Characterization of flavonoid glycosides from rapeseed bee pollen using a combination of chromatography, spectrometry and nuclear magnetic resonance with a step-wise separation strategy.

To identify the structures of flavonoid glycosides in bee pollen collected from rapeseed plants (Brassica napus L.), we utilised an approach that combined liquid chromatography-diode array detector-electrospray ionization-mass spectrometry (LC-DAD-ESI-MS) and nuclear magnetic resonance (NMR) technology with a step-wise separation strategy. We identified four constituents of high purity in rape bee pollen samples: (1) quercetin-3-O-β-D-glucosyl-(2→l)-β-glucoside, (2) kaempferol-3, 4'-di-O-β-D-glucoside, (3) 5, 7, 4'-trihydroxy-3'-methoxyflavone-3-O-β-D-sophoroside and (4) kaempferol-3-O-β-D-glucosyl-(2→l)-β-D-glucoside. This study will also provide useful reference standards for qualification and quantification of four flavonoid glycosides in natural products.