In-source decay and fragmentation characteristics of peptides using 5-aminosalicylic acid as a matrix in matrix-assisted laser desorption/ionization mass spectrometry.

The use of 5-aminosalicylic acid (5-ASA) as a new matrix for in-source decay (ISD) of peptides including mono- and di-phosphorylated peptides in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) is described. The use of 5-ASA in MALDI-ISD has been evaluated from several standpoints: hydrogen-donating ability, the outstanding sharpness of molecular and fragment ion peaks, and the presence of interference peaks such as metastable peaks and multiply charged ions. The hydrogen-donating ability of several matrices such as alpha-cyano-4-hydroxycinnamic acid (CHCA), 2,5-dihydroxybenzoic acid (2,5-DHB), 1,5-diaminonaphthalene (1,5-DAN), sinapinic acid (SA), and 5-ASA was evaluated by using the peak abundance of a reduction product [M + 2H + H](+) to that of non-reduced protonated molecule [M + H](+) of the cyclic peptide vasopressin which contains a disulfide bond (S-S). The order of hydrogen-donating ability was 1,5-DAN > 5-ASA > 2,5-DHB > SA = CHCA. The chemicals 1,5-DAN and 5-ASA in particular can be classified as reductive matrices. 5-ASA gave peaks with higher sharpness for protonated molecules and fragment ions than other matrices and did not give any interference peaks such as multiply-protonated ions and metastable ions in the ISD mass spectra of the peptides used. Particularly, 1,5-DAN and 5-ASA gave very little metastable peaks. This indicates that 1,5-DAN and 5-ASA are more "cool" than other matrices. The 1,5-DAN and 5-ASA can therefore be termed "reductive cool" matrix. Further, it was confirmed that ISD phenomena such as N-Calpha bond cleavage and reduction of S-S bond is a single event in the ion source. The characteristic fragmentations, which form a- and (a + 2)-series ions, [M + H - 15](+), [M + H - 28](+), and [M + H - 44](+) ions in the MALDI-ISD are described. Copyright 2010 American Society for Mass Spectrometry. Published by Elsevier Inc. All rights reserved.