High-purity D-xylulose > 97% was prepared from D-xylose in gram quantities using a simple three-step procedure; enzymatic isomerization with commercial glucose isomerase (xylose isomerase EC. 5.3.1.5), ethanol precipitation, and ion-exchange chromatography on an anionic resin in sulfite form. In the chromatographic step, 0.7-1.4 g xylulose was produced within 10-15 h. Impurities in the preparation are of sugar origin and may consist of glucose, ribulose, and xylose. Fractions of high purity, > 99%, but in small amounts, could be produced in the chromatographic step. When the procedure was simplified by performing the precipitation in a saturated isomerization solution, almost identical results were obtained. If thee precipitation step was excluded, it resulted in poorer separation in the chromatographic step, but xylulose could still be produced with a purity of 96-97%, though in smaller amounts, 0.35-0.74g.