An instrumental quantitative high-performance thin-layer chromatographic (HPTLC) method has been developed for the determination of vinca-alkaloids (antineoplastic compounds) in chemotherapeutic infusion bags prepared in a hospital pharmacy. The method uses automated band application onto silica gel plates containing a fluorescent indicator and scanning densitometry of fluorescence-quenched zones of samples and standards. Samples were analyzed to check the content of the active substance against the label declaration of the preparation. The four compounds were separated using the following solvent system CH(2)Cl(2)-CH(3)OH (93:7, v/v). Vincristine (VCR) and vinorelbine (NVB) were assessed in the same run whilst vinblastine (VLB) and vindesine (VDS) were analyzed in a second run. HPTLC allows the identification and the quantitation of more than 20 samples in the same chromatographic run. The analysis of the samples requires 30 min compared with more than 2 h using a typical HPLC method. Moreover, there is no need for a conditioning step, as with HPLC, and each analysis by HPTLC is less expensive.