Gender differences in nicotine response with regard to nicotine sensitivity and withdrawal symptomatology have been reported, with the suggestion that ovarian hormones play a role. Few studies, however, have directly assessed hormonal influences on nicotine response. This study focused on the effects of a transdermal nicotine patch in women during acute smoking abstinence when tested in different phases of their menstrual cycle. Thirty women were randomized to order of menstrual cycle phase (late luteal or follicular) and patch condition (active or placebo). Two 7-day outpatient-testing periods were conducted with 2 days of ad lib baseline smoking and 5 days of smoking abstinence. Dependent measures included scores from the Minnesota Nicotine Withdrawal Scale, Questionnaire on Smoking Urges, and Premenstrual Assessment Form, as well as weight. The severity of both premenstrual symptomatology and nicotine withdrawal symptoms was greater in the late luteal phase. Correlation coefficients confirmed overlap between premenstrual and withdrawal symptomatology, especially for the affect subscale. A significant patch effect was observed, showing diminished craving and premenstrual affect on pain subscale scores for women on active patch. Results showed that nicotine craving and premenstrual pain and water retention symptoms were diminished in women on transdermal patch, and that this effect was greatest in the late luteal phase. In addition, the greatest weight gain was demonstrated for participants in the late luteal phase, placebo condition. In summary, during short-term smoking abstinence in women, transdermal nicotine appears to have a more pronounced effect in the late luteal phase than in the follicular phase in reducing craving and certain premenstrual symptoms.