Background: The rodent carcinogen acrylamide was discovered in 2002 in commonly consumed foods. Epidemiologic studies have observed positive associations between acrylamide intake and endometrial, ovarian, and breast cancer risks, which suggest that acrylamide may have sex-hormonal effects. Methods: We cross-sectionally investigated the relationship between acrylamide intake and plasma levels of sex hormones and sex hormone-binding globulin (SHBG) among 687 postmenopausal and 1,300 premenopausal controls from nested case-control studies within the Nurses' Health Studies. Results: There were no associations between acrylamide and sex hormones or SHBG among premenopausal women overall or among never-smokers. Among normal-weight premenopausal women, acrylamide intake was statistically significantly positively associated with luteal total and free estradiol levels. Among postmenopausal women overall and among never-smokers, acrylamide was borderline statistically significantly associated with lower estrone sulfate levels but not with other estrogens, androgens, prolactin, or SHBG. Among normal-weight women, (borderline) statistically significant inverse associations were noted for estrone, free estradiol, estrone sulfate, DHEA, and prolactin, whereas statistically significant positive associations for testosterone and androstenedione were observed among overweight women. Conclusions: Overall, this study did not show conclusive associations between acrylamide intake and sex hormones that would lend unequivocal biologic plausibility to the observed increased risks of endometrial, ovarian, and breast cancer. The association between acrylamide and sex hormones may differ by menopausal and overweight status. We recommend other studies investigate the relationship between acrylamide and sex hormones in women, specifically using acrylamide biomarkers. Impact: The present study showed some interesting associations between acrylamide intake and sex hormones that urgently need confirmation.