Document ev2VY83ZDYaqLzjJDkxaeNOoq

a.a 6 -- TOXICITY TO TERRESTRIAL PLANTS 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 86.9% by LC/MS, 1H-HMR, 19F-NMR and elemental analyses techniques. METHOD__________________________________________________ Method: The study was conducted using a protocol based on procedures outlined in U.S. EPA OPPTS Guidelines 850.4100 and 850.4225 and the Organization for Economic Cooperation and Development (OECD) proposed revision of Guideline No. 208. Test type: Artificial soil GLP: Yes Year Completed: 2003 Species Monocots: Allium cepa (onion - Texas Grano') Lolium perenne (ryegrass - `Manhattan 3') Dicots: Medicago sativa (alfalfa) Linum usitatissimum (flax) Lactuca sativa (lettuce - `Summertime') Glycine max (soybean - `Green Envy') Lycopersicon esculentum (tomato - `Rutgers') Analytical monitoring: Test substance concentrations were measured by LCMS in soil at Days 0 and test termination and in plant and (when possible) fruit tissues at test termination. Statistical methods: EC values calculated, when possible, by non-linear regression analysis of Bruce and Versteeg, or with non-linear interpolation using the ICp approach of Norberg-King. Treatment group means were compared to control means with Dunnett's test (a = 0.05) using SAS Version 8 to determine if significant differences in emergence, survival, weight or height from the control occurred. Test organism source: Onion: Ryegrass: Alfalfa: Flax: Territorial Seed Co., Cottage Grove, OR Meyer Seed Co., Baltimore, MD Frontier Natural Products, Norway, IA Arrowhead Mills, Hereford, TX Page 1 o f 12 002181 Lettuce: Territorial Seed Co., Cottage Grove, OR Soybean: Johnny's Selected Seeds, Albion, ME Tomato: Meyer Seed Co., Baltimore, MD Test organism age at study initiation: seed Test conditions Test media: Artificial loam soil of the following composition: Kaolin clay, industrial quartz sand and peat in a ratio of 2:25:1 by weight. Crushed limestone (to buffer the pH) and a slow-release fertilizer were also added. Final mix consisted of 49% sand, 30% silt, 21% clay, and 2.1% organic matter. Soil pH was 7.79. Environmental conditions: Study conducted in a temperature- controlled greenhouse. Artificial lighting supplemented natural sunlight to provide a uniform 14-hour photoperiod. Stock and test solutions preparation: Approximately 70 kg of each test concentration and the negative control was prepared separately. The test substance was first mixed with a 1000 g aliquot of the 70 kg of test soil (soil moisture of 15%). The test substance- containing aliquot was then added back to the 70 kg of bulk soil and mixed for 10 minutes. The negative control soil was prepared in the same manner, but no test substance was added. Exposure vessels: Plastic pots, approximately 16 cm wide and 12 cm deep. Number of replicates: four Number of seeds per replicate: ten Number of concentrations: five plus a negative control Watering procedure: subirrigation Test conditions parameters: Temperature range: 14.1 - 32.0C Relative humidity range: 8 - 9 1 % Light intensity range: 12.8 - 26.6 moles PAR Exposure period (germination and growth test): 21-Days Exposure period (extended test): up to 205-days (variable by species) After observations on day 21, all but one plant was clipped from each pot. The remaining