Abstract
Lipids are essential components of all living cells. In an oenological context, the supply of unsaturated lipids in grape juice allows the yeasts to grow and ferment, despite very low levels of oxygen. The current study proposes a systematic optimization procedure for the analysis of fatty acids and sterols relevant to the grape fermentation process, including both extracellular and intracellular (i.e., yeast cells) lipids. Even though it was extensive, the sample preparation yielded reproducible results for all compounds of interest. The stability of the analyzed compounds was also tested to offer some implementation flexibility for the extensive procedure. The performance parameters (i.e., selectivity, linearity, limit of detection and quantitation, accuracy, and precision) indicated that the method was suitable for future practical implementation. The proof of concept also suggests that the list of compounds of interest can be expanded if additional peaks are identified. Given the large variation in concentrations, the dilution of the matrix needs to be carefully considered in order to ensure that the lipids of interest are still within the dynamic range and not below the limit of detection and/or quantification.
Highlights
Lipids are defined as hydrophobic molecules that are soluble in organic solvents and can be classified according to their molecular structure as well hydrolysis products/chemical backbone
Ergosterol is the main sterol produced in yeasts and, to fatty acids, this sterol is synthesized with acetyl-CoA as the main substrate [16]
As lipids are one of the main classes of molecules considered ‘building blocks’ of all biological organisms, their analysis has been developed for a variety of applications
Summary
Lipids are defined as hydrophobic molecules that are soluble in organic solvents and can be classified according to their molecular structure as well hydrolysis products/chemical backbone. Lipids can be regarded as simple (sterols, fatty acids, etc.) or complex (glycerophospholipids, glycolipids); the former yields, at most, two products after hydrolysis, and the latter results in three or more products [2,3]. These hydrophobic compounds are found in microorganisms [4,5], plants [6], and animals [7]. The synthesis of fatty acids is a complex process mediated by a variety of enzymes with acetyl-CoA as the main building block [15]. The presence of unsaturated acids and sterols in the surrounding environment is necessary for yeast growth in oxygen-limiting conditions such as those occurring during the alcoholic fermentation of grape must [19]
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