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Capillary electrochromatography (CEC) has been shown to be a promising technique for miniaturized enantioseparations. Here we present the preparation and application of positively charged cellulose derivative-based chiral stationary phases (CSPs) for the CEC separation of enantiomers. The cellulose derivatives were chemically immobilized onto diethylenetriaminopropylated silica gel using tolylene-2,4-diisocyanate as a spacer. The separation of enantiomers in CEC can be performed on the positively charged CSPs with nonaqueous and aqueous mobile phases.