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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 /> Cerioda hnia dubia <br />CLIENT NAME: Twentymile Coal Company SAMPLE I.D.: 101509-6 <br />50 % MORTALITY TEST: Fail TEST RESULT LC50: 70.136% <br />% SURVIVAL IN 100% DILUTION: 0% <br />SAMPLE DATE: 10-14-09 TIME: 15:OOPM CLIENT ID: Site 62 <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 10-15-09 11:50AM TEST FINISH: 10-17-09 11:50AM <br />TYPE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />MODERATELY HARD FRESH SYNTHETIC DILUTION WATER CONSTITUENT <br />AMMONIA: ND D.O.: 7.06 mg/L <br />HARDNESS: 92.00 mg/L CHLORINE: ND <br />ALKALINITY: 54.42 mg/L TEMPERATURE: 19.7 deg C <br />pH: 8.10 <br />SURVIVAL DATA DILUTIONS (%) <br />DATE (HOURS) CONTROL 6.25 12.5 25 50 75 100 <br />10/15/2009 INITIAL 20 20 20 20 20 20 20 <br />10/16/2009 24 20 20 20 20 20 20 16 <br />10/17/2009 48 20 20 20 20 18 11 0 <br />MORTALITY: 0 0 0 0 2 9 20 <br />% SURVIVAL: 100% 100% 100% 100% 90% 55% 0% <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (microS) 314/329 628/641 945/965 1497/1507 2650/2710 3490/3630 4340/4520 <br />D.O. (mg/L) 7.1/7.2 7.1/7.2 7.2/7.3 7.2/7.4 7.2/7.7 7.2/8.2 7.2/8.6 <br />pH 8.0/8.1 8.0/8.1 8.1/8.2 8.1/8.3 8.0/8.4 8.0/8.2 7.9/8.1 <br />TEMPERATURE (deg C) 19.7/20.4 19.7/20.4 19.7/20.4 19.7/20.4 19.7/20.4 19.7/20.4 19.7/20.4 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 8.61 mg/L <br />HARDNESS: 2618.00 mg/L CHLORINE: ND <br />ALKALINITY: 451.19 mg/L TEMPERATURE: 19.7 deg C <br />pH: 7.94 D.0 AFTER AERATION: NA <br />ANOEC _CALC?JLATION METH METHOD REFERENCE > <br />25% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS: <br />Analysis <br />Conducted by: - i o •a y Reviewed by: 4,-? <br />Date: Date: M 1 a9