Multiple Hydrogen Loss from [M + H]+ and [a]+ ions of Peptides in MALDI In-Source Decay Using a Dinitro-Substituted Matrix.

Abstract

The formation and radical-directed dissociation of multiple hydrogen-abstracted peptide cations [M + H - mH]·+ has been reported using MALDI-ISD with dinitro-substituted matrices. The MALDI-ISD of synthetic peptides using 3,5-dinitrosalicylic acid (3,5-DNSA) and 3,4-dinitrobenzoic acid (3,4-DNBA) as matrices resulted in multiple hydrogen abstraction from the analyte [M + H]+ and fragment [a]+ ions, i.e., [M + H - mH]+ and [a - mH]+ (m = 1-8). All of the ISD spectra showed unusually intense [a]+ ions originating from cleavage at the Cα-C bond of the Leu-Xxx residues when peptides without Phe/Tyr/His/Cys residues were used. The intensity of the [an]+ series ions generated using 3,5-DNSA and 3,4-DNBA rapidly decreased with increasing residue number n, suggesting cleavage at multiradical sites of [M + H - mH]•+. It was suggested that multiple hydrogen abstraction from protonated peptides [M + H]+ mainly takes place from the backbone amide nitrogen.