title: Amino acid analysis by high-performance liquid chromatography after derivatization with diethyl ethoxymethylenemalonate.
abstract: Amino acids were determined by precolumn derivatization with diethyl ethoxymethylenemalonate and reversed-phase high-performance liquid chromatography (HPLC) with spectrophotometric detection at 280 nm. The reaction time was 50 min and the derivatives were stable at room temperature. Chromatographic resolution of a mixture of the derivatives of seventeen amino acids, including proline and cystine, was achieved within 35 min using a binary gradient system. The detection limit was 3 pmol. Amino acid analyses of acid hydrolysates of two proteins gave results equivalent to those obtained by conventional ion-exchange-based amino acid analysis. The simplicity of the procedure allows its use on any multi-purpose HPLC system.
authors.affiliation: Instituto de la Grasa y sus Derivados (CSIC), Seville, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Alaiz
authors.forename: J L
authors.initials: JL
authors.lastname: Navarro
authors.forename: J
authors.initials: J
authors.lastname: Girón
authors.forename: E
authors.initials: E
authors.lastname: Vioque
mesh_terms: D000596:Amino Acids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D006868:Hydrolysis; D008314:Malonates / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
chemical_list: D000596:Amino Acids; D008314:Malonates; C044472:diethyl (ethoxymethylene)malonate