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A single-step purification for glycine betaine determination in plant extracts by isocratic HPLC
M A Bessieres1, Y Gibon, J C Lefeuvre
1Groupe de Physiologie et Biochimie Végétales "Métabolismes de Stress" and Laboratoire d'Evolution des Systèmes Naturels et Modifiés, UMR CNRS 6553, Université de Rennes I, Campus de Beaulieu, 35042 Rennes Cedex, France.
Abstract:
The glycine betaine content of higher plants can be determined by rapid and simple isocratic high-performance liquid chromatography on a C(18) column. The method described is particularly suitable for a large number of samples. Starting from a complex dry plant, it consists of a single aqueous extraction followed by a purification over a sole cationic ion-exchange resin (AG1, OH(-)) before simple HPLC quantification. The single-step purification was allowed because the AG1, OH(-) resin removed 80% of the total amino acids contained in the extracts. Proline, which interferes with GB HPLC quantification, was completely retained by this column. The optimized method for GB quantification was then validated by comparaison with an (1)H NMR method.