Integration of Rapid Derivatization and Gradient Elution Techniques for Enhanced High‐Performance Liquid Chromatography Analysis of Key Amino Acids in Wheat Flour

Abstract

A rapid pre-column derivatization process before ultraviolet chromatographic analysis using a PicoTag C 18 column was developed in combination with modified gradient elution techniques for the analysis of key amino acids (proline and glutamine) in wheat flour. With use of a vacuum concentrator at 45°C, the pre-column derivatization process was significantly accelerated before entering a reverse-phase high-performance liquid chromatography. When running at an optimized scheme-in which the gradient elution started at a flow rate of 1 mL/ min and then gradually increased to 1.3 mL/min in 22 min, followed by reset to 1 mL/min at 45 min-the profiled mobile phase mixture was able to yield chromatograms with desirable retention factor and resolution.

The effect of heat treatment on the soluble protein content of oats

“…Amino acid analysis of defatted and non-defatted samples of both KHT and K oats were achieved with cation exchange chromatography coupled with UV detection at 280 nm using a Biochrom 30 series analyzer (Fermin et al, 2003). The amino acids were labeled with UV active compound prior to HPLC analysis and prepared in the following way.…”

Properties of Bread Dough with Added Fiber Polysaccharides and Phenolic Antioxidants: A Review

“…When dough is washed in running water, the starch is removed and a viscoelastic rubbery mass is obtained that is called “gluten” ( Hargreaves and others 1995 ; Bloksma and Bushuk 1998 ). Gluten is a protein complex, with proline (10%), glycine (20%), and glutamine (approximately 35%) being the most abundant amino acids responsible for gluten development ( Fermin and others 2003 ; Pommet and others 2005 ; Wellner and others 2005 ). Gluten proteins can be categorized based on their solubility into gliadins (alcohol–water soluble) and glutenins (insoluble) ( Wieser 2007 ).…”

“…Two cystein residues, either derived from the same protein via intramolecular bonding or different protein chains via intermolecular bonding, can form a loop within the protein ( Hoseney 1994 ). During dough development, these disulphide bonds can be mobilized through disulphide interchange reactions ( Bushuk 1998 ; Fermin and others 2003 ; Abang Zaidel and others 2008 ). When gluten proteins are heated up to 75 °C, sulphydryl-disulphide interchanges are accelerated ( Sun and others 2008 ).…”

Physicochemical Properties of Bread Dough and Finished Bread with Added Pectin Fiber and Phenolic Antioxidants

“…The major ingredients of wheat flour are gluten‐forming monomeric gliadins and polymeric glutenins that account for 80% to 90% of the total flour proteins, with albumins and globulins occurring in small amounts (Shewry and others 2002). Gliadins and glutenins play vital roles in the functional properties of gluten (Fermin and others 2003), with gliadins providing viscosity for dough development and glutenins imparting strength and elasticity (Toufeili and others 1999). Gliadins and glutenins are also involved in the physical interactions and chemical reactions such as Maillard, caramelization, and gelatinization reactions during dough development and bread baking.…”