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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 - 1st Accelerated <br />LIENT NAME: Twentvmile Coal Companv SAMPLE I.D.: 082809-1 <br />% MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />SURVIVAL IN 100% DILUTION: 55% <br />AMPLE DATE:-8-27-09 TIME: 11:30AM CLIENT ID: Site 62 <br />RGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />=ST START: 8-28-09 09:30AM TEST FINISH: 8-30-09 09:30AM <br />(PE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />AMMONIA: ND <br />HARDNESS: 93.20 mg/L <br />ALKALINITY: 55.19 mg/L <br />pH: 8.00 <br />DATE HOURS CONTROL 6.25 <br />8/28/2009 INITIAL <br />8/29/2009 24 <br />8/30/2009 48 <br />MORTALITY: <br />% SURVIVAL: <br />D. 0.: 7.10 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.3 deg C <br />DILUTIONS (%) <br />12.5 25 50 75 100 <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 20 20 <br />20 20 20 19 19 15 11 <br />0 0 0 1 1 5 9 <br />100% 100% 100% 95% 95% 75% 55% <br />II DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />n (325 12 S 2.ri -r; n 7ti inn <br />CONDUCTIVITY (micros; <br />D.O. (mg/L) <br />pH <br />TEMPERATURE (deg C) <br />315/327 513/541 724/734 1089/1105 1742/1850 2560/2670 3160/3270 <br />7.1/7.2 7.1/7.2 7.1/7.2 7.1/7.2 7.1/7.2 7.0/7.2 7.0/7.3 <br />8.0/8.1 8.0/8.3 8.0/8.3 8.0/8.4 8.0/8.6 7.9/8.4 7.9/8.2 <br />19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 7.30 mg/L <br />HARDNESS: 1650.00 mg/L CHLORINE: ND <br />ALKALINITY: 339.96 mg /L TEMPERATURE: 20.3 deg C <br />pH: 7.91 D.O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />50% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL <br />Analysis <br />Conducted by: <br />Date: <br />Reviewed by: <br />Date: