A high performance liquid chromatographic method with quaternary gradient elution and fluorometric detection was developed for profiling of tryptophan (TRP), 5-hydroxytryptophan, serotonin (5-HT) and 5-hydroxyindole-3-acetic acid (5-HIAA) in urine, platelet-rich plasma and (tumour) tissue of patients with carcinoid tumours. Prior to injection, urine samples were diluted and filtered. Platelet-rich plasma and tissue homogenates were prepurified by C18 solid phase extraction. Detection limits were approx. 2 pmol. Results of urinary 5-HT and 5-HIAA compared favourably with those of single component analyses. No consistent diurnal variations were found for TRP, 5-HT and 5-HIAA in 12-h urine samples from 15 healthy adults. Abstinence of 5-HT-rich foods reduced urinary levels of 5-HT and 5-HIAA. C18 extraction of indoles from protein-containing matrices was studied in platelet-rich plasma. Although time-consuming and complicated for daily routine use, the present approach offers particular advantages over single component analyses in the study of TRP metabolism in patients with carcinoid tumours.