高效液相色谱-蒸发光散射法检测酶促催化反应中底物D-半乳糖

目的：建立酶促催化反应中底物D-半乳糖的测定方法。方法：采用高效液相色谱-蒸发光散射法（HPLC-ELSD法）测定酶促催化反应中底物D-半乳糖的含量,色谱条件：色谱柱为Diamonsil C18（2）柱（250mm×4.6mm,5μm）,流动相为乙腈-水,流速为1.0ml·min-1,柱温为35℃,检测器为SEDEX-75型蒸发光散射检测器（漂移管温度为45℃,载气压力为3.0bar）。结果：D-半乳糖在0.5~10.0mg·ml-1范围内呈良好的线性关系（r=0.9995）,平均回收率分别为100.3%,RSD为1.6%。结论：该方法准确、重复性好,适用于酶促催化反应中底物D-半乳糖的测定。

HPLC-ELSD determination of D-galactose as a substrate in enzymatic reaction

Abstract：Objective：To establish the analysis methods for determination of D-galactose as a substrate in enzymatic reaction. Methods： HPLC-ELSD method is use to determine the content of D-galactose as a substrate in enzymatic reaction. Chromatographic conditions included Diamonsil C18（2） column（250 mm×4.6 mm, 5μm）, the mobile phase consisting of a mixture of acetonitrile-water. The flow rate was 1.0ml·min-1, the column temperature was 35℃ and the detector was SEDEX-75 （Eva：45℃, Gas flow： 3.0bar）.Results ：The linear range of D-galactose was 0.5-10.0mg·ml-1 （r=0.9995）.The average recovery was 100.3%,RSD was 1.6%.Conclusion：The method is sensitive and with good repeatability, and it can be applicable in the detection of D-galactose as a substrate in enzymatic reation.