A multi-segment chromatographic column comprises a main liquid storage bottle and a plurality of auxiliary liquid storage bottles connected the main liquid storage bottle. The bottoms of the main liquid storage bottle and the plurality of auxiliary liquid storage bottles are all provided with throttling valves. Packing columns are arranged between the main liquid storage bottle and the auxiliary liquid storage bottles and among the auxiliary liquid storage bottles. T-shaped three-way tubes are arranged below the packing columns, each t-shaped three-way tube is provided with a three-way valve, each auxiliary liquid storage bottle is provided with a dissolvent filling port, a tail-segment packing column is arranged below the last auxiliary liquid storage bottle, and a valve is arranged at the lower end of the tail-segment packing column. According to actual needs, the quantity of the packing columns, packing, and packing of the ail-segment packing column can be changed flexibly, multi-stage separation of a complex mixture can be achieved, the packing columns have separation functions, the liquid storage bottles can bring convenience to addition of eluent, and the eluent does not need to be added manually and constantly. The multi-segment chromatographic column is simple in structure, convenient in use and good in effect.