Abstract: This paper mainly discussed the extraction of metalothionein from the viscera of Patinopecten yessoensis. Based on the content of metallothionein in the extract, the concentration of extracting agent, pH and heating temperature were optimized by single factor experiment and response surface experiment.The molecular sieve pre-packed column was used to separated the crude extra-ction. The results showed that the optimum extraction conditions of metallothi-onein in the viscera of Patinopecten yessoensis were as follows:the concentrate-ion of extractant was 0.01 mol/L, the pH of extractant was 8.23, and the heating temperature was 70℃. Under these conditions, the content of MT ext-racted was 0.115 mg/g. The mixture of MT-I and MT-II two isomers were purified by molecular sieve column chromatography. The purity of the MT-I and MT-II mixture were 81%. The extraction technology obtained in this study could obtain high purity of visceral metallothionein from Patinopecten yessoensis.