To determine the most useful index of pubertal gonadotropin secretion we measured spontaneous LH and FSH levels every 20 min for 24 h and the LH and FSH responses to LHRH in a total of 37 girls and 30 boys representing each of the 5 stages of puberty. Mean 24-h LH and FSH levels rose significantly with increasing pubertal stage in both girls and boys. LH peak amplitude increased significantly with increasing pubertal stage for both sexes, whereas FSH peak amplitude did not. LH and FSH peaks were present throughout the 24-h period in all children, but the frequency did not change significantly with increasing pubertal stage. Mean gonadotropin levels, peak amplitudes, and peak frequencies tended to be higher at night from pubertal stages 1-4 of puberty. There were no significant sex differences in mean LH, LH peak amplitude, or LH peak frequency. The LHRH-stimulated peak LH to peak FSH ratio was greater in boys than girls during pubertal stages 1-3 and was less useful in distinguishing pubertal from prepubertal boys. For girls, the most accurate index of pubertal gonadotropin secretion was a LHRH-stimulated peak LH to peak FSH ratio greater than 0.66, which detected 96% of the pubertal girls with no false positives. For boys, the most accurate index was a maximum spontaneous nighttime LH level of 12 IU/L or more, which detected 90% of the pubertal boys with no false positives. We conclude that there are important sex differences in the gonadotropin responses to LHRH during puberty, and that criteria for the onset of pubertal gonadotropin secretion should be sex specific.