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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 - 3rd Accelerated <br />LIENT NAME: Twentymile Coal Company SAMPLE I.D.: 091809-1 <br />% MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />SURVIVAL IN 100% DILUTION: 90% <br />AMPLE DATE: 9-17-09 TIME: 15:30PM CLIENT ID: Site 62 <br />RGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />=ST START: 9-18-09 10:05AM TEST FINISH: 9-20-09 10:05AM <br />t'PE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br />RATE <br />AMMONIA: ND <br />HARDNESS: 92.80 mg/L <br />ALKALINITY: 54.42 mg/L <br />pH: 8.16 <br />rION WATER CONSTITUENT <br />D.O.: 7.10 mg/L <br />CHLORINE: ND <br />TEMPERATURE: 20.4 deg C <br />DATE (HOURS) CONTROL 6.25 <br />9/18/2009 INITIAL <br />9/19/2009 24 <br />9/20/2009 48 <br />MORTALITY: <br />% SURVIVAL: <br />DILUTIONS (%) <br />125 25 5n 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 20 18 <br />0 0 0 0 0 0 2 <br />100% 100% 100% 100% 100% 100% 90% <br />STRY - (MIN/MAX) DILUTIONS (%) <br />R R 29; 19 5 9r, ?n 7r Inn <br />CONDUCTIVITY (microS) <br />D.O. (mg/L) <br />pH <br />TEMPERATURE (deg C) <br />343/354 491/491 626/641 847/865 1333/1375 2850/3010 3530/3720 <br />7.1/7.2 7.1/7.2 7.1/7.2 7.1/7.2 7.0/7.1 7.0/7.1 7.0/7.1 <br />8.1/8.2 8.1/8.2 8.1/8.2 8.2/8.4 8.2/8.4 8.1/8.4 8.0/8.2 <br />20.1/20.4 20.1/20.4 20.1/20.4 20.1/20.4 20.1/20.4 20.1/20.4 20.1/20.4 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: ND D.O.: 7.03 mg/L <br />HARDNESS: 2080.00 mg/L CHLORINE: ND <br />ALKALINITY: 379.16 mg /L TEMPERATURE: 2p.4 deg C <br />pH: 7.99 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: <br />Date: <br />Reviewed by: <br />Date: