We have previously shown expression of CRH and POMC genes and peptides in the human skin. To ascertain the identity of those peptides, we used methods of peptide extraction and purification combined with the highly specific technique of liquid chromatography-mass spectrometry. Testing extracts of human skin, we identified endogenous peptides with masses and retention times corresponding to CRH, ACTH 1-39, ACTH 1-13, and alpha-MSH standards. Thus, conclusive evidence is provided for the presence of CRH and the POMC-derived ACTH 1-39, ACTH 1-13, and alpha-MSH peptides in human skin. Direct identification of these peptides is consistent with translation of the corresponding genes, and it also suggests intermediate pituitary lobe-like POMC peptide processing.