plant was maintained in the extended test, until (if possible) fruit formation. At test termination, the one plant was harvested and the fruit and vegetative tissues were analyzed separately for PFOS. Element Basis: EC25, EC50, N O EC - emergence, survival, shoot height, shoot weight Other observations: seedling condition Method of reporting test concentrations: nominal RESULTS________________________________________________________ Nominal concentrations: Bk control, 3.91, 15.6, 62.5, 250, 1000 mg/kg Initial measured concentrations: <LOQ, 3.61, 11.1, 50.8, 276, 998 mg/kg 002182 Page 2 of 12 Element values: 21-Day Emergence, mg/kg Species Onion Ryegrass Alfalfa Flax Lettuce Soybean Tomato LOEC 250 250 1000 1000 1000 >1000 1000 NOEC 62.5 62.5 250 250 250 1000 250 e c 25 51 200 370 400 390 >1000 310 ECso 210 340 745 600 560 >1000 470 21-Day Survival, mg/kg Species Onion Ryegrass Alfalfa Flax Lettuce Soybean Tomato LOEC 62.5 250 250 250 250 >1000 62.5 NOEC 15.6 62.5 62.5 62.5 62.5 1000 15.6 e c 25 47 170 250 140 260 >1000 69 ECso 57 310 450 230 390 >1000 105 21-Day Shoot Height, mg/kg Species Onion Ryegrass Alfalfa Flax Lettuce Soybean Tomato LOEC 62.5 15.6 250 250 3.91 250 62.5 NOEC 15.6 3.91 62.5 62.5 <3.91 62.5 15.6 e c 25 29 46 100 98 6.8 280 22 ECso 47 130 250 140 40 460 94 002183 c0GnOs 21-Day Shoot Weight, mg/kg Species Onion Ryegrass Alfalfa Flax Lettuce Soybean Tomato LOEC 15.6 15.6 250 250 3.91 250 62.5 NOEC 3.91 3.91 62.5 62.5 <3.91 62.5 15.6 EC25 13 7.5 53 82 8.9 160 12 28 54 150 120 20 330 29 All element values were calculated using nominal concentrations. Statistical Evaluation Variables: Emergence: Number of emerged seedlings per 10 planted seeds per pot. Survival: Number of living emerged seedlings per 10 seeds per pot. Weight: Mean shoot wet weight of living emerged seedlings in each pot Height: Mean height of living emerged seedlings in each pot. Biological observations: Onion - Day 21_______________ Nominal Soil Concentration, mo/ka Mean Emergence % Mean Survival % Mean Seedling Fresh W eight, g M ean Seedling Height cm Mean Seedling Condition S c o re <1) Subiethal Effects Noted T# p lants w ith effectl Neg. Control 88 85 0.10 8.1 2.9 None 3.91 93 90 0.09 7.3 3.8 Chlorosis [1] 15.6 83 80 0.07* 7.3 4.8 Necrosis [1] 62.5 63 33* 0.02* 2.6* 250 40* 0* . . 1000 0* 0* _ (1,A score of 0 = normal seedling; 100 = dead seedling Statistically significantly different from the control (p<0.05) 48 100 _ None _ Ryegrass - Day 21 Nominal Soil Concentration, mg/kg Mean E m ergence % Mean Survival % Mean Seedling Fresh W eight, 9 Mean Seedling Height cm Mean Seedling C o n d itio n S c o re 111 Subiethal Effects Noted [# plants with effect] Neg. Control 80 80 0.12 16.9 0 None h* CO 00 lO 3.91 15.6 62.5 250 1000 93 90 85 7.5* 93 90 83 53* 7.5* 0.11 0.07* 0.08* 0.01* 0.03* 15.4 13.7* 13.6* 2.0* 0 None 0 None 2.9 None 22 Necrosis f11] 13 Necrosis [2] Statistically significantly different from the control (p<0.05) Page 4 of 12 Alfalfa - Day 21 Nominal Soil Concentration, mgfkg Neg. Control Mean Emergence % 90 Mean Survival % Mean Seedling Fresh W eight, 9 88 0.12 Mean Seedling Height cm 6.5 Mean Seedling C o n d itio n S co re<1) 4.2 Sublethal Effects Noted [# plants w ith effect] Necrosis [1] 3.91 85 83 0.11 6.4 2.9 None 15.6 80 80 0.10 5.4 5.9 Necrosis [3] 62.5 80 80 0.10 