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5 isoflavones, puerarin, daidzin, daidzein, 3′-hydroxy-puerarin and 3′-methoxy-puerarin had been isolated and purified concurrently from Pueraria lobata for the primary time by high efficiency counter-present chromatography (HPCCC) using a system consisting of hexane-ethyl acetate-n-butanol-ethanol-water (0.5 : 2 : 1 : 0.5 : 3.5, v/v/v/v/v). In whole, 155 mg of puerarin, 41 mg of daidzin, 12 mg of 3′-hydroxy-puerarin, 6 mg of 3′-methoxy-puerarin and 106 mg of daidzein have been purified from 5.1 g of the ethyl acetate extract of P. lobata; the purities of the five isoflavones as decided by excessive efficiency liquid chromatography (HPLC) had been 98.77%, 96.53%, 97.59%, 90.21% and 98.36%, respectively. The structures of the 5 compounds were recognized by their retention time and the electrospray ionization multi-stage tandem mass spectrometry (ESI-MSn) in the optimistic ion mode, and confirmed by 1H-NMR experiments. The characteristic ESI-MS fragmentation patterns of the five compounds are mentioned. Trade Promotion Authority Act
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