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3/26/2013 INITIAL <br />3/27/2013 24 <br />3/2812013 48 <br />MORTALITY <br />% SURVIVAL <br />Aqua-Toy, Inc. <br />20 <br />4880 Robb Street, Suite 8 <br />20 <br />Wheat Ridge, CO 80031 <br />_ 20 <br />(303) 621 -4880 www aq _ia -tox com <br />AMMONIA <br />20 <br />20 <br />ACUTE TOXICITY REPORT <br />20 <br />Ceriodaphnia dubia <br />CLIENT NAME Twentymile Coal Company <br />SAMPLE I -D. 032613 -1 <br />50 % MORTALITY TEST Pass <br />TEST RESULT LC50- >100% <br />% SURVIVAL IN 100% DILUTION: 100% <br />20 <br />SAMPLE DATE: 03 -25 -13 TIME: 13:25P <br />CLIENT ID 2000 -84 <br />ORGANISM Cenodal)hma dubia AGE, _ <" 24 hours old <br />_ <br />TEST START 03 -26 -13 13,20 PM <br />TEST FINISH. 03 -28 -13 13.20 PM <br />TYPE OF TEST- LC50 48 hours static, renewpl, 1110deratelYh8rd fresh synthetic dilt2tion,water, <br />MODERATELY HARD FRESH SYNTHETIC DILUTION WATER CONSTITUENT <br />AMMONIA ND <br />D,O.. 6 84 mg /L <br />HARDNESS 86.23 mg /L <br />CHLORINE. ND <br />ALKALINITY 58 29 mg /L <br />TEMPERATURE 19.3 deg C <br />pH 8.08 <br />SURVIVAL DATA <br />DILUTIONS ( %) <br />I DATE HOURS CONTROL 6 25 <br />12 5 25 50 75 100 i <br />3/26/2013 INITIAL <br />3/27/2013 24 <br />3/2812013 48 <br />MORTALITY <br />% SURVIVAL <br />20 <br />20 <br />20 <br />20 <br />20 <br />_ 20 <br />20 <br />AMMONIA <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />20 <br />0 <br />20 <br />q <br />20 <br />0 <br />0 <br />0 <br />0 <br />0 <br />100% <br />100% <br />100% <br />100% <br />100% <br />_ 100% <br />100% <br />DAILY CHEMISTRY - (MIN /MAX) DILUTIONS (c ,) <br />0 6.215 12 5 25 50 75 100 <br />7ONDUC FIVITY (micros) <br />DO (mg /L, <br />TEMPERATURE (deg 4:1-9 <br />318/332 <br />619/648 <br />988/1027 <br />1474/1632 <br />2530/2910 <br />3840/3960 <br />4870%5020( <br />AMMONIA <br />6,8/77 <br />6,9/77 <br />7 0/7,6 <br />7 3/7.6 <br />7 3/7 6 <br />7,6/77 <br />7,5/76 <br />ALKALINITY" <br />432 64 mg /L TEMPERATURE <br />_ <br />19 3 deg C <br />pH <br />80/8.8 <br />AERATION <br />03/20.1 <br />3/20 1 <br />19 3120.1 <br />19 3/20 1119 <br />3/20 1 <br />19 3/20 1 <br />19,03/20.1 <br />19 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA <br />0 91 iTlg /L <br />DO. <br />7 63 mg /L <br />HARDNESS <br />1065.88 mg /L <br />CHLORINE <br />ND <br />ALKALINITY" <br />432 64 mg /L TEMPERATURE <br />_ <br />19 3 deg C <br />pH <br />7,95 D O AFTER <br />AERATION <br />NA <br />ANOEC <br />CALCULATION METHOD <br />METHOD REFERENCE <br />100% <br />l <br />SPEARMAN- KARBER <br />EPA - 821- R -02 -012 October 2002 pg.78 <br />ADDITIONAL COMMENTS <br />i Anaiysi* <br />I <br />Conducted by <br />l <br />Reviewed b;, <br />Date' <br />Date. <br />