HPLC-DAD法同时测定茵黄合剂中绿原酸、栀子苷、黄芩苷和大黄酸的含量

Simultaneous determination of the content of chlorogenic acid, jasminoidin, baicalin and rheinic acid in Yinhuang Mixture by HPLC-DAD

目的:建立了同时测定茵黄合剂中绿原酸、栀子苷、黄芩苷和大黄酸4种成分含量的RP-HPLC方法。方法:采用Kromasil C_(18)(4.6mm×250mm,5μm)色谱柱,流动相为甲醇-0.1%甲酸水溶液,梯度洗脱,流速1.0m L/min,DAD检测器,检测波长200-400nm(254nm),进样体积为20μL。结果:绿原酸、栀子苷、黄芩苷和大黄酸4种成分色谱峰分离良好,浓度分别在5.25-105、11.8-236、5.35-107和0.89-17.8μg/m L范围内呈良好的线性关系;精密度试验的RSD分别为0.07%、0.04%、0.15%和0.09%,重复性试验的RSD为0.99%、1.37%、1.59%和1.88%,平均回收率分别为99.85%(RSD为1.75%)、98.97%(RSD为1.29%)、100.6%(RSD为1.80%)和99.20%(RSD为1.81%)。结论:该方法简便、灵敏,结果准确,可用于茵黄合剂的质量控制。

Objective: To establish the method for simultaneously determining the content of chlorogenic acid, jasminoidin, baicalin and rheinic acid in Yinhuang Mixture by RP-HPLC. Methods: Kromasil C_(18)(4.6mm×250mm, 5μm) column was used. The detection wavelength was 200-400 nm with DAD(254nm). The mobile phase was consisted of 0.1% formic acidmethanol with gradient elution, at the flow rate of 1.0m L/min. Injection volume was 20μL. Results: The chlorogenic acid, the jasminoidin, the baicalin and the rheinic acid were separated completely from impurities. The linear range of chlorogenic acid, jasminoidin, baicalin and rheinic acid was 5.25-105, 11.8-236, 5.35-107 and 0.89-17.8μg/m L, respectively. The content RSD of the precision were 0.07%, 0.04%, 0.15% and 0.09%, respectively. The content RSD of reproducibility were 0.99%, 1.37%, 1.59% and 1.88%, respectively. The average recovery rate were 99.85%(RSD =1.75%), 98.97%(RSD=1.29%), 100.6%(RSD=1.80%) and 99.20%(RSD=1.81%) respectively. Conclusion: This method is exclusive and sensitive, which is reliable for the determination of Yinhuang Mixture.