固相萃取-高效液相色谱法测定银杏叶提取物中芦丁、槲皮素、山柰酚和异鼠李素

Determination of rutin,quercetin,kaempferol,and isorhamnetin in extract of Ginkgo biloba leaves by HPLC with solid phase extraction

目的建立固相萃取-反相高效液相色谱法同时测定银杏叶提取物中芦丁、槲皮素、山柰酚和异鼠李素的方法。方法采用Waters Sep-Park-C18固相萃取小柱对银杏叶提取物样品进行处理,流动相:甲醇-0.1%甲酸水溶液,线性梯度洗脱;检测波长:360nm;柱温:40℃;进样量:20μL;体积流量:1.0mL/min。结果芦丁、槲皮素、山柰酚、异鼠李素的定量限分别为20、6、5、5mg/mL,芦丁、槲皮素、山柰酚、异鼠李素分别在14～198、15～205、3～40、3～41mg/L时与峰面积线性关系良好,回收率为93.7%～104.7%,日内及日间精密度的RSD分别小于2%、3%。结论方法简单、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 determine four flavonol components as markers, rutin, quercetin, kaempferol, and isorhamnetin for quality control of extract of Ginkgo biloba （EGb） by a validated high performance liquid chromatographic （HPLC） method with solid phase extraction. Methods The four flavonols of EGb were extracted with Waters Sep-Park-C18 solid-phase extraction column. Separation was achieved using a Kromasil C18 column （250 mm×4.6 mm, 5 μm ） at 40 ℃ with a step linear gradient using methanol ： 0.1% formic acid at a flow rate of 1.0 mL/min, wavelength was 360 nm, sample amount was 20μL. Results The limits of quantitation for rutin, quercetin, kaempferol, and isorhamnetin were 20, 6, 5, and 5 mg/mL, respectively. This method was linear over concentration ranges of 14--198, 15--205, 3--40 and 3--41 mg/L for rutin, quercetin, kaempferol, and isorhamnetin, respectively. The solid phase extraction recoveries for rutin, quercetin, kaempferol, and isorhamnetin were 93.7%-104.7%. The precision of intra-day and inter-day were less than 2 % and 3% for all compounds, repectively. Conclusion The validated method is simple, accurate, and able to provide the basis for the quality control of flavonols in EGb.