液相色谱法测定柑橘果实中类黄酮的方法研究

Study on the HPLC Method for Determination of Flavonoids in Citrus Fruits by High Performance Liquid Chromatography

文中以柑橘中常见的、含量丰富的柚皮素-7-β-芸香糖苷等10种类黄酮为研究对象,建立了同时测定柑橘中黄烷酮和多甲氧基黄酮含量的高效液相色谱分析方法。采用Waters Symmetry C_(18)色谱柱(250 mm×4.6 mm,5μm),流动相为0.2%乙酸水溶液和乙腈,梯度洗脱,二极管阵列检测器,检测波长为287 nm和330 nm,柱温25℃,流速1.0 mL/min。外标法定量,柚皮素-7-β-芸香糖苷、柚皮苷、橙皮苷、新橙皮苷、香风草苷、柚皮素、橙皮素、甜橙黄酮、川皮苷和桔黄酮等10种类黄酮的线性范围为0.01～27.00 mg/L(r≥0.99),检出限为0.01～0.02 mg/L。果汁的平均加标回收率为在94.20%～106.58%之间,相对标准偏差为1.15%～5.41%;果皮的平均加标回收率为95.15%～101.19%,相对标准偏差为1.47%～3.78%。

Flavonoids, which are rich in citrus fruits, are good to health. Therefore, studies on how to determine the amount of flavonoids had become a hot research field. A method was established for the identification and quantification of flavonoids including flavanones and polymethoxyflavones in citrus fruits by reverse--phase HPLC coupled with photodiode array detection using Waters Symmetry C18 column. The mobile phase used was 0.2M acetic acid solvent and acetonitrile, in a concentration gradient, and the column temperature was 25℃, the UV detection wavelengths were 287 nm and 330 nm. The flavonoids were quantified by external standard method. The linear ranges of 10 flavonoids were 0.01-27.00 mg/L（r≥0.99）, and the detection limits were 0.01-0.02 mg/L. The average recoveries of flavonoids in juice and dry peel powder were 94. 20%-106.58% and 95.15%-101.19%, RSD were 1. 15%-5.41% and 1.47%-3.78% respectively. The analytic parameters showed that the method was sensitive, reliable, accurate and reproducible.