Methylation in combination with temperature programming enables rapid identification of polysaccharides by ambient micro-fabrication glow discharge plasma (MFGDP) desorption ionization mass spectrometry.

Obtaining the fingerprints of polysaccharides was known to be notoriously difficult by ambient mass spectrometry due to their resistance for desorption and ionization. Ambient mass spectrometry technology has recently been recognized as a quick analysis tool for obtaining fingerprints, which is attributed to its characteristics of no sample pretreatment and easy operation under atmospheric pressure. However, it still remains a challenge for accurate identification of the fingerprints of macromolecular polysaccharides by ambient micro-fabrication glow discharge plasma (MFGDP). In this study, a simplified methylation method was introduced to realize rapid analysis of polysaccharide mixture by MFGDP with the assistance of a temperature-programmed system (TPS). At the optimal temperature of TPS, oligosaccharides, plant polysaccharides and polysaccharide mixtures were all well characterized by TPS-MFGDP. In the proposed method, the characteristic [M + NH] adduct ions of oligosaccharides ions of oligosaccharides that are difficult for other ambient mass spectrometric methods were abundantly produced, making it possible to simultaneously identify a mixture of five or more polysaccharides. In addition, a supervised classification model which based on MS spectra, was used to classify seven typical hypoglycemic polysaccharides with excellent sensitivity, specificity and accuracy, indicating the good classification performance of the RF model constructed. Thus, the proposed mass spectroscopic method provides a cost effective, accurate and high throughput tool for identification and classification of polysaccharides, which is beneficial for studying the biological activity of polysaccharides.