Wheat gluten amino acid composition analysis by high-performance anion-exchange chromatography with integrated pulsed amperometric detection

作者：Rombouts Ine^*; Lamberts Lieve; Celus Inge; Lagrain Bert; Brijs Kristof; Delcour Jan A

来源：Journal of Chromatography A, 2009, 1216(29): 5557-5562.

DOI：10.1016/j.chroma.2009.05.066

• Amino acid
• Gluten
• Wheat
• High-performance anion-exchange chromatography with integrated
• pulsed amperometric detection
• HPAEC-IPAD

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 (6.0 M hydrochloric acid solution at 110 degrees C for 24 h), evaporation of hydrolyzates (110 degrees C), and chromatographic separation of the liberated amino acids. Correction factors W accounted for incomplete cleavage of peptide bonds involving Val (f = 1.07) and Ile (f = 1.13) after hydrolysis for 24 h and for Set (f = 1.32) losses during evaporation. Gradient conditions including an extra eluent (0.1 M 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).

• 出版日期2009-7-17