Objective: To establish a quantitative analysis of multi-components by single marker(QAMS) method for the determination of 8 isoflavones in Weixuening mixture, and establish the HPLC fingerprint of “intermediate finished product”, so as to provide reference and basis for its quality control. Methods: The Acclaim 120-C18 column (250 mm×4.6 mm,5 μm) was used with methanol (A)-0.2% phosphoric acid aqueous solution(B)-0.1% phosphoric acid(C) as mobile phase. Gradient elution was performed at a flow rate of 1.0 mL·min-1, with the detection wavelength of 210 nm and the column temperature of 30 ℃. Using polydatin as the reference substance, relative correction factors of catechin, epicatechin, resveratrol, quercetin, luteolin, genistein and formononetin were calculated, respectively. The relative correction factors were used to calculate the contents and fingerprints of 7 substances to be tested in 3 batches of Weixuening mixture, and the differences between the two methods were compared to verify the accuracy and feasibility. Results: The linear ranges of catechin, epicatechin, polydatin, resveratrol, quercetin, luteolin, genistein and formononetin were 1.066-53.300, 1.062-53.100, 4.384-219.200, 1.118-55.900, 1.036-51.800, 1.110-55.400, 1.040-52.000, 0.530-53.000 μg·mL-1, respectively. The relative correction factors of catechin, epicatechin, resveratrol, quercetin, luteolin, genistein and formononetin were 0.375 4, 0.348 6, 0.585 3, 0.597 2, 0.517 6,0.588 7, 0.659 9, respectively. The repeatability was good under different experimental conditions. At the same time, the content range of 8 components in the extracts, clear extracts and mixture was polydatin (45.26-90.45 μg·mL-1), catechin (21.29-38.18 μg·mL-1), epicatechin (15.06-21.46 μg·mL-1), resveratrol (6.26-8.57 μg·mL-1), quercetin (1.22-2.13 μg·mL-1), luteolin (3.35-4.44 μg·mL-1), genistein (1.97-3.17 μg·mL-1), and anthocyanin (0.65-1.13 μg·mL-1). There were 27 common peaks in the fingerprint of three batches of Weixuening mixture, and 8 peaks of polydatin, quercetin, catechin, epicatechin, luteolin, anthocyanin and resveratrol were identified, most of which were more than 0.9 similar. Conclusion: The established system is simple, time-saving and efficient, which is conducive to provide reference for the quality control of Weixuening mixture.
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