HPLC-UV-ELSD法同时测定痰热清注射液中主要成分含量___

Objective: To establish an HPLC-UV-ELSD method for the simultaneous 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 curve in the range of 0.221-4.42 mg·L^-1 for chlorogenic acid, 20.3-406 mg·L^-1 for baicalin, 19.8-396 mg·L^-1 for ursodeoxycholic acid and 17.9-179 mg·L^-1 for chenodeoxycholic acid was 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.