The present paper describes the protocol for the preparation of a home made chiral micro-HPLC system and the chiral packing material with cellulose-based chiral stationary phase. The column packing technique was investigated and the key factors which may influence the performance of the chiral separation system were evaluated with several chiral compounds. The composition of the mobile phase and its flow rate were optimized. Finally, five enantiomers were successfully separated. It was illustrated that the system was characterized by simplicity, high resolution, low consumption of the mobile phase and high speed. The system cauld be easily prepared at the laboratory for enantioseparation.