Methadone is widely used for the treatment of opioid dependence. HPLC and CE are widespread methods for drug monitoring and metabolism studies. Although the methods are widely used for methadone and its main metabolite EDDP [1, 2], a definite direct peak identification for EDDP enantiomers is not described. This study describes a method for specific identification of each peak in the chromatogram and electropherogram of methadone analysis. The result of the study demonstrates differences in the elution order of the enantiomers of methadone and EDDP due to the technique used for analysis. The elution order of EDDP using HPLC is interchanged with respect to the order of the peaks in the electropherogram.