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Propolis is a resinous product collected by honey bees from plant-derived materials, with high content of polyphenols associated to various bioactivities. In this study, a powerful separation and identification method of polyphenol compounds from Chinese propolis samples were investigated using off-line two-dimensional reversed-phase liquid chromatography (2D-RPLC/RPLC) in combination with Q-TOF-MS. A Click OEG and an XTerra C18 column were employed for the first and second dimensional separation, respectively. The identification efficacy of 2D-RPLC/RPLC and unidimensional LC (1D-LC) were compared, and 81 components involving 60 polyphenol compounds were characterized by the former method. In contrast, only 36 peaks were detected by the 1D-LC method. The proposed method was demonstrated to be more efficient for the existence or absence of target compounds identification, and that would provide a promising means for accurate determination and quality evaluation of Chinese propolis.

Separation and identification of polyphenol compounds in propolis using offline 2D-RPLC/RPLC coupled with Q-TOF-mass spectrometry