Hollow fiber liquid phase microextraction (HF-LPME) coupled with high performance chromatography (HPLC) was applied to research the binding of four furocoumarins (psoralen, oxypeucedanin, imperatorin and isoimperatorin) to bovine serum albumin. The results suggest that the percent of four furocoumarins-protein binding should not rely on the concentrations of furocoumarins and protein, and four target drugs scarcely compete for binding bites of protein. Hence, furocoumarin-protein binding percents can be determined simultaneously and rapidly by the approach.