Although widely cited as strong evidence that sexual selection has shaped human facial attractiveness judgments, evidence that preferences for masculine characteristics in men's faces are related to women's hormonal status is equivocal and controversial. Consequently, we conducted the largest ever longitudinal study of women's preferences for facial masculinity (N=584). Analyses showed no evidence that preferences for facial masculinity were related to changes in women's salivary steroid hormone levels. Furthermore, both within-subject and between-subject comparisons showed no evidence that oral contraceptive use decreased masculinity preferences. However, women generally preferred masculinized over feminized versions of men's faces, particularly when assessing men's attractiveness for short-term, rather than long-term, relationships. Our results do not support the hypothesized link between women's preferences for facial masculinity and their hormonal status.