. Split-ejaculate samples were provided at 0, 2, 5, and 8 h after drug ingestion, on separate days after 3 days of sexual abstinence. NVP and IDV showed time-dependent concentrations in seminal plasma, with peak concentrations in both fractions at 2 and 2 to 5 h, respectively, after drug ingestion. The NVP concentrations were not significantly different between the first and second fractions of the ejaculate at all time points measured and were in the therapeutic range, except for the predose concentration in two patients. The median (range) predose IDV concentrations in the first and second fractions of the ejaculate were 448 (353 to 1,015) ng/ml and 527 (240 to 849) ng/ml, respectively (P ؍ 0.7). In conclusion, NVP and IDV concentrations in seminal plasma are dependent on the time after drug ingestion. Furthermore, our data suggest that NVP and IDV achieve therapeutic concentrations in both the testes and prostate and the seminal vesicles throughout the dosing interval.Penetration of antiretroviral drugs into all body compartments is important in the treatment of human immunodeficiency virus (HIV) infection. The central nervous system and the male genital tract are considered anatomical reservoirs for HIV, as the blood-brain barrier and the blood-testis barrier may prevent antiretroviral drugs from entering these organs. Suboptimal antiretroviral drug concentrations in the male genital tract could allow continuing production of HIV-1 and the emergence of drug-resistant HIV type 1 (HIV-1) strains (4-6, 22).Data available on drug concentrations in semen show that the penetration of the protease inhibitors nelfinavir, ritonavir (RTV), and saquinavir is poor (14; M. Reijers, R. van Heeswijk, H. Schuitemaker, P. Portegies, G. J. Weverling, J. Lange, and R. Hoetelmans, 7th Conf. Retrovir. Opportunistic Infect., abstr. 316, 2000). The nucleoside analogues zidovudine c Plasma HIV-1 RNA was measured using the NucliSens HIV-1 QT assay (Organon Teknika). d Plasma HIV-1 RNA was measured using the quantiplex bDNA assay (Bayer Corporation).