Use of a 2,5-dihydroxybenzoic acid/aniline MALDI matrix for improved detection and on-target Derivatization of glycans: A preliminary report.
N-Linked glycans derived from human and bovine alpha1-acid glycoprotein, as well as chicken egg white albumin, were analyzed by MALDI-TOF mass spectrometry using a novel MALDI matrix consisting of 2,5-dihydroxybenzoic acid (DHB) and aniline. A significant increase in signal was observed for these oligosaccharides relative to the signal obtained when unmodified DHB was used as a matrix for the same set of samples. The use of aniline/DHB matrix also led to facile on-target derivatization of the glycans via nonreductive amination, as aniline was found to form a stable Schiff base with the reducing end GlcNAc residue without the need for prolonged incubation periods and elevated temperatures. Both native and derivatized glycans ionized as sodium adducts and had similar MS/MS fragmentation patterns consisting mainly of Y/B-cleavage ions. In our experiments, we obtained evidence for persistence of the derivatization reaction in the solid phase; i.e., the reaction appeared to be taking place even after the sample-matrix spot had dried. This is the first report on such solid-phase on-target derivatization of carbohydrates for subsequent analysis by MALDI mass spectrometry.
Snovida
Sergei I
SI
Department of Chemistry, University of Manitoba, Winnipeg, Manitoba, Canada.
Chen
Vincent C
VC
Perreault
Hélène
H
D000418:Albumins / Q000737:chemistry; D000586:Amination; D000814:Aniline Compounds / Q000737:chemistry*; D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D002645:Chickens; D005841:Gentisates / Q000737:chemistry*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009961:Orosomucoid / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013696:Temperature
D000418:Albumins; D000814:Aniline Compounds; D005841:Gentisates; D009961:Orosomucoid; D011134:Polysaccharides; C023650:aniline; C010925:2,5-dihydroxybenzoic acid