A novel LC-MS/MS method for complete composition analysis of polysaccharides by aldononitrile acetate and multiple reaction monitoring

Carbohydrate analysis has always been a challenging task due to the occurrence of high polarity and multiple isomers. Aldoses are commonly analyzed by gas liquid chromatography (GLC) following aldononitrile acetate derivatization (AND). However, the GLC technique cannot be applied for the simultaneous determination of aldoses, ketoses, and uronic acids. In this study, a new method based on the combination of liquid chromatography-tandem mass spectrometry (LC-MS/MS) and AND is developed for the complete characterization of monosaccharide composition (i.e., aldoses, ketoses, alditols, amino sugars, and uronic acids) in plant derived polysaccharides. In addition to discussing the possible byproducts, the study optimizes the multiple reaction monitoring (MRM) parameters and LC conditions. The final separation of 17 carbohydrates is performed on a BEH Shield RP18 column (150 mm x 2.1 mm, 1.7 mu m) within 25 min, without using any buffer salt. Notably, the complex polysaccharides extracted from Ligusticum chuanxiong, Platycodon grandiflorum, Cyathula officinalis Kuan, Juglans mandshurica Maxim, and Aralia elata (Miq.). Seem bud can be successfully characterized using the developed method. Overall, the results demonstrated that the newly established LC-MS/MS MRM method is more effective and powerful than the GLC-based methods reported previously, and it is more suitable for the analysis of highly complex natural polysaccharides, including complex pectins, fructosans, and glycoproteins.

Automated summary

A new method combining liquid chromatography-tandem mass spectrometry (LC-MS/MS) and aldononitrile acetate derivatization (AND) has been developed for the comprehensive characterization of monosaccharide composition in plant derived polysaccharides. The optimized method can effectively separate 17 carbohydrates and is more powerful than traditional GLC-based methods for analyzing complex natural polysaccharides.