6.1 0.9 Necrosis [1] 250 73 63* 0.03* 2.8* 17 Necrosis [1] 1000 33* 15* 0.02* 1.0* 75 Necrosis 141 (1,A score of 0 = normal seedling; 100 = dead seedling ` Statistically significantly different from the control (p<0.05) Flax - Day 21 Nominal Soil Concentration, mg/kg Mean Emergence % Neg. Control 73 Mean Survival % Mean Seedling Fresh W eight, 9 Mean Seedling Height cm Mean Seedling Condition S c o re '111 73 0.19 8.9 1.4 Sublethal Effects Noted [# plants with effect] Unshed seed coat [1] 3.91 73 68 0.18 8.7 9.7 L eaf curl [1] 15.6 80 80 0.18 8.8 2.5 Unshed seed coat [1] 62.5 85 85 0.16 8.2 1.2 Necrosis [1] 250 73 33* 0.02* 1.3* 1000 0* 0* _ (1,A score of 0 = normal seedling; 100 = dead seedling ` Statistically significantly different from the control (p<0.05) 91 Unshed seed coat f131 __ Lettuce - Day 21 Nominal Soil Concentration, mgfcg Mean Emergence % Mean Survival % Mean Seedling Fresh W eight, a M ean Seedling Height cm Mean Seedling Condition S c o re '111 Sublethal Effects Noted T# plants w ith effectl Neg. Control 88 88 0.38 6.4 0 None 3.91 90 90 0.25* 5.0* 0 None 15.6 90 90 0.24* 5.0* 1.1 Necrosis [1] 62.5 85 85 0.05* 2.6* 33 Necrosis [29] 250 83 68* 0.01* 1.0* 42 Necrosis [17] Leaf curl [1] 1000 13* 5.0* Insufficient sample* 1.0* 94 Necrosis [1] Unshed seed coat [1] ` Statistically significantly different from the control (p<0.05) 002185 Page 5 of 12 Soybean - Day 21 Nominal Soil Concentration, mg/kg Neg. Control Mean Emergence % 100 Mean Survival % 100 Mean Seedling Fresh W eight, 9 3.63 Mean Seedling Height cm 28.3 Seedling C o n d itio n S co re(1) 0 Sublethal Effects Noted T# plants w ith effectl None 3.91 98 98 3.64 26.8 0.8 Leaf curl [1] 15.6 95 95 3.63 26.9 0 None 62.5 100 100 4.00 30.7 1.8 Leaf curl [2] 250 100 98 2.07* 22.5* 7.0 Chlorosis [2] Leaf curl [3] Necrosis [1] 1000 100 100 0.57* 4.1* 1(A score of 0 = normal seedling; 100 = dead seedling 'Statistically significantly different from the control (p<0.05) 51 Unshed seed coat [2] Stem curl [7] Leaf curl [11] Necrosis [25] Tomato - C'ay 21_______________________________________ ______ Nominal Soil Concentration, mg/kg Mean Emergence % Mean Survival % Mean Seedling Fresh W eight, 9 Mean Seedling Height cm Mean Seedling Condition S c o re (1) Sublethal Effects Noted [# plants with effect] Neg. Control 93 93 0.35 5.2 0 None 3.91 83 83 0.40 5.6 0 None 15.6 83 80 0.28 4.8 6.7 Leaf curl [1] Necrosis [3] 62.5 85 68 0.08* 2.6* 40 Necrosis [19] *o cvi 250 73 7.5 0.03* 97 Necrosis [3] 1000 10* 0* - - (1,A score of 0 = normal seedling; 100 = dead seedling 'Statistically significantly different from the control (p<0.05) 100 . Control response: Satisfactory for all species Extended test observations: There were no additional findings of phytotoxicity at the termination of the extended growth portion of the test. Analytical Methodology: Analyses of test soils and plants were performed at Wildlife International Ltd. using high performance liquid chromatography with triple quadrupole mass spectrometric detection (HPLC/MS/MS). When determining the concentration of the test substance in the samples, 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) in soil was 1.18 mg/kg. For vegetative tissue, the LOQ 0 0 2 1 8 6 Page 6 of 12 ranged from 2.4 - 7.6 mg/kg. For fruit, the LOQ ranged from 0.12 - 6.4 mg/kg. The