IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 LC-HRMS data analysis of natural polymer homologue series Application on wine neutral oligosaccharides

LC-HRMS data analysis of natural polymer homologue series Application on wine neutral oligosaccharides

Abstract

Although oligosaccharides have much impact both on health (prevention of diabetes, cardiovascular disease), and on the perception of wine (sweetness, astringency, acidity or bitterness), information on their composition in wine is still limited. In a previous work, neutral oligosaccharide fractions isolated from wine were analyzed. The results present a composition of different monosaccharide units (hexose, pentose, uronic acid and deoxyhexose) and show the presence of several structures of oligo-rhamnogalacturonan type I substituted through the rhamnose moieties by arabinan and/or galactan chains.
The aim of this work is to explore new approaches for processing LC-HRMS data to identify these compounds containing repeating units (homologous series) such as arabinans or galactans.  The presented approach allows visualization of these series in the form of a Kendrick mass defect (KMD) plot to facilitate their characterization.
The chromatographic profiles obtained by LC-HRMS analysis of these fractions showed a poorly resolved bump, and the mass spectra were very complex consisting of mono, di, and tricharged ions peaks over a mass range between 500 and 2500. They allowed however to visualize numerous series formed by separated monocharged peaks of 132 m/z, or dicharged peaks of 66 m/z, i.e. a pentose unit.
The construction of the KMD plot is done with a change of scale for which the mass taken into account of (C5H8O4) is 132.0000 (nominal mass) instead of 132.0423 Da (exact mass). All masses of the spectrum are thus recalculated and called Kendrick masses (KM). The mass defect (KMD) for each peak of the spectrum is the difference between its Kendrick mass and its nominal mass. All compounds of the form R-(C5H8O4)n, R being a common radical, will have the same mass defect. The graphical representation, thus makes it possible to visualize the set of compounds that differ only by their number of pentose units on the same line.
In our case, the accuracy of the measurement at masses above m/z 1000 does not allow this calculation. This limitation was resolved by processing the data with the Compound DiscovererTM software (ThermoScientific) to obtain a list of monocharged masses, for which several crude formulas were proposed. A sorting of these crude formulas was carried out considering the possible ratios between number of carbons, oxygens and hydrogens. The exact masses of the 2045 remaining formulas were then calculated and allowed to draw the Kendrick mass defect plot.
Finally, the Kendrick diagram approach allows visualization of the homologous series of arabinoses. Identification hypotheses were proposed for 555 compounds attributed to oligo-rhamnogalacturonan type I, and its arabinans/galactans side chains degradation products. This study demonstrated the relevance of this analytical approach for the determination of the structure of wine oligosaccharides.

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Poster

Authors

Meudec Emmanuelle1, Vallverdu-Queralt Anna2,3, Sommerer Nicolas1, Cheynier Véronique1, Williams Pascale1 and Doco Thierry1

1SPO, INRAE, Univ Montpellier, Institut Agro, Montpellier, France
INRAE, PROBE research infrastructure, PFP Polyphenol Analytical Facility, Montpellier, France <<

2Department of Nutrition, Food Science and Gastronomy, XIA, Faculty of Pharmacy and Food Sciences, Institute of Nutrition and Food Safety (INSA-UB), University of Barcelona, Barcelona, Spain
3CIBER Physiopathology of Obesity and Nutrition (CIBEROBN), Institute of Health Carlos III, Madrid, Spain 

Contact the author

Keywords

HRMS, oligosaccharides, homologue series, Kendrick mass defect plot, KMD

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Elucidating vineyard site contributions to key sensory molecules: Identification of correlations between elemental composition and volatile aroma profile of site-specific Pinot noir wines

The reproducibility of elemental profile in wines produced across multiple vintages has been previously reported using grapes from a single scion clone of Vitis vinifera L. cv. Pinot noir. The grapevines were grown on fourteen different vineyard sites, from Oregon to southern California in the U.S.A., which span distances from approximately hundreds of meters to 1450 km, while elevations range from near sea level to nearly 500 m. In addition, sensorial (i.e. aroma, taste, and mouthfeel) and chemical (i.e. polyphenolic and volatile) differences across the different vineyard sites have also been observed among these wines at two aging time points. While strong evidence exists to support that grapes grown in different regions can produce wines with unique chemical and sensorial profiles, even when a single clone is used, the understanding of growing site characteristics that result in this reproducible differentiation continues to emerge. One hypothesis is that the elemental profile that a vineyard site imparts to the grape berries and the resulting wine is an important contributor to this differentiation in chemistry and sensory of wines. For example, various classes of enzymes that catalyze the formation of key aroma compounds or their precursors require specific metals. In this work, we begin to report correlations between elemental and volatile aroma profiles of site-specific Pinot noir wines, made under standardized winemaking conditions, that have been previously shown to be distinguished separately by these chemical analyses.

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

The use of rootstock as a lever in the face of climate change and dieback of vineyard

As viticulture faces challenges such as climate change or vineyard dieback, the choice of the variety and rootstock becomes more and more crucial. To study rootstock levers in the Bordeaux region, a parcel of Cabernet Sauvignon (CS) was planted with four rootstocks in 2014. Twenty repetitions of each of the following four rootstocks were set up: 101-14 MGt, Nemadex AB, 420A MGt and Gravesac. The number of bunches, yields and pruning weights of the vine shoots were measured individually on 240 vines from 2017 to 2021. Since 2020, nitrogen status assessed by assimilable nitrogen level, hydric status assessed by δ13C and berry maturity were measured on 80 samples taken from 20 repetitions of the four rootstocks. A lower yield was measured for CS grafted onto Nemadex AB due to the lower number of bunches and the lower weight of berries. The differences between the other three rootstocks are small, but CS grafted onto 420A MGt was the most productive. The CS grafted onto Nemadex AB had the lowest pruning weight while 101-14 MGt had the highest. In 2020, δ13C showed a more moderate water stress with 101-14 MGt and 420A MGt than with Nemadex AB. Surprisingly, the Gravesac was under more stress than the 101-14 MGt. The nitrogen status in the berries was better for Nemadex AB but this was perhaps due to the significantly lower weight of the berries.Rootstock 101-14 MGt attained the highest accumulation of sugars in the berries while 420A MGt allows to preserve higher acidity. The parcel is still young which may explain some of the results. These measures must therefore be continued over the next several years to fully assess the effects of these rootstocks on the development of the vines and the quality of the production under new climatic conditions.

Green berries on Gewürztraminer (Vitis vinifera L.) in South Tyrol (Italy)

The grape variety Gewürztraminer is known to be affected by two physiological disorders namely berry shrivel and bunch stem necrosis. During the season 2014 we noticed a new symptomatology type of ripening disorder on the variety. The new symptom showed not all berries fallowing the normal maturation stages, but single berries remaining at a soft but green stage till harvest. The broad distribution of these so called “green berries” symptoms in different production sites of our region, caused huge damage due to the difficulty of eliminating single berries per bunch before harvesting. Therefore, the Research Centre Laimburg began to investigate the reasons and origins of this new symptom. This work shows the results of first attempts to find causes for the symptom as well as the resulting approach to mitigate symptoms. Applications of magnesium leaf fertilizer showed first promising results against this putative disorder. To study the causal effect of the green berries 30 symptomatic vineyards in 2014 have been selected for a monitoring during the season 2016. To evaluate the foliar nutrient treatment two vineyards have been selected for application of magnesium sulfate and magnesium chloride. Leaf and berry nutrient analysis, as well as the main quality parameters during ripening have been performed. As soon as “green berries” symptoms appeared, incidence and severity have been evaluated. Most of the symptomatic vineyards of the 2016 monitoring showed light to clear magnesium deficit symptoms on their foliage. Only during the seasons 2020 and 2021 “green berries” symptoms could be found in the leaf fertilizer treatment vineyards. Both seasons showed a significant effect of the magnesium treatments to reduce the incidence and severity of the symptom. It seems that the appearance of the “green berries” symptom on Gewürztraminer is correlated to a disturbed uptake of magnesium of the vines.

Analysis of Cabernet Sauvignon and Aglianico winegrape (V. vinifera L.) responses to different pedo-climatic environments in southern Italy

Water deficit is one of the most important effects of climate change able to affect agricultural sectors. In general, it determines a reduction in biomass production, and for some plants, as in the case of grapevine, it can endorse fruit quality. The monitoring and management of plant water stress in the vineyard