Fingerprint of Fufang Kushen Changyankang Tablets and the Content Determination of Its Multi-components by HPLC

OBJECTIVE To establish a method for the qualitative analysis of fingerprints and quantitative analysis of 8 components of Fufang Kushen Changyankang tablets by HPLC, and to provide a more reliable method and basis for the quality control of Fufang Kushen Changyankang tablets. METHODS The determination was performed on AgilentTC-C₁₈(250 mm×4.6 mm, 5 μm) column with mobile phase consisted of acetonitrile(A)-0.05% phosphoric acid(gradient elution) at the flow rate of 1.0 mL·min^-1. The detection wavelength was set at 220 nm. The column temperature was set at 30℃ and the injection volume was 10 μL. Baicalin was selected as the reference peak. The HPLC fingerprints were analyzed and the similarity was evaluated with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine. RESULTS The linear range was 1.080-9.723 μg·mL^-1 (r=0.999 8)for geniposidic acid, 5.213-46.92 μg·mL^-1 (r=0.999 6) for chlorogenic acid, 18.95-170.5 μg·mL^-1(r=0.999 4) for matrine, 31.79-286.1 μg·mL^-1(r=0.999 7) for oxymatrine, 49.53-445.8 μg·mL^-1(r=0.999 4) for paeoniflorin, 113.5-102 2 μg·mL^-1(r=0.999 5) for baicalin, 70.77-636.9 μg·mL^-1(r=0.999 6)for berberine hydrochloride, 10.69-96.21 μg·mL^-1(r=0.999 5) for ammonium glycyrrhizinate, limits of quantitation were 0.867 0, 1.051, 1.123, 1.021, 0.903 1, 0.896 1, 1.063, 1.216 μg·mL^-1; average recoveries were 98.6%, 99.0%, 98.4%, 98.7%, 98.8%, 99.1%, 98.9%, 98.8% (RSD<2.0%, n=6), respectively; RSDs of precision, reproducibility and stability tests (48 h) were <2.0%(n=6). Compared with the common pattern of fingerprints, the similarities of 14 batches of samples were >0.9 with 22 common peaks, 8 of which were identified and assigned. CONCLUSION The HPLC fingerprints established have a good separation effect with strong characteristics. The method is stable and simple. The better quality control of Fufang Kushen Changyankang tablets can be achieved by multi-index composition determination combined with fingerprint analysis.