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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 />M <br /> ACUTE TOXICITY REPORT <br /> Ceriodaphnia dubia - 1st Accelerated <br />CLIENT NAME: Twentymile Coal Companx SAMPLE I.D.: 110609-1 <br />50 % MORTALITY TEST: Fail TEST RESULT LC50: 88.704% <br />% SURVIVAL IN 100% DILUTION: 25% <br />SAMPLE DATE: 11-5-09 TIME: 15:20PM CLIENT ID: Site 62 <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 11-6-09 13:OOPM TEST FINISH: 11-8-09 13:OOPM <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.: 8.00 mg/L <br />HARDNESS: 92.89 mg/L CHLORINE: ND <br />ALKALINITY: 55.31 mg/L TEMPERATURE: 20.0 deg C <br />pH: 7.97 <br />SURVIVAL DATA DILUTIONS (%) <br />DATE (HOURS)CONTROL 6.25 12.5 25 50 75 100 <br />11/6/2009 INITIAL 20 20 20 20 20 20 20 <br />11/7/2009 24 20 20 20 20 20 20 19 <br />11/8/2009 48 20 20 20 20 20 17 5 <br />MORTALITY: 0 0 0 0 0 3 15 <br />% SURVIVAL: 100% 100% 100% 100% 100% 85% 25% <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (micros) 319/338 596/634 843/862 1301/1330 2300/2340 3020/3140 3810/3930 <br />D.O. (mg/L) 7.1/7.2 7.1/7.3 7.2/7.2 7.2/7.3 7.2/7.5 7.3/7.8 7.3/8.0 <br />pH 8.0/8.1 8.1/8.1 8.2/8.3 8.2/8.4 8.1/8.5 8.1/8.3 8.0/8.1 <br />TEMPERATURE (deg C) 20.0/20.7 20.0/20.7 20.0120.7 20.0/20.7 20.0/20.7 20.0/20.7 20.0/20.7 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 8.00 mg/L <br />HARDNESS: 2222.22 mg/L CHLORINE: ND <br />ALKALINITY: 449.90 mg/L TEMPERATURE: 20.0 deg C <br />pH: 8.04 D.0 AFTER AERATION: NA <br />AN;OEC CALCULATION METHOD:, METHODiREFERENCE "y <br />50% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS- <br />Analysis <br />Conducted by: Reviewed by: <br />Date: Date: % r, iG 101,