Characterization of free and glycosidically bound simple phenols in hybrid grape varieties using liquid chromatography coupled to high resolution mass (q-orbitrap)

Grape by-products (including skins, seeds, stems and vine shoots) are rich in health promoting polyphenols. Their extraction from winery waste and their following purification are of special interest to produce extracts with high added value compounds. Meanwhile, the growing concern over environmental problems associated with economic constraints, require the development of environmentally sustainable extraction technologies. The extraction using semi-continuous subcritical water, as a natural solvent at high temperature and high pressure a technology is promising “green” technology that is environmentally friendly, energy efficient and improve the extraction process in plant tissues.

Chitosan is a polysaccharide produced from the deacetylation of chitin extracted from crustaceous and fungi. In winemaking chitosan is mainly used in the clarification of grape juice and wine, stabilization of white wines, removal of metals and to prevent wine spoilage by undesired microorganisms. The addition of chitosan to model wine systems was able to retard browning, reduce levels of metallic ions (Fe and Cu) and to protect varietal thiols due to its antiradical activity1. The present experiment was planned in order to evaluate the use of chitosan as a secondary antioxidant at three different stages of Sauvignon blanc fermentation and winemaking. Sauvignon blanc juices from three different locations were obtained at a commercial winery in Marlborough, New Zealand. One lots of grapes was collected from a receival bin and pressed into juice with a water-bag press, and a further juice sample was collected from a commercial pressing operation. Chitosan (1 g/L, low molecular weight, 75 – 85% deacetylated) was added to the juice after pressing, after cold settling, after fermentation, or at all these stages. Controls without any chitosan additions were also prepared.

Light-struck taste is a defect prevalent in white wines bottled in clear glass light-exposed for a considerable amount of time leading to a loss of color and appearance of sulfur-like odors. The reaction involves riboflavin (RF), a highly photosensitive compound that undergoes to intermolecular photoreduction by the uptake of two electron equivalents from an external donor, the methionine. The reaction includes different steps forming methional which is extremely unstable and decomposes to methane thiol and acrolein. The reaction of two molecules of methane thiol yields dimethyl disulfide. Methane thiol is highly volatile, has a low perception threshold (2 to 10 µg/L in wine) and confers aroma-like rotten eggs or cabbage.

A critical aspect of wine quality from a consumer perspective is the overall impression of wine flavour, which is formed by the interplay of volatile aroma compounds, their precursors, and taste and matrix components. Grapes contribute some potent aroma compounds, together with a large pool of non-volatile precursors (e.g. glycoconjugates and amino acid conjugates). Aroma precursors can break down through chemical hydrolysis reactions, or through the action of yeast or enzymes, significantly changing the aroma profile of a wine during winemaking and storage. In addition, glycoconjugates of monoterpenes, norisoprenoids and volatile phenols, together with sulfur-conjugates in wine, provide a reservoir of additional flavour through the in-mouth release of volatiles which may be perceived retro-nasally.

All techniques usually used to assay proteins was not reliable in vegetable extract due to interferences with the components included in extracts like polyphenols, tanins, pectines, aromatics compounds. Absorbance at 280nm, Kjeldhal assay, Biuret and Lowry methods, Acid Bicinchonique technique and Bradford assay give the results depending on the composition of extract, on the presence or not of detergent and on the raw material (Marchal, 1995). Another difficulty in these extracts for the quantification of proteins comes from the large amount of water included in vegetable and the low concentration of proteins. Thus in red wines, proteins are usually not taken into account due to their low concentration (typically below 10 mgL-1) and to the presence of anthocyanis and polyphenols.