Application of Fingerprint Combined with QAMS in Quality Evaluation of Gegenqinlian Tablets

OBJECTIVE To establish the fingerprint of Gegenqinlian tablets and determine the content of eleven components(3’-hydroxy puerarin, puerarin, 3’-methoxy puerarin, daidzin, baicalin, palmatine hydrochloride, berberine hydrochloride, scutellarin, baicalein, ammonium glycyrrhizinate and wogonin) by quantitative analysis of multi-components by single-marker(QAMS).METHODS Ten batches of Gegenqinlian tablets were determined by HPLC and a common fingerprint was established. Baicalin was selected as internal reference. The relative correction factors of the component with 3’-hydroxy puerarin, puerarin, 3’-methoxy puerarin, daidzin, palmatine hydrochloride, berberine hydrochloride, scutellarin, baicalein, ammonium glycyrrhizinate and wogonin were calculated and their contents were calculated. The feasibility and scientificity of QAMS was evaluated by comparison on the results between the measured value and calculation value by external standard method and QAMS. The chromatographic separation was performed on ananalytical column of Waters Xbridge-C₁₈(250 mm× 4.6 mm, 5 μm) with gradient elution, the mobile phase was acetonitrile 0.1% phosphoric acid aqueous solution, at a flow rate of 1.0 mL·min^-1. The column temperature was 30 ℃ and the detection wavelength was 260 nm.RESULTS There were 20 peaks in 10 batches of Gegenqinlian tablets and 11 chemical constituents were identified. The similarity of 10 batches of Gegenqinlian tablets was >0.97. The linear range of 3’-hydroxy puerarin, puerarin, 3’-methoxy puerarin, daidzin, baicalin, palmatine hydrochloride, berberine hydrochloride, scutellarin, baicalein, ammonium glycyrrhizinate and wogonin were 0.056 6-2.830 2, 0.241 2-12.058 6, 0.128 0-6.401 0, 0.059 7-2.983 5, 0.242 7-12.134 9, 0.045 7-2.285 7, 0.192 8-9.641 0, 0.043 3-2.167 0, 0.018 0-0.900 2, 0.021 0-1.048 4, 0.011 5-0.575 4 μg (r²= 0.999 6-1.000) respectively. The average recovery were 100.23%, 102.01%, 101.66%, 102.73%, 100.17%, 98.45%, 98.41%, 100.95%, 101.85%, 97.97%, 100.09%(RSD=1.24%-2.57%, n=6) respectively. The relative correction factors of 3’-hydroxy puerarin, puerarin, 3’-methoxy puerarin, daidzin, palmatine hydrochloride, berberine hydrochloride, scutellarin, baicalein, ammonium glycyrrhizinate and wogonin were 0.860 4, 0.605 3, 0.850 9, 0.582 8, 0.557 1, 0.498 6, 0.767 2, 0.652 1, 2.608 1, 0.545 2 respectively. RAD between QAMS method and external standard method were 0.03%-2.12%. CONCLUSION The combination of QAMS and fingerprint can provide reference for the quantitative determination and quality control of Gegenqinlian tablets.