recovery of matrix fortifications analyzed concurrently during sample analysis ranged from 83.9% - 89.0% i n the soil, 86.3% - 117% in the fruit and 88.0% - 110% in the vegetative tissue. Soil samples collected at test initiation had measured values from 71.2% to 11 0 % of nominal. Measured values for samples taken at test termination ranged from 22.8% to 62.8% of nominal. Values reported as <LOQ were excluded from all calculations of mean concentration. Summaries of Analytical Chemistry Data (All results reported as dry weight): Soil, Day 0: Nominal Test Concentration, mg/kg Measured Replicate Values, mg/kg Negative Control All < LOQ(1) 3.91 3.43, 2.83, 4.56 15.6 11.1, 11.2 62.5 50.0, 44.7, 57.6 250 415,244, 170 1000 (l)LOQ = 1.18 mg/kg 1070, 983, 942 Mean Concentration, mg/kg <LOQ Percent of Nominal _ 3.61 92.3 11.1 71.2 50.8 81.3 276 110 998 99.8 Soil, termina tion (Day 205): Nominal Test Concentration, mg/kg Measured Replicate Values, mg/kg Negative Control 3.91 15.6 62.5 250 1000 `\ 0 Q = 1.18 mg/kg All < LOQ(1) 1.21,1.36 4.91,2.21 16.3, 16.1 153,161 432, 515 Mean Concentration, mg/kg Percent of Nominal <LOQ 1.29 3.56 16.2 157 474 33.0 22.8 25.9 62.8 47.4 002187 Page 7 of 12 Onion Vegetative Tissue, Termination (67-days): Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Mean tissue Concentration, mg/kg Negative Control 3.91 All < LOQ(1) All < LOQ(1) <LOQ <LOQ 15.6 62.5 5.86, 15.4, <LOQ Insufficient sample 10.6 . 250 No plants living 1000 (1)LOQ = 4 .7 mg/kg No plants living Onion Fruit, Termination (67-days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control 0.517, 0.487, insufficient sample 3.91 5 .8 1 ,0 .4 5 3 , insufficient sample 15.6 <LOQ(1), 19.7, 24.6 62.5 Insufficient sample 250 Insufficient sample 1000 (1,LOQ = 0.42 mg/kg No plants living Mean tissue Concentration, mg/kg 0.502 3.13 22.2 . _ _ Ryegrass Vegetative Tissue, Termination (205 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control All <LOQ(1> Mean tissue Concentration, mg/kg <LOQ 3.91 7.13, 8.70, 8.70 8.18 15.6 13.8, 49.0, <LOQ 31.4 62.5 42.5, 67.0, 36.7 48.7 250 1000 (1,LOQ = 3.3 mg/kg 31.9, 37 .5,129 Insufficient sample 66.1 Ryegrass fruit - No sample (did not mature by 205 days) 002188 Alfalfa Vegetative Tissue, Termination (141 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg M ean tissue Concentration, mg/kg Negative Control 3.91 All <LOQ(1) <LOQ, <LOQ, 6.16 <LOQ 6.16 15.6 5.66, 2.74, <LOQ 4.2 62.5 250 12.9, 9.66, 11.1 14.2, 15.8,17.3 11.2 15.8 1000 (l'LOQ = 2.7 mg/kg Insufficient sample _ Alfalfa Fruit, Termination (141 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control All <LOQ(1) 3.91 15.6 Insufficient sample, insufficient sample, <LOQ <LOQ (composite) 62.5 <LOQ (composite) 250 1000 (,)LOQ = 6.4 mg/kg <LOQ (composite) Insufficient sample Mean tissue Concentration, mg/kg <LOQ <LOQ <LOQ <LOQ <LOQ . Flax Vegetat ve Tissue, Termination 94 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Mean tissue Concentration, mg/kg Negative Control 3.91 15.6 62.5 250 1000 uLOQ = 2.4 mg/kg All <LOQ(1> 4.56, 6.43, 3.85 13.1,26.4, 17.0 54.7, 72.7, 37.5 Insufficient sample No plants living <LOQ 4.95 18.8 55.0 . - 002lb9 Page 9 of 12 Flax Fruit, Termination (94 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control 3.91 All <LOQ(1) 0.217, 0.277, 0.195 15.6 1.07, 0.622, 