The measurement of cortisol in saliva has become a reliable tool for both thc scientist and the clinician for studying adrenal cortical function in the adult. We have measured salivary cortisol in samples from 138 healthy infants, children, and adolescents, and from 14 adults. Saliva samples were obtained at home using a cotton swab and a saliva-collecting tube at 800, 1300, and 1800 h before meals. Cortisol was measured using a time-resolved fluorcsccnt irnmunoassay. Cortisol levels in saliva ranged from less than 2 nmol/L up to more than 100 nmol/L. Cortisol levels were age-dependent. Intcrcstingly, after thc age of 6 y, cortisol levcls corrclated significantly with pubertal stages (analysis of variance). No sex difference was found. In addition, cortisol morning levels and daily cortisol levels (area under the curve from three measurements) increased with body weight and body mass index. The highest cortisol levels were measured in saliva of children younger than 1 y. No circadian variation was evident before thc agc of 9 mo. After 1 y of age, salivary cortisol levels varied in a circadian fashion. The measurement of salivary cortisol levels is an attractive way of testing adrenal function in infants and children. It provides a rcliable tool for the determination of the physiology and developmental characteristics of cortisol metabolism. (Pediatr Res 37: 502-506, 1995)The measurement of cortisol in saliva provides a reliable cents and from 1 4 adults to obtain more insight into regulatory tool for investigations of hypothalamus-pituitary-adrenal axis mechanisms of adrenal function throughout childhood and activity (1-1 1). Saliva samples can be obtained stress-free. In adolescents. addition, salivary cortisol reprcsents the free fraction of cortisol in plasma and does not depend on salivary sccretion rate (flow rate) nor on salivary protein content (4, 5, 9, 11). About METHODS 5-10% of total plasma cortisol is not bound to cortisol-binding Subjects and study protocol. Saliva samples were obtained globulin (= free plasma and diffuses into in the home setting at 800, 1300, and 1800 h before meals. saliva. Salivary cortisol measurements have been extensively Saliva samples were sent to the laboratory by surface mail. A used in psychobiologic, psychiatric, and endocrine re-complete medical examination was performed, and height, search. Surprisingly, relatively little information is available on weight, and pubertal stages were recorded by means of a the use of salivary cortisol determinations in children and standardized examination form. adolescents. Importantly, relatively few studies deal with the Height, weight, and body mass index values of the subjects diurnal variation of cortisol levels in children, developmental were all within the third to 97th percentile range (26). A total and physiologic aspects of function in young age, and of 178 healthy subjects were recruited from an inner city influ' ' ' ' of and pubertal On neighborhood (Munich, Germany), the local kindergarden, and cortisol levels (12)(13)(14)...