HPLC测定不同产地砂仁中原儿茶酸和香草酸的含量

Simultaneous Determination of Protocatechuic Acid and Vanillic Acid in Amomi Fructus by HPLC

目的:建立高效液相色谱法(HPLC)同时测定砂仁中原儿茶酸和香草酸的含量,并对不同产地砂仁的原儿茶酸和香草酸含量进行比较。方法:采用Phenomenex Luna C_(18)色谱柱(4.6 mm×250 mm,5μm),以甲醇-0.1%甲酸水为统计相进行梯度洗脱,柱温35℃,流速0.8 mL·min^(-1),进样体积15μL,检测波长260 nm。结果:原儿茶酸在0.000 7~0.008 9 g·L^(-1)呈良好的线性关系,平均回收率100.4%(RSD 3.2%);香草酸在0.000 6~0.030 3 g·L^(-1)呈良好的线性关系,平均回收率99.3%(RSD 2.8%)。对24批不同产地砂仁样品的原儿茶酸及香草酸含量测定结果进行聚类分析,结果显示不同产地砂仁样品分为两大类,一类为阳春砂与绿壳砂,另一类为进口壳砂仁、艳山姜伪品及加工方式为生晒的云南产阳春砂。结论:该方法简便、准确、重复性好,可用于不同产地砂仁中原儿茶酸和香草酸含量测定,为鉴别不同产地的砂仁样品提供参考。砂仁中香草酸含量受其加工方式影响含量变化明显。

Objective： To determinate the contents of protocatechuic acid and vauillic acid in Amomi Fructus by High Performance Liquid Chromatography （HPLC） and compare the contents of protocatechuic acid and vanillic acid in Amomi Fructus from different regions. Method： The determination was performed on Phenomenex Luna C18 （4.6 mm × 250 mm, 5 μm） with methanol-0. 1% formic acid solution as mobile phase for gradient elution, and the wavelength was set at 260 nm. The flow rate was 0.8 mL·min^-1; column temperature was 35℃ and injection volume was 15 μL. Result： Protocatechuic acid showed good linearity in the range of 0. 000 7- 0. 008 9 g·L^-1 , and the average recovery was 100.4% （RSD 3.2% ）. Vanillic acid showed good linearity in the range of 0. 000 6-0. 030 3 g·L^-1 , and the average recovery was 99.3% （RSD 2.8% ）. The 24 batches of Amomi Fructus samples were divided into two main groups by clustering analysis. One was Amomum villosum and A. villosum var. xanthioides. The other was the imported Amomi Fructus, Alpinia zerumbet and A. villosum from Yunnan that directly dried without fire curing. Conclusion： The established method is simple, accurate, and with good repeatability, which can be used to determine the contents of protocatechuic acid and vanillic acid in Amomi Fructus from different regions, providing reference to distinguishing the samples from different regions. In addition,the content of vanillic acid in Amomi Fructus was affected significantly by the drying method.