In 20 women with proven fertility, one menstrual cycle was monitored by ovarian ultrasonography, laparoscopy and estimation of 17 beta-estradiol (E2), estrone (E1), progesterone, testosterone, androstenedione (Adion), dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), and sex hormone binding globulin (SHBG) in serum and peritoneal fluid. Three groups were studied, in which the peritoneal fluid was collected within 1, 3 and 5 days after ovulation. E1, E2 and progesterone levels in peritoneal fluid were highest shortly after ovulation and decreased with time. Testosterone and Adion in peritoneal fluid showed no changes, but peritoneal fluid levels were always higher than serum levels. No differences were found between the peritoneal fluid and serum levels of DHEA and DHEAS. SHBG in serum was always higher than in peritoneal fluid. The results are compared with reported steroid levels in follicular fluid from the literature and factors complicating the interpretation of steroid levels in peritoneal fluid are discussed.