Abstract
Introduction. Gliadin proteins are one of the gluten fractions. They are soluble in alcoholic solution and divided into four groups (α + β, γ, ω1.2, and ω5-gliadins). In this paper gliadins were extracted from wheat flour, and optimal conditions for their separation were determined.
 Study objects and methods. The separation was performed by capillary gel electrophoresis on Agilent apparatus, CE 7100 (a capillary with an inner diameter of 50 μm, a total length of 33 cm, and an effective length of 23.50 cm). In order to determine the optimal conditions, different solvent concentrations (50, 60, and 70% ethanol), capillary temperatures (20, 25, 30, 35, and 40°C), and electrode voltages (–14.5, –16.5, –17.5 and –18.5 kV) were applied. Migration time and relative concentration of each protein molecules within gliadin fractions in the electrophoregram were analysed using Agilent ChemStation Software.
 Results and discussion. The optimal conditions for gliadin separation were: solvent 70% (v/v) ethanol, capillary temperature of 25°C, and electrode voltage of –16.5 kV. Under these conditions, the total proteins were indetified as Xav = 23.50, including α + β gliadin fraction (Xav = 7.50 and relative concentration RC = 28.29%), γ-gliadins (Xav = 5.00, RC = 26.66%), ω1.2-gliadins (Xav = 4.33, RC = 14.93%), and ω5-gliadins (Xav = 6.67, RC = 30.98%).
 Conclusion. The results of the research can be of fundamental importance in the study of gluten proteins and the influence of technological procedures on their change and the possibility of reducing the allergic effect of gluten during processing.
Highlights
Capillary gel electrophoresis (CGE) is a widely used tool for the size-based analysis of protein
In terms of relative migration time (RMT), results obtained with the gel from Beckman Coulter were in the range of the data published by Nunnally et al.[4] for proteins ~ 50 kDa and slightly increased for proteins } 50 kDa (Tables 1 and 3)
Results obtained with the Beckman Coulter gel were in terms of RMT very close to, and in terms of RMT repeatability with 0.02 – 0.07 RSD% in the range of the data reported by Nunnally et al.[4] (Table 4)
Summary
Capillary gel electrophoresis (CGE) is a widely used tool for the size-based analysis of protein. New gels for CGE with UV detection were introduced onto the market by Advanced Analytical. These gels were originally developed for multi-capillary instruments[5]. SDS-MW Size Standard, IgG Control Standard, 10 kDa Internal Standard, SDS Sample Buffer, SDS Gel Buffer, 0.1 M HCl and 0.1 N NaOH were from Beckman Coulter (Fullerton, CA, USA). Sample preparation The SDS-MW Size Standard, BSA and reduced IgG Control Standard were prepared as described in Nunnally et al[4]. Non-reduced IgG Control Standard samples spiked with bLG were prepared as follows: 1. Non-reduced IgG Control Standard samples spiked with bLG were prepared as follows: 1. Thaw a 47.5 μL aliquot of the IgG Control Standard at room temperature
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