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Aqua-Tox, Inc. <br /> 4880 Robb Street, Suite 8 <br /> <br />y Wheat Ridge, CO 80033 <br /> x? <br />(303) 621-4880 aqua-toxsco <br /> ACUTE TOXICITY REPORT <br /> Cerioda hnia dubia <br />CLIENT NAME: Twentymile Coal Company SAMPLE I.D.: 061809-1 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: >100% <br />% SURVIVAL IN 100% DILUTION: 70% <br />SAMPLE DATE:-6-17-09 TIME: 13:31 PM CLIENT ID: Site 115 <br />ORGANISM: Ceriodaphnia dubia AGE: < 24 hours old <br />TEST START: 6-18-09 12:40PM TEST FINISH: 6-20-09 12:40PM <br />TYPE OF TEST: LC50, 48 hours, static, renewal, moderately hard fresh synthetic dilution water. <br /> <br />MODERATELY HARD FRESH SYNTHETIC DILUTION WATER CONSTITUENT <br />AMMONIA: ND D.O.: 7.10 mg/L <br />HARDNESS: 92.49 mg/L CHLORINE: ND <br />ALKALINITY: 57.13 mg/L TEMPERATURE: 20.0 deg C <br />pH: 7.95 <br /> <br />SURVIVAL DATA <br />DILUTIONS (%) <br />-11 <br />DATE HOURS CONTROL 6.25 12.5 25 50 75 100 <br />6/18/2009 INITIAL 20 20 20 20 20 20 20 <br />6/19/2009 24 19 20 20 19 .20 19 18 <br />6/20/2009 48 19 20 20 19 20 17 14 <br />MORTALITY: 1 0 0 1 0 3 6 <br />% SURVIVAL: 95% 100% 100% 95% 100% 85% 70% <br />I ? <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (microS) 3301337 651/655 982/997 1584/1595 294012970 4130/4180 5270/5320 <br />D.O. (mg/L) 7.0/7.2 7.0/7.2 7.1/7.2 7.1/7.2 7.1/7.3 7.1/7.3 7.1/7.4 <br />pH 8.0/8.0 8.1/8.3 8.1/8.5 8.1/8.6 8.1/8.6 8.1/8.6 8.0/8.4 <br />TEMPERATURE (deg C) 19.9/20.0 19.9/20.0 19.9/20.0 19.9/20.0 19.9/20.0 19.9/20.0 19.9/20.0 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: 2.00 mg/L D.O.: 7.40 mg/L <br />HARDNESS: 426.43 mg/L CHLORINE: ND <br />ALKALINITY: 831.03mg/L TEMPERATURE: 20.0 deg C <br />pH: 8.04 D.O AFTER AERATION: NA <br />w ` 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: v • z s- o? Reviewed by: <br />Date: Date: Z-? <br />/ ?, <br />I // v If/