The synthesis of a number of N-acyl-5-aminosalicylic acids and their derivatives is described. These compounds allow the sensitive and specific mass spectrometric determination of unlabelled or deuterium-labelled N-acetyl-5-aminosalicylic acid in biological samples. An in vivo study shows that the labelled compound, administered to rats, is excreted isotopically unchanged to at least 97%, and that no significant deacetylation/acetylation mechanism exists for this metabolite N-acetyl-5-aminosalicylic acid of salicylazosulphapyridine.