In the non-aqueous solvent system, a ternary complex was synthesized with o-vanillin-glycine Schiff base, 8-hydroxyquinoline and anhydrous bismuth chloride, which was confirmed to be [Bi(C10H10NO4)(C9H6NO)2], by elemental analysis, thermal analysis, spectral analysis, etc. Phenolic hydroxyl oxygen and the nitrogen of carbon-nitrogen double bond in the Schiff base were bonded to Bi(III) ion, and the oxygen atom and heterocyclic nitrogen atom of 8-hydroxyquinoline were bidentate-coordinated with Bi(III) ion, forming a five-member chelate ring. In particular, the interaction between the complex and bovine serum albumin (BSA) was investigated by fluorescence spectroscopy at different temperatures. It was proved that the fluorescence quenching of BSA by Schiff base-complex was a result of the formation of a complex with a stoichiometric ratio of 1:1. The quenching constant was calculated to be 6.21×104 L/mol. And the positive ΔH and ΔS values indicated that hydrophobic force might play a major role during the binding process. Finally, synchronous fluorescence spectroscopy was applied to examine the effect between complex and BSA. The tryptophan residue was found to decrease with a red shift of emission wavelength, which demonstrated that the complex changed the conformation of BSA.