2.41 62.5 250 1.26, 2.2 0 ,4 .2 3 Insufficient sample 1000 (1)LOQ = 0.12 mg/kg No plants living Mean tissue Concentration, mg/kg <LOQ 0.230 1.36 2.56 _ _ Lettuce Vegetative Tissue, Termination (67 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control All <LOQ(1) 3.91 12.2, 6.16, 7.52 15.6 15.5, 7.06, 9.26 Mean tissue Concentration, mg/kg <LOQ 8.63 10.6 62.5 250 1000 (,,L O Q = 5 .5 m g/kg 39.8, 58.9, 27.0 Insufficient sample Insufficient sample 41.9 _ _ Soybean Vegetative Tissue, Terminaltion (67 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control All <LOQ(1) Mean tissue Concentration, mg/kg <LOQ 3.91 17.7, 10.0, 19.1 15.6 15.6 29.3, 48.9, 29.3 35.8 62.5 250 63.9, 70.7, 55.4 64.2, 175, 103 63.3 114 1000 l l,LOQ = 6.0 mg/kg Insufficient samp le _ 0021 0 Page 10 of 12 Soybean Fruit, Termination (67 days Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control All <LOQ<1) 3.91 0.947, 0.339, 2.91 15.6 0.464,1.63, 0.528 62.5 250 1.14, 1.14, 1.33 1.57, 5.56, 2.49 1000 (1)LOQ = 0.18 mg/kg Insufficient sample Mean tissue Concentration, mg/kg <LOQ 1.40 0.874 1.20 3.21 _ Tomato Vegetation, Termination (day 94) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control All <LOQ(1) 3.91 All <LOQ 15.6 12.5, 27.9, 61.1 Mean tissue Concentration, mg/kg <LOQ _ 33.8 62.5 70.9, 44.5, 34.9 50.1 250 1000 (1,LOQ = 7.6 mg/kg Insufficient sample No plants living . Tomato Fruit, Termination (day 94) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Negative Control 3.91 All <LOQ<1) <LOQ, <LOQ, insufficient sample 15.6 62.5 <LOQ, 1.51, 0.58 0.946, <LOQ, <LOQ 250 Insufficient sample 1000 (1,LOQ = 0.47 mg/kg No plants living Mean tissue Concentration, mg/kg <LOQ < LOQ 1.05 0.946 _ - 002191 Samples of soil and plant tissues from all species were also obtained at Day 21 and analyzed for PFOS. However, plant tissues and soil obtained on Day 21 had considerably higher than expected PFOS concentrations. Day 21 soil samples were collected by scraping the surface of the soil, Page 11 of 12 whereas a soil core sampler was used for samples collected at test termination. The laboratory concluded that PFOS concentrated in the top layer of the soil as a result of watering by subirrigation. Plant tissues collected at Day 21 were very small in size, and likely contaminated with surface soil due to improper collection techniques. Therefore, Day 21 values were considered to be not representative, and were not reported. CONCLUSIONS The potassium perfluorooctanesulfonate 21-day EC50range for seven species of terrestrial plants for the four study endpoints were found to be: Emergence: 210 - > 1000 mg/kg Survival: 57 - >1000 mg/kg Height: 40 - 460 mg/kg Shoot Weight: 2 0 - 330 mg/kg The 21-day no observed effect concentration ranges were found to be: Emergence: 6 2 .5 -1 0 0 0 mg/kg Survival: 1 5 .6 - 1000 mg/kg Height: <3.91 - 62.5 mg/kg Shoot Weight: <3.91 - 62.5 mg/kg Submitter: 3M Corporation, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133 DATA QUALITY_________________________________________________ Reliability: Klimisch ranking 1. REFERENCES__________________________________________________ This study was conducted at Wildlife International, Ltd. Easton, MD at the request of the 3M Company. OTHER_________________________________________________________ Last changed: 6/3/03 OUk.132