Female infertility after occupational exposure to inhaled anesthetic agents has attracted critical attention, but systematic studies focusing on the impact of inhaled anesthetics on the female reproductive system have not been well-established. We used a murine model to study the effect of isoflurane exposure on infertility in female adult mice and investigated the potential underlying mechanism. One hundred adult female C57 mice were randomly allocated into 5 groups exposed in air containing 0, 2500, 5000, 10 000 or 20 000 ppm isoflurane for 15 consecutive days. Estrous cycle length was measured based on vaginal smear examination, ovarian histopathologic enumeration of follicles, and serum estradiol (E-2), anti-Mullerian hormone (AMH), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) levels to assess the effect of isoflurane on ovarian reserve. Compared to the control group, significant prolongation of the estrous cycle of the adult female mice was observed in the 20 000 ppm isoflurane exposure group. Serum AMH was significantly decreased, and FSH and LH levels profoundly increased in the 5000, 10 000, and 20 000 ppm isoflurane exposure groups compared to the control group. The histopathologic examination revealed a reduced number of developing follicles and an increased number of atretic follicles after isoflurane exposure, but the difference was not statistically significant. Thus, exposure to a higher concentration of isoflurane might have an adverse effect on ovarian reserve in sexually-mature female mice.