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Aqua-Tox, Inc. <br /> 4880 Robb Street, Suite 8 <br /> <br />7 7 <br />° Wheat Ridge, CO 80033 <br />=fir. xt (303) 621-4880 aqua-tox.com <br /> <br /> ACUTE TOXICITY REPORT <br /> Ceriodaphnia dubia <br />CLIENT NAME: Twentymile Coal Company SAMPLE I.D.: 090908-1 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 95% <br />SAMPLE DATE: 9-8-08 TIME: 11:35AM CLIENT ID: Site 62 <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 9-9-08 16:22PM TEST FINISH: 9-11-08 16:22PM <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.: 7.20 mg/L <br />HARDNESS: 90 mg/L CHLORINE: ND <br />ALKALINITY: 54.96 mg/L TEMPERATURE: 19.7 deg C <br />pH: 8.00 <br />SURVIVAL DATA DILUTIONS (%) <br />DATE HOURS CONTROL 6.25 12.5 25 50 75 100 <br />9/9/2008 INITIAL 20 20 20 20 20 20 20 <br />9/10/2008 24 20 20 20 20 20 20 20 <br />9/11/2008 48 20 20 20 20 20 20 19 <br />MORTALITY: 0 0 0 0 0 0 1 <br />% SURVIVAL: 100% 100% 100% 100% 100% 100% 95% <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (micros) 340/369 626/645 839/926 1332/1428 12270/2390 3000/3220 3810/3910 <br />D.O. (mg/L) 7.2/7.3 7.317.3 7.2/7.3 7.2/7.4 7.2/7.7 7.2/7.9 7.2/8.2 <br />pH 8.0/8.2 8.1/8.2 8.1/8.3 8.1/8.4 8.1/8.5 8.0/8.3 8.0/8.1 <br />TEMPERATURE (deg C) 19.5/19.7 19.5/19.7 19.5/19.7 19.5/19.7 19.5/19.7 19.5/19.7 19.5/19.7 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 8.20 mg/L <br />HARDNESS: 2,230.5 mg/L CHLORINE: ND <br />ALKALINITY: 401.24 mg/L TEMPERATURE: 19.7 deg C <br />pH: 7.98 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 by: Reviewed by: <br />Date: Date: