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~G~ Table of Contents ADDRESS ALL CORRESPONDENCE TO: COMMERCIAL TESTING & ENGINEERING CO. 4665 PARIS STREET, SURE B•200 DENVER, CO 80239 TEL: (303) 3734772 Abstract Page 2 Methods Pg. 2-4 Results Pg. 5-7 Narrative & Discussion Pg. 8-9 Table 1 -Colorado Test Conditions Pg. 10 Abstract Table 2 -Methods and Equipment Page 10 References Page 10 Reference Toxicity Data Chart Page 11 Supplement Forms' Page 12 SGS Commercial Testing and Engineering, Co. conducted a Preliminary Toxicity Identification (PTI) on Twentymile Coal Company effluent discharged at outfall 62 at Fish Creek Tipple (FCT). The Twentymile Coal Company discharge permit states that the facility must conduct either a PTI evaluation or a Toxicity Identification Evaluation (TIE) once a "pattern of toxicity" has been established. CT&E opted, with permission by Twentymile Coal Company, to conduct an acute PTI on effluent discharged at ouffall 62 at Fish Creek Tipple. The purpose of this PTI was to determine the possible cause(s) of the acute toxicity to Ceriodaphnia dubia. The results were conclusive. The evidence strongly suggests that the acute toxicity is due to total dissolved solids (TDS), caused by calcium sulfate (CaSO4), based on the results of the effluent manipulations. Methods A Preliminary Toxicity Identification evaluation was conducted with effluent from outfall 62 at Fish Creek Tipple. An effluent grab sample was collected on June 23, 2003, stored on ice, and delivered to SGS Commercial Testing and Engineering, Co. (CT&E) on the following day. The effluent sample was received at a temperature of 14.0 ° C and assigned CT&E sample tracking number 072-5445-001. The Initial Toxicity Test was initiated approximately 24 hours after sample collection. The characterization tests were initiated approximately 48 hours after sample collection and 24 hours after sample manipulation. Testing was conducted in accordance with the most recent protocols specified by the Water Quality Control Division of the Colorado Department of Health (1998), see Table 1, and with reference to the EPA's Methods for Aquatic Toxicity Identification Evaluations. Phase /Toxicity Characterization Procedures. (EPA/600/3-88/034 September 1988). Basic characteristics of the effluent and dilutions were measured by the methods and equipment outlined in Table 2. Statistically, the Binomial and Probit methods determined the LCSp s2. Significant differences in mortality were analyzed using the Steel's Many-One Rank tests. ' Supplemental forms include statistical analysis and chain-of-custody forms 2 LCso - an estimate of the concentration lethal to 50°/D of the organisms in the time prescribed by the test Commercial Testing & Engineering Co. Minerals Services -Corporate Office 1919 5. Highland Ave., Suita 210B, Lombard, IL 60146 t (6301953-9300 116301953-9306 www.sgs.com ' MDmEar W 1M 565 6,ouD ISDf,Yt06AMreN °e SUrvelSencal Fd25