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317/325 <br />431/440 <br />552/561 <br />- <br />773/787 <br />1170/1184 <br />10 <br />1515/1537 <br />100 <br />1905/1915 <br />7 2/7 6 <br />7 3/7 6 <br />7.3/7 7 <br />7 4/7.7 <br />7.4/7 8 <br />7.4/7.9 <br />7.4/8.0 <br />8 0/8.1 <br />8.0/8 2 <br />8 0/8.2 <br />8.0/8 2 <br />7.9/8 2 <br />7.9/8.3 <br />7.8/8.4 <br />20 1/20.4 <br />20.2/20 8 <br />20 2/20 8 <br />20 2/20 8 <br />20 2/20 8 <br />20 2/20 8 <br />20 1/20 4 <br />20 <br />20 <br />20 20 <br />20 <br />20 20 <br />20 <br />20 <br />20 20 <br />20 <br />20 20 <br />20 <br />20 <br />20 20 <br />20 <br />20 20 <br />0 0 <br />I <br />0 0 <br />I <br />0 <br />0 0 <br />100% 100% <br />100% 1% <br />100% <br />100% I 100% <br />1 <br />1 <br />1 <br />1 <br />1 <br />1 <br />1 <br />1 <br />1 <br />1 <br />1 <br />Aqua -Tox, Inc. <br />4880 Robb Street, Suite 8 <br />Wheat Ridge, CO 80033 <br />(303) 621 -4880 www aqua -tox com <br />ACUTE TOXICITY REPORT <br />Ceriodaphnia dubia <br />CLIENT NAME: Twentymile Coal Company <br />50 % MORTALITY TEST. Pass <br />% SURVIVAL IN 100% DILUTION 100% <br />SAMPLE I.D.: 032812 -2 <br />TEST RESULT LC50 >100% <br />SAMPLE DATE: 03 -27 -12 TIME. 15 CLIENT ID. 2000 -87 <br />ORGANISM. Ceriodaphnia dubia AGE. < 24 hours old <br />TEST START: 03 -28 -12 1200 TEST FINISH. 03 -30 -12 12 00 <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. 7.58 mg /L <br />HARDNESS: 83 01 mg /L CHLORINE ND <br />ALKALINITY: 56 56 mg /L TEMPERATURE: 20 2 deg C <br />pH 8.00 <br />SURVIVAL DATA <br />DATE (HOURS) CONTROL <br />3/28/2012 INITIAL <br />3/29/2012 24 <br />3/30/2012 48 <br />MORTALITY. <br />% SURVIVAL: <br />6.25 <br />DILUTIONS ( %) <br />12.5 25 <br />100 <br />DAILY CHEMISTRY - (MIN /MAX) <br />0 <br />CONDUCTIVITY (microS) <br />D 0 (mg /L) <br />pH <br />TEMPERATURE (deg C) <br />6 25 <br />DILUTIONS ( %) <br />1 <br />ADDITIONAL COMMENTS <br />Analysis <br />Conducted by <br />Date <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA 0.02 mg /L <br />HARDNESS 968 42 mg /L <br />ALKALINITY 116.76 mg /L <br />pH 7.84 <br />D.0 . 7.99 mg /L <br />CHLORINE. ND <br />TEMPERATURE 20.2 deg C <br />D 0 AFTER AERATION. NA <br />ANOEC <br />100% <br />CALCULATION METHOD <br />SPEARMAN - KARBER <br />METHOD REFERENCE <br />EPA - 821 -R -02 -012 October 2002 pg.78 <br />Reviewed by. <br />Date <br />4414t <br />V1/1 <br />