HPLC method for determination of sophoricoside was established and was applied in the teaching process of instrumental analysis experiment for medical laboratory students. By introduction of the principle of HPLC, experiment operation, experiment report writing, laboratories visiting, animation demo of components of HPLC and discussion on relevant knowledge,students could not only master the basic theory and operation of HPLC, but also improve interests and independence in learning. It also improved teaching quality of instrumental analysis experiment.