The bioanalysis of vinca‐alkaloids has been investigated extensively. High‐performance liquid chromatography coupled to ultraviolet, fluorescence or electrochemical detection have been described. During recent years liquid chromato‐graphy coupled with mass spectrometry (LC‐MS) has become the first choice for the quantitative bioanalysis of the vinca anticancer agents. This paper reviews recent methods for the bio‐analysis of vinca‐alkaloids using LC‐MS, supplemented with our own experience. We will focus on sample pre‐treatment, chromatography and MS detection and pay attention to problems which can occur during the bioanalysis of vinca‐alkaloids. These problems encounter carry‐over and absorption effects and solutions will be provided how to circumvent these problems. Copyright © 2009 John Wiley & Sons, Ltd.