In this study, it is shown that preparative density gradient ultracentrifugation can be applied to separate racemate crystals from excess enantiomer crystals. This was demonstrated for alanine crystals separated in a 50 wt % Nycodenz gradient The recovery rates of the pure enantiomers from the mixtures were good (70-90%), and the separation method showed accuracy in the third digit of the density. As such density differences between racemate and enantiomers can be found for a wide range of crystalline chiral compounds, the presented separation method should be applicable to a large variety of substances on a preparative scale.