1. By performing microdialysis, this study investigated the pharmacokinetics of unbound camptothecin in rat blood, brain and bile in the presence of P-glycoprotein mediated transport modulators (cyclosporin A, berberine, quercetin, naringin and naringenin). Pharmacokinetic parameters of camptothecin were assessed using a non-compartmental model. 2. Camptothecin rapidly crosses the blood-brain barrier (BBB) within 20 min after camptothecin administration. The disposition of camptothecin in rat bile appeared to have a slow elimination phase and a peak concentration after 20 min of camptothecin administration. The area under the concentration versus time curve (AUC) for camptothecin in bile significantly surpassed that in blood, suggesting active transport of hepatobiliary excretion. 3. In the presence of cyclosporin A camptothecin AUC, in the brain, was significantly elevated but no significant change in the presence of berberine, quercetin, naringin and naringenin. 4. With treatment by smaller doses of quercetin (0.1 mg x kg(-1)), naringin (10 mg x kg(-1)) and naringenin (10 mg x kg(-1)), they significantly diminished the camptothecin AUC in bile, but was not altered by the treatment of berberine (20 mg x kg(-1)), a higher dose of quercetin (10 mg x kg(-1)), and cyclosporin A treated (20 mg x kg(-1)) and pretreated groups. 5. The distribution ratio (AUC(bile)/AUC(blood)) of camptothecin in bile was decreased in the cyclosporin A, quercetin, naringin and naringenin treated groups. However, the distribution ratio in the brain was increased in the cyclosporin A groups, but was decreased in the groups treated with quercetin, naringin and naringenin. These results revealed that P-glycoprotein might modulate hepatobiliary excretion and BBB penetration of camptothecin.