An h.p.l.c. method for the simultaneous determination of naphthol, naphthyl glucuronide and naphthyl sulphate in urine is described. This procedure is based on the selective formation of an ion pair between the sulphate conjugate and tetrabutyl lammonium, which allows its resolution from the glucuronide on a reverse phase column. 2. The h.p.l.c. method is sed to assess the selective enzyme hydrolysis method which is frequently employed in drug conjugate assays. Results suggest that the use of sulphatase and beta-glucuronidase may give data which are quantitatively and qualitatively erroneous.