The bio-analysis and pharmacokinetics of vinca alkaloids have been the subject of many investigations. In most cases radiolabelled compounds have been used for quantification purposes. Although this method lacks selectivity, it has provided valuable information on tissue distribution of unchanged drug and metabolites in an early stage of clinical and preclinical investigations. During the last few years, methods based on high-performance liquid chromatography have been presented. This paper reviews the methods described in the literature for the bio-analysis of vinca alkaloids, supplemented with our own experience in this field. Special attention is paid to the problems that may arise during the analytical processes.