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A cool and high salt-tolerant ionic liquid matrix for preferential ionization of phosphopeptides by negative ion MALDI-MS

Abstract

Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) is commonly used to characterize peptides and proteins. However, MALDI-MS analysis of phosphorylated peptides remains a challenge because of low ionization efficiency. In this study, an ionic liquid matrix that consisted of 1,1,3,3-tetramethylguanidium (TMG) and 2,4,6-trihydroxyacetophenone (THAP) was optimized for preferential ionization of phosphopeptides in the negative ion mode of MALDI-MS. The results indicated that the ionic liquid matrix with a TMG-to-THAP molar ratio of 3:1 had the best performance towards phosphopeptide analysis; this matrix was denoted as G3THAP. Furthermore, the addition of phosphoric acid (PA) to G3THAP significantly reduced the background noise because PA competes with the phosphate group of the phosphopeptide for capturing alkali-ions. The resultant matrix denoted as G3THAP/PA showed great reproducibility and significantly improved the detection intensity of phosphopeptides in the presence of high concentrations of salts. The selective analysis of phosphopeptides in peptide mixtures and in proteolytic digests by negative ion MALDI-MS was also demonstrated and compared to the results obtained using the 3-aminoquinoline (3-AQ)/α-cyano-4-hydroxycinnamic acid (CHCA)/ammonium dihydrogen phosphate (ADP) and 2,5-Dihydroxybenzoic acid (DHB)/PA matrices. In general, G3THAP/PA is a cool and high salt-tolerant ionic liquid matrix for the preferential ionization of phosphopeptides by negative ion MALDI-MS.

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Publication details

The article was received on 17 May 2017, accepted on 04 Sep 2017 and first published on 04 Sep 2017


Article type: Paper
DOI: 10.1039/C7NJ01706E
Citation: New J. Chem., 2017, Accepted Manuscript
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    A cool and high salt-tolerant ionic liquid matrix for preferential ionization of phosphopeptides by negative ion MALDI-MS

    L. ling, C. Xiao, S. Wang, Y. Li, X. Chen and X. guo, New J. Chem., 2017, Accepted Manuscript , DOI: 10.1039/C7NJ01706E

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