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Aqua-Tox, Inc. <br /> 4380 Robb Street, Suite 8 <br /> Wheat Ridge, CO 80033 <br /> (303) 621-4880 aqua-tox.com <br /> ACUTE TOXICITY REPORT <br /> Cerioda hnia dubia <br />CLIENT NAME Twentymile Coal Company SAMPLE I.D.. 040209-1 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 100% <br />SAMPLE DATE: 4-1-09 TIME: 13:35PM CLIENT ID: Site 10 RT <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START 4-2-09 10:40AM TEST FINISH: 4-4-09 10:40AM <br />TYPE OF TEST: LC50 48 hours static renewal moderately hard fresh synthetic dilution water. <br /> <br />MODERATELY HARD FRESH SYNTHETIC DILUTION WATER CONSTITUENT <br />AMMONIA ND D.O. 7.40 mg/L <br />HARDNESS: 98.15 mg/L CHLORINE: ND <br />ALKALINITY 55.56 mg/L TEMPERATURE: 20.2 deg C <br />pH: 8.01 <br />SURVIVAL DATA DILUTIONS (%) <br />DATE LHOURS) CONTROL 6.25 12.5 25 50 75 100 <br />4/2/2009 INITIAL 20 20 20 20 20 20 20 <br />4/3/2009 24 20 20 20 20 20 20 20 <br />4/4/2009 48 20 20 20 20 20 20 20 <br />MORTALITY: 0 0 0 0 0 0 0 <br />% SURVIVAL 100% 100% 100% 100% 100% 100% 100% <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (micros) 319/329 560/571 796/803 1184/1191 1995/2280 2880/3030 3700/3830 <br />DO. (mg/L) 7 2/7.4 7.2/7.4 7.1/7.5 7.1/7.6 7.1/8.0 7.1/8,1 7.1/8.9 <br />pH 8.0/81 8 1/83 8.1/8.3 8.2/8.4 8.2/8.6 8.2/8.4 8.1/8.3 <br />TEMPERATURE (deg C) 19.7/20.3 19.7/20.3 197/20.3 19.7/203 19.7/20.3 19.7/20.3 19.7/20.3 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA. 0.42 mg/L D.O . 8.88 mg/L <br />HARDNESS: 1093.64 mg/L CHLORINE: ND <br />ALKALINITY: 477.69 mg/L TEMPERATURE: 20.2 deg C <br />pH: 8.12 D O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />100% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS. <br />Analysis <br />Conducted b ?- <br />y: Reviewed by <br />1- A <br />Date: <br />Date: <br />f