An analytical micellar electrokinetic chromatographic method was developed and validated for the determination of the L-enantiomer of nateglinide. Separations were carried out in a 50 μm, 64.5/56 fused-silica capillary. The optimized conditions included 75 mM borate buffer, pH 9.2, containing 50 mM of sodium dodecyl sulfate and 25 mg/mL of methyl-β-cyclodextrin as background electrolyte, an applied voltage of 20 kV and a temperature of 15, UV detector at 210 nm. The assay was validated for the L-enantiomer of nateglinide. The limit of detection and quantification were 0.07 and 0.2% respectively. Intraday precision was ranged between 0.12 and 1.7%. Interday precision ranged between 0.73 and 1.73%. The assay was applied to the determination of the L-enantiomer of nateglinide in pharmaceutical formulations.