Title: Wheat gluten amino acid composition analysis by high-performance anion-exchange chromatography with integrated pulsed amperometric detection
Authors: Rombouts, Ine ×
Lamberts, Lieve
Celus, Inge
Lagrain, Bert
Brijs, Kristof
Delcour, Jan #
Issue Date: Jul-2009
Publisher: Elsevier
Series Title: Journal of Chromatography A vol:1216 issue:29 pages:5557-5562
Abstract: A simple accurate method for determining amino acid composition of wheat gluten proteins and their gliadin and glutenin fractions using high-performance anion-exchange chromatography with integrated pulsed amperometric detection is described. In contrast to most conventional methods, the analysis requires neither pre- or post-column derivatization, nor oxidation of the sample. It consists of hydrolysis (6.0M hydrochloric acid solution at 110 degrees C for 24h), evaporation of hydrolyzates (110 degrees C), and chromatographic separation of the liberated amino acids. Correction factors (f) accounted for incomplete cleavage of peptide bonds involving Val (f=1.07) and Ile (f=1.13) after hydrolysis for 24h and for Ser (f=1.32) losses during evaporation. Gradient conditions including an extra eluent (0.1M acetic acid solution) allowed multiple sequential sample analyses without risk of Glu contamination on the anion-exchange column. While gluten amino acid compositions by the present method were mostly comparable to those obtained by a conventional method involving oxidation, acid hydrolysis and post-column ninhydrin derivatization, the latter method underestimated Tyr, Val and Ile levels. Results for the other amino acids obtained by the different methods were linearly correlated (r>0.99, slope=1.03).
ISSN: 0021-9673
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Centre for Food and Microbial Technology
× corresponding author
# (joint) last author

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