Identification of isomeric disaccharides in mixture by the 1-phenyl-3-methyl-5-pyrazolone labeling technique in conjunction with electrospray ionization tandem mass spectrometry
1-Phenyl-3-methyl-5-pyrazolone (PMP) labeling technique has hitherto proved to be a convenient and sensitive method for separating and detecting oligosaccharides. However, the detailed fragmentation of the derivatives by tandem mass spectrometry has been reported limitedly and no characteristic fragment ions for isomers have been detected. In this study, eight disaccharide isomers were labeled with PMP and analyzed by positive ion electrospray ionization multi-stage tandem mass spectrometry (ESI-MS n ). In comparison with the native disaccharides, PMP labeled disaccharides gave rise to more fragment ions in the tandem mass spectra. The distinctive diagnostic fragment ions formed from cleavage of C C bonds have been detected in the fragmentation of PMP-labeled disaccharide linkage isomers, allowing unambiguous assignment of the position of the glycosidic linkages. This feature is particularly useful for the structural determination of unknown isomeric disaccharides mixed together. In addition, the anomeric configurations can also be easily assigned based on the relative abundance ratios of the selected ion pairs. To verify the feasibility of the method used in the analysis of natural product, water soluble Panax Ginseng extract has been further investigated to identify its unknown disaccharides. The results confirmed that the PMP labeling technique in conjunction with ESI-MS n could offer a powerful and convenient tool for differentiation of structurally closely related isomers, even the unknown mixtures of isomeric disaccharides with different linkage types.
危险品化学品经营许可证（带存储）