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Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a widely used tool for the analysis of carbohydrates. However, the detection of carbohydrates by MALDI-TOF MS is often limited by the unsatisfactory ionization efficiency, instability and the matrix interference in low molecular weight region. Here, we demonstrate that a reactive matrix, 2-hydrazinoquinoline (2-HQ), can be used to detect neutral, sialic and low molecular weight carbohydrates sensitively both in the positive and negative ion mode. Since 2-HQ reacts efficiently with the reducing end of carbohydrate to form stable hydrazone, the ionization efficiency of derived carbohydrates is significantly enhanced. Using 2-HQ, the sensitivity for analyzing glycans has been improved 10-fold and 100-fold compared with those using 3-aminquinoline (3-AQ) and 2,5-dihydroxybenzoic acid (DHB) as matrix, respectively. Moreover, quantitative analysis of neutral, acidic and low molecular weight carbohydrates has been achieved because of the good reproducibility by using 2-HQ as matrix. As a result, up to 50 glycans in a single sample spot of human fresh serum without any prior purification and enrichment have been successfully detected. Therefore, 2-HQ as a new reactive matrix has shown great potentials in widespread applications for sensitive, selective, quantitative, high speed and high throughput analysis of carbohydrates in complex samples by MALDI-TOF MS. [Display omitted] • A dual-mode reactive matrix, 2-hydrazinoquinoline (2-HQ) was used to detect carbohydrates sensitively. • Using 2-HQ as matrix, the sensitivity for analyzing glycans has been improved 100-fold. • Up to 50 glycans in a single sample spot of human fresh serum have been successfully detected. • 2-HQ was used for quantitative analysis of neutral, acidic and low molecular weight carbohydrates. [ABSTRACT FROM AUTHOR]