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Malignant melanoma (MM) is a cancerous transformation of melanocytes. It is a disease with the worst response to therapy and, compared to other malignancies, presents much earlier with metastases. MM still belongs to relatively late-detected malignant diseases. Even so, the MM mortality rate is up to 96% for a relatively small incidence (5%). The gold standard for MM diagnosis is a histopathological examination that requires invasive surgery. An invasive sampling method of a biological material can be a stressful factor for the patient, which is often the reason why patients do not seek medical assistance as soon as possible. Our goal was to find a link between metabolites in urine and the stage of MM. Two excitation peaks at 360–370 nm and 450 nm were characterised in spectra of urine samples. The emission spectra have shown one significant peak at 410–460 nm. After addition of glutathione reductase to the samples, fluorescence dropped down only in patient samples and hidden fluorophores appeared. Malignant diseases are associated with the presence of specific metabolites that can be detected fluorescently in biological material such as urine, which can be a suitable alternative for an early detection of cancer or for tracking changes during and after treatment.