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Aqua-Tox, Inc. <br />4880 Robb Street, Suite 8 <br />Wheat Ridge, CO 80033 <br />(303) 621-4880 aqua-tox.com <br />ACUTE TOXICITY REPORT <br />Ceriodaphnia dubia - 2nd Accelerated <br />;LIENT NAME: Twentymile Coal Company SAMPLE I.D.: 090509-1 <br />0 % MORTALITY TEST: Fail TEST RESULT LC50: 86.06% <br />o SURVIVAL IN 100% DILUTION: 30% <br />AMPLE DATE:-9-4-09 TIME: 10:OOAM CLIENT ID: Site 62 <br />)RGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />EST START: 9-5-09 11:55AM TEST FINISH: 9-7-09 11:55AM <br />YPE OF TEST: LC50. 48 hours, static, renewal, moderatelv hard fresh synthetic dilution water. <br /> <br />IODERATELY HARD FRESH SYNTHETIC DILUTION WATER CONSTITUENT <br />AMMONIA: ND D.O.: 8.03 mg/L <br />HARDNESS: 94.00 mg/L CHLORINE: ND <br />ALKALINITY: 55.19 mg/L TEMPERATURE: 19.9 deg C <br />pH: 8.03 <br />URVIVAL DATA DILUTIONS (%) <br />DATE HOURS CONTROL 6.25 12.5 25 50 75 100 <br />9/5/2009 INITIAL 20 20 20 20 20 20 20 <br />9/6/2009 24 20 20 20 20 20 20 20 <br />9/7/2009 48 20 20 20 20 18 14 6 <br />MORTALITY: 0 0 0 0 2 6 14 <br />% SURVIVAL: 100% 100% 100% 100% 90% 70% 30% <br />41LY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (microS) <br />D.O. (mg/L) <br />PH <br />TEMPERATURE (deg C) <br />334/340 600/619 839/850 1263/1290 2260/2270 3020/3110 3820/3840 <br />6.9/7.3 6.9/7.3 6.9/7.3 7.0/7.3 7.0/7.3 7.1/7.3 7.1/7.3 <br />8.0/7.9 8.1/8.1 8.1/8.3 8.1/8.4 8.0/8.5 8.0/8.3 7.9/8.1 <br />19.8/20.3 19.8/20.3 19.8/20.3 19.8/20.3 19.8/20.3 19.8/20.3 19.8/20.3 <br />HEMISTRY - SAMPLE <br />AMMONIA: ND <br />HARDNESS: 2176.00 mg/L <br />ALKALINITY: 404.96 mg /L <br />pH: 7.93 <br />D.O.: 7.13 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 19.9 deg C <br />D.O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />25% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS: <br />Analysis <br />Conducted by: Reviewed by: <br />Date: Date: