THE EFFECT OF THE DURATION OF SEPARATION AND WAVELENGTHS OF ABSORBANCE’S MEASUREMENT ON THE EFFECTIVENESS OF ELECTROPHORETIC SEPARATION OF GLIADINS BY GCE
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Датум
2022
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Апстракт
Gliadins are one of the gluten components which are present in most wheat products. Based on their mobility in acidic polyacrylamide gel electrophoresis (A-PAGE), gliadins are divided into α+β, γ, ω1.2, and ω5 classes. Gliadins are allergenic proteins which play a key role in celiac disease pathogenesis. Considering that the number of people with celiac disease is increasing, the aim of this study was to examine how different duration of analysis (30, 35 and 40 minutes) and wavelengths (200, 210 and 230 nm) effect the gliadin separation effectiveness by glassy carbon
electrode (GCE). We examined the effect of duration of capillary gel electrophoresis (GCE) separation (30, 35 and 40 minutes) and measurement of absorbance at different wavelengths (200, 210, and 230 nm) on the effectiveness of electrophoretic separation by capillary gel electrophoresis, with the aim of faster identification and quantification of gliadin proteins. Separation of gliadin proteins by GCE was performed on an Agilent, CE 7100 apparatus. SDS-MW analysis kit, PA 800 plus (2015 Beckman Coulter, USA) was used. Based on the obtained results the optimal duration of GCE separation was 30 minutes, because the number of observed proteins is the highest and the effectiveness of electrophoretic separation is the best (Xav = 23.50). A wavelength of 200 nm proved to be the best wavelength. By measuring the absorbance at this wavelength, the total number of observed proteins is Xav=18.33. In this study a duration of 30 minutes and a wavelength of 200 nm proved to be optimal. As the duration of analysis increases, the number of proteins decreases
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Кључне речи
Gliadins, Different duration of GCE separation, Wavelengths, GCE