Rutin is an effective constituent of some chinese medicinal herb. Interaction study of the medicament with albumin not only can expound the relation of structure and function of the protein and also the action mechanism of the medicament. By using fluorescence polarization and energy transfer technique, we studied on the interaction of human serum albumin (HSA) with medicaments vitamin B₆ and rutin. The dissociation constants of medicaments with albumin were obtained. The distances between the number 214 tryptophane and binding sites of vitamin B₆ and rutin on the protein are 23.4Å and 24.02Å respectively.