A capillary electrophoresis (CE) method for the quantitative determination of R(+) and S(−) rogletimide in serum was developed. Stereoselective resolution of the rogletimide enantiomers was achieved using 60 mM alpha-cyclodextrin (α-CD) in 10 mM sodium dihydrogen phosphate/5 mM sodium tetraborate buffer of pH 2.5. A 72 cm uncoated fused-silica capillary at a constant voltage 15kV was used for the analysis and 0.03 mM hexadecyltrimethyl ammonium bromide (HTAB) was added to the buffer to decrease the adsorption of endogenous substances onto the silica wall. The analytes of interest were extracted from serum using a solid phase extraction (SPE) procedure. An octadecyl cartridge gave good recoveries in excess of 85% for both R(+) and S(−) rogletimide without any interference.