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Aqua- i ox, 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.: 070209-1 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 95% <br />SAMPLE DATE: 6-30-09 TIME: 16:OOPM CLIENT.I'D: Site 10 RT <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 7-2-09 10:45AM TEST FINISH: 7-4-09 10:45AM <br />TYPE OF TEST: LC50 48 hours static renewal moderately hard fresh synthetic dilution water. <br />AMMONIA: ND D.O.: 6.34 mg/L <br />HARDNESS: 94.45 mg/L CHLORINE: ND <br />ALKALINITY: 56.76 mg/L TEMPERATURE: 20.3 deg C <br />pH: 7.96 <br />URVIVAL DATA DILUTIONS (%) <br />RS CONTROL 6 25 125 25 50 75 100 <br />DATE r-1 <br />7/2/2009 INITIAL <br />7/3/2009 24 <br />7/4/2009 48 <br />MORTALITY: <br />% SURVIVAL: <br />20 20 20 20 20 20 20 <br />20 20 20 20 20 20 20 <br />20 20 20 20 19 20 19 <br />0 0 0 0 1 0 1 <br />100% 100% 100% 100% 95% 100% 95% <br />ULY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6 25 125 25 50 75 100 <br />CONDUCTIVITY (microS) <br />D.O. (mg/L) <br />pH <br />TEMPERATURE (deg C) <br />319/325 546/553 747/753 1114/1120 1777/1784 282012870 3620/3700 <br />7.2/7.2 7.2/7.2 7.2/7.2 7.117.2 6.9/7.2 6.6/7.1 6.3/7.1 <br />8.0/8.1 7.9/8.1 7.9/8.2 7.8/8.2 7.8/8.2 7.8/8.3 7.7/8.3 <br />19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 19.7/20.3 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: 1.06 mg/L D.O.: 6.34 mg/L <br />HARDNESS: 550.28 mg /L CHLORINE: ND <br />ALKALINITY: 485.29 mg/L TEMPERATURE: 20.3 deg C <br />pH: 7.73 D.O AFTER AERATION: NA <br />ANOEC CALCULATION METHOD METHOD, REFERENCE <br />100% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENT <br />Analysis <br />Conducted by: ?•za.a Reviewed by Date: Dat <br />f <br />i