This study was conducted to investigate optimal time of artificial insemination (AI) to Hanwoo female after natural estrus. AI was occurred 12 and 24 hours after natural estrus in both heifer and multiparous recipient then pregnancy and parturition rates were estimated. Results indicated that AI performed at 24 hours after natural estrus showed significant (p<0.05) higher pregnancy rate in both heifer and multiparous recipient groups with significantly (p<0.05) higher abortion rate. However, there are no significant differences of parturition rate, twin birth and sex ratio in both heifer and multiparous recipient groups. Therefore, our results may suggest that performance of AI at 24 hours after natural estrus promise higher pregnancy rate than AI at 12 hours after natural estrus in both heifer and multiparous recipient.