Objective To analyze the disparities in social, emotional, and cognitive behaviors between male and female mice with autism spectrum disorder (ASD) induced by various valproic acid (VPA) exposure method, and to establish a foundation for selecting experimental animal sex and induction method in the VPA-induced ASD model. Methods At 12. 5 days of gestation, mice received 600 mg / kg VPA injected intraperitoneally or an equal volume of 0. 9% sodium chloride solution. The offspring were designated as the prenatal (Preg) VPA group and Preg control group, respectively. At 14 days post-birth, mice were administered 400 mg / kg VPA injected subcutaneously (SC VPA group), with the corresponding control group receiving an equal volume of 0. 9% sodium chloride solution (SC control group). Additionally, on postnatal day 14, another group of mice received intraperitoneal injection of 400 mg / kg VPA (IP VPA group), and its control group received an equal volume of 0. 9% sodium chloride solution (IP control group). Mortality and abortion rates were monitored post-administration, and the impact of various VPA exposure method on the social, emotional, and cognitive behaviors of male and female mice was examined through a series of three-box social behavior experiments, self-grooming assessments, open field tests, Y-maze trials, and water maze evaluations. Results The mortality rate among pregnant rats at 12. 5 days of gestation following VPA injection was 50%, and the abortion rate was 25%. No fatalities occurred following postnatal 14 d mice subcutaneous or intraperitoneal VPA administration. The three-box social experiment, self-grooming experiment, and open field experiment indicated that both sexes of VPA mice displayed social impairments, novelty-induced social deficits, repetitive stereotyped behaviors, and anxiety-like behaviors in comparison to the control group, irrespective of the injection method. Y-maze spontaneous alternation and water maze experiments indicated that male mice in the Preg VPA group and the SC VPA group both exhibited cognitive deficits, whereas those injected with IP VPA group demonstrated a propensity for cognitive impairment without statistical significance; female mice exposed to VPA through various method did not exhibit cognitive impairments. Conclusions The mortality and abortion rates of pregnant mice exposed to VPA before birth were higher, as no deaths occurred after subcutaneous or intraperitoneal injection of postnatal 14 d mice. Male mice in the Preg VPA group and the SC VPA group had autism-like characteristics, including diminished social interaction, repeated stereotyped actions, heightened anxiety, and cognitive impairment; Results in female mice were unstable. The IP VPA group led to moderate social and social novelty deficits with slight anxiety-like behaviors, but no significant cognitive impairment.