梯度洗脱高效液相色谱法测定桑叶中3种活性成分的含量

建立同时测定桑叶中绿原酸、芦丁和槲皮素含量的分析方法。色谱柱为NUCLEODUR C18 RP(250 mm×4.6 mm,5μm),光电二极管检测器,流动相为甲醇-质量分数0.5%磷酸水溶液,梯度洗脱程序为0 min(30:70)-15min(30:70)-25min(50:50)-35min(85:15)-40min(30:70);流速0.8 ml.min-1,检测波长为350 nm。绿原酸、芦丁和槲皮素的线性范围分别为0.1144~1.0296μg(r=0.9996)、0.0436~0.3924μg(r=0.9999)和0.0452~0.4068μg(r=0.9997),平均回收率分别为97.7%(RSD=1.7%)、98.4%(RSD=2.2%)和100.6%(RSD=1.5%)。方法快速简便,专属性强,重复性好,可作为桑叶中绿原酸、芦丁和槲皮素的定量分析方法。

Determination of Three Kinds of Active Ingredients in Mulberry Leaves by RP-HPLC with Gradient Elution

A method for simultaneous determination of chlorogenic acid,rutin and quercetin in mulberry leaves was established.A NUCLEODUR C18 RP (4.6 mm×250 mm,5 μm) column was used with a mobile phase consisted of phase A(methanol)-phase B(0.5 % phosphoric acid water solution) in gradient elution 0 min(30﹕70)-15 min(30﹕70)-20 min(50﹕50)-35 min(85﹕15)-35 min(30﹕70) by flow rate of 0.8 mL·min-1 in this method,the detection wavelength was set at 350 nm and the column temperature was 25 ℃.Photodiode array detector was used.The calibration curve was linear at a range of 0.1144～1.0296 μg for chlorogenic acid(r=0.9996).The average recovery was 97.7 % and RSD was 1.7 %.The calibration curve was linear at a range of 0.0436～0.3924 μg for rutin(r=0.9999).The average recovery was 98.4 % and RSD was 2.2 %.The calibration curve was linear at a range of 0.0452～0.4068 μg for quercetin(r=0.9997).The average recovery was 100.6 % and RSD was 1.5 %.The method is rapid,simple and reproducible.It can be used for determination of chlorogenic acid,rutin and quercetin in mulberry leaves.