HPLC法同时测定甘草及甘草头中4种成分含量

Simultaneous Determination of Four Constituents in Roots and Knotty Rhizome of Glycyrrhiza uralensis by HPLC

目的：建立HPLC法同时测定甘草饮片及甘草头中甘草苷、异甘草苷、甘草素和甘草酸的含量。方法：采用 Diamonsil C18色谱柱（250 mm×4.6 mm，5μm），0.1%磷酸水-乙腈梯度洗脱，流速1.0 mL· min-1，检测波长为276 nm（0-18 min）、360 nm（18-24 min）、276 nm（24-30 min）、250 nm（30-65 min），柱温30℃。结果：在该色谱条件下，甘草苷、异甘草苷、甘草素、甘草酸的进样量分别在0.1085-1.085、0.0168-0.168、0.00494-0.0494、0.407-4.07μg范围内线性关系良好。加样回收率均在96.61%-100.89%，RSD值均小于0.81%。5个不同批次的甘草饮片中4种成分的含量分别为0.513%、0.0729%、0.0484%、1.945%；2个批次甘草头中4种成分含量分别为0.456%、0.0636%、0.0362%、1.630%。结论：4种成分的响应与浓度之间呈良好的线性关系；甘草头中4种成分的含量与甘草饮片基本相当，可作为活性成分提取的原料加以利用。

This study was aimed to establish an HPLC method for the determination of liquiritin, isoliquiritin, liquiritigenin and glycyrrhizic acid in roots and knotty rhizome of Glycyrrhiza uralensis. The analysis was performed on a Diamonsil C18 column （250 mm × 4.6 mm, 5 μm） by using a gradient elution with mobile phase of water, phosphoric acid, acetonitrile at the flow rate of 1.0 mL·min-1. The detection wavelength was 276 nm （0-18 min）, 360 nm （18-24 min）, 276 nm（24-30 min）, and 250 nm （30-65 min）. The column temperature was set at 30℃. The results showed that the linear range of iquiritin, isoliquiritin, liquiritigenin, glycyrrhizic acid was 0 . 108 5-1 . 085、0 . 016 8-0 . 168、0 . 0049 4-0 . 049 4、0 . 407-4 . 07μg , respectively . The average recoveries of four constituents were 96.61%-100.89%, with RSD ≤ 0.81%. The contents of four constituents in roots of five batches were 0.513%, 0.072 9%, 0.048 4%, and 1.945%, respectively. Contents of four constituents in knotty rhizome from two batches were 0.456%, 0.063 6%, 0.036 2%, and 1.630%, respectively. It was concluded that there was good linear relationship between the response and concentration. Contents of four constituents in knotty rhizome were basically the same as those in the roots. The knotty rhizome can be used as raw material for the extraction of active components.