HPLC-UV/ELSD Fingerprint of Liushen Pills

OBJECTIVE HPLC-UV/ELSD fingerprint was developed to assign and identify the main characteristic peaks in Liushen Pills. METHODS High performance liquid chromatography: Kromasil C₁₈ column (250 mm × 4.6 mm， 5 μm) was used with a gradient mobile phase system of acetonitrile-water containing 0.05% trifluoroacetic acid. Ultraviolet detector: the detection wavelength was 296 nm; Evaporative light scattering detector: The temperature of drift tube was maintained at 70 ℃， sprayer parameter was cooling mode and nitrogen pressure was 25 psi. The injection volume was 20 μL; The flow rate was 1 mL/min. The similarities between the HPLC-UV/ELSD fingerprints of 14 batches of Liushen Pills were calculated by similarity evaluation software， and the common peaks were assigned and identified simultaneously. RESULTS The chromatography fingerprint of Liushen Pills from 14 different origins was established. In the chromatography fingerprint with HPLC-UV of Liushen Pills， 14 common peaks were damarcated and the similarities of Liushen Pills were between 0.940-0.988， and the chromatography fingerprint with HPLC-ELSD of Liushen Pills， 13 common peaks were damarcated and the similarities of Liushen Pills were between 0.968-0.998. Gamabufotalin， arenobufagin， telocinobufagin， bufotalin， cinobufotalin， bufalin， cholic acid， chenodeoxycholic acid， deoxycholic acid were identified. CONCLUSION It is the first time to establish the HPLC-UV/ELSD fingerprint of Liushen Pills. The method is simple， accurate， and reproducible， which can be used to characterize the chemical composition information of Liushen Pills and provide a scientific evidence for the quality control of Liushen Pills.