Liquid chromatographic enantiomeric resolution of amino acids with β-cyclodextrin bonded phases and derivatization with o-phthalaldehyde

Abstract

The reaction of amino acids with o-phthalaldehyde (OPA) and 2-mercaptoethanol is evaluated as a precolumn derivatization technique in order to achieve the chiral separation and final fluorimetric detection of l,d-glutamic acid in β-cylodextrin bonded phases. The pertinent chromatographic parameters were determined and the effect of organic modifiers, pH, ionic strength and temperature on retention and resolution were evaluated. The chromatographic method finally developed allowed the optical resolution of OPA-derivatized l- and d-glutamic acid using methanol—1% triethylammonium acetate (pH 7.4) (30 + 70, v/v) as eluent.