In nonaqueous medium, a series of troxerutin amidation analogues were synthesized by the reaction of vinyl-troxerutin esters with fatty amines. The influence parameters including enzyme source, the amount of the enzyme, reactive medium, water content of solvents and reaction time were investigated. The results showed that the amidation occurred regioselectively at vinyl-ester bond. Using pyridine as solvent (water content <1%), the amidation reaction catalyzed by lipase LS-10 (20 mg/mL) reached plateau after 21 h at 50 ℃.