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Immunoaffinity chromatographic columns were prepared for direct enantiomeric determination of racemic methamphetamine and amphetamine in this study. The stationary phase was synthesized by covalently bonding an anti-D-methamphetamine monoclonal antibody onto a pre-activated support (e.g. silica, sepharose 4B). Chromatographic results revealed that the immunoaffinity columns achieved enantiomeric separation of racemic amphetamine and methamphetamine. The immunoaffinity columns also have the ability to directly extract D-methamphetamine from urine by changing the pH of the mobile phase, this ability making it practical for the columns to determine a very low concentration of D-methamphetamine in urine.