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 (13)C-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.
authors.affiliation: AIMMS Division of BioAnalytical Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081HV Amsterdam, The Netherlands; Systems Bioinformatics/AIMMS/NISB, VU University Amsterdam, De Boelelaan 1085, 1081HV Amsterdam, The Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Krumpochova
authors.affiliation: AIMMS Division of BioAnalytical Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081HV Amsterdam, The Netherlands.
authors.forename: B
authors.initials: B
authors.lastname: Bruyneel
authors.affiliation: Systems Bioinformatics/AIMMS/NISB, VU University Amsterdam, De Boelelaan 1085, 1081HV Amsterdam, The Netherlands.
authors.forename: D
authors.initials: D
authors.lastname: Molenaar
authors.affiliation: AIMMS Division of BioAnalytical Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081HV Amsterdam, The Netherlands.
authors.forename: A
authors.initials: A
authors.lastname: Koukou
authors.affiliation: AIMMS Division of BioAnalytical Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081HV Amsterdam, The Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Wuhrer
authors.affiliation: AIMMS Division of BioAnalytical Chemistry, VU University Amsterdam, De Boelelaan 1083, 1081HV Amsterdam, The Netherlands; Hyphen MassSpec, De Wetstraat 8, 2332 XT Leiden, The Netherlands.
authors.forename: W M A
authors.initials: WM
authors.lastname: Niessen
authors.affiliation: Systems Bioinformatics/AIMMS/NISB, VU University Amsterdam, De Boelelaan 1085, 1081HV Amsterdam, The Netherlands; Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2300 RC Leiden, The Netherlands. Electronic address: m.a.giera@lumc.nl.
authors.forename: M
authors.initials: M
authors.lastname: Giera
mesh_terms: D000596:Amino Acids / Q000032:analysis* / Q000737:chemistry; D002138:Calibration; D002247:Carbon Isotopes / Q000737:chemistry; D002623:Chemistry Techniques, Analytical; D002853:Chromatography, Liquid / Q000379:methods*; D005410:Flame Ionization / Q000379:methods; D005561:Formates / Q000032:analysis; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D057927:Hydrophobic and Hydrophilic Interactions; D007477:Ions; D057230:Limit of Detection; D009555:Ninhydrin / Q000737:chemistry; D011492:Protein Hydrolysates; D015203:Reproducibility of Results; D009764:o-Phthalaldehyde / Q000737:chemistry
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D005561:Formates; D007477:Ions; C101090:Primatone RL; D011492:Protein Hydrolysates; C083354:chlorocarbonic acid; D009764:o-Phthalaldehyde; D009555:Ninhydrin
keywords: Amino acids; GC–MS; HILIC; LC–MS/MS