The human metabolism of 4‐hydroxyanisole was investigated by the analysis of urine samples from melanoma patients treated with this substance. The samples were hydrolyzed with glucoronidase and/or arylsulphatase, extracted with ethyl acetate, and, after derivatization with pentafluoropropionylanhydride, analyzed by gas chromatography‐mass spectrometry. We were able to identify peaks by their retention times and mass spectra corresponding to 4‐hydroxyanisole, 3,4‐dihydroxyanisole, and two of its o‐methyl derivatives, namely, 3‐hydroxy‐4‐methoxy‐ and 4‐hydroxy‐3‐methoxyanisole. We also detected a peak of hydroquinone, which may have originated (at least partly) from 4‐hydroxyanisole. All the above‐mentioned compounds were excreted predominantly as sulphates and glucuronides. Only a small proportion of the substances was present in urine in an unconjugated form. Our results demonstrate that 3,4‐dihydroxyanisole is the most important m
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