The concentration of melatonin and LH were determined in plasma samples obtained at 10-min intervals during 4 h of darkness (00.00-04.00 h) from 4 normal women, age 23-27 years, in the early follicular phase of the menstrual cycle and in 6 normal men, age 23-31 years. Additionally, melatonin concentration was determined in samples obtained from the men at 10-min intervals for 4 h during the day (10.00-14.00 h). A pulsatile pattern of melatonin secretion was found for all the subjects during darkness. There was no significant difference between women and men as to the number of pulses (2.8 ± 0.5 vs 5.2 ± 1.0 per 4 h), amplitude of pulses (51.3 ± 28 vs 27.2 ± 6 ng/l), concentration per 4 h (32.5 ± 13 vs 31.0 ± 5 ng/l), or apparent half-life of melatonin (19.3 ± 2.3 vs 15.3 ± 7.5 min). The mean amplitude of the melatonin pulse correlated (r = 0.863, p<0.001) with the mean melatonin concentration per 4 h. A pulsatile LH secretion pattern was found for the 10 subjects and did not correlate significantly with the melatonin secretion pattern. The results are consistent with an independent signal for the demonstrated nyctohemeral pulsatile melatonin and LH secretions.