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This study was conducted to examine the effect of mothers' perception of birth on separation anxiety, a rarely studied but clinically important topic in perinatal mental health. A descriptive and correlational design was used to identify relationships without intervention. The sample consisted of 305 postpartum women between the seventh day and sixth week, recruited from a private hospital. Data were collected using the Introductory Information Form, the Perception of Birth Scale, and the Adult Separation Anxiety Questionnaire. Statistical analyses included independent <i>t</i>-tests, ANOVA, Pearson correlation, and multiple regression. The mean age of participants was 29.09 ± 6.48 years. Separation anxiety was found to be significantly negatively correlated with the total score of the Perception of Birth Scale (<i>r</i> = -0.43, <i>p</i> ⩽ 0.001). Regression analysis showed that experiences during the pain period (β = -0.32, <i>p</i> < 0.001) and spousal involvement (β = -0.12, <i>p</i> < 0.05) negatively predicted separation anxiety. These two factors explained 21% of the variance. The findings suggest that positive childbirth perceptions, particularly effective pain management and partner support, may reduce maternal separation anxiety. This study contributes an innovative perspective by linking birth perception with separation anxiety, highlighting the need for interventions that promote positive childbirth experiences in order to protect maternal mental health.