Rutin and its hydrolysis products (isoquercitrin and quercetin) are widely used as important materials in food and pharmaceutical industry. In this study, the effects of various acids and enzymes as catalysts on the hydrolysis reaction of rutin were studied. In comparison with acidic and enzymatic catalysis of rutin, the research results indicated that there was a sharp difference in the selectivity of hydrolysis product between the methods. When 2.5% H3PO4, 1% HCl and 0.5% H2SO4 were used as catalysts, transformation yields of isoquercitin hydrolyzed from rutin were 9.60, 0.69 and 1.25%, but those of quercetin were 11.13, 100 and 2.57%, respectively. When hesperidinase, snailase and cellulase-T2440 were used as catalysts, transformation yields of isoquercitin hydrolyzed from rutin were 43.21, 3.07 and 0.00%, but those of quercetin were 58.10, 96.39 and 30.89%, respectively. In conclusion, the aglycon of rutin was deglycosolated easily under mild acidic hydrolysis conditions at appropriate temperatures, but its secondary glucoside was difficult to be obtained. Contrarily, the prepared isoquercitrin by enzymatic hydrolysis of rutin was preferable to the acidic hydrolysis, especially for hesperidinase.Key words: Rutin, isoquercitrin, quercetin, hydrolysis, acid, enzym