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Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns
Published on: April 26, 2016
Simultaneous determination of thiamine and its phosphate esters by a liquid chromatographic method based on
Tomás Pérez-Ruiz1, Carmen Martínez-Lozano, María Dolores García-Martínez
1Department of Analytical Chemistry, Faculty of Chemistry, University of Murcia, 30071 Murcia, Spain. tpr@um.es
Abstract:
A sensitive method for the post-column detection of thiamine (T) and its phosphate esters is described. The procedure is based on the on-line photolysis of the analytes into photoproducts which have a strong enhancing effect on the CL permanganate-luminol reaction. The complete separation of the thiamines was obtained on a RP-amide C(16) column in isocratic elution with an analysis time of less than 7 min. Under the optimum conditions, analytical curves, based on standard solutions, were linear over the range 10-1000 nM for thiamine and 100-2000 nM for its mono- and di-phosphate esters. Intra- and inter-day precision values of less than 1.14% relative standard deviation (RSD) (n=10) and 1.86% RSD (n=15), respectively, were obtained. The method was successfully applied to the determination of the thiamines in pharmaceutical preparations and baby foods.
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