Objective: To establish the fingerprint and multi-component determination method of Bazhen Yizhi prescription by using ultra-high performance liquid chromatography (UPLC). Methods: ACQUITY UPLC HSS T3(100 mm×2.1 mm, 1.8 μm) column was used, the mobile phase was acetonitrile (A)-0.2% formic acid aqueous solution (B) with gradient elution at a flow rate of 0.4 mL·min -1, the detection wavelengths were 254 nm (glycyrrhizic acid) and 300 nm (3,4-dihydroxybenzoic acid, liquiritin, hesperidin, isoliquiritin, nobiletin, tangeretin). The common peaks were identified by comparison with reference substances and by Q-TOF/MS technology, and UPLC-PAD was used for content determination. Results: The similarity of fingerprints of 10 batches of Bazhen Yizhi prescription samples to reference fingerprint were all above 0.936. A total of 19 common peaks in the fingerprint of Bazhen Yizhi prescription were assigned, and 14 components were identified. The contents of 3,4-dihydroxybenzoic acid, liquiritin, hesperidin, isoliquiritin, nobiletin, tangeretin and glycyrrhizic acid were determined. The 7 substances showed good linear relationship within certain concentration ranges. The correlation coefficients were all above 0.999 9. The methodological investigation results showed that all indicators meet the requirements, the recovery rates were between 92.8%-106.7%, and the RSDs were less than 4.23%. The content ranges of 3,4-dihydroxybenzoic acid, liquiritin, hesperidin, isoliquiritin, nobiletin, tangeretin and glycyrrhizic acid in 10 batches of samples were 0.006-0.017,0.035-0.220,0.622-2.113,0.015-0.045,0.007-0.022,0.003-0.010 and 0.237-0.642 mg·g-1, respectively. Conclusion: The fingerprint and the determination method of multi-index components are stable, feasible and specific, and can be used for the quality control of Bazhen Yizhi prescription, providing a basis for new drug development and clinical safe drug use.
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