This article describes the development of a polybutylmethacrylate-based monolithic capillary column as a chiral stationary phase. The chiral monolithic column was prepared by polymerization of butyl methacrylate (BMA), ethylene dimethacrylate (EDMA), and N-methacryloyl-l-glutamic acid (MAGA) in the presence of porogens. The porogen mixture included N,N-dimethyl formamide and phosphate buffer. MAGA was used as a chiral selector. The effect of MAGA content was investigated on electrochromatographic enantioseparation of d,l-histidine, d,l-tyrosine, d,l-phenyl alanine, and d,l-glutamic acid. The effect of acetonitrile (ACN) content in mobile phase on electro-osmotic flow was also investigated. It was demonstrated that the poly(BMA-EDMA-MAGA) monolithic chiral column can be used for the electrochromatographic enantioseparation of amino acids by capillary electrochromatography (CEC). The mobile phase was ACN/10?mM phosphate buffer (45:55%) adjusted to pH 2.7. It was observed that l-enantiomers of the amino acids migrated before d-enantiomers. The separation mechanism of electrochromatographic enantioseparation of amino acids in CEC is discussed. Chirality 24:606609, 2012. (c) 2012 Wiley Periodicals, Inc.