Study on HPLC fingerprint of Youguiyin and its multi-component quantitative analysis / 中草药

ABSTRACT

Objective: To establish the HPLC fingerprint and determine nine components (geniposidic acid, morroniside, chlorogenic acid, geniposide, loganin, pinoresinol diglucoside, liquiritin, rutin, and glycyrrhizic acid) of Youguiyin, so as to provide a scientific basis for the quality control. Methods: HPLC analysis was performed on Pursuit XRs 5 C18 column (250 mm × 4.6 mm, 5 μm). The gradient elution was performed by the mobile phase consisting of acetonitrile and 0.1% formic acid aqueous with the flow rate of 1.0 mL/min, the detection wavelength was set at 230 nm, and the column temperature was 30 ℃. Fingerprints of ten batches of Youguiyin were determined, and the similarities among fingerprints were evaluated. Attributive analysis and identification of common peaks were performed by comparing the retention time and UV spectra among 10 batches of Youguiyin. The formula ingredients and reference substance, and nine components content were determined. Results: The similarities of fingerprints of 10 batches of Youguiyin and reference fingerprints were all greater than 0.904. There were 30 mutual peaks marked in total, which were eight mutual peaks from Eucommia ulmoides, eight mutual peaks from Glycyrrhiza uralensis, six mutual peaks from Cornus officinalis, six mutual peaks from Aconitum carmichaeli, one mutual peaks from Cinnamomum cassia, and none of mutual peaks were originated from Dioscorea opposita and Lycium barbarum, and three of them cannot be originated. Based on the the identification of the common peaks, nine components [geniposidic acid (peaks 7), morroniside (peaks 9), chlorogenic acid (peaks 11), gardenoside (peaks 12), loganin (peaks 13), pinoresinol diglucoside (peaks 14), liquorice glycosides (peaks 18), rutin (peaks 21), and glycyrrhizic acid (peaks 30)] were identified and quantified. The quality faction of nine components were 87.6—119.1 μg/g, 323.6—365.6 μg/g, 108.3—124.1 μg/g, 79.5—85.0 μg/g, 171.7—188.0 μg/g, 163.0—238.3 μg/g, 64.5—53.3 μg/g, 159.8—168.5 μg/g, and 72.8—83.6 μg/g in raw material. Conclusion: The method established in this study is simple, accurate and highly reproducible, and can provide basis for quality control of Youguiyin.