In this study,the detection method of ATP related compounds of fishery products by using HPLC was researched.The optimal conditions such as pH value,flow rate and concentration of mobile phase were determined.The accuracy,precision and limit of quantitation were surveyed and evaluated.The results showed that when the pH value was at 6.5 and the flow rate 0.70 mL/min the seven chromatographic peaks were well separated.When the concentration of seven ATP related compounds were from 2 μg/mL to 500μg/mL,the concentration of standard substance and peak area were kept good linear relationship.The RSD% is between 0.08% and 6.81%,the coefficient of recovery reached 98.11%~102.25%.The limits of quantitation were from 52ng/mL to 53ng/mL.According to the result this method was better to detecting the ATP related compounds of fishery products.