company, Environmental Laboratory, P. O. Box 33331 St. Paul, Minnesota 55133, USA.

DATA QUALITY Reliability: Klimisch ranking = 3. The study lacks analytical measurement of test substance ENV/ JM/ RD( 2002) 17/ FINAL 153 concentrations in the test solutions and sample purity is not sufficiently characterized. Additionally, data are for a mixture and toxicity cannot be positively attributed to PFOS as the diethylene glycol butyl ether could also contribute to the toxicity. The basic water quality parameters ( hardness, alkalinity and calcium/ magnesium ratio) were not included in the final report.