Colorimetric [14], spectrophotometric [15] and high-performance liquid chromatography (HPLC) methods have been used for the analysis of L-canavanine [16-18]. Chromatographic separation in the HPLC methods that have been described so far provides a much higher selectivity, but these methods in general are time consuming and are also hampered by the limited stability of reagents or derivatized amino acid. The goal of this research was to determine whether HPLC chromatography of the ethoxymethylenemalonate (DEEMM) derivatives of free amino acids can be used for determina- tion of L-canavanine as well as other free amino acids in the seeds of V disperma. DEEMM is a universal reagent for amino groups and has been used in amino sugar [19] and amino acid [20] chemistry, as well as for amino acid analysis [21,22]