高效液相色谱法对枳实提取物中黄酮类药效组分的定量分析

目的:建立同时对枳实提取物中柚皮苷、橙皮苷、新橙皮苷和枸橘苷进行定量分析的HPLC方法。方法:采用BDS液相色谱柱(5μm,4.6mm×250mm),流速1mL/min,检测波长283nm,流动相条件为乙腈和0.05%磷酸水梯度洗脱系统。0～5min为乙腈:0.05%磷酸水(19:81);5～45min之间乙腈由19%等度梯度上升至40%。结果:柚皮苷在0.03200～0.07466μg(r=0.9995),橙皮苷在0.00095～0.00475μg(r=0.9994),新橙皮苷在0.04888～1.4664μg(r=0.9993),枸橘苷在0.01700～0.34900μg(r=0.9999)之间线性关系良好;精密度和重现性试验中待测的各黄酮类组分RSD值均小于2%;稳定性试验中样品在32h内各黄酮类组分RSD值均小于3%;柚皮苷、橙皮苷、新橙皮苷和枸橘苷的加样回收率结果分别为100.51%(RSD=2.90%,n=5)、99.86%(RSD=2.79%,n=5)、102.83%(RSD=2.17%,n=5)、98.86%(RSD=1.97%,n=5)。结论:检测方法稳定,精密度、重现性和回收率好,可作为控制枳实提取物质量的依据之一。

Quantitative Analysis of Some Flavonoids in Extract of Aurantii Fructus Immaturus by High Performance Liquid Chromatography

Objectives:A high performance liquid chromatography method was established to determine the contents of naringin,hesperidin,neohesperidin and poncirin in extract of Aurantii Fructus Immaturus simultaneously.Methods:The extract was analyzed with a Alltech BDS(5μm,4.6mm×250mm)column by a gradient elution using(A)acetonitrile-(B)0.05% phosphoric acid solution as the mobile phase.0min(19%A)→5min(19%A)→45min(40%A).The detection wavelength was 283nm and the flow rate was 1mL/min.Results:Naringin,hesperidin,neohesperidin and poncirin showed a good linear relationshipat ranges of 0.03200～0.07466μg(r=0.9995),0.00095～0.00475μg(r=0.9994),0.04888～1.4664μg(r=0.9993)and 0.01700～0.34900μg(r=0.9999)respectively.The RSD of naringin,hesperidin,neohesperidin and poncirin were less than 2% in precision and duplicate tests.The RSD of naringin,hesperidin,neohesperidin and poncirin were less than 3% in 32 hours.The recoveries of naringin,hesperidin,neohesperidin and poncirin were 100.51%(RSD=2.90%,n=5),99.86%(RSD=2.79%,n=5),102.83%(RSD=2.17%,n=5),98.86%(RSD=1.97%,n=5)respectively.Conclusions:The method of quantitative analysis is stable.The precision,duplicate and recovery of the method are good.This method can be used as one of scientific evidences to control quality of extract from Aurantii Fructus Immaturus.