title: Determination of L-canavanine and other free amino acids in Vicia disperma (Fabaceae) seeds by precolumn derivatization using diethyl ethoxymethylenemalonate and reversed-phase high-performance liquid chromatography.
abstract: A method for determination of the non-protein amino acid l-α-amino-γ-(guanidinooxy)-n-butyric acid (L-canavanine) and other free amino acids in Vicia disperma is presented. Seed extracts were derivatized by reaction with diethyl ethoxymethylenemalonate and analyzed by reverse-phase high-performance liquid chromatography. Calibration curves showed very good linearity of the response. The limit of detection and quantification were 0.15 and 0.50 μM, respectively. The method has a high intra- (RSD=0.35%) and inter-repeatability (RSD=2.86%), and a remarkable accuracy with a 99% recovery in spiked samples. The method is very easy to carry out and allows for ready analysis of large number of samples using very basic HPLC equipment because the derivatized samples are very stable and have very good chromatographic properties.
authors.affiliation: Instituto de la Grasa (C.S.I.C.), Avda. Padre García Tejero 4, 41012 Sevilla, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Megías
authors.affiliation: Instituto de la Grasa (C.S.I.C.), Avda. Padre García Tejero 4, 41012 Sevilla, Spain.
authors.forename: Isabel
authors.initials: I
authors.lastname: Cortés-Giraldo
authors.affiliation: Instituto de la Grasa (C.S.I.C.), Avda. Padre García Tejero 4, 41012 Sevilla, Spain.
authors.forename: Julio
authors.initials: J
authors.lastname: Girón-Calle
authors.affiliation: Instituto de la Grasa (C.S.I.C.), Avda. Padre García Tejero 4, 41012 Sevilla, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Vioque
authors.affiliation: Instituto de la Grasa (C.S.I.C.), Avda. Padre García Tejero 4, 41012 Sevilla, Spain. Electronic address: alaiz@ig.csic.es.
authors.forename: Manuel
authors.initials: M
authors.lastname: Alaiz
mesh_terms: D000596:Amino Acids / Q000032:analysis*; D002172:Canavanine / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000379:methods*; D008314:Malonates / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry*; D031306:Vicia / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D008314:Malonates; D002172:Canavanine; C044472:diethyl (ethoxymethylene)malonate
keywords: Diethyl ethoxymethylenemalonate; Free amino acids; HPLC; Vicia disperma; l-canavanine; l-α-amino-γ-(guanidinooxy)-n-butyric acid