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Aqua-Tox, Inc. <br /> 4880 Robb Street, Suite 8 <br /> Wheat Ridge, CO 80033 <br />4p ? _ (303) 621-4880 aqua-tox.com <br /> <br /> ACUTE TOXICITY REPORT <br /> Pimephales promelas - Fathead Minnow <br />CLIENT NAME: 20-Mile Coal Comp any SAMPLE I.D.: 111610-2 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 80% <br />SAMPLE DATE: 11-15-10 TIME: 16:15P CLIENT ID: Site 62 <br />ORGANISM: Fathead Minnow AGE: DOH 11-11-10 <br />TEST START: 11-16-10 14:10P TEST FINISH: 11-20-10 14:10P <br />TYPE OF TEST: LC50, 96 hours static renewal moderately hard fresh s nnthetic dilution water. <br /> <br />MODERATELY HARD FRESH SYNTHETIC DILUTION WATER CONSTITUENT <br />AMMONIA: ND D.O.: 6.94 mg/L <br />HARDNESS: 98.91 mg/L CHLORINE: ND <br />ALKALINITY: 58.26 mg/L TEMPERATURE: 19.5 deg C <br />pH: 8.04 <br />SURVIVAL DATA DILUTIONS (%) <br />DATE HOURS CONTROL 6.25 12.5 25 50 75 100 <br />11/16/2010 INITIAL <br />11/17/2010 24 <br />11/18/2010 48 <br />11/19/2010 72 <br />11/20/2010 96 <br />MORTALITY: <br />% S URVIVAL: <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 20 19 <br />20 20 20 20 20 19 18 <br />20 20 20 20 20 19 17 <br />20 20 20 20 20 19 16 <br />0 0 0 0 0 1 4 <br />100% _TJ <br />100% 100% 100% 100% 95% 80% <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />CONTROL 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (micros) 323/334 945/959 1578/1590 2030/2060 2470/2510 3310/3340 4280/4320 <br />D.O. (mg/L) 6.6/7.0 6.6/7.1 6.5/7.1 6.5/7.2 6.6/7.3 6.7/7.4 6.7/7.5 <br />pH 7.7/8.2 7.7/8.3 7.7/8.4 7.6/8.5 7.6/8.6 7.6/8.7 7.6/8.8 <br />TEMPERATURE (deg C) 19.5/20.1 19.5/20.1 19.5/20.1 19.5/20.1 19.5/20.1 19.5/20.1 19.5/20.1 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: 0.05 mg/L D.O.: 7.62 mg/L <br />HARDNESS: 2,438.45 mg/L CHLORINE: ND <br />ALKALINITY: 515.00 mg/L TEMPERATURE: 19.5 deg C <br />pH: 7.87 D.O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />75% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS: <br />Analysis <br />Conducted by: Reviewed b . <br />Date: it- 21-1o Dat.