A separation and purification method for vinca alkaloids comprises the following steps: (1) the pretreatment of coarse crude products: putting vinca alkaloids crude product on an aluminum oxide column, eluting and collecting alkaloid fraction, combining and then decompressing and concentrating till being dry to obtain a pretreated product; (2) high performance liquid chromatography separation: firstly preparing chromatographic column stationary phase, filling being silica gel, sample handling, gradient elution, following the situation of the preparative chromatography separation by a detector,thin layer chromatography or high efficient liquid phase detection, and combining enriched vinca alkaloids fraction; (3) recrystallization: decompressing and concentrating alkaloid fraction till being dry, dissolving lower alkyl alcohols, laying aside, crystallizing, filtering and drying. The method is suitable for the separation and purification of the vinca alkaloids, does not need particular filling, also does not need to use high toxicity solvent, is simple to operate, has high automatic degree, good separation effect; the filling used by stationary phase is packed into the column at onetime and used for many times. Mobile phase can be recycled, is environmental friendly, economic and suitable for industrialization production.