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AR226-3214 TRADE SECRET Study Title H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum Laboratory Project ID: DuPont-8863 AUTHOR: Terry Lee Sloman, B.S. STUDY COMPLETED ON: January 4, 2002 PERFORMING LABORATORY: E.I. du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050 .Company Sanitized. Does not contain TSCA CB1 Page 1 of 11 H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 CERTIFICATION I, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. Issued by Stndy Director: B.S.Date ^J^u ^JUL. ^^T^OL^_________ /JTerry Lw Sloman, Associate Scientist Q'-^'JAfU- 3.00.L e9apawsanslsM-^^^^ 2- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastfum capricornutum STUDY INFORMATION Substance Tested: Synonyms/Codes: Submitter's Notebook Numberfs): Haskell Number: 25134 DuPont-8863 Physical Characteristics: Dark brown solid Stability: The test substance appeared to be stable under the conditions of the study; no evidence of instability was observed. Sponsor: E.I. du Font de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: December 12, 2001 / (see report cover page) In-Life Initiated/Completed: December 12, 2001 / December 15, 2001 .C^^,^|fted.Ot,ntcnntalnT8CACB .3- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 SUMMARY A study was conducted to determine the effect of H-25134 on the growth and growth rate of the green alga Selenastrum capricornutum. The algae were exposed to nominal concentrations of 0.01, 0.1, 1, 10,100, and 1000 mg H-25134/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 ECso and the NOEC8 based on nominal concentrations are as follows:1''0''1 Healthy Cell Count ECso NOEC >1000 mg/L >1000 mg/L H-25134 exhibited low concern under TSCA in a 72-hour, acute test using the green alga Selenastrum capricornutum. a The EC5o is defined as the "effective concentration" producing a 50% inhibition of growth relative to the control. The NOEC is defined as the highest concentration of test substance that has no significant effect on the measured parameter relative to the control. b BBN Software Products Corp., (1998). An RS/1 RPL procedure conducting Williams' test and probit analysis. c McCullogh, L.P. and Nelder, J.A. (1989). Generalized Linear Models, 2nd edition. 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 of data. Environmental Toxicology and Risk Assessment, ASTMSTP 1179. (W.G. Landis, J.S. Hughes, and M.A. Lewis, Eds.), pp 22-39. American Society for Testing and Materials, Philadelphia. Company SanHteed. Dow not contain TSCA CBI "T H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 RESULTS Based on visual observations, the blank (normal culture medium) control solution had no color before addition of algal cells. However, the 0.01, 0.1, 1, 10,100, and 1000 mg/L test solutions contained undissolved test substance 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 8066 lumens/m2 (= lux). The shaking speed was 105 revolutions per minute (rpm). The pH measurement of the culture medium was approximately 7.5 at study start. The pH measurements of the definitive test solutions at test initiation ranged from 7.44 to 7.81, and at test termination ranged from 7.78 to 8.02. >Doenot contain TSCA CBf 5- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 TABLES cAp^^"'-0^'"'"1""1 6- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 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 7.44 7.78 7.81 7.77 7.68 7.65 7.67 7.78 7.86 7.97 7.84 7.91 7.87 8.02 Company SMiMnd. Does not contain SSCA01 7- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricomutum DuPont-8863 TABLE 2 DATA SUMMARY Nominal Concentration Day 0 Healthy Cell Count ____(cells/mL) Day 3 Healthy Cell Count (ceUs/mL) Blank Control Replicate 1 Replicate 2 10,000 10,000 2,100,000 2,420,000 0.01 mg/L Replicate 1 Replicate 2 10,000 10,000 1,970,000 2,270,000 0.1 mg/L Replicate 1 Replicate 2 10,000 10,000 2,310,000 2.010.000 1 mg/L Replicate 1 Replicate 2 10,000 10,000 1,940,000 1,450,000 10 mg/L Replicate 1 Replicate 2 10,000 10,000 1,800,000 1,870.000 100 mg/L Replicate 1 Replicate 2 10,000 10,000 2,750,000 2.470.000 1000 mg/L Replicate 1 Replicate 2 10,000 10,000 2,040,000 1,790,000 Company Sanitized. Does not containlSQlK^I 8- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 APPENDIX Company Sanitized. Does not contain TSCA CB{ -9- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 APPENDIX A 72-Hour Healthy Cell Count Statistics -10- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 72-HOUR HEALTHY CELL COUNT STATISTICS W.T. Rank 0 0 0 0 0 0 0 Dose 0 .00 0 .01 0 .10 1 .00 10 .00 100 .00 1000 .00 WILLIAMS TEST OF ff@ALGAE@SCREENH25134 72 Hr Healthy Cell Count (#cells/mL) Mean Std Response Error Transform Transform Mean Std Error n 2260000 160000 6.353017 0.030798 2 2120000 150000 6.325246 0.030780 2 2160000 150000 6.333404 0.030208 2 1695000 245000 6.224585 0.063217 2 1835000 35000 6.263557 0.008285 2 2610000 140000 6.416015 0.023318 2 1915000 125000 6.281242 0.028389 2 A m 6 .329325 6 .329325 6 .301386 6 .301386 6 .301386 6 .281242 t 0 .488792 0 .488792 1 .065206 1 .065206 1 .065206 1 .480793 W.T. Rank 0 0 0 0 0 0 0 t crit. 1.895 2.002 2.039 2.058 2.069 2.076 One-sided teat at the 95% CL tor a decreasing trend. Common logarithm transformation, MSE = 0.002349 with 7 df. i.e. Y = loglO(X), applied to the data. Ref; Williams, D. A. Biometrics 28: 519-531 (1972). 5om^any Sanitized. Does wi ^iwn TSCA CB? 11-