title: MALDI-TOF MS analysis of native and permethylated or benzimidazole-derivatized polysaccharides.
abstract: MALDI-TOF MS provides rapid and sensitive analyses of larger biomolecules. However, MS analyses of polysaccharide have been reported to have lower sensitivity compared to peptides and proteins. Here, we investigated some polysaccharides chemically derivatized by permethylation and ortho-phenylene diamine (OPD) tagging. Methylated glycan is obviously able to improve the sensitivity for mass spectrometry detection. Oxidative condensation by UV-activation tagging to saccharides by OPD and peptide-OPD also improve the sensitivity of MALDI-TOF MS analyses. Polysaccharides including dextran, glucomannan, arabinoxylan, arabinogalactan and beta-1,3-glucan, isolated from nutritional supplements of Ganoderma lucidum and Saccharomyces pastorianus were measured using MALDI-TOF MS with 2,5-dihydroxybenzoic acid (2,5-DHB) as the matrix. These glycans were also derivatized to methylated and benzimidazole-tagged glycans by chemical transformation for molecular weight analysis. The derivatized polysaccharides showed excellent MALDI-TOF MS signal enhancement in the molecular weight range from 1 to 5 kDa. Here, we demonstrate an efficient method to give glycan-benzimidazole (glycan-BIM) derivatives for polysaccharide determination in MALDI-TOF MS. Therefore, permethylated or benzimidazole-derivatized polysaccharides provide a new option for polysaccharide analysis using MALDI-TOF MS.
authors.affiliation: Genomics Research Center, Academia Sinica, 128, Academia Road Sec. 2, Taipei 115, Taiwan.
authors.forename: Wei-Ting
authors.initials: WT
authors.lastname: Hung
authors.forename: Shwu-Huey
authors.initials: SH
authors.lastname: Wang
authors.forename: Yi-Ting
authors.initials: YT
authors.lastname: Chen
authors.forename: Hui-Ming
authors.initials: HM
authors.lastname: Yu
authors.forename: Chung-Hsuan
authors.initials: CH
authors.lastname: Chen
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Yang
mesh_terms: D001562:Benzimidazoles / Q000737:chemistry*; D005841:Gentisates / Q000737:chemistry; D008745:Methylation; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry; D010655:Phenylenediamines / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis*; D020940:Reishi / Q000737:chemistry; D012440:Saccharomyces / Q000737:chemistry; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014466:Ultraviolet Rays
chemical_list: D001562:Benzimidazoles; D005841:Gentisates; D010455:Peptides; D010655:Phenylenediamines; D011134:Polysaccharides; C034193:1,2-diaminobenzene; C031000:benzimidazole; C010925:2,5-dihydroxybenzoic acid