Quantification of red wine phenolics using ultraviolet-visible, near and mid-infrared spectroscopy combined with chemometrics

Phenolic compounds are secondary metabolites of grapes. Plants produce a wide variety of this type of metabolites through diverse biosynthesis pathways and their production is sometimes a response to external stimuli, either environmental or biotic stresses. Some of them may act as chemical defenses against pathogens or herbivores and their synthesis is increased when the attack exists. However, it is remarkable that the synthesis of these interesting compounds can be activated even when the stimulus is not present, with the use of elicitors. These are substances that when applied exogenously trigger the biosynthetic pathways conducting to the synthesis of these defense compounds.

Bacterial malolactic fermentation (MLF) has a considerable impact on wine quality. The yeast strain used for primary fermentation can consistently stimulate (MLF+ phenotype) or inhibit (MLF- phenotype) malolactic bacteria and the MLF process as a function of numerous winemaking practices, but the molecular evidence behind still remains a mystery. In this study, such evidence was elucidated by the direct comparison of extracellular metabolic profiles of MLF+ and MLF- yeast phenotypes. Untargeted metabolomics combining ultrahigh-resolution FT-ICR-MS analysis, powerful machine learning methods and a comprehensive wine metabolite database, discovered around 800 putative biomarkers and 2500 unknown masses involved in phenotypic distinction.

Proteins constitute one of the three main components of grape juice and white wine, phenolic compounds and polysaccharides being the others. A specific group of the total grape-derived proteins resists degradation or adsorption during the winemaking process and remains in finished wine if not removed by the commonplace commercial practice of bentonite fining. While bentonite is effective in removing the problematic proteins, it is claimed to adversely affect the quality of the treated wine under certain conditions, through the removal of colour, flavor and texture compounds. A number of studies have indicated that different protein fractions require distinct bentonite concentrations for protein removal and consequent heat stabilization.

The color of a red wine is one of the most important parameters of its quality, giving much information on its status, such as the grape variety used or the winemaking style. As the result of a complex equilibrium between different forms of anthocyanins and polymerization reactions which occur over the course of time, color can also serve as an indication of a wines’ age. For this purpose the “chemical age” i and ii indexes have been introduced by Somers in 1977. The chemical age index i measures the color absorbance after the addition of acetaldehyde while chemical index ii provides an indication of how much of the total red pigments are resistant to SO2 bleaching.

A method of suspect screening analysis to study grape metabolomics, was developed [1]. By performing ultra-high performance liquid chromatography (UHPLC) – high-resolution mass spectrometry (HRMS) analysis of the grape extract, averaging 320-450 putative grape compounds are identified which include mainly polyphenols. Identification of metabolites is performed by a new HRMS-database of putative grape and wine compounds expressly constructed (GrapeMetabolomics) which currently includes around 1,100 entries.