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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 <br />CLIENT NAME: 20-Mile Coal Company SAMPLE I.D.: 052610-1 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 70% <br />SAMPLE DATE: 05-25-10 TIME: 08:32A CLIENT ID: Site 62 <br />ORGANISM: Ceriodanhnia dubia AGE: < 24 hours old <br />TEST START: 05-26-10 10:20A TEST FINISH: 05-28-10 10:20A <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.: 7.16 mg/L <br />HARDNESS: 92.19 mg/L CHLORINE: ND <br />ALKALINITY: 57.81 mg/L TEMPERATURE: 19.7 deg C <br />pH: 8.01 <br />'URVIVAL DATA DILUTIONS (%) <br />DATE HOURS CONTROL 6.25 12.5 25 50 75 100 <br />5/2612010 INITIAL <br />5/27/2010 24 <br />5/28/2010 48 <br />MORTALITY: <br />% SURVIVAL: <br />20 20 20 20 20 20 20 <br />20 20 20 19 20 20 17 <br />20 20 20 19 20 19 14 <br />0 0 0 1 0 1 6 <br />100% 100% 100% 95% 100% 95% 70% <br />AILY 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 />311/331 544/576 846/878 1361/1395 2470/2510 3550/3590 4320/4360 <br />7.2/7.4 7.2/7.4 7.2/7.4 7.2/7.3 7.1/7.3 7.1/7.3 7.1/7.2 <br />8.0/8.3 8.0/8.4 7.9/8.5 7.9/8.6 7.8/8.7 7.8/8.8 7.7/8.9 <br />19.7/19.9 19.7/19.9 19.7/19.9 19.7/19.9 19.7/19.9 19.7/19.9 19.7/19.9- <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 7.07 mg/L <br />HARDNESS: 2,479.98 mg/L CHLORINE: ND <br />ALKALINITY: 536.08 mg/1 TEMPERATURE: 19.7 deg C <br />pH: 7.68 D.O AFTER AERATION: NA <br />ANOEC ZALCULATION.METHOD METHOD.-REFERENCE <br />75% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL CO NTS' <br />Analysis <br />Conducted by: Reviewed by: <br />Dat W],1 ?a Date: ..,?/»