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Integration of Rapid Derivatization and Gradient Elution Techniques for Enhanced High‐Performance Liquid Chromatography Analysis of Key Amino Acids in Wheat Flour
Author(s) -
FERMIN B. C.,
RADINSKY J. A.,
KRATOCHVIL R. J.,
HALL J. E.,
O Y. M. L
Publication year - 2003
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1365-2621.2003.tb05786.x
Subject(s) - derivatization , chromatography , chemistry , elution , high performance liquid chromatography , resolution (logic) , glutamine , gradient elution , yield (engineering) , proline , amino acid , biochemistry , materials science , artificial intelligence , computer science , metallurgy
A rapid pre‐column derivatization process before ultraviolet chromatographic analysis using a Pico‐Tag C18 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 chro‐matography. 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. Keywords: proline, glutamine, glutamic acid, reverse‐phase HPLC, resolution