高效离子交换色谱法测定蒜氨酸的含量和有关物质___

摘要 目的：建立测定蒜氨酸含量和有关物质的高效离子交换色谱法。方法：采用Hypersil SCX柱(250 mm×4.6 mm，5 μm，氢型阳离子交换色谱柱)，以磷酸盐缓冲液(取磷酸二氢钾13.6 g，加水溶解并稀释成1000 mL，用85%磷酸调pH至2.0)为流动相，流速1.0 mL·min^-1，检测波长214 nm，柱温30 ℃，进样量20 μL。结果：蒜氨酸及其有关物质色谱分离良好，蒜氨酸浓度在0.06～1.8 mg·mL^-1的范围内线性关系良好(r=0.9999)，平均加样回收率(n=9)为100.2%，精密度良好(日内RSD=1.2%，n=5；日间RSD=1.4%，n=5)，检测限(S/N=3)为0.16 μg·mL^-1。结论：本方法准确、快速，重复性好，可用于蒜氨酸含量和有关物质测定，适合蒜氨酸制备过程的质量控制。

关键词：蒜氨酸；含量；有关物质；高效离子交换色谱法

Abstract   Objective:To establish a high performance ion exchange chromatography(HPIC）method for assay and related substance test of alliin.Methods:Analysis was carried out on a SCX column(250 mm×4.6 mm,5 μm,strongly acidic cation exchange chromatographic column）with a phosphate buffer solution(dissolve 13.6 g of KH2PO4 into 1000 mL water and adjust to pH 2.0 with 85% H₃PO₄）as mobile phase,column temperature was 30 ℃,the flow rate was 1.0 mL·min^-1,UV detection wavelength was 214 nm，injection volume was 20 μL.Results:The resolution of alliin with its related substances was good and linear calibration was found for alliin within the range of 0.06-1.8 mg·mL^-1,the precision of method was good with RSD(n=5） of within-day and between-day within 1.2% and 1.4% respectively,the limit of detection was 0.16 μg·mL^-1.Conclusions:The method is simple,accurate,reproducible and suitable both for assay and related substances test of alliin,especially suitable for process quality control of separating or purifying alliin from garlic.

Key words：alliin；assay；related substances；high performance ion exchange chromatography