Wastewater generated in the secondary battery production process contains lithium and high-concentration sulfate. Recently, as demand as demand for high-Ni precursors with high-energy density has surged, nickel emission is also a concern. Lithium and sulfate are not included in the current water pollutant discharge standard, so if they are not properly processed and discharged, the negative effect on future environment may be great. Therefore, in this study, the ecotoxicity of lithium, nickel, and sulfate, which are potential contaminants that can be discharged from the secondary battery production process, was evaluated using water flea (Daphnia magna) and luminescent bacteria (Aliivibrio fischeri). As a result of the ecotoxicity test, 24-hour and 48-hour D. magna EC50 values of lithium were 18.2mg/L and 14.5mg/L, nickel EC50 values were 7.2mg/L and 5.4mg/L, and sulfate EC50 values were 4,605.5mg/L and 4,345.0mg/L, respectively. In the case of D. magna, it was found that there was a difference in ecotoxicity according to the contaminants and exposure time (24 hours, 48 hours). Comparing the EC50 of D. magna for lithium, nickel, and sulfate, the EC50 of nickel at 24h and 48h was 39.6-37.2% compared to lithium and 0.1-0.2% compared to sulfate, which was the most toxic among the three substances. The difference appeared to be at a similarlevelregardless of the exposure time. The EC50 of sulfate was 253.0-299.7% and 639.5-804.6%, respectively, compared to lithium and nickel, showing the least toxicity among the three substances. The 30-minute EC50 values of luminescent bacteria forlithium, nickel, and sulfate were 2,755.8mg/L, 7.4mg/L, and 66,047.3mg/L,respectively. Unlike nickel, it was confirmed that there was a difference in sensitivity between D. magna and A. fischeri bacteria to lithium and sulfate. Studies on the mixture toxicity of these substances are needed.