Analysis of Underivatized Amino Acids: Zwitterionic Hydrophilic Interaction Chromatography Combined with Triple Quadrupole Tandem Mass Spectrometry

Abstract

Analysis of underivatized amino acids is challenging regarding both separation and detection of this small, polar, and largely UV-inactive compounds. Additives for reversed phase chromatography such as ion-pairing reagents can hamper mass spectrometric detection. Zwitterionic hydrophilic interaction chromatography using MS compatible eluents together with tandem mass spectrometry in multiple reaction monitoring mode for selective detection is an attractive approach to overcome the above mentioned issues.

An UPLC-ESI-MS/MS Assay Using 6-Aminoquinolyl-N-Hydroxysuccinimidyl Carbamate Derivatization for Targeted Amino Acid Analysis: Application to Screening of Arabidopsis thaliana Mutants

“…Despite of that, column care ( i.e. , installation of in-line filter and guard column [ 23 ]) and long equilibration times (usually about 10 min in order to ensure retention time repeatability [ 23 ]) are essential in HILIC analysis. Furthermore, HILIC columns suffer of poor separation efficiency compared to the RPLC technique [ 24 , 25 ].…”

Kynurenine to tryptophan ratio as a biomarker of acute stress in fish

“…Furthermore, by switching to HILIC from RP-LC, common disadvantages including peak fronting and tailing, column bleed, irreversible sorption, and slow equilibration times can be avoided (Walker et al, 2012). In the past decade there has been an increase in the use of HILIC promoted by the need to analyze difficult polar compounds at low concentrations in increasingly complex biological and environmental matrixes (Hemstrom & Igrum, 2006;Boersema et al, 2008;Appelblad et al, 2009;Spagou et al, 2010;Kawachi et al, 2011;Buszewski & Noga, 2012;Dell'mour et al, 2012;Tang et al, 2016;Kathriorachchia et al, 2018;Garcia et al, 2019;Werneth et al, 2019). Hydrophilic interaction chromatography mass spectrometry-based metabonomic studies has highlighted the potential for using HILIC as a complement…”

Direct determination of amino acids by hydrophilic interaction liquid chromatography with charged aerosol detection

“…For example, a limited set of underivatized amino acids were partially separated by HILIC using a 20 minute analysis time and further resolved by the use of triple quadrupole MS using the multiple reaction monitoring (MRM) mode with positive electrospray ionization (ESI) [21,22]. However, this analysis did not include tryptophan, asparagine, glutamine, histidine or cysteine.…”

“…In the literature, many attempts have been made in the last 25 years to separate and detect amino acids by several chromatographic techniques. Variations to the separation have been attempted, including the use of ion pairing reagents [4][5][6][7], derivatization of the analytes [7][8][9][10][11][12][13][14], variations of both stationary phase and mobile phase [15][16][17][18][19][20][21][22][23][24] as well as the use of an alternate detector such as a mass spectrometer (MS) [5,7,15,18,19,[21][22][23][24] , pulsed amperometric detection (PAD) [16,17] and combination fluorescence/laboratory modified multiple electrochemical detector [20]. Ultimately, the analysis of amino acids to date has been complex and/or time-consuming process requiring multiple steps and expensive equipment.…”