Early loss experience (ELE) due to death or separation is a major risk factor for the development of several psychiatric and physical disorders in adulthood. Few studies have focused on the effects of ELE on neuroendocrine systems, which might mediate this risk in part. The goal of this study was to evaluate salivary cortisol responses to awakening in individuals with and without ELE. A total of 95 healthy college students (29 men, 66 women) completed a questionnaire on ELE and were instructed to collect saliva immediately after awakening and 30 min later. Fifty-five of the 95 subjects reported having experienced the separation or divorce of their parents and/or the death of a close relative before the age of 14 years. Subjects with such ELE exhibited decreased salivary cortisol responses to awakening compared to subjects without ELE (net increase: 4.78 nmol/l versus 9.83 nmol/l; t93 = 2.88, p = 0.005). The effect was most pronounced in individuals who experienced multiple types of ELE, while there were no sex differences. In conclusion, ELE appears to be associated with decreased salivary cortisol responses to awakening. Low cortisol awakening responses are believed to reflect altered dynamics of the hypothalamic-pituitary-adrenal (HPA) axis, possibly conferring risk for certain stress-related disorders.