Vitiligo is characterized by a progressive loss of inherited skin color. The cause of the disease is still unknown. To date, there is accumulating in vivo and in vitro evidence for massive oxidative stress via hydrogen peroxide (H₂O₂) and peroxynitrite (ONOO^–) in the skin of affected individuals. Autoimmune etiology is the favored theory. Since depletion of the essential amino acid L‐tryptophan (Trp) affects immune response mechanisms, we here looked at epidermal Trp metabolism via tryptophan hydroxylase (TPH) with its downstream cascade, including serotonin and melatonin. Our in situ immunofluorescence and Western blot data reveal significantly lower TPH1 expression in patients with vitiligo. Expression is also low in melanocytes and keratinocytes under in vitro conditions. Although in vivo Fourier transform‐Raman spectroscopy proves the presence of 5‐hydroxytryptophan, epidermal TPH activity is completely absent. Regulation of TPH via microphthalmia‐associated transcription factor and L‐type calcium channels is severely affected. Moreover, dopa decarboxylase (DDC) expression is significantly lower, in association with decreased serotonin and melatonin levels. Computer simulation supports H₂O₂/ONOO^–‐mediated oxidation/nitration of TPH1 and DDC, affecting, in turn, enzyme functionality. Taken together, our data point to depletion of epidermal Trp by Fenton chemistry and exclude melatonin as a relevant contributor to epidermal redox balance and immune response in vitiligo.—Schallreuter, K. U., Salem, M. A. E. L., Gibbons, N. C. J., Martinez, A., Slominski, R., Lüdemann, J., Rokos, H. Blunted epidermal L‐tryptophan metabolism in vitiligo affects immune response and ROS scavenging by Fenton chemistry, part 1: epidermal H₂O₂/ONOO^–‐mediated stress abrogates tryptophan hydroxylase and dopa decarboxylase activities, leading to low serotonin and melatonin levels. FASEB J. 26, 2457‐2470 (2012). www.fasebj.org