A new approach is reported, based on Proton Nuclear Magnetic Resonance (H-1 NMR) spectral data, to study qualitatively and quantitatively the hydrolysis level in complex lipid mixtures. It is based on a detailed study of the spectra of tri-, di-, mono-glycerides and fatty acids. It has been validated by using mixtures of standard compounds which simulate the complex mixtures formed in lipid digestion. To this aim the 1H NMR spectra of pure compounds, as well as of mixtures containing different known proportions of triglycerides, 1,2-diglycerides, 1,3-diglycerides, 1-monoglycerides, 2-monoglycerides and fatty acids, were acquired. These mixtures cover a very broad range of concentrations and simulate edible oils and fats of vegetable and animal origins, including fish, with different levels of hydrolysis. Equations based on spectral data are proposed to determine the molar percentage of the different kinds of compounds that may be formed in lipid digestion, thus allowing quantification of the advance of lipolysis. To the best of our knowledge, this is the first time that a method based on 1H NMR spectral data to quantify triglycerides, as well as partial glycerides and fatty acids in complex lipid mixtures coming from lipolysis is proposed. This methodology offers many advantages and is useful not only in the fields of food technology and nutrition, but also in others like enzymology, pharmacology, medicine and petrochemistry, among others. (C) 2014 The Authors. Published by Elsevier Ltd.