Analysis of native amino acid and peptide enantiomers by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry
Article first published online: 18 FEB 2004
Copyright © 2004 John Wiley & Sons, Ltd.
Journal of Mass Spectrometry
Volume 39, Issue 2, pages 177–187, February 2004
How to Cite
Desai, M. J. and Armstrong, D. W. (2004), Analysis of native amino acid and peptide enantiomers by high-performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. J. Mass Spectrom., 39: 177–187. doi: 10.1002/jms.571
- Issue published online: 18 FEB 2004
- Article first published online: 18 FEB 2004
- Manuscript Accepted: 10 OCT 2003
- Manuscript Received: 21 MAY 2003
- National Institutes of Health. Grant Number: R01 GM53825-07.
- native amino acids;
- liquid chromatography/mass spectrometry;
- atmospheric pressure chemical ionization;
High-performance liquid chromatography (HPLC) coupled to atmospheric pressure chemical ionization (APCI) mass spectrometry was used for the separation and detection of amino acid and peptide enantiomers. With detection limits as low as 250 pg, 25 amino acids enantiomers were baseline resolved on a Chirobiotic T chiral stationary phase. APCI demonstrated an order of magnitude better sensitivity over electrospray ionization (ESI) for free amino acids and low molecular mass peptides at the high LC flow-rates necessary for rapid analysis. As the peptide chain length increased (peptides with Mr ≥ 300 Da), however, ESI proved to be the more ideal atmospheric pressure ionization source. A mobile phase consisting of 1% (w/w) ammonium trifluoroacetate in methanol and 0.1% (w/w) formic acid in water increased the sensitivity of the APCI method significantly. A step gradient was then used to separate simultaneously all 19 native protein amino acid enantiomers in less than 20 min using extracted ion chromatograms. Copyright © 2004 John Wiley & Sons, Ltd.