The relationship between tumor physiology and the pharmacokinetics of 2',2' difluorodeoxycytidine [gemcitabine (dFdC)] in ex vivo perfused human small cell lung cancer was examined. Two small cell lung cancer sublines, 54A and 54B, with known in vivo sensitivity to dFdC, were grown as tissue-isolated tumors in athymic mice and perfused ex vivo with or without 20-40 micrometer dFdC. Arteriovenous differences in gases, pH, and metabolites were determined before and during drug infusion. The geometric flow resistance (FR) of individual tumors was calculated, and dFdC and its inactive metabolite 2',2' difluorodeoxyuridine were determined by high-performance liquid chromatography of consecutive samples from the output line. Both tumors had prominent lactate production concurrent with a significant O2 consumption. The arteriovenous pH drop was approximately 0.3 in both tumor lines. Significant metabolic differences between 54A and 54B tumors were found that elucidated previously described differences further. Pharmacokinetic analysis showed that the initial tumor uptake of dFdC was flow limited, and a significant inverse correlation between the geometric FR and initial drug uptake was found. The rate constant for recovery of the drug in the tumor outflow was greater in 54B tumors (P < 0.05), and the geometric FR was greater in 54A tumors (P < 0.01). The drug conversion rate was independent of physiological parameters. Attempts to modify the delivery of dFdC should be directed at the tumor blood flow distribution. More generally, our experimental model provides useful new insight into metabolism and intratumor pharmacokinetics of chemotherapeutic agents in solid tumors.