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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: Twentymile Coal Company SAMPLE I.D.: 102108-2 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 80% <br />SAMPLE DATE: 10-20-08 TIME: 14:55PM CLIENT ID Site 62 <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 10-21-08 10:10AM TEST FINISH: 10-23-08 10 10AM <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.: 7.44 mg/L <br />HARDNESS 91.05 mg/L CHLORINE: ND <br />ALKALINITY: 55.96 mg/L TEMPERATURE: 19.8 deg C <br />pH: 8.03 <br />SURVIVAL DATA DILUTIONS (%) <br />DATE (HOURS) CONTROL 6.25 12.5 25 50 75 inn <br />10/21/2008 INITIAL 20 20 20 20 20 20 20 <br />10/22/2008 24 20 20 20 20 20 20 20 <br />10/23/2008 48 20 20 19 20 19 18 16 <br />MORTALITY: 0 0 1 0 1 2 4 <br />% SURVIVAL: 100% 100% 95% 100% 95% 90% 80% <br />I <br />DAILY CHEMISTRY - (MIN/MAX) <br />0 6.25 <br />12.5 DILUTIONS (%) <br />25 <br />50 <br />75 100 <br />CONDUCTIVITY (micros) 330/350 585/623 829/860 1225/12751 2240/2260 2910/2950 3610/3710 <br />D.0 (mg/L) 7.3/7.4 7.3/7.5 7.3/7.6 7.3/7.9 7.4/8.3 74/7.9 7.4/8.0 <br />pH 8.0/8.3 8.0/8.2 8.1/8.3 8.1/8.4 8.0/8.4 8.0/8.3 8.0/8.2 <br />TEMPERATURE (deg C) 19.6/20.1 19.6/20.1 19.6/201 19.6/20.1 19.6/20.1 19.6/201 19.6/201 <br />I <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA. ND D.O.: 7.95 mg/L <br />HARDNESS 1.820.91 mg/L CHLORINE ND <br />ALKALINITY: 435.72 mg/L TEMPERATURE: 19.8 deg C <br />pH. 7.98 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: Reviewed by: t C. -3c.o$ <br />Dat : Ic o Date- <br />IzI17 7