HPLC法测定4个活血化瘀方中6种芳香酸的含量

HPLC determination of six aromatic acids in four traditional Chinese medicine formulae for promoting blood circulation to remove stasis

目的:建立高效液相色谱法测定血府逐瘀汤、少腹逐瘀汤、膈下逐瘀汤、身痛逐瘀汤4个活血化瘀方中没食子酸、原儿茶酸、香草酸、咖啡酸、绿原酸、阿魏酸6个芳香酸的含量。方法:应用SunFireTM C18柱(4.6mm×250mm,5μm),以0.5%醋酸水溶液和甲醇为流动相,梯度洗脱,流速为1.0mL·min-1,柱温30℃,二极管阵列检测器,检测波长254和323nm,外标法定量。结果:6个芳香酸的线性关系良好(r=0.9997～0.9999),线性范围分别为8.898～1139,3.016～386.0,2.477～317.1,2.320～296.9,3.219～412.1,5.180～663.2μg.mL-1;精密度、重复性、稳定性的RSD都低于4%;加样回收率为95.6%～100.9%。结论:该方法前处理简单、重复性好、结果准确,可用于芳香酸类成分对活血化瘀方的作用研究。

Objective：A high performance liquid chromatographic method was established for simultaneous determination of six aromatic acids including gallic acid,protocatechui acid,vanlillic acid,caffeic acid,chlorogenic acid and ferulic acid in four formulae named as Xuefu Zhuyu decoction,Shaofu Zhuyu decoction,Gexia Zhuyu decoction and Shentong Zhuyu decoction for promoting blood circulation to remove stasis.Methods：These six compounds were analyzed simultaneously with a SunFireTM C18 column（4.6 mm×250 mm,5 μm）by gradient elution using 0.5% acetic acid-methanol as the mobile phase,the flow rate was 1.0 mL·min-1,the column was maintained at 30 ℃ and detected with diode array detector（DAD）at 254 and 323 nm.The quantified analysis of external standard calibration curves was used.Results：All the aromatic acid compounds showed good linearity（r =0.9997~0.9999）in the range of the test concentration,the linear ranges were 8.898-1139,3.016~386.0,2.477~317.1,2.320~296.9,3.219~412.1 and 5.180~663.2 μg·mL-1,respectively.The RSDS of the precision,reproducibility and stability test were less than 4%;the average recoveries of the method were in the range of 95.6 %-100.9 %.Conclusion：The established method has advantages of good simplicity reproducibility and precision,the method can be applied to study the effect of aromatic acid in Traditional Chinese Medicine Formulae for promoting blood circulation to remove stasis.