Hollow fiber liquid-liquid phase microextraction followed by high-performance liquid chromatography was employed to research the binding of four furocoumarin compounds (psoralen, oxypeucedanin, imperatorin and isoimperatorin) to bovine serum albumin. The results reveal that the percent of drug-protein binding did not rely on the furocoumarins and protein concentrations, and four furocoumarins scarcely competed for protein binding sites. The protein-binding percents of psoralen, oxypeucedanin, imperatorin and isoimperatorin were 55.1 ± 4.5, 14.4 ± 2.4, 44.1 ± 6.7, and 41.3 ± 3.8%, respectively, the numbers of binding sites were 1.2, 1.6, 0.6 and 0.3, and the binding constants were 1,801, 212, 937 and 4,025 L mol -1 , respectively. The proposed new approach is simple, rapid, and effective for the study of the interaction between protein and furocoumarin compounds which is either individual or coexistent with the similar ones.