Amino acid analysis using chromatography–mass spectrometry: An inter platform comparison study

P. Krumpochova, B. Bruyneel, D. Molenaar, A. Koukou, M. Wuhrer, W.M.A. Niessen, M.A. Giera

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

The analysis of amino acids has become a central task in many aspects. While amino acid analysis has traditionally mainly been carried out using either gas chromatography (GC) in combination with flame ionization detection or liquid chromatography (LC) with either post-column derivatization using ninhydrin or pre-column derivatization using o-phthalaldehyde, many of today's analysis platforms are based on chromatography in combination with mass spectrometry (MS). While derivatization is mandatory for the GC-based analysis of amino acids, several LC platforms have emerged, particularly in the dawn of targeted metabolite profiling using hydrophilic interaction liquid chromatography (HILIC) coupled to MS, allowing the analysis of underivatized amino acids. Among the numerous analytical platforms available for amino acid analysis today, we here compare three prominent approaches, being GC-MS and LC-MS after amino acid derivatization using chloroformate and HILIC-MS of underivatized amino acids. We compare and discuss practical issues as well as performance characteristics, e.g., the use of 13C-labeled internal standards, of the different platforms and present data on their practical implementation in our laboratory. Finally, we compare the real-life applicability of all three platforms for a complex biological sample. While all three platforms are very-well suited for the analysis of complex biological samples they all show advantages and disadvantages for some analytes as discussed in detail in this manuscript.
Original languageEnglish
Pages (from-to)398-407
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume114
DOIs
Publication statusPublished - 2015

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Spectrometry
Spectrum Analysis
Liquid chromatography
Liquid Chromatography
Amino Acids
Mass spectrometry
Mass Spectrometry
Gas chromatography
Hydrophobic and Hydrophilic Interactions
Gas Chromatography
o-Phthalaldehyde
Ninhydrin
Flame Ionization
Metabolites
Chromatography
Gas Chromatography-Mass Spectrometry
Ionization

Cite this

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title = "Amino acid analysis using chromatography–mass spectrometry: An inter platform comparison study",
abstract = "The analysis of amino acids has become a central task in many aspects. While amino acid analysis has traditionally mainly been carried out using either gas chromatography (GC) in combination with flame ionization detection or liquid chromatography (LC) with either post-column derivatization using ninhydrin or pre-column derivatization using o-phthalaldehyde, many of today's analysis platforms are based on chromatography in combination with mass spectrometry (MS). While derivatization is mandatory for the GC-based analysis of amino acids, several LC platforms have emerged, particularly in the dawn of targeted metabolite profiling using hydrophilic interaction liquid chromatography (HILIC) coupled to MS, allowing the analysis of underivatized amino acids. Among the numerous analytical platforms available for amino acid analysis today, we here compare three prominent approaches, being GC-MS and LC-MS after amino acid derivatization using chloroformate and HILIC-MS of underivatized amino acids. We compare and discuss practical issues as well as performance characteristics, e.g., the use of 13C-labeled internal standards, of the different platforms and present data on their practical implementation in our laboratory. Finally, we compare the real-life applicability of all three platforms for a complex biological sample. While all three platforms are very-well suited for the analysis of complex biological samples they all show advantages and disadvantages for some analytes as discussed in detail in this manuscript.",
author = "P. Krumpochova and B. Bruyneel and D. Molenaar and A. Koukou and M. Wuhrer and W.M.A. Niessen and M.A. Giera",
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Amino acid analysis using chromatography–mass spectrometry: An inter platform comparison study. / Krumpochova, P.; Bruyneel, B.; Molenaar, D.; Koukou, A.; Wuhrer, M.; Niessen, W.M.A.; Giera, M.A.

In: Journal of Pharmaceutical and Biomedical Analysis, Vol. 114, 2015, p. 398-407.

Research output: Contribution to JournalArticleAcademicpeer-review

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AU - Krumpochova, P.

AU - Bruyneel, B.

AU - Molenaar, D.

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AU - Wuhrer, M.

AU - Niessen, W.M.A.

AU - Giera, M.A.

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