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Aqua -Tox, Inc.i <br />4880 Robb Street, Suite 8 <br />Wheat Ridge, CO 80033 <br />(303) 621 -4880 www aqua -tox com <br />ACUTE TOXICITY REPORT <br />CLIENT NAME. Twentymile Coal Company SAMPLE I.D. 051013 -1 <br />50 % MORTALITY TEST- Pass 'TEST RESULT LC50- >100% <br />% SURVIVAL IN 100% DILUTION: 100% , - <br />SAMPLE DATE: 05 -09 -13 TIME: 13*10P CLIENT ID12000 -87 - (6 7 Vj <br />,ORGANISM: Daphnia magna AGE. < 24 hours old <br />TEST START, 05 -10 -13 09 45A TEST FINISH: 05 -12 -13 09:45A <br />TYPE OF TEST, LC50, 48 hours, static, renewal moderately hard fresh svnthetic dilution water, <br />I -Kt:bH sYN I Hh11G DILUTION WATER CONSTITUENT <br />AMMONIA: ND D.O 7.38 mg /L <br />HARDNESS: 89,47 mg /L CHLORINE: ND <br />ALKALINITY 54,05 mg /L. TEMPERATURE 20.6 deg C <br />pH. 7.98 <br />SURVIVAL DATA DILUTIONS ( %) <br />DATE (HOURS) CONTROL 6.25 12.5 25 50 75 100 <br />5/10/2012 INITIAL <br />5/11/2012 24 <br />5/12/2012 48 <br />MORTALITY' <br />SURVIVAL <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />19 <br />20 <br />_ <br />20 <br />20 <br />20 <br />20 <br />20 <br />19 <br />20 <br />20 <br />20 <br />0 <br />0 <br />0 <br />1 <br />0 <br />0 <br />0 <br />100% <br />100% <br />1 100% <br />95% <br />100% <br />100% <br />1 100% <br />LILY CHEMISTRY - (MIN /MAX) DILUTIONS ( %) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (microS) <br />DO (mg /L) <br />pH <br />TEMPERAI URE (deg C) <br />325/342 <br />5041514 <br />641/650 <br />887/900 <br />1357/1391 <br />1681/1761 <br />197512260 <br />7.0/7.4 <br />7,0/7.5 <br />7.0/7.9 <br />70/8.1 <br />7 0/8 4 <br />7 0/8 7 <br />71/9.1 <br />8.0/8.1 <br />8.0/8.2 <br />8.0/85 <br />8.0/8.8 <br />8.1/9.1 <br />82/9.2 <br />83/9.4 <br />20,0/20,9 <br />20.0/20.9 <br />20 0120,9 <br />20.0/20.9 <br />20 0/20.9 <br />20.0/20.9 <br />20 0/20 9 <br />CHEMISTRY - SAMPLE <br />AMMONIA <br />0,04 mg /L <br />D.0 <br />9.08 mg /L <br />HARDNESS. <br />1,142 43 mg /L <br />CHLORINE- <br />ND <br />ALKALINITY' <br />102.08 mg /L <br />TEMPERATURE, <br />20,6 deg C <br />pH: <br />939 <br />D.0 AFTER AERATION: <br />NA <br />ANOEC CALCULATION METHOD METHOD REFERENCE <br />100% SPEAR MAN- KARBER EPA - 821 -R -02 -012 October 2002 pg.78 <br />ADDITIONAL COMMENTS. I <br />Analysis <br />Conducted by: <br />Date <br />Reviewed by� ' <br />Dater <br />L <br />i <br />