HPLC法同时测定返魂草颗粒中6种活性成分

目的 利用HPLC法同时测定返魂草颗粒中同型原儿茶酸、原儿茶酸、对羟基苯乙酸、绿原酸、咖啡酸和金丝桃苷.方法 以Agilent Zorbax SB-C18色谱柱(4.6 mm×250 mm,5 μm),乙腈-0.1％三氟乙酸溶液梯度洗脱,体积流量1.0 mL/min,柱温30℃,检测波长280 nm.结果 6种成分均达到基线分离,同型原儿茶酸、原儿茶酸、对羟基苯乙酸、绿原酸、咖啡酸和金丝桃苷的质量浓度与峰面积,分别在1.14～114 μg/mL (r=0.9999)、0.74 ～186μg/mL(r =0.9992)、5.26～526 μg/mL(r =0.999 8)、2.88～288 μg/mL(r=0. 9998)、1.50～148 μg/mL(r=0.9999)、1.42～205 μg/mL(r=0.9999)范围呈良好的线性关系；平均加样回收率分别为100.4％、99.4％、98.4％、99.5％、96.2％、97.6％; RSD分别为1.3％、2.2％、2.2％、3.6％、1.8％、3.4％.结论 该方法准确,简便,灵敏,为返魂草颗粒的质量评价提供了依据.

Simultaneous determination of six active components in Fanhuncao Granule by HPLC

AIM To develop an HPLC method for simultaneously determining homoprotocatechuic acid,protocatechuic acid,p-hydroxy benzeneacetic acid,chlorogenic acid,caffic acid and hyperoside in Fanhuncao Granule(Senecio cannabifolius Less or Senecio cannabifolius var.integrilifolius).METHODS Agilent Zorbax SB-C18 column(4.6 mm ×250 mm,5 μm) was used with acetonitrile-1% trifluoroacetic acid as the mobile phase in gradient elution.The flow rate was 1 mL/min.The column temperature was maintained at 30 ℃,and the detection wavelength was set at 280 nm.RESULT Six active components were in baseline separation.Homoprotocatechuic acid was linear in the range from 1.14 to 114 μg/mL(r=0.999 9) and the average recovery was 100.4%(RSD=1.3%).Protocatechuic acid was linear in the range from 0.74 to 186 μg/mL(r=0.999 2) and the average recovery was 99.4%(RSD=2.2%).p-Hydroxy benzeneacetic acid was linear in the range from 5.26 to 526 μg/mL(r=0.999 8) and the average recovery was 98.4%(RSD=2.2%).Chlorogenic acid was linear in the range from 2.88 to 288 μg/mL(r=0.999 8) and the average recovery was 99.5%(RSD=3.6%).Caffic acid was linear in the range from 1.50 to 148 μg/mL(r=0.999 9) and the average recovery was 96.2%(RSD=1.8%).Hyperoside was linear in the range from 1.42 to 205 μg/mL(r=0.999 9) and the average recovery was 97.6%(RSD=3.4%).CONCLUSION The method is accurate,simple and feasible.It can be used as a quality evaluation for Fanhuncao Granules.