Study on HPLC and HPLC-MS/MS Determination Methods of Five Chemical Components in Jinshengyuan Capsules

OBJECTIVE To establish a method for the determination of five constituents(liquiritin, glycyrrhizic acid, salvianolic acid B, cinobufagin and resibufogenin) in Jinshengyuan capsules by HPLC and HPLC-MS/MS. METHODS HPLC was used for imultaneous determination of liquiritin, glycyrrhizic acid and salvianolic acid B. The analysis was performed on SHIMADZU C₁₈ ODS(250 mm×4.6 mm, 5 μm), with the mobile phase comprising of acetonitrile(A)-0.05% phosphoric acid(B) flowing at 1.0 mL·min^-1 in a gradient elution manner, and the detection wavelength was set at 276, 250 and 286 nm by diode array detector. Cinobufagin and resibufogenin was analyzed by HPLC-MS/MS method, ESI source as ion source, positive ion multiple reaction monitoring mode, which performed on 30 ℃ thermostatic ZORBAXSB-C₁₈(250 mm×4.6 mm, 5 μm), with the mobile phase comprising of acetonitrile(A)-0.1% formic acid(B) flowing at 1.0 mL·min^-1. RESULTS The liner ranges of liquiritin, glycyrrhizic acid, salvianolic acid B, cinobufagin and resibufogenin were within 3.14-62.8, 4.18-83.6, 7.00-140.0, 0.495-9.90, 0.520-10.4 μg·mL^-1. The average recovery rates were 99.13%, 98.96%, 97.96%, 99.51% and 96.22% with the RSD of 1.43%, 2.76%, 1.68%, 2.54% and 1.39%. CONCLUSION The method has good separation, accuracy and reliability, high specificity and sensitivity, which can provide experimental basis for the detection and quality control of the trace components of dried toad skin in compound traditional Chinese medicine.