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一测多评法同时测定山楂叶中6种有效成分的含量

         

摘要

目的:建立同时测定山楂叶中绿原酸、牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、芦丁、金丝桃苷含量的方法。方法:采用高效液相色谱法,以牡荆素葡萄糖苷为基准峰,分别计算其与绿原酸、牡荆素鼠李糖苷、牡荆素、芦丁、金丝桃苷的相对校正因子(RCF),通过RCF计算山楂叶中上述5种成分的含量。色谱柱为Agilent ZORBAX SB C18,流动相为0.1%甲酸-乙腈-四氢呋喃(梯度洗脱),流速为1.0 ml/min,检测波长为350 nm,柱温为30℃,进样量为10µl。结果:绿原酸、牡荆素葡萄糖苷、牡荆素鼠李糖苷、牡荆素、芦丁、金丝桃苷检测进样量线性范围分别为12.50~400.0µg(r=0.9998)、25.00~800.0µg(r=0.9999)、31.25~1000.0µg(r=0.9999)、6.470~260.0µg(r=0.9999)、2.50~80.0µg(r=0.9998)、9.375~300.0µg(r=0.9999);精密度、稳定性、重复性试验的RSD<2.0%;加样回收率分别为99.2%~103.9%(RSD=1.6%)、97.9%~100.8%(RSD=1.2%)、99.2%~100.8%(RSD=0.5%)、97.3%~101.3%(RSD=1.5%)、98.0%~103.0%(RSD=1.9%)、95.5%~101.5%(RSD=2.2%),n均为6。牡荆素葡萄糖苷与绿原酸、牡荆素鼠李糖苷、牡荆素、芦丁、金丝桃苷间的RCF分别为1.119、1.009、0.706、1.063、0.830。结论:该方法操作简便,精密度、稳定性、重复性好,可用于6种有效成分含量的同时测定。%OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid,vitexin glucoside,vi-texin rhamnoside,vitexin,rutin and hyperoside in Crataegus pinnatifida. METHODS:With reference peak of vitexin glucoside, HPLC was conducted to calculate the relative correction factor(RCF)of chlorogenic acid,vitexin glucoside,vitexin rhamnoside, vitexin,rutin and hyperoside,then the contents of above-mentioned 5 components in C. pinnatifida were calculated. The column was Agilent ZORBAX SB C18 with mobile phase of 0.1% formic acid-acetonitrile-tetrahydrofuran (gradient elution) at a flow rate of 1.0 ml/min,the detection wavelength was 350 nm,column temperature was 30 ℃,and the injection volume was 10 μl. RE-SULTS:The linear range was 12.50-400.0 µg for chlorogenic acid(r=0.999 8),25.00-800.0 µg for vitexin glucoside(r=0.999 9), 31.25-1 000.0 µg for vitexin rhamnoside(r=0.999 9),6.470-260.0 µg for vitexin(r=0.999 9),2.50-80.0 µg for rutin(r=0.999 8) and 9.375-300.0 µg for hyperoside(r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2.0%;re-coveries were 99.2%-103.9%(RSD=1.6%,n=6),97.9%-100.8%(RSD=1.2%,n=6),99.2%-100.8%(RSD=0.5%,n=6), 97.3%-101.3%(RSD=1.5%,n=6),98.0%-103.0%(RSD=1.9%,n=6)and 95.5%-101.5%(RSD=2.2%,n=6). RCFs of vitex-in glucoside with chlorogenic acid,vitexin rhamnoside,vitexin,rutin and hyperoside were 1.119,1.009,0.706,1.063 and 0.830, respectively. CONCLUSIONS:The method is simple with good precision,stability and reproducibility,and it can be sued for the simultaneous determination of 6 components in C. pinnatifida.

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