The interaction between mitiglinide calcium and bovine serum albumin (BSA) is studied by fluorescence emission spectroscopy, UV-visible absorption spectroscopy and synchronous fluorescence spectroscopy. Experimental results show that mitiglinide calcium leads to the fluorescence quenching of BSA, and the quenching mechanism is a dynamic quenching procedure according to the Stern-Volmer equation. The binding distance between mitiglinide calcium and BSA is calculated to be about 5.461 nm based on F rster theory. The effect of mitiglinide calcium on the conformation of BSA is studied by synchronous fluorescence spectroscopy. The results show that a slight blue shift is observed when the difference between excitation wavelength and emission wavelength is 60 nm. The blue shift indicates that the peripheral microenvironment near tryptophan residue of BSA is affected by the mitiglinine calcium molecule, which makes the polarity of the microenvironment decrease and the hydrophobicity of BSA increase.