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Inhibition of lipase activity is one of the approaches to reduced fat intake with nutritional prevention promoting healthier diet. The food industry is very interested in the use of natural extracts, hence reducing the side effects of commercial drugs inhibiting lipolysis. In this work we propose a novel methodology to rapidly assess lipolysis/inhibition in a single droplet by interfacial tension and dilatational elasticity. The evolution of the interfacial tension of lipase in simplified duodenal fluid in the absence and that in the presence of the pharmaceutical drug Xenical are the negative (5 ± 1 mN/m) and positive (9 ± 1 mN/m) controls of the inhibition of lipolysis, respectively. Then, we correlate the inhibition with the reduction of the interfacial activity of lipase and further identify the mode of action of the inhibition based on dilatational response (conformational changes induced in the molecule/blocking of adsorption sites). This …