高效液相色谱法分析山楂中的四种黄酮成分

Analysis of Four Flavonoids in Hawthorn Fruit by HPLC

建立了山楂中四种黄酮成分(芦丁、牡荆素、金丝桃苷和槲皮素)的高效液相色谱分析方法。采用Agilent 5 TC-C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)和0.2%醋酸水溶液(B),流速为1 mL/min,进样量为10μL,检测波长为355nm,柱温为40℃。线性梯度洗脱程序:0~15min,20%~30%A;15~30min,30%~60%A。四种成分线性关系良好;日内精密度保留时间RSD均<0.5%,峰面积RSD均<2.5%;日间精密度保留时间RSD均<0.5%,峰面积RSD均<5.0%;平均加样回收率为103.4%~106.7%,RSD均<2.5%。60%乙醇为最佳提取溶剂。6批山楂样品中芦丁和金丝桃苷的含量范围分别为51.8~264.5μg/g和148.0~441.6μg/g,槲皮素只在两批样品中检出,分别为11.0μg/g和12.7μg/g,牡荆素在所有山楂样品中均未检出。

A HPLC method was developed to determine four flavonoids in hawthorn fruit,including rutin,vitexin,hypericin and quercetin.An Agilent 5 TC-C18 column(250 mm×4.6 mm,5μm)was used with the mobile phases consisting of acetonitrile(A)and 0.2%acetic acid(B)at the flow rate of 1 mL/min.The injection volume was 10μL,while the detection wavelength was set at 355 nm and the column temperature was 40℃.The linear gradient elution procedure was 20%~30%A in 0~15 min,and 30%~60%A in 15~30 min.Methodological results showed that the four components have a good linear relationship.The intraday precision retention time RSD was<0.5%,and the peak area RSD was<2.5%.The RSD of interday precision retention time was<0.5%and the RSD of peak area was<5.0%.The average recoveries ranged from 103.4%to 106.7%,and the RSD was<2.5%.The optimal extraction solvent was 60%ethanol.The contents of rutin and hypericin in 6 batches of hawthorn samples were in the range of 51.8~264.5μg/g and 148.0~441.6μg/g,respectively.Quercetin was detected in 2 batches of hawthorn samples,with contents of 11.0 and 12.7μg/g,and vitexin was undetectable in all samples.