Objective: To establish a quality evaluation method of Huaiqin ointment based on HPLC fingerprints, quantitative analysis of multi-components and chemometrics analysis. Methods: The chromatographic column was Agilent Extend C18column (250 mm×4.6 mm, 5 μm), and the column temperature was set at 25 ℃. The detection wavelength was set at 230 nm. The mobile phase was composed of acetonitrile-0.2% phosphate acid solution in gradient elution manner at a flow rate of 1.0 mL·min-1. The HPLC fingerprint of Huaiqin ointment was established, the common peaks were identified by similarity evaluation system for chromatographic fingerprint of Tradition Chinese Medicine (Version 2012), and the similarity was calculated. The content determination methods for paeoniflorin, rutin, enistin, sophoricoside, naringin, neohesperidin, baicalin, wogonoside and paeonolwere validated. The chemometrics methods such as cluster analysis and principal component analysis were used to evaluate the quality of Huaiqin ointment with different batches based on the results of fingerprint common peak area. Results: The fingerprint of Huaiqin ointment was established. Twenty-one common peaks were identified. The similarities of fingerprints of 10 batches of samples were above 0.95. Nine components had good linear relationship in the range of mass concentration, and the average recoveries were 97.4%-99.2% with the RSDs of 0.86-1.4%. The concentrations of paeoniflorin, rutin, genistin, sophoricoside, naringin, neohesperidin, baicalin, wogonoside and paeonolin 10 batches of samples were 5.34-6.31, 3.67-4.81, 0.76-1.27, 10.95-12.70, 6.70-7.14, 6.78-7.15, 17.45-18.92, 4.08-4.36 and 0.57-0.69 mg·g-1, respectively. The results of cluster analysis showed that 10 batches of Huaiqin ointment were clustered into three groups, and the results of principal component analysis showed that the principal components 1-4 were the main factor affecting the quality evaluation of Huaiqin ointment. Conclusion: The method is simple, accurate and reproducible, which can be used for the quality control and evaluation of Huaiqin ointment.
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