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Determination of lipase activity is also important in toxicology, when lipase activity can be both increased and decreased by organophosphates and other pesticides and in medicine for diagnosis of heart diseases. The standard method for lipase activity determination is based on cleaving ester bonds in lipase buffer containing Tween. Our aim was to find a method with faster and more sensitive response. It is known that acetylcholinesterase belongs to the same group of hydrolases enzymes as lipases and it cleaves indoxyl acetate, so we assume indoxyl acetate could report a similar reaction with lipase. Our method is based on indoxyl acetate as a substrate for lipase, where indoxyl acetate is cleaved by lipase to indoxyl and acetate moiety and blue indigo is created. The method was optimized for different times and amount of enzyme and compared with the standard Tween assay. The calibration curve measured in reaction time 20 minutes with 10\u2009<jats:italic>\u03bc<\/jats:italic>l of lipase exhibited the best analytical parameters, and it showed Michaelis\u2013Menten response with the Michaelis\u2013Menten constant equal to 8.72\u2009mmol\/l. The indoxyl acetate-based method showed faster and more sensitive response than the standard method for lipase activity determination, so it has great potential in biosensor construction and it could be used in industry, medicine, toxicology, and common practice where the activity of lipases is need to be measured.<\/jats:p>","DOI":"10.1155\/2019\/8538340","type":"journal-article","created":{"date-parts":[[2019,12,2]],"date-time":"2019-12-02T14:16:30Z","timestamp":1575296190000},"page":"1-7","source":"Crossref","is-referenced-by-count":10,"title":["Indoxyl Acetate as a Substrate for Analysis of Lipase Activity"],"prefix":"10.1155","volume":"2019","author":[{"given":"Tomas","family":"Valek","sequence":"first","affiliation":[{"name":"Faculty of Military Health Science, University of Defense, Trebesska 1575, 50001 Hradec Kralove, Czech Republic"},{"name":"Faculty of Pharmacy, Charles University, Akademika Heyrovskeho 1203, 50005 Hradec Kralove, Czech 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