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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 />IENT NAME: Twentymile Coal Company SAMPLE I.D.: 072809-1 <br />% MORTALITY TEST: Fail TEST RESULT LC50: 96.8% <br />SURVIVAL IN 100% DILUTION: 40% <br />AMPLE DATE: 7-27-09 TIME: 11:OOAM CLIENT ID: Site 62 <br />;GANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />ST START: 7-28-09 20:30PM TEST FINISH: 7-30-09 20:30PM <br />PE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />AMMONIA: ND <br />HARDNESS: 91.95 mg/L <br />ALKALINITY: 55.93 mg/L <br />pH: 7.90 <br />DATE (HOURS) CONTROL 6.25 <br />7/28/2009 INITIAL <br />7/29/2009 24 <br />7/30/2009 48 <br />MORTALITY: <br />% SURVIVAL: <br />TION WATER CONSTITUENT <br />D.O.: 6.60 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 21.0 deg C <br />DILUTIONS (%) <br />12.5 25 rin 75 1 nn <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 15 8 <br />0 0 0 0 0 5 12 <br />100% 100% 100% 100% 100% 75% 40% <br />EMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 625 125 25 ,in 75 inn <br />CONDUCTIVITY (microS; <br />D.O. (mg/L) <br />pH <br />TEMPERATURE (deg C; <br />330/337 605/614 845/855 1310/1337 2320/2380 3100/3210 3890/3980 <br />6.6/7.0 6.7/7.0 6.7/7.1 6.8/7.0 7.0/7.0 7.1/7.1 7.1/7.3 <br />7.9/8.1 8.0/8.1 8.0/8.3 8.0/8.4 8.0/8.4 8.0/8.2 7.9/8.1 <br />20.2/21.0 20.2/21.0 20.2/21.0 20.2/21.0 20.2/21.0 20.2/21.0 20.2/21.0 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 7.30 mg/L <br />HARDNESS: 2248.88 mg/L CHLORINE: ND <br />ALKALINITY: 322.02 mg /L TEMPERATURE: 21.0 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 COMMENTS: <br />Analysis <br />Conducted by: <br />Date: <br />Reviewed by: <br />Date: