Quantitative Methods for Assessing True Lipase Activity Using Long-Chain Triglycerides and Complex Lipid Substrates

Lipases (E.C. 3.1.1.3) are enzymes that catalyze the hydrolysis of triacylglycerols (TAGs) into glycerol and fatty acids (FAs), making them essential for various industrial applications, including food processing, detergents, pharmaceuticals, and biofuels. This chapter focuses on methods used to assess true lipase activity, specifically those for measuring the release of FAs from long-chain triglycerides with varying saturation levels (e.g., C8:0, C10:0, C12:0, C14:0, C16:0, C18:0, C18:1), as well as complex, standardized lipid substrates such as oils, mayonnaise, lipstick, sebum, beef fat, and butterfat. These are assessed using the non-esterified free fatty acid (NEFA/FFA) colorimetric assay. The combined use of triglycerides, standardized test materials (e.g., stained fabrics), and the NEFA assay provides a more practical and realistic evaluation of true lipase activity, particularly relevant for industrial applications such as detergents.