复合酶法结合液相色谱技术测定山药酒中薯蓣皂苷的含量

Determination of diosgenin content in yam wine by complex enzyme method combined with liquid chromatography technology

对复合酶法结合高效液相色谱(HPLC)测定山药酒中薯蓣皂苷的方法进行优化,通过单因素和正交试验确定酶法提取优化条件为采用复合酶(纤维素酶∶淀粉酶为1∶1)对山药酒进行酶解,复合酶用量为2 U/mL,酶解时间为12 h,酶解温度为40℃,酶解初始pH值为5.0。高效液相色谱法最优检测条件为采用Inertsil~?ODS-3色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-水(50∶50,V/V);流速1.0 mL/min;柱温30℃;检测波长203 nm。采用该方法对市售山药酒进行测定,薯蓣皂苷含量为0.106~1.156 mg/L,不同品牌产品含量水平存在差异。

The determination method of diosgenin in yam wine was optimized by complex enzyme method combined with HPLC. By single factor and orthogonal experiments, the extraction optimization conditions of enzyme method were determined as complex enzyme(cellulase∶amylase=1∶1) 2 U/ml,hydrolysis time 12 h, temperature 40 ℃, initial pH 5.0. The optimum detection conditions of HPLC were Inertsil~?ODS-3 chromatographic column(250 mm×4.6 mm, 5 μm), the mobile phase: acetonitrile-water(50∶50, V/V), flow rate 1.0 ml/min, column temperature 30 ℃, determine wavelength 203 nm.The method was used for the determination of diosgenin in commercial yam wine, the diosgenin content was 0.106-1.156 mg/L and there were differences in the diosgenin content of different brands products.