Enzyme-catalyzed Michael addition for the synthesis of warfarin and its determination via fluorescence quenching of l-tryptophan

A sensitive fluorescence sensor for warfarin was proposed via quenching the fluorescence of l -tryptophan due to the interaction between warfarin and l -tryptophan. Warfarin, as one of the most effective anticoagulants , was designed and synthesized via lipase from porcine pancreas (PPL) as a biocatalyst to catalyze the Michael addition of 4-hydroxycoumarin to α, β-unsaturated enones in organic medium in the presence of water. Furthermore, the spectrofluorometry was used to detect the concentration of warfarin with a linear range and detection limit (3σ/k) of 0.04–12.0   μmol   L −   1 (R 2   =   0.994) and 0.01   μmol   L −   1 , respectively. Herein, this was the first application of bio-catalytic synthesis and fluorescence for the determination of warfarin. The proposed method was applied to determine warfarin of the drug in tablets with satisfactory results.