Salivary testosterone measurements: reliability across hours, days, and weeks

Salivary testosterone measurements would appear to be useful in behavioral research, where subjects are often reluctant to provide serum samples. The usefulness of salivary measurements depends upon their reliability, however, which was the focus of the present investigation. In four studies, 270 male and 175 female subjects collected saliva samples at times ranging from 30 min to 8 weeks apart. Subjects collected samples on at least two days, at time of awakening, midmorning, late afternoon, and late evening. Mean testosterone concentration dropped about 50% from morning to evening for both sexes, with largest drops early in the day. Mean reliability was r = .64 across two days and r = .52 across seven-eight weeks. Menstrual cycle effects were negligible. Reliability can be increased by using more than one measurement, and it is probably desirable to combine measurements taken several weeks apart. Salivary assays offer a practical way of measuring testosterone in free-ranging subjects outside the laboratory.