By measuring the yield of parvalbum, a kind of extraction solution of parvalbumin from Pseudosciaena crocea was screened out, and the relative extracting process was optimized. The results showed that when the extraction time was 42 h, the extract concentration was 72 mmol/L; the pH of the extract was 8, and the ratio of material to liquid was 1∶180 (g/mL); the extraction temperature was 50 ℃, and the extraction rate of parvalbumin reached 0.174 2%. Parvalbumin was then separated and purified by anion gel column chromatography, and its molecular weight was determined to be 12 kDa by Western blot analysis.