Document oM0Y6yea4yj6aOZKpjRkeBJw

TRADE SECRET AR226-3170 DuPont-448 Study Title H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum Laboratory Project ID: DuPont-4488 AUTHOR: Terry Lee Sloman, B.S. STUDY COMPLETED ON: November 16, 2000 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 WORK REQUEST NUMBER: r ^ ^ l wj^^^^^^^^B~ 1 STUDY CODE NUMBER(:--lB--BBIl Company Saniffzed. Does nof contain TSCA CBf 1 - H-24616: Influence on Growth and Growth Rate of the Green Alga Selenasti-um capricornutum DuPont-4488 STUDY INFORMATION Substance Tested: UUBBBt J Synonvms/Cojdes: H-24616 1 Haskell Number: 24616 Purity: Known | Impurities Physical Characteristics: Sponsor: E. I. du Pont de Nemours and Company Wilmington, Delaware 19898 U.S.A. Study Initiated/Completed: November 6, 2000 / (see report cover page) m-Life Initiated/Completed: November 6, 2000 / November 9, 2000 company Sanitized. Does not contain TSCA CBI 2- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-4488 SUMMARY A study was conducted to determine the effect of H-24616 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-24616/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 EC5o, including the 95% confidence intervals (C.I.), and the NOEC1 based on nominal concentrations are as follows:2'3'4 Healthy Cell Count ECso NOEC 30 mg/L (95% C.I. not calculated) 100 mg/L H-24616 exhibited medium concern5 under TSCA and assigned "acute toxic" to aquatic organisms (R52)6 and classified as dangerous for the environment under the EU Directive in a 72-hour, acute test using the green alga Selenastrum capricornutum. ' The ECso 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. 2 BBN Software Products Corp., 1988. An RS/1 RPL.procedure conducting Williams' test and probit analysis. 3 McCullogh, L.P. and J.A. Nelder. 1989. Generalized Linear Models 2nd Ed. Chapman and Hall, London 511 pp. (See Chapter 9, p. 328). 4 Snedecor, G.W. and W.G. Cochren. 1980. Statistical Methods, 711'Ed. Iowa Stat. Univ. Press, Ames, Iowa. 507 pp. (See Table A 4, p. 469). 5 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. 6 Official Journal of the European Communities Vol. 36, 4 May 1993, p. 68, section 5.2.1. Company Sanitized. Does not contain TSCA CBI -^- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-4488 CERTIFICATION I, the undersigned, declare that this report provides an accurate evaluation of data obtained from this study. bsaed by Study Director: B.S.Date LAAAU ^UL. )^cr^-o^.______ ffTerry Lee Sloman, Associate Scientist tt>-)OV-a-000 Company Sanitized. Does no? contain TSCA CBI -4- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-4488 RESULTS Based on visual observations, the blank (normal culture medium) control solution and the 0.01, 0.1,1,10, and 100 mg/L test solutions were clear and had no color before addition of algal cells. However, the 1000 mg/L test solution was slightly cloudy 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 8440 lumens/m2 (= lux). The shaking speed was 110 revolutions per minute (rpm). The pH measurement of the culture medium was 7.45 at study start. The pH measurements of the definitive test solutions at test initiation ranged from 6.86 to 7.45, and at test termination ranged from 7.09 to 7.54. Company Sanitized. Does no! contain TSCA CBI 5- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-4488 TABLES Company Sanitized. Doss not contain TSCA CB1 6- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastmm capricornutum DuPont-4488 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 7.45 6.87 6.87 6.86 6.86 6.86 6.86 7.18 7.09 7.28 7.38 7.25 7.49 7.54 Company Sanitized. Does not contain TSCA CB8 7- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricomutum DuPont-4488 TABLE 2 DATA SUMMARY Nominal Day 0 Healthy Cell Count Day 3 Healthy CeU Count Concentration ____(cells/mL)____ ____(cells/mL)____ Blank Control Replicate 1 Replicate 2 10,000 10,000 2,190,000 2,590,000 0.01 mg/L Replicate 1 Replicate 2 10,000 10,000 2,130,000 2.280,000 0.1 mg/L Replicate 1 Replicate 2 10,000 10,000 2,090,000 2,590,000 1 mg/L Replicate 1 Replicate 2 10,000 10,000 2,160,000 2,270,000 10 mg/L Replicate 1 Replicate 2 10,000 10,000 2,340,000 2,490,000 100 mg/L Replicate 1 Replicate 2 10,000 10,000 2,170,000 2,470,000 1000 mg/L Replicate 1 Replicate 2 10,000 10,000 680,000 770,000 :3ffipany Sanitized. Does not contain TSCA CBI -8- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricornutum DuPont-4488 FIGURES wpany Sanitized. Does not contain TSCA CBI -9- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricomutum DuPont-4488 FIGURE 1 72-HOUR HEALTHY CELL COUNT DOSE RESPONSE CURVE TOLERANCE DISTRIBUTION BASSO ON HftXIBOM QUASI-LlBBrSH_O_O_D_ USTED FOR BACKGROUND RESPONSE TON OF KOHMIT COKVE PABAMBTBRS 72 Hr Healthy Cell Count (cells/inL> 110 100- 90- so lo-- 60-- 50- 40- 30- Obs. Pitted 0.001 0.010 1.000 lo.ooo ioo.ooo looo.ooo Dose (ing/Ll Quasi-likelihood aSBuned; Variance = 1.11793e+12*ean^0. Noraitft) 15.083342 t -l0.248316*LoglO(Dose) with 95% confidence limits. loooo.ooo "*CTnpany SanRi'zed. Does not eonfafn TSCA CBT 10. H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastmm capricornutum DuPont-4488 APPENDICES company Saniffze3. Does not conIaTn'TSCACBI 11- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastrum capricomutum DuPont-4488 APPENDIX A 72-Hour Healthy Cell Count company Sanitized. Does not contain TSCA CBI 12- H-24616: Influence on Growth and Growth Rate of the Green Alga Selenastiiim capricornutum DuPont-4488 M.T. Rank 0 0 0 0 0 0 1 Dose 0 .00 0 .01 0 .10 1 .00 10.00 100 .00 1000 .00 WILLIAMS TEST OF ftMiGaBBSC 72 Hr Healthy Cell Count (ftcefT--e/,m--L--) Mean Std Response Error Tr,ansform Transform Me.an Std Error n 2390000 200000 6 .376872 0.036428 2 2205000 75000 6 .343157 0.014778 2 2340000 250000 6 .366723 0.046577 2 2215000 55000 6 .345240 0.010786 2 2415000 75000 6.382708 0.013492 2 2320000 150000 6 .364578 0.028119 2 725000 45000 5 .859500 0.026991 2 m 6.360481 6.360481 6.360481 6.360481 6.360481 5.B59500 t 0 .412269 0 .412269 0 .412269 0 .412269 0 ,412269 13 .013216 W.I. _ Rank t crit. 1.895 2.002 2.039 2.058 2.069 2.076 One-sided test at the 95t CL for a decreasing trend. Common logarithm transformation, i.e. Y = logLO(X), applied to the data. MSE a 0.001581 with 7 df. Ref: Williams, D. A. Biometrics 28: 519-531 (1972). Parameter Background Intercept Slope Statistic: Value QUASI-LIKELIHOOD E5T1HATES OF NQRHIT PARAMETERS FO Estiiniate 23900 00.000000 15.083342 10.248316 72 Hr Healthy Standard Error 200000.000000 910.830377 420.889905 Cell Count (feel Is/Hit) Corr. Kith Background 1.000000 Chisa uare df Significance cispersion 1. 11793e-f13 10.000000 0.001 1.11793e+12 Corr. ufth Intercept 0.000000 1.000000 t-yatue 2.228139 Corr. Kith Slope 0 .000000 -1 .029*16 .000000 <t*Cs lope)*2 8173 .682287 Ouantile (X) 1 5 10 20 2S 30 40 50 60 70 rs 80 90 95 99 E ftimate 49.976753 1.2 883207 39 521856 35 802244 34 482763 33 339324 31 369545 Z9 633801 Z7 994100 26 340131 25 466700 24 528133 22 219660 20 477997 17 570710 Loner 95%CI Upper 95XCI auati-likelihood assumed; Variance = 1.117S3e*12l^eanAO. Convergence achieved. ReLative change in quas i - I iicel i hood 5.235931e-0fi Variances and covariances have been multiplied by the d i .pert ion parameter of 1.11793e+12. Company Sanitized. Does not contain TSCA CB1 13-