Acute toxicity test of heavy metal (Cd, Cu, Zn) were examined using the hatching rates of fertilized eggs in the oliver flounder, Paralichthys olivaceus. Eggs were exposed to Cd, Cu, Zn (0, 10, 100, 500, 1000, 2500, 5000 ppb) and then normal hatching rates were investigated after 48 h. The normal hatching rates in the control condition (not including Cd, Cu and Zn) were greater than 80%, but suddenly decreased with increasing of heavy metal concentrations. Cd, Cu and Zn reduced the normal hatching rates in concentration-dependent way and a significant reduction occurred at concentration grater than 1000, 100, 100 ppb, respectively. The ranking of heavy metal toxicity was Zn>Cu>Cd, with $EC_{50}$ values of 584, 1015 and 1282 ppb, respectively. The no-observed-effect-concentration (NOEC) and the lowest-observed-effect-concentration (LOEC) showed each 100 and 500 ppb of normal hatching rates in exposed to Cu and Zn. The NOEC and LOEC of normal hatching rates in Cd were 500 ppb and 1000 ppb, respectively. From these results, the normal hatching rates of P. olivaceus have toxic effect at greater than the 100 ppb concentrations in Cu, Zn and the 500 ppb concentrations in Cd in natural ecosystems. These results suggest that biological assay using the normal hatching rates of P. olivaceus are very useful test method for the acute toxicity assessment of a toxic substance as heavy metal in marine ecosystems.