Document OzgjVELe8JJ0ynqkj2Mv2RMkK

AR226-3176 DuPont-6011 TRADE SECRET Study Title H-24516: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum Laboratory Project ID: DuPont-6011 Author: Terry Lee Sloman, B.S. Study Completed on: March 12,2001 Performing Laboratory: E.I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 ' Company Sanitized. Does not contain Page 1 of 12 ^ H-24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum li (pr STUDY INFORMATION DuPont-6011 cKnown Impurities: Physical Characteristics: Pale to medium tan waxy solid Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed. CoBpsrsy Sanitized' Doss not contain &C i?3l H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 STUDY INFORMATION (Continued) Stability: The test substance appeared to be stable under the conditions of the study; no evidence o f instability was observed. Sponsor: E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Tnitiated/Completed: March 1, 2001 / (see report cover page) In-Life Initiated/Completed: March 1, 2001 / March 4,2001 Company Sanitized. Does not contain TSCA CBt -3 - H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 SUMMARY A study was conducted to determine the effect o f 16) on the growth arid growth rate of the green alga Selenasfrum capricornutum. The algae were exposed to nominal concentrations of 0.01,0.1,1,10,100, and 1000 mg H-24516/Liter of nutrient medium (ppm). The algae were exposed for 72 hours (3 days) without test medium renewal. The effect was expressed as percent inhibition in growth based on healthy cell count (also referred to as cell density) relative to the blank (culture medium) control for the 72-hour (day 3) interval of the test. The 72-hour EC50, including the 95% confidence intervals (C.I.), and the NOECa based on nominal concentrations are as follows:b'c,d Healthy Cell Count EC50 >1000 mg/L H-24516 exhibited low concern under TSCA in a 72-hour, acute test using the green alga Selenastrum capricornutum. a The EC50 is defined as the "effective concentration" producing a 50% inhibition o f growth relative to the control. The NOEC is defined as the highest concentration o f test substance that has no significant effect on the measured parameter relative to the control. b BBN Software Products Coip., (1998). An RS/1 RPL procedure conducting Williams' test and probit analysis, c McCullogh, L.P. and Nelder, J.A. (1989). Generalized Linear Models, 2ndedition, Chapter 9, pp 328. Chapman and Hall, London. d Snedecor, G.W. and Cochran, W.G. (1967). Statistical Methods, 6th edition, Table A4, pp 469. The Iowa State University Press, Ames. e Smrchek, J., Clements, R., Morcock, R., and Rabert, W. (1993). Assessing ecological hazard under TSCA: methods and evaluation o f data. Environmental Toxicology and Risk Assessment, ASTM STP 1179. (W.G. Landis, J.S. Hughes, and M.A. Lewis, Eds.), pp 22-39. American Society for Testing and Materials, Philadelphia. pompany Sanitized. Does not contain TSCA CBt H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricomutum D u P o n t- 6 0 11 CERTIFICATION I, the undersigned, declare that this report provides an accurate evaluation o f data obtained from this study. Issued by Study Director: Jju. Terry Lee Stornati, B.S. Associate Scientist [X fhAH a-oof Date Sanitized. Does not contain TSCA c e l Company -5 - H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 RESULTS Based on visual observations, the blank (normal culture medium) control solution and the 0.01, 0.1, 1,10; 100, and 1000 mg/L test solutions were clear and had no color before addition of algal cells. After the addition of algal cells, the test and blank control flasks were placed on a shaker table in a chamber with a temperature of approximately 25C. The algae were incubated for 72 hours (3 days) without medium renewal. Illumination was supplied by cool-white fluorescent tubes. The mean light intensity in the chamber was 9718 lumens/m2 (= lux). The shaking speed was 108 revolutions per minute (rpm). The pH measurement o f the culture medium was 7.50 at study start. The pH measurements of the definitive test solutions at test initiation ranged from 6.94 to 7.50, and at test termination ranged from 6.97 to 7.83. Company Sanitized, Does not contain T8CA CBi -6 - H-24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 TABLES Company Sanitized. Does not contain TSCA CBi -7 - H-24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricomutum DuPont-6011 TABLE 1 pH MEASUREMENTS OF TEST SOLUTIONS Nominal Concentration (mg/L) Test Initiation (0 hours) pH Test Termination (72 hours) pH Blank Control 0.01 0.1 1 10 100 1000 7.50 7.21 7.15 7.04 7.03 7.00 6.94 6.97 7.17 7.42 7.52 7.64 7.83 7.78 Company Sanitized. Does not contain t s c a CBt H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 TABLE 2 DATA SUMMARY Nominal Day 0 Healthy Cell Count Day 3 Healthy Cell Count Concentration (cells/mL) (cells/mL) Blank Control Replicate 1 Replicate 2 10,000 10,000 2,760,000 2,340,000 0.01 mg/L Replicate 1 Replicate 2 10,000 10,000 2,570,000 2,820,000 0.1 mg/L Replicate 1 Replicate 2 10,000 10,000 3,020,000 3,280,000 1 mg/L Replicate 1 Replicate 2 10,000 10,000 2,550,000 2,310,000 10 mg/L Replicate 1 Replicate 2 10,000 10,000 3,250,000 3,120,000 100 mg/L Replicate 1 Replicate 2 10,000 10,000 2,580,000 2,960,000 1000 mg/L Replicate 1 Replicate 2 10,000 10,000 2,770,000 1,260,000 Company Sanitized. Does not contain TSCA C8I H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 APPENDIX J) - 10- H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricornutum DuPont-6011 APPENDIX A 72-Hour Healthy Cell Count Statistics not contain jppmpany Sanitized. Does -11 - H -24516: Influence on Growth and Growth Rate o f the Green Alga Selenastrum capricomutum 72-HOUR HEALTHY CELL COUNT STATISTICS DuPont-6011 WILLIAMS TEST OF #@ALGAESCREEW@H24516 72 Hr Healthy Cell Count (#cells/mL) W.T. Rank Dose 0 0.00 0 0.01 0 0.10 0 1.00 0 10.00 0 100.00 0 1000.00 Mean Response 2550000 2695000 3150000 2430000 3185000 2770000 2015000 Std Error 210000 125000 130000 120000 65000 190000 755000 Transform Mean 6.405062 6.430091 6.497940 6.385076 6.503019 .441456 6.271425 Transform Std Error 0.035847 0.020158 0.017933 0.021464 0.008864 0.029836 0.171055 nm 2 2 6.464016 2 6.464016 2 6.444048 2 6.44404B 2 6.441456 2 6.271425 t -0.609894 -0.609894 -0.403315 -0.403315 -0.376502 1.382528 W.T. Rank t crit. 0 0 1.895 0 2.002 0 2.039 0 2.058 0 2 .069 0 2 .076 One-eided test at the S5% CL for a decreasing trend. Common logarithm transformation, i.e. Y = loglO(X), applied to the data. MSE 0.009343 with 7 df. Ref: Williams, D. A. Biometrics 28: 519-531 (1972). Company Sanitized. Does not contain TSCA CBt - 12-