Separation of amino acids in plant tissue extracts by capillary zone electrophoresis with indirect UV detection using aromatic carboxylates as background electrolytes

Abstract

Amino acids in extracts of plant tissue were separated and detected by capillary zone electrophoresis (CZE) with indirect UV detection. Various aromatic carboxylates such as salicylate, benzoate, phthalate and trimellitate were investigated as background electrolytes (BGEs). A BGE of benzoate gave the best resolution and detector response. Amino acids were separated at a highly alkaline pH to charge amino acids negatively. Separation was achieved by the co-electroosmotic flow (Co-EOF) by the addition of the cationic surfactant myristyltrimethyl-ammonium bromide (MTAB) to the electrolyte. The condtions affecting the separation of amino acids, including electrolyte pH, concentrations of both benzoate and MTAB, were investigated and optimised. Separation of a mixture of 17 amino acids at pH 11.20 with indirect UV detection at 225 nm was achieved with a BGE of 10 mM benzoate containing 1.0 mM MTAB at pH of 11.20. Detection limits ranged between 10 and 50 μM. The proposed method was demonstrated by the determination of amino acids in extracts of Eucalypt leaves with direct injection of samples.