高效液相色谱法测定刺苋中芦丁、槲皮素和山奈酚

Determination of Rutin, Quercetin, Kaempferol in Amaranthus spinosus L. by High Performance Liquid Chromatography

本文建立了以甲醇溶液为萃取剂，结合超声辅助萃取，利用高效液相色谱法同时分离测定刺苋中的芦丁、槲皮素和山奈酚三种黄酮的方法。色谱条件为：Phenomenexc18色谱柱（250mm×4．6mm，5μm）；流动相为甲醇-0．2％冰醋酸水溶液，并采用梯度洗脱程序；流速1mL／min；紫外检测波长360nm；柱温为35℃。结果表明该方法对芦丁、槲皮素和山奈酚均在0．0025—62．5000μg/mL范围内线性关系良好，相关系数（r）分别为0．99999、0．99997、0．99999；最低检测限（S／N=3）依次为0．1411ng/mL，0．0795ng/mL，0．0188ng／mL，最低定量限（S／N=10）依次为0．4703ng/mL，0．2652ng／mL，0．0627ng／mL；样品回收率为95．79％-101．41％。本法灵敏度高、定量准确且操作简便，为刺苋中芦丁、槲皮素和山奈酚的分离检测提供了有效方法。

A simple high performance liquid chromatography （HPLC） method was developed for the simultaneous determination of rutin, quercetin, kaempferol in Amaranthus spinosus L. using methanol as extraetant with ultrasonic assisted extraction. Chromatographic separation was carried out using a Phenomencx Cn column with a gradiem dution,the mobile phase was methanol-0.2% acetic acid solution with the UV detection wavelength set at 360 nm and the flow rate of 1.0 mL/min. The linearity range for rutin, quercetin, kaempferol were found to be 00.0025-62.5000 μg/mL, and the correlation coefficients were 0. 99999, 0. 99997, 0. 99999, respectively. The limits of detection （ LOD, S/N = 3 ） were 0. 1411 ng/mL,0.0795 ng/mL,0.0188 ng/mL,respeetively. The limit of quantification （ LOQ,S/N = 10） were 0. 4703 ng/mL,0. 2652 ng/mL,0.0627 ng/mL,respectively. Accuracy of the method was checked by conducting recovery studies at three different concentration levels for all the three compounds and the average recoveries were ranged from 95. 79% to 101.41%. The developed HPLC method was found to be simple,precise,specific,sensitive and accuracy. It provided a new and scientific mean for routine quality control of Amaranthus spinosus L.