HPLC法同时测定雪松果中槲皮素、山柰酚和异鼠李素的含量

Research of contents determination of quercetin, kaempferol and isorhamnetin in pine cones by HPLC

目的:建立同时测定雪松果中槲皮素、山柰酚和异鼠李素含量的高效液相色谱(HPLC)法。方法:雪松果用60%甲醇回流提取,提取液加10%盐酸水解。采用Agilent TC-C18(250mm×4.6mm,5μm)色谱柱,流动相为[甲醇-0.1%磷酸(50∶50)]-[乙腈-0.1%磷酸(40∶60)](80∶20),流速1.0m L/min,检测波长360nm,柱温为25℃。结果:雪松果中槲皮素和异鼠李素在0.2-2.4μg/m L、山柰酚在0.6-7.0μg/m L浓度范围内呈良好线性关系(r≥0.9991),加样回收率RSD(n=6)依次为2.86%、1.24%、1.61%。雪松果中槲皮素、山奈酚和异鼠李素的含量依次为0.054、0.301、0.161mg/g。结论:该方法简单、快速、高效,可用于雪松果中3个黄酮类化合物的含量测定。

Objective： To develop an HPLC method for contents determination of quercetin, kaempferoi, isorhamnetin in pine cones of cedrus deodara~ Methods： The sample was pretreated, including reflux extraction with 60% methanol for lh, and then extracting solution hydrolysis with 10%hydrochloric acid for another lh. Agilent TC-C18 （250mm×4.6mm, 5μm） chromatographic column was adopted. The mobile phase was [methanol-0.1% phosphoric acid （50 ： 50）]-[ acetonitrile-0.1%phosphoric acid （40 ： 60）] （80 ： 20） solution with the gradient elution at the flow rate of 1.0mL/min. The column temperature was 25℃ and the detection wavelength was 360nm. Results： Good linear correlation were found in the ranges of 0.2-2.4 μg/mL for quercetin and isorhamnetin and 0.6-7.0μg/mL for kaempferol （r〉0.9991）. The average recoveries RSD （n=6） of the three flavonoids were 2.86%, 1.24%, 1.61%, respectively, The quercetin, kaempferol, isorhamnetin in pine cones of cedrus deodara were 0.054, 0.301, 0.161mg/g, Conclusion： The method is simple, rapid and efficient, and applicable for determination of three flavonoids in pine cones of cedrus deodara.