Document ZBx1D4w0547Ornen6d8Rvv3M7

BACK TO MAIN DIETARY ACUTE MALLARD DUCK STUDY REVISED Tissue analysis is now included. TEST SUBSTANCE______________________ Identity: Perfluorooctanesulfonate; may also be referred to as PFOS or FC-95. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8heptadecafluoro-, potassium salt, CAS # 2795-39-3) Remarks: 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 850.2200, OECD 205, and FIFRA Subdivision E. Section 71-2 Type: Dietary acute GLP: Yes Year completed: 2000 Species: Anas platyrhynchos Supplier: Whistling Wings, Inc., Hanover, IL 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: LC50values 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. Identification: 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 BACK TO MAIN 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. 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 Ferm (Fermatco) 1 0.25 Salt Iodized 0.10 'Fermentation by-products (source of unidentified growth factors) Vitamin and Mineral Premix Vitamin or Mineral Vitamin D3 Vitamin A Riboflavin Niacin Pantothenic Acid Vitamin B12 Folic Acid Biotin Pyridoxine Thiamine Vitamin E Vitamin K (Menadione dimethylpyrimidinol bisulfite) Manganese Zinc Copper Iodine Iron Selenium Amount Per Ton 2,000,000 I.C.U. 7,000,000 I.U. 6 grams 40 grams 10 grams 8 mg 600 mg 64 mg 1.2 grams 1.2 grams 20,000 I.U. 5.8 grams 102 grams 47 grams 6.8 grams 1.5 grams 51 grams 182 grams Prophylaxis: None Brooding compartment mean temperature: 38 + 2oC BACK TO MAIN 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. 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: Bk 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 BACK TO MAIN 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 Percent of Concentration, Nominal ppm 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 19.5 107 1171 1239, 1221,1118, 1301,1163, 1133 1196 102 Verification in Avian Diet Nominal Test Concentration, ppm Mean Measured Percent of Measured Duplicate Concentrations at Day 0, ppm Concentration, Nominal ppm Negative Control 36.6 < LOQ 45.7, 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 Ambient Stability in Avian Diet Day 0 Mean Measured Concentration, ppm Negative Control Mean Measured Mean Measured Duplicate Concentrations at Day 5, ppm Concentration, Percent of ppm Day 0 < 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 77.8 104 174 291 537 1196 167, 160 297, 293 552, 530 1150, 1122 164 295 541 1136 94.3 101 101 95 BACK TO MAIN 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. BACK TO MAIN Percent Cumulative Mortality Nominal Concentration, ppm Negative Control Day 1 0 Day 2 0 Day 3 0 Day 4 0 Day 5 0 9.1 0 0 0 0 0 18.3 0 0 0 0 0 36.6 0 0 0 0 0 73.2 0 0 0 0 0 146 0 0 0 0 0 293 0 0 0 0 0 586 0 0 0 0 10 1171 0 0 0 20 60 *No mortalities occurred in any of the treatment levels from Day 8 to Day 22 Day 6 0 0 0 0 0 0 0 20 80 Day 7 0 0 0 0 0 0 10 30 90 Day 8* 0 0 0 0 0 0 20 30 90 Summary of Analytical Results for Liver Tissue1(ug/g wet) during Day 1 through Day 7 Test Period2 - Individual Values Reported Nom inal Conc., Day 4 Liver Day 5 Liver Day 6 Liver Day 7 Liver PPm 293 Conc. Conc. Conc. Conc. 148 586 216 197 180 1171 116,122 172, 137, 147, 131, 121 178 185 1Concentrations were not corrected for purity of PFOS standard used (lot 171). No attempt was made to correct for moisture content. 2Reported results are only for concentrations where mortality occurred. During this time period, the negative control and concentrations <293 ppm exhibited no mortality. There was also no mortality in concentrations > 293 ppm during days 1 through 3. Summary of Analytical Results for Liver Tissue (ug/g wet) and Serum (ug/mL) at Day 8 and Day 221 - Average Value Reported Nom inal Conc., Day 8 Liver Day 8 Serum Day 22 Liver Day 22 Serum PPm Conc. Conc. Negative Control <LOQ (~0.030) <LOQ (0.005) Conc. Conc. <LOQ (~0.030) <LOQ (0.010) 9.1 4.67 7.38 1.03 1.49 18.3 6.65 13.8 2.04 4.32 36.6 15.3 30.5 3.36 6.82 73.2 29.7 48.1 9.21 11.3 146 38.8 53.9 12.7 18.9 293 55.6 65.8 9.2 13.4 586 59.5 107 20.2 34.0 1171 NS NS Not applicable LA "'Concentrations were not corrected for purity of PFOS standard used (lot 171). No attempt was made to correct for moisture content in relevant tissue, liver. NS = No Sample analyzed. LA = Lab Accident BACK TO MAIN 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 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 Grams Grams Grams Feed/Bird/Day Feed/Bird/Day Feed/Bird/Day Feed/Bird/Day Days 0-5 Days 6-8 Days 8-15 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 BACK TO MAIN Gross Pathological Observations from Birds that Died in Study Finding 293 ppm 586 ppm 1171 ppm N N=2 N=3 =9 Crop empty Emaciated 02 11 7 4 G.I. Tract, primarily empty Gizzard contents bile stained 11 02 1 4 Gizzard, empty 00 1 Intestinal contents, black and tar-like Keel prominent 00 00 1 2 Kidneys, pale Loss of muscle mass 00 22 1 5 Spleen, grey Spleen, small and pale 00 11 1 2 Spleen, pale Thin 02 11 3 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 chisquare 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 DATA QUALITY______________________________________________ Reliability: Klimisch ranking = 1 REFERENCES This study was conducted at Wildlife International Ltd., Easton, MD at the request of the 3M Company. The tissue analysis was conducted by 3M Environmental Laboratory, St. Paul, MN. OTHER Last changed: 5/1/00, 6/28/01 BACK TO MAIN