Document JrqjzZDBR5mMDbRxgXke6gz2B

TOXICITY TO TERRESTRIAL PLANTS TEST SUBSTANCE Identity: P e rflu o ro o cta n e su lfo n a te ; m ay also be referred to as PFO S or FC-95. (1-O ctanesulfonic 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 te st su b sta n ce is a w h ite pow der. S a m ple w a s taken from 3M lot num ber 217. Sam ple w as stored under am bient conditions prior to testing. P u rity d e te rm in e d to be 86.9% by LC /M S, 1H-HM R, 19F -N M R and elemental analyses techniques. M ETH O D ______________________________________________________________ Method: T h e stu d y w a s con d u cte d using a protocol based on proced ure s outlined in U.S. E P A O P P T S G uide lines 8 5 0 .4 1 0 0 and 8 5 0 .4 2 2 5 and the O rganization for Econom ic Cooperation and Developm ent (OECD) proposed revision of G uideline No. 208. Test type: A rtificia l soil GLP: Yes Year Completed: 2003 Species M onocots: Allium cepa (onion - `T exa s G ra n o ') Lolium perenne (ryegrass - `M a n h a tta n 3 ') Dicots: Medicago sativa (alfalfa) Linum usitatissimum (flax) Lactuca sativa (lettu ce - `S u m m e rtim e ') Glycine max (soybean - `G reen E n vy') Lycopersicon esculentum (tom ato - `R u tg e rs') Analytical monitoring: T e st substance co nce ntratio ns w e re m easured by LC M S in soil at D ays 0 and te st te rm in a tio n and in p lant and (w hen possible) fruit tissues at test term ination. Statistical methods: EC values calculated, w h en possible, by non-linear regression analysis of Bruce and Versteeg, or with non-linear interpolation using the ICp approach of Norberg-King. Treatm ent group m eans were com pared to control means with Dunnett's test (a = 0.05) using SAS V e rsio n 8 to de te rm in e if sig n ifica n t d iffe re n ce s in em ergence, 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 of 12 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: A rtificia l loam soil o f the fo llo w in g com position: K aolin clay, in dustrial q u a rtz sand and peat in a ratio o f 2:25:1 by w eight. Crushed lim estone (to buffer the pH) and a slow-release fertilizer w ere also added. Final mix consisted of 49% sand, 30% silt, 21% clay, and 2.1% organic m atter. Soil pH w as 7.79. Environmental conditions: S tudy conducted in a te m p era ture- controlled greenhouse. Artificial lighting supplem ented natural sunlight to provide a uniform 14-hour photoperiod. Stock and test solutions preparation: A p p ro xim a te ly 70 kg of each test concentration and the negative control was prepared separately. The test substance was first m ixed with a 1000 g aliquot of the 70 kg of test soil (soil m oisture of 15%). The test substance- containing aliquot was then added back to the 70 kg of bulk soil and m ixed for 10 minutes. The negative control soil w as prepared in the sam e m anner, but no te st substa n ce w a s added. Exposure vessels: P lastic pots, a p p ro xim a te ly 16 cm w id e and 12 cm deep. Number of replicates: fo u r Number of seeds per replicate: ten Number of concentrations: five plus a neg ative control Watering procedure: subirrigation Test conditions parameters: Temperature range: 14.1 - 32.0C Relative humidity range: 8 - 91% Light intensity range: 12.8 - 2 6 .6 m oles PAR Exposure period (germination and growth test): 21-Days Exposure period (extended test): up to 205-days (variable by species) A fter observations on day 21, all but one plant w as clipped from each pot. T he rem a ining plant w a s m a intained in the extended test, until (if possible) fruit form ation. At test term ination, the one plant was harvested and the fruit and vegetative tissues were analyzed separately for PFOS. Element Basis: E C 25, EC 50, N O EC - em ergence, survival, shoot height, shoot weight Other observations: seedling condition Method of reporting test concentrations: nom inal RESU LTS_____________________________________________________________ Nominal concentrations: Bk control, 3.91, 15.6, 62.5, 250, 1000 m g/kg Initial measured concentrations: <LO Q, 3.61, 1 1 .1 ,5 0 .8 , 276, 998 m g /kg Page 2 of 12 mm 00 001 001 Element values: 21-Day Emergence, mg/kg Species O nion Ryegrass Alfalfa Flax Lettuce Soybean Tom ato LOEC 250 250 1000 1000 1000 >1000 1000 NOEC 62.5 62.5 250 250 250 1000 250 EC25 51 200 370 400 390 >1000 310 210 340 745 600 560 >1000 470 21-Day Survival, mg/kg Species O nion Ryegrass Alfalfa Flax Lettuce Soybean Tom ato LOEC 62.5 250 250 250 250 >1000 62.5 NOEC 15.6 62.5 62.5 62.5 62.5 1000 15.6 EC25 47 170 250 140 260 >1000 69 57 310 450 230 390 >1000 105 21-Day Shoot Height, mg/kg Species O nion Ryegrass Alfalfa Flax Lettuce Soybean Tom ato 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 EC25 29 46 100 98 6.8 280 22 EC50 47 130 250 140 40 460 94 Page 3 of 12 21-Day Shoot Weight, mg/kg Species O nion Ryegrass Alfalfa Flax Lettuce Soybean Tom ato 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 EC50 28 54 150 120 20 330 29 All elem ent values were calculated using nominal concentrations. Statistical Evaluation Variables: Em ergence: N um ber of em erged seedlings per 10 planted seeds per pot. Survival: N um ber of living em erged seedlings per 10 seeds per pot. W eight: M ean sh o o t w e t w e ig h t o f living em e rg e d se e d lin g s in each pot Height: M ean height o f living e m erged se e d lin g s in each pot. Biological observations: Onion - Day 21 Nominal Soil Mean Mean MeanSeedling MeanSeedling MeanSeedling Sublethal Concentration, Emergence Survival FreshWeight, Height Condition Effects Noted mg/kg %% g cm Score111 [#plantswitheffect] 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* 48 None 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) 100 _ _ Ryegrass - Day 21 Nominal Soil Mean Mean MeanSeedling MeanSeedling MeanSeedling Sublethal Concentration, Emergence Survival FreshWeight, Height Condition Effects Noted mg/kg %% g cm Score111 [#plantswitheffect] Neg. Control 80 80 0.12 16.9 0 None 3.91 93 93 0.11 15.4 0 None 15.6 90 90 0.07* 13.7* 0 None 62.5 85 83 0.08* 13.6* 2.9 None *00 LO 250 53* 0.01* 3.7* 22 Necrosis [11] 1000 7.5* 7.5* 0.03* 2.0* (1)A score of 0 = normal seedling; 100 = dead seedling ` Statistically significantly different from the control (p<0.05) 13 Necrosis [2] Page 4 of 12 Alfalfa - Day 21 Nominal Soil Mean Mean MeanSeedling MeanSeedling MeanSeedling Sublethal Concentration, Emergence Survival FreshWeight, Height Condition Effects Noted mg/kg %% g cm Score111 [#plantswitheffect] Neg. Control 90 88 0.12 6.5 4.2 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` (1)A score of 0 = normal seedling; 100 = dead seedling ` Statistically significantly different from the control (p<0.05) 75 Necrosis [4] Flax - Day 21 Nominal Soil Concentration, mg/kg Neg. Control Mean Emergence % 73 Mean Survival % 73 MeanSeedling MeanSeedling MeanSeedling FreshWeight, Height Condition g cm Score111 0.19 8.9 1.4 Sublethal Effects Noted [#plantswitheffect] Unshed seed coat [1] 3.91 73 68 0.18 8.7 9.7 Leaf 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 [13] __ Lettuce - Day 21 Nominal Soil Mean Mean MeanSeedling MeanSeedling MeanSeedling Sublethal Concentration, Emergence Survival FreshWeight, Height Condition Effects Noted mg/kg %% g cm Score111 [#plantswitheffect] 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 1.0` sample* (1)A score of 0 = normal seedling; 100 = dead seedling ` Statistically significantly different from the control (p<0.05) 94 Necrosis [1] Unshed seed coat [1] Page 5 of 12 Soybean - Day 21 Nominal Soil Mean Mean MeanSeedling MeanSeedling Seedling Sublethal Concentration, Emergence Survival FreshWeight, Height Condition Effects Noted mg/kg %% g cm Score111 [# plantswitheffect] Neg. Control 100 100 3.63 28.3 0 None 3.91 98 98 3.64 26.8 0.8 Leafcurl [1] 15.6 95 95 3.63 26.9 0 None 62.5 100 100 4.00 30.7 1.8 Leafcurl [2] 250 100 98 2.07* 22.5* 7.0 Chlorosis [2] Leafcurl [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 - Day 21 Nominal Soil Mean Mean MeanSeedling MeanSeedling MeanSeedling Sublethal Concentration, Emergence Survival FreshWeight, Height Condition Effects Noted mg/kg %% g cm Score111 [#plantswitheffect] 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] 250 73 7.5 0.03* 2.0* 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: S atisfactory fo r all species Extended test observations: T he re w e re no additional findings of phytotoxicity at the term ination of the extended growth portion of the test. Analytical Methodology: A n alyse s o f test soils and plants w e re perform ed at W ildlife International Ltd. using high perform ance liquid chrom atography with triple quadrupole mass spectrom etric detection (HPLC/MS/MS). W hen determ ining the concentration of the test su b sta n ce in the sam ples, the sam e and m ost p ro m in e n t p ea k respon se for perfluorooctanesulfonate was used. No attem pt was made to quantify on the basis of individual isom eric com ponents. The LOQ (lim it of quantitation) in soil w a s 1.18 m g/kg. For v e g e ta tive tissue, the LOQ Page 6 of 12 ranged from 2.4 - 7.6 mg/kg. For fruit, the LOQ ranged from 0.12 - 6.4 m g/kg. The recovery of m atrix fortifications analyzed concurrently during sam ple a na lysis ranged from 83.9% - 89.0% i n the soil, 86.3% - 117% in the fru it and 88.0% - 110% in the v e g e ta tive tissue. Soil sam p le s co lle cte d at test initiation had m easured values from 71.2% to 110% of nominal. M easured values for sam ples taken at test term ination ranged from 22.8% to 62.8% of nominal. V alues reported as <LOQ w ere 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 (1)LOQ = 1.18 mg/kg 1070, 983, 942 Mean Percent of Concentration, Nominal mg/kg <LOQ _ 3.61 92.3 11.1 71.2 50.8 81.3 276 110 998 99.8 Soil, termination (Day 205): Nominal Test Concentration, mg/kg Measured Replicate Values, mg/kg Negative Control 3.91 15.6 62.5 250 1000 (1)LOQ = 1.18 mg/kg All < LOQ(1) 1.21,1.36 4.91,2.21 16.3, 16.1 153, 161 432, 515 Mean Percent of Concentration, Nominal mg/kg <LOQ 1.29 3.56 16.2 157 474 _ 33.0 22.8 25.9 62.8 47.4 Page 7 of 12 Onion Vegetative Tissue, Termination (67-days): Nominal Test Measured Replicate TissueValues, Concentration, mg/kg mg/kg Mean tissue Concentration, mg/kg Negative Control All < LOQ(1) <LOQ 3.91 All < LOQ(1) <LOQ 15.6 62.5 250 1000 (1)LOQ = 4.7 mg/kg 5.86, 15.4, <LOQ Insufficient sample No plants living No plants living 10.6 _ . _ Onion Fruit, Termination (67-days) Nominal Test Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Negative Control 0.517, 0.487, insufficient sample 3.91 5.81,0.453, 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 Measured Replicate TissueValues, Concentration, mg/kg mg/kg Mean tissue Concentration, mg/kg Negative Control All <LOQ(1) <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) Page 8 of 12 Alfalfa Vegetative Tissue, Termination (141 days) Nominal Test Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Mean tissue Concentration, mg/kg Negative Control All <LOQ(1) <LOQ 3.91 <LOQ, <LOQ, 6.16 6.16 15.6 5.66, 2.74, <LOQ 4.2 62.5 12.9, 9.66, 11.1 11.2 250 1000 (1)LOQ = 2.7 mg/kg 14.2, 15.8, 17.3 Insufficient sample 15.8 _ Alfalfa Fruit, Termination (141 days) Nominal Test Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Negative Control All <LOQ(1) 3.91 Insufficient sample, insufficient sample, <LOQ 15.6 <LOQ (composite) 62.5 <LOQ (composite) 250 <LOQ (composite) 1000 (1)LOQ = 6.4 mg/kg Insufficient sample Mean tissue Concentration, mg/kg <LOQ <LOQ <LOQ <LOQ <LOQ _ Flax Vegetative Tissue, Termination 94 days) Nominal Test Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Mean tissue Concentration, mg/kg Negative Control All <LOQ(1) <LOQ 3.91 4.56, 6.43, 3.85 4.95 15.6 13.1,26.4, 17.0 18.8 62.5 250 1000 (1)LOQ = 2.4 mg/kg 54.7, 72.7, 37.5 Insufficient sample No plants living 55.0 _ - Page 9 of 12 Flax Fruit, Termination (94 days) Nominal Test Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Negative Control All <LOQ(1) 3.91 0.217, 0.277, 0.195 15.6 1.07, 0.622, 2.41 62.5 1.26, 2.20, 4.23 250 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 Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Mean tissue Concentration, mg/kg Negative Control All <LOQ(1) <LOQ 3.91 12.2, 6.16, 7.52 8.63 15.6 15.5, 7.06, 9.26 10.6 62.5 39.8, 58.9, 27.0 250 Insufficient sample 41.9 _ 1000 (1)LOQ = 5.5 mg/kg Insufficient sample - Soybean Vecjetative Tissue, Terminaltion (67 days) Nominal Test Concentration, mg/kg Measured Replicate Tissue Values, mg/kg Mean tissue Concentration, mg/kg Negative Control All <LOQ(1) <LOQ 3.91 17.7, 10.0, 19.1 15.6 15.6 29.3, 48.9, 29.3 35.8 62.5 63.9, 70.7, 55.4 63.3 250 64.2, 175, 103 114 1000 (1)LOQ = 6.0 mg/kg Insufficient sample _ Page 10 of 12 Soybean Fruit, Termination (67 days) Nominal Test Measured Replicate Tissue Values, Concentration, mg/kg 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 1.14, 1.14, 1.33 250 1.57, 5.56, 2.49 1000 Insufficient sample (1)LOQ = 0.18 mg/kg 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 Mean tissue Concentration, mg/kg Negative Control 3.91 All <LOQ(1) All <LOQ <LOQ _ 15.6 12.5, 27.9, 61.1 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 Measured Replicate Tissue Values, Concentration, mg/kg mg/kg Negative Control All <LOQ(1) 3.91 <LOQ, <LOQ, insufficient sample 15.6 <LOQ, 1.51,0.58 62.5 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 _ _ Sam ples of soil and plant tissues from all species w ere also obtained at D ay 21 and an a lyze d fo r PFO S. H ow ever, plant tissu e s and soil obtained on D ay 21 had co n sid e ra b ly h igher than e xpe cted PFO S conce ntratio ns. D ay 21 soil sa m p le s w e re co lle cte d by scra p in g the su rfa ce o f the soil, Page 11 of 12 w hereas a soil core sam pler was used for sam ples collected at test term ination. T he laboratory conclude d that PFOS co n ce n tra te d in the top layer of the soil as a result of w atering by subirrigation. Plant tissues co lle cte d at D ay 21 w e re ve ry sm all in size, and likely co n ta m in a te d w ith surfa ce soil due to im p ro p e r collectio n techniques. T he refore, D ay 21 values were considered to be not representative, and w ere not reported. C O N C L U S IO N S T he potassium p e rflu o ro o cta n e su lfo n a te 2 1 -d a y E C 50 range fo r seven species of terrestrial plants for the four study endpoints w ere found to be: Emergence: 210 - > 1000 mg/kg Survival: 57 - >1000 mg/kg Height: 40 - 460 m g/kg Shoot W eight: 20 - 330 mg/kg The 21-day no observed effect concentration ranges w ere found to be: Emergence: 62.5 - 1000 mg/kg Survival: 15.6 - 1000 mg/kg Height: <3.91 - 62.5 m g/kg Shoot W eight: <3.91 - 62.5 mg/kg Submitter: 3M C o rporation, E n vironm ental Laboratory, P.O. Box 33331, St. Paul, M innesota, 55133 DATA Q UALITY Reliability: K lim isch ranking 1. R E F E R E N C E S _______________ This study w as conducted at W ildlife International, Ltd. Easton, MD at the request of the 3M Company. O T H E R ______________________________________________________________ Last changed: 6/3/03 Page 12 of 12