In order to enhance the substrate concentration of lipase catalyzed resolution of DLmenthol in non-aqueous phase, a solvent-free system was established using vinyl acetate as acyl donor. The effects of substrate concentration, enzyme dosage, temperature and reaction time on kinetic resolution of DL-menthol were investigated. The optimum conditions were determined as follows: substrate concentration 2.8 mol/L, enzyme dosage 0.5 g, reaction temperature 30 ℃ , reaction time 6 h. Under above conditions, the conversion reached 22.7% with e.e.p value of 99.1%. In addition, the lipase reserved 38.7% of its activity with stable enantioselectivity after 20 recycles