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…

Impact on leaf morphology of Vitis vinifera L. cvs Riesling and Cabernet Sauvignon under Free Air Carbon dioxide Enrichment (FACE)

Atmospheric carbon dioxide (CO2) concentration has continuously increased since pre-industrial times from 280 ppm in 1750, and is predicted to exceed 700 ppm by the end of 21st century. For most of C3 plant species elevated CO2 (eCO2) improve photosynthetic apparatus results in an increased plant biomass production. To investigate the effects of eCO2 on morphological leaf characteristics the two Vitis vinifera L. cultivars, Riesling and Cabernet Sauvignon, grown in the Geisenheim VineyardFACE (Free Air Carbon dioxide Enrichment) system were used. The FACE site is located at Geisenheim University (49° 59′ N, 7° 57′ E, 94 m above sea level), Germany and was implemented in 2014 comparing future atmospheric CO2-concentrations (eCO2, predicted for the mid-21st century) with current ambient CO2-conditions (aCO2). Experiments were conducted under rain-fed conditions for two consecutive years (2015 and 2016). Six leaves per repetition of the CO2 treatment were sampled in the field and immediately fixed in a FAA solution (ethanol, H2O, formaldehyde and glacial acetic acid). After 24 h leaf samples were transferred and stored in an ethanol solution. Subsequently, leaf tissue was dehydrated using ethanol series and embedded in paraffin. By using a rotary microtomesections of 5 µm were prepared and fixed on microscopic slides. Subsequent the samples were stained using consecutive staining and washing solutions. Afterwards pictures of the leaf cross-sections were taken using a light microscope and consecutive measurements were conducted with an open source image software. Differences found in leaf cross-sections of the two CO2 treatments were detected for the palisade parenchyma. Leaf thickness, upper and lower epidermis and spongy parenchyma remained less affected under eCO2 conditions. The observed results within grapevine leaf tissues can provide first insights to seasonal adaptation strategies of grapevines under future elevated CO2 concentrations.

VineyardFACE: Investigation of a moderate (+20%) increase of ambient CO2 level on berry ripening dynamics and fruit composition

Climate change and rising atmospheric carbon dioxide concentration is a concern for agriculture, including viticulture. Studies on elevated carbon dioxide have already been on grapevines, mainly taking place in greenhouses using potted plants or using field grown vines under higher CO2 enrichment, i.e. >650 ppm. The VineyardFACE, located at Hochschule Geisenheim University, is an open field Free Air CO2 Enrichment (FACE) experimental set-up designed to study the effects of elevated carbon dioxide using field grown vines (Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon). As the carbon dioxide fumigation started in 2014, the long term effects of elevated carbon dioxide treatment can be investigated on berry ripening parameters and fruit metabolic composition.
The present study aims to investigate the effect on fruit composition under a moderate increase (+20%; eCO2) of carbon dioxide concentration, as predicted for 2050 on both Riesling and Cabernet Sauvignon. Berry composition was determined for primary (sugars, organic acids, amino acids) and secondary metabolites (anthocyanins). Special focus was given on monitoring of berry diameter and ripening rates throughout three growing seasons. Compