高效液相色谱法同时测定银杏叶提取物中5种黄酮类成分

Simultaneous determination of five flavonols in extract of Ginkgo biloba by high performance liquid chromatography

目的建立反相高效液相色谱法同时测定银杏叶提取物中5种黄酮类成分芦丁、槲皮苷、槲皮素、山奈酚、异鼠李素的含量。方法采用70%甲醇提取样品,以Kromasil C18为固定相,甲醇-0.1%甲酸水溶液为流动相,线性梯度洗脱,流速1.0 ml/min;柱温：40℃;检测波长：360 nm;结果芦丁、槲皮苷、槲皮素、山奈酚、异鼠李素的定量限分别为0.03、0.02、0.008、0.006、0.005 mg/L,芦丁、槲皮苷、槲皮素、山奈酚、异鼠李素分别在2.02-50.5、1.932-48.3、2.01-50.4、0.49-12.3、0.39-9.8 mg/L浓度范围内其峰面积与浓度的线性关系良好（r〉0.9992）,回收率为86.5%-99.2%,日内精密度及日间精密度的变异系数分别小于2%、3%。结论本方法简单、准确,为银杏叶提取物中黄酮类成分的质量控制提供了依据。

Objective To describe a high performance liquid chromatographic （HPLC） method to quantitate five flavonol components in the extract of Ginkgo biloba （EGb）. Methods The five flavonols of EGb were extracted with 70% methanol and separated by passing through a kromasil C18 column （250 ×4.6 mm, 5μm ） at 40℃ with a step linear gradient using methanol： 0. 1% formic acid at a flow rate of 1.0 ml/min. Results The limits of quantitation for rutin, quereitrin, quereetin, kaempferol and isorhamnetin were 0.03, 0.02, 0.008, 0.006 and 0.005 mg/L, respectively. This method was linear over the concentration ranges of 2.02 - 50.5, 1. 932 - 48.3, 2.01 - 50.4, 0.49 - 12, 3 and 0.39 - 9.8 mg/L respectively（ r 〉0. 9992 ）. The rates of recovery for the five components ranged 86.5% - 99.2%, with the coefficient of precision being less than 2% within one day and less than 3% between two days. Condusion The method is simple and accurate, and hence, can be validated as a means offering the basis for the quality control of flavonols in EGb .