The aim to assess quality of life (QoL) and profile of mood states in patients with childhood-onset hypopituitarism and growth hormone (GH) deficiency during the discontinuation of recombinant growth hormone (rGH) replacement therapy at the transition from adolescence to adulthood. Methods mood states and QoL were assessed in 136 (85 men, 51 women, age 16.8 +/- 1.1 years) patients during discontinuation of rGH replacement and 82 (40 men, 42 women, age 16.5 +/- 1.7 years) control individuals using the Profile of Mood States2 (POMS2) and the Quality of Life - Assessment of Growth Hormone Deficiency in Adults (QoL-AGHDA) questionnaires. Results No difference in QoL was between patients and controls, but patients had higher levels of tension-anxiety (median 9.0 score vs. 6.0 score, p < 0.001), depression-dejection (median 9.0 score vs. 8.0 score, p = 0.042), fatigue-inertia (median 9.0 score vs. 6.0 score, p < 0.001), confusion-bewilderment (median 4.0 score vs. 3.0 score, p = 0.003) than controls. Basal cortisol concentration correlated negatively with QoL-AGHDA score (r = -0.288, p = 0.012), depression-dejection score (r = -0.317, p = 0.005). Height correlated negatively with anger-hostility (r = -0.223, p = 0.010), fatigue-inertia (r = -0.188, p = 0.030) scores. Body mass index (BMI) correlated positively with fatigue-inertia score (r = 0.181, p = 0.040). Age at the discontinuation of rGH replacement correlated negatively with QoL-AGHDA score (r = -0.204, p = 0.026), depression-dejection (r = -0.460, p = 0.021), anger-hostility (r = -0.190, p = 0.040), fatigue-inertia (r = -0.205, p = 0.026) scores. Conclusion mood states (higher tension-anxiety, depression-dejection, fatigue-inertia, confusion-bewilderment), but not disease-specific QoL are impaired in patients with childhood-onset growth hormone deficiency during the discontinuation of rGH replacement therapy at the transition from adolescence to adulthood.