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Aqua-Tox, Inc. <br /> 4880 Robb Street, Suite 8 <br /> Wheat Ridge, CO 80033 <br />LL (303) 6214880 aqua-tox.com <br /> <br /> ACUTE TOXICITY REPORT <br /> Daphnia magna <br />CLIENT NAME: 20-Mile Coal Company SAMPLE I.D.: 1001 10-1 <br />50 % MORTALITY TEST: Pass TEST RESULT LC50: 22100% <br />% SURVIVAL IN 100% DILUTION: 75% <br />SAMPLE DATE: 09-30-10 TIME: 16:OOP CLIENT ID: Site 84 - Pond D <br />ORGANISM: Daphnia magna AGE: < 24 hours old <br />TEST START: 10-01-10 10:10A TEST FINISH: 10-03-10 10:10A <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.: 7.03 mg/L <br />HARDNESS: 97.22 mg/L CHLORINE: ND _ <br />ALKALINITY: 58.47 mg/L TEMPERATURE: 20.1 deg T <br />pH: 7.99 <br />SURVIVAL DATA DILUTIONS (%) <br />nATF (HOURS) CONTROL 6.25 12.5 25 50 75 100 <br />10/1/2010 INITIAL 20 20 20 20 20 20 20 <br />10/2/2010 24 20 20 20 20 E] <br />20 Et 18 <br />10/3/2010 48 20 20 20 20 20 15 <br />MORTALITY: 0 0 0 0 0 1 5 <br />% SURVIVAL: 100% 100% 100% 100% 100% 95% 75% <br />DAILY CHEMISTRY - (MIN/MAX) DILUTIONS (%) <br />0 6.25 12.5 25 50 75 100 <br />CONDUCTIVITY (microS) 306/319 855/889 1530/1560 2420/2440 3620/3650 5150/5180 6630/6680 <br />D.O. (mg/L) 7.0/7.3 7.1/7.3 7.1/7.4 7.2/7.4 7.2/7.4 7.3/7.4 7.3/7.4 <br />pH 8.0/8.4 7.9/8.3 7.7/8.3 7.5/8.4 7.4/8.4 7.2/8.4 7.0/8.5 <br />TEMPERATURE (deg C) 20.1/20.5 20.1/20.5 20.1120.5 20.1/20.5 20.1/20.5 20.1/20.5 20.1/20.5 <br />GENERAL CHEMISTRY - SAMPLE <br />AMMONIA: 0.07 mg/L D.O.: 7.31 mg/L <br />HARDNESS: 603.21 mg/L CHLORINE: ND <br />ALKALINITY: 740.99 mg/L TEMPERATURE: 20.1 deg C <br />pH: 7.04 D.O AFTER AERATION: NA <br />ANOEC' CALCULATION'METHOD.- ` METHOD. REFERENCE <br />75% SPEARMAN-KARBER EPA -821-R-02-012 October 2002 pg.78 <br />ADDITIONAL COMMENTS: <br />? , <br />Analysis / I I ? '- Zlv <br />? Reviewed by: <br />Conducted by: . <br />; %!'.ti `; <br />Date: Datte: <br />e ` /