UPLC法同时测定红旱莲中5个成分含量

Simultaneous determination of five components in Hypericum ascyron L.by UPLC

目的:建立超高效液相色谱法(UPLC)同时测定红旱莲中芦丁、金丝桃苷、槲皮素、异槲皮苷、山柰苷5个有效成分含量方法。方法:采用Waters Acuity UPLC HSS T3色谱柱(2.1mm×150 mm,1.8μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,流速为0.2 mL·min^(-1),检测波长为360 nm,柱温为35℃。结果:芦丁、金丝桃苷、槲皮素、异槲皮苷、山柰苷线性范围分别为1.34~21.52(r=0.9993)、1.07~17.1(r=0.9996)、1.34~21.388(r=0.9995)、1.64~26.18(r=0.9998)、0.88~14.16μg·mL^(-1)(r=0.9999),平均加样回收率(n=9)分别为103.72%、98.27%、98.45%、103.05%、100.25%,RSD均小于1.5%。测定6个批次的供试品中5个成分的含量。结果:槲皮素含量最高,其次为金丝桃苷和异槲皮素,山柰苷和芦丁的含量较低。结论:UPLC可用于红旱莲中5个有效成分的含量测定,且方法简单、有效、结果准确,可用于其质量控制。

Objective:To establish a UPLC method for the simultaneous determination of five components in Hypericum ascyron L.including rutin,hyperoside,quercetin,isoquercetrin,kaempferitrin and study the quantification of active ingredient content.Methods:The UPLC analysis was carried out on a Waters Acquity UPLC HSS T3 column(2.1 mm×150 mm,1.8μm),the mobile phase composed of acetonitrile(A)-0.1%phosphoric acid(B)solution with gradient elution was at a flow rate of 0.2 mL·min^(-1),the detection wavelength was set at 360 nm,column temperature was set at 35℃.Results:The liner ranges of rutin,hyperoside,quercetin,isoquercetrin and kaempferitrin were at 1.34-21.52(r=0.9993),1.07-17.1(r=0.9996),1.34-21.388(r=0.9995),1.64-26.18(r=0.9998),0.88-14.16μg·mL^(-1)(r=0.9999),respectively.The average recovery rates were 103.72%,98.27%,98.45%,103.05%and 100.25%respectively,with RSDs less than 1.5%.The contents of 5 components in 6 batches of test samples were determined.The results showed that the content of quercetin was the highest,followed by hyperoside and isoquercetrin,and kaempferitrin and rutin were lower.Conclusions:The UPLC method can be used for the determination of five active ingredient content in Hypericum ascyron L.,and the method is simple,effective and accurate,and which can be used for quality control of Hypericum ascyron L.