反相高压液相色谱法测定鲜蒜中的蒜氨酸

为准确、快速、方便的测定大蒜中的蒜氨酸,采用反相高效液相色谱法(RP-HPLC),HypersilODSC18(250mm×4·6mm)色谱柱,检测波长200nm,5%甲醇+95%pH=5磷酸缓冲液为流动相,流速为0·5mL/min,柱温为20℃等条件下进行测定。测定结果为:蒜氨酸浓度与峰面积呈良好的线性关系,线性范围为10~350μg/mL;利用蒜氨酸酶反应样品的HPLC行为差异或纯蒜氨酸均可对蒜氨酸进行定性测定。测定结果表明,该方法经济、简便、快速、重现性好。

Determination of Alliin from Fresh Garlic by Reversed-phase High-performance Liquid Chromatography

The aim of the paper is to fast, conveniently, accurately determine the alliin in the fresh garlic (Allium sativum L.). The conditions by the reversed-phase high-performance liquid chromatography(RP-HPLC) were: Hypersil ODS C_~18 (250×4.6 mm) column, UV detector at 200 nm, 5% methanol and 95% phosphatic buffer (pH=5) as mobile phase at the flow rate of 0.5 mL/min, column temperature at 20℃. The calibration graphs were linear in the range of 10～350 μg/mL alliin. The retention time were determined based on both pure alliin and the peak difference between samples with the alliinase reaction and without alliinase reaction. The variation coefficient was 3.39%. The recovery rate was 97±2.0%. Alliin in the fresh garlic was 1.2815±0.0697%. We concluded that this is a sensitive, specific, simple and accurate method.