Zidovudine 3'-azido-3'-deoxythymidine is the drug chosen for the treatment of patients suffering from AIDS; zidovudine being a potent inhibitor of HIV replication. The drug is extensively metabolized by conjugation with glucuronic acid into an inactive compound, and 30-40% of the dose is eliminated presystemically. We studied the variability and characterized the frequency distribution of the activity of zidovudine glucuronosyl transferase in 93 specimens of human liver. A rapid and reproducible radiometric assay for the glucuronidation of 14C-zidovudine is reported. The method is based on the extraction of the unreacted zidovudine into organic solvents and the radioactivity of the unextractable zidovudine glucuronide was measured in the aqueous phase residue. The rate of zidovudine glucuronidation was neither sex- nor age-dependent, ranged over 1 order of magnitude, and was positively skewed. The possibility that endogenous bilirubin should interact with glucuronidation of zidovudine was explored and the endogenous concentration of bilirubin was measured in the microsomal preparations of 59 liver samples. The final concentration of bilirubin in the assay mixture for zidovudine glucuronidation ranged between 2.2 and 13.2 microM and did not interact with the rate of zidovudine glucuronidation. The kinetics of glucuronosyl transferase towards zidovudine was studied in 20 livers, Michaelis-Menten kinetics were observed and the K(m) estimate ranged over 2-fold with an average of 2.89 mM. These in vitro results are consistent with the view that the rate of glucuronidation varies over 1 order of magnitude in the human liver and its distribution is positively skewed. This variability may modulate the patient's exposition to zidovudine and thereby the efficacy of therapy.