The effectiveness of oral contraceptives with concomitant use of ampicillin has been questioned. Eleven volunteer subjects were studied during 2 consecutive menstrual cycles; they were given Demulen with placebo or ampicillin during each of the cycles. Serum estradiol (E2), progesterone (P4), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were evaluated on days 13 and 19 of each cycle. Testosterone-binding globulin (TeBG) capacity and ferning of cervical mucus were evaluated on day 13 of each cycle. Subjective symptoms were recorded. All cycles appeared to be anovulatory. Steroid suppression was present in 10 of 11 patients during both cycles. Estrogen-dependent TeBG was not affected by ampicillin. It is possible that cervical mucus ferning was improved by concurrent ampicillin administration. Ampicillin appears unlikely to diminish the effectiveness of the oral contraceptive studied.