A new experimental system was applied to obtain information about the local disposition of lipophilic mitomycin C (MMC) prodrugs after intra-arterial injection. Rabbit hind leg was perfused in situ using a single-pass technique and the outflow concentration-time curves of drugs following pulse injection were evaluated by using statistical moment analysis. Moment parameters (moments) were transformed to disposition parameters which express local disposition characteristics together with respect to elimination, distribution, and dispersion. Six lipophilic derivatives of MMC were tested together with the parent drug. Dimethylsulfoxide (DMSO) was used as a model injection vehicle that can be mixed with blood. Using this system, the following results were obtained. 1) Local disposition of the drug is decided by the balance of its affinities for intravascular and extravascular components. 2) Disposition can be modified by derivation of the drug to a more lipophilic prodrug. 3) Higher lipophilicity of a drug results in a larger organ clearance in the absence of albumin, but a smaller clearance in albumin-containing perfusate. 4) In the case of DMSO injection, intermediate clearance values are obtained regardless of the lipophilicity of drugs, and it was suggested that DMSO decreases both partitioning from the injection vehicle to the tissue and plasma protein binding.