OBJECTIVE To establish the HPLC characteristic chromatogram, simultaneously determinate the contents of allantoin, gallic acid, geniposidic acid and berberine in the substance benchmark of Yihuang Decoction, and clarify the quality value transmitting rules from decoction pieces to water decoction to substance benchmarks.
METHODS Agilent ZORBAX SB-Aq chromatographic column (250 mm×4.6 mm, 5 μm) was used, eluted with acetonitrile-0.1% acetic acid in gradient elution, and the detection wavelength was 205, 254, and 280 nm, respectively. The transfer rates of the above four components were used as the indexes for quality evaluation to study the quality value transmitting rule from decoction pieces to water decoction to substance benchmarks.
RESULTS The HPLC characteristic chromatogram of Yihuang Decoction was established, the similarity of 15 batches of samples was greater than 0.90, and a total of 15 common peaks were found and five of them allantoin (peak 2), gallic acid (peak 4), geniposidic acid (peak 6), phellodendrine (peak 7), berberine (peak 15) were identified. The contents of allantoin, gallic acid, geniposidic acid and berberine in 15 batches of benchmarks were 0.801%-1.087%, 0.047%-0.057%, 0.037%-0.081%, 0.250%-0.387%, respectively. The transfer rates (from water decoction to substance benchmarks) of allantoin, gallic acid, geniposidic acid and berberine were 73.09%-91.37%, 74.99%-89.32%, 71.77%-94.38%, 76.54%-88.43%, respectively.
CONCLUSION The HPLC characteristic chromatogram combined with the analytical method of multicomponents in simultaneous quantitation established in this experiment are stable and reliable, which can provide a theoretical guidance for the quality evaluation of Yihuang Decoction. The quality value transmitting results shows that the preparing technology of substance benchmarks is scientific and stable, which can lay a foundation for the subsequent development of classic prescription Yihuang Decoction preparation.
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