In recent years, the ecological risks of plastics to marine environments and organisms have attracted increasing attention, especially the leachates from plastics. However, a comprehensive knowledge about the leaching characteristics and subsequent toxicological effects of leachates is still sparse. In this study, 15 different plastic products were immersed in simulated seawater and fish digest for 16 h. The leachates were analyzed through non-target and target analyses and their toxicological signatures were assessed by bioassays. In total, 240 additives were identified from the plastic leachates, among which plasticizers represented the most (16.7%), followed by antioxidants (8.7%) and flame retardants (7.1%). Approximately 40% of plastic leachates exhibited significant inhibitory effects on the bioluminescence using a recombinant luminescent assay. In addition, both the hyperactive and hypoactive behaviors were displayed in the larvae of marine medaka (Oryzias melastigma) exposed to some plastic leachates. In general, the number and amount of identified compounds under simulated fish digest were less than those under simulated seawater. However, the simulated fish digest leachates triggered higher toxicity. Redundancy analysis demonstrated that identified additives did not adequately explain the toxicological effects. Future research should focus on the identification of more additives in the plastic leachates and their potential ecological risks.