Here, we report a new application using liquid chromatography-electrospray mass spectrometry (UHPLC-ESI-MS) using aldononitrile acetate derivatives for simultaneous baseline separation and detection of eight neutral saccharides, two uronic acids, one ketose, and eight alditols within 14 min. The separation was performed on a Cortecs C₁₈ column using acetonitrile (A) and water (B) as the mobile phase with gradient elution. The target components were detected in selected ion monitoring (SIM) mode by mass spectrometry with an electrospray ionization (ESI) source operating in positive ionization mode. A comparison with traditional methods was used to determine the validity of the results. The UHPLC-ESI-MS method was used for quantitative analysis of free carbohydrates in water extracts of Crataegus pinnatifida as well as determination of Polygonatum cyrtonema and Glossy ganoderma monosaccharides in polysaccharides. The results demonstrate that this protocol is a comprehensive and effective technique for qualitative and quantitative analysis of plant polysaccharides from TCMs.Copyright © 2021. Published by Elsevier Ltd.