HPLC-UV-ELSD法同时测定痰热清注射液中主成分含量

HPLC-UV-ELSD simultaneous determination of major component contents in Tanreqing injection

目的:建立同时测定痰热清注射液中绿原酸、黄芩苷、熊去氧胆酸含量和鹅去氧胆酸限量的高效液相色谱法。方法:色谱条件:Agilent Zorbax SB-C18柱;流动相为0.15%醋酸溶液-0.15%醋酸乙腈,线性梯度洗脱,流速为0.8 mL.min-1;柱温为30℃;紫外检测波长:绿原酸326 nm,黄芩苷276 nm;蒸发光散射检测器漂移管温度为100℃,雾化气流速为1.5 L.min-1。结果:绿原酸、黄芩苷、熊去氧胆酸和鹅去氧胆酸分别在0.221～4.42,20.3～406,19.8～396,17.9～179 mg.L-1质量浓度范围内呈良好的线性关系,r均大于0.9995;平均加样回收率分别为101%,98.4%,99.4%,102%;4种化合物含量重复性试验RSD小于1.8%。结论:该方法简便、准确,重复性好,在对痰热清注射液中主要有效成分绿原酸、黄芩苷、熊去氧胆酸进行定量的同时,实现对鹅去氧胆酸限量的测定。

Objective ：To establish an HPLC - UV - ELSD method for the simuhaneous determination of chlorogenic acid,baicalin,ursodeoxycholic acid and chenodeoxycholic acid in Tanreqing injection. Methods：The chromatographic separation was performed on an Agilent Zorbax SB -C18 column. The mobile phase was mixtures of 0. 15% acetic acid - water and 0. 15% acetic acid - acetonitrile with linear gradient elution at the flow rate of 0. 8 mL · min^-1. The column temperature was maintained at 30 ℃. The UV detection wavelength of chlorogenic acid and baicalin were 326 nm and 276 nm,respectively. The drift tube temperature of ELSD was set at 100 ℃ and the nitrogen flow rate at 1.5 L · min^-1. Results：The calibration curves in the range of 0. 221 -4.42 mg·L^-1 for chlorogenic acid,20.3 -406 mg· L^-1 for haicalin,19. 8 -396 mg·L^-1 for ursodeoxycholic acid and 17.9 -179 mg · L^-1 for chenodeoxycholic acid were good linear,respectively. The values of correlation coefficient were higher than 0. 9995 for all analytes. The average recoveries of chlorogenic acid, baicalin, ursodeoxycholic acid and chenodeoxycholic acid were 101% ,98.4% ,99.4% , 102% , respectively. Repeatability experiments showed that relative standard deviation （RSD） values of their contents were less than 1.8%. Conclusion：The method is simple, accurate and repeatable. It can be used to determinate the content of chlorogenic acid,baicalin and ursodeoxycholic acid which are the major effective components in Tanreqing injection, and monitor the limit content of chenodeoxycholic acid at the same time.