Document v14bJpavYNXBma5jaXyKvv0yZ

AR226-3214 DuPont-8863 TRADE SECRET Study Title H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum Laboratory Project ID: DuPont-8863 A u t h o r : Terry Lee S lo m a n , B .S . S t u d y C o m p l e t e d o n : January 4,2002 P e r f o r m in g L a b o r a t o r y : 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 o f data obtained from this study. Issued by Study Director: b iX - Terry Lee Sloman, B.S. Associate Scientist Date % Does HO*Contain TscA Cfij -2 - H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum STUDY INFORMATION Substance Tested: Submitter's Notebook Nnmhe.rfsV Haskell Number: 25134 CAS Registry Number: Com position^ Purity: jjp Known Impurities: DuPont-8863 Physical Characteristics: Dark brown solid Stability: The test substance appeared to be stable under the conditions o f 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 Initiated/Completed: December 12, 2001 / (see report cover page) ImLife Initiated/Completed: December 12, 2001 / December 15, 2001 Oow TSCACW 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 o f H-25134 on the growth and growth rate o f 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 o f 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 o f the test. The 72-hour EC50and the NOECa based on nominal concentrations are as follows:b,c,d Healthy Cell Count EC50 NOEC >1000 mg/L >1000 mg/L H-25134 exhibited low concern6under TSCA in a 72-hour, acute test using the green alga Selenastrum capricornutum. a The EC50is defined as the "effective concentration" producing a 50% inhibition of 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 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 o f data. Environmental Toxicology and Risk Assessment, A ST M ST P 1179. (W.G. Landis, J.S. Hughes, and M.A. Lewis, Eds.), pp 22-39. American Society for Testing and Materials, Philadelphia. Company SanWzed. Does not contain T SC A CBI -4 - H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 m 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 o f algal cells. After the addition o f algal cells, the test and blank control flasks were placed on a shaker table in a chamber with a temperature o f 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 o f the culture medium was approximately 7.5 at study start. The pH measurements o f 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. % % -5 - H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 TABLES Does notcontain Company 8anWzed' -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 S ^H hed. Does noVconiato T SC A d Company -7- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 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, 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 Company8anttized. Does not containH Q icSgi -8- H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 APPENDIX m Company Sanitized. Does not contain TSCA C B I H-25134: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-8863 APPENDIX A 72-Hour Healthy Cell Count Statistics Company Sanitized. Does not contain T 6 A @BI - 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 #@ALGAE@SCREEN@H25134 72 Hr Healthy Cell Count (#cells/mL) Mean Std Response Error Transform Transform Mean Std Error n 2260000 2120000 2160000 1695000 1835000 2610000 1915000 160000 150000 150000 245000 35000 140000 125000 6.353017 6.325246 6.333404 6.224585 6.263557 6.416015 6.281242 0.030798 0.030780 0.030208 0.063217 0.008285 0.023318 0.028389 2 2 2 2 2 2 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 t crit. 0 0 1.895 0 2.002 0 2.039 0 2.058 0 2.069 0 2.076 One-sided test at the 95% CL for a decreasing trend. Common logarithm transformation, i.e. Y = loglO(X), applied to the data. MSB = 0.002349 With 7 df. Ref: Williams, D. A. Biometrics 28: 519-531 (1972). Company Sanitized. Does no.' .-oniain TSCA CB! -11 -