First,preparative high-pressure liquid chromatography was used to isolate and purify the naringin palmitate enzymatically synthesized by immobilized lipase Novozym 435 in the t-amyl alcohol system.Next,IR spectrum,mass spectrum and 13C NMR were used to identify the purified product,and the mechanism of regioselectivity of the reaction was discussed.Then,the feasible operation conditions were determined as follows： a C18 column（21.5mm×250.00mm,10μm） with methanol as the mobile phase at a flowrate of 20mL/min,an injection volume of 3mL with the concentration of 30mg/mL.In these conditions,the product with a purity of more than 99% was obtained.Finally,the purified product was identified as monoester of naringin and palmitic acid,and the regioselective acylation was found preferring the 6″-hydroxyl group of naringin glucose.