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Piperazines for peptide carboxyl group derivatization: effect of derivatization reagents and properties of peptides on signal enhancement in matrix‐assisted laser desorption/ionization mass spectrometry
Author(s) -
Qiao Xiaoqiang,
Sun Liangliang,
Chen Lingfan,
Zhou Yuan,
Yang Kaiguang,
Liang Zhen,
Zhang Lihua,
Zhang Yukui
Publication year - 2011
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.4903
Subject(s) - chemistry , derivatization , piperazine , chromatography , trifluoroacetic acid , peptide , hydrochloride , mass spectrometry , reagent , matrix assisted laser desorption/ionization , organic chemistry , desorption , biochemistry , adsorption
Abstract Piperazine‐based derivatives, including 1‐(2‐pyridyl)piperazine (2‐PP), 1‐(2‐pyrimidyl)piperazine (2‐PMP), 1‐(4‐pyridyl)piperazine (4‐PP), and 1‐(1‐methyl‐4‐piperidinyl)piperazine (M‐PP), were used for the derivatization of carboxyl groups on peptides with 1‐(3‐dimethylaminopropyl)‐3‐ethylcarbodiimide hydrochloride (EDC) and 1‐hydroxy‐7‐azabenzotriazole (HOAt) as coupling reagents, and trifluoroacetic acid (TFA) as activator. Taking synthetic peptides RVYVHPI (RI‐7) and APGDRIYVHPF (AF‐11) as samples, the yields of derivatized peptides by 2‐PP, 2‐PMP and 4‐PP were higher than 94%. The effect of piperazine derivatives on the signals of tryptic digests of α‐transferrin and bovine serum albumin (BSA) was investigated, and it was found that peptides derivatized by 2‐PP and 2‐PMP exhibited obviously improved ionization efficiency. Furthermore, comparison of identified peptides before and after derivatization showed that peptides with low molecular weight (MW) and high pI value were preferably detected after derivatization. In addition, after derivatization with 2‐PP and 2‐PMP, protein myelin basic protein S, 20 kDa protein, and histone H were confidently identified from the tryptic digests of two fractions of rat brain protein separated by reversed‐phase high‐performance liquid chromatography (HPLC), indicating the potential application of 2‐PP and 2‐PMP for the highly sensitive determination of peptides in comprehensive proteome analysis. Copyright © 2011 John Wiley & Sons, Ltd.