Analytical development of seamless procedures on cation-exchange chromatography and ion-pair chromatography with high-precision mass spectrometry for short-chain peptides

Yoshinori Takano, Yasuhiro Oba, Satoshi Furota, Hiroshi Naraoka, Nanako O. Ogawa, Thomas M. Blattmann, Naohiko Ohkouchi

研究成果: Contribution to journalArticle査読

抄録

We report comprehensive analytical procedures for characterization of underivatized short-chain peptides by wet chemical treatment and ion-pair liquid chromatography with electrospray ionization–mass spectrometry (LC/ESI–MS). The chromatographic separation via a porous graphitic carbon stationary column eluted using an acetonitrile solvent gradient revealed a positive correlation between the retention time and the degree of hydrophobicity reflected in a distinct relationship between methylene (–CH2–) elongation of amino acids. We also validated the chromatographic separation utilizing an InertSustain PFP column and high-resolution mass spectrometry of the Orbitrap system, which supports the analytical accuracy for identification of monoisotopic mass. Using the quantitative analysis on a Corona charged aerosol detector, the recovery of the underivatized short-chain peptide fraction was >94.0 ± 4.9% (n = 10) after purification via cation-exchange chromatography. By eliminating the inorganic and organic matrix effect, we can avoid ion suppression and enhancement to demonstrate accurate analytical evaluations with robust mass spectroscopic responses. The developed method provides new insight into short-chain peptide chemistry that can be utilized for numerous interdisciplinary research applications.

本文言語英語
論文番号116529
ジャーナルInternational Journal of Mass Spectrometry
463
DOI
出版ステータス出版済み - 5 2021

All Science Journal Classification (ASJC) codes

  • 器械工学
  • 凝縮系物理学
  • 分光学
  • 物理化学および理論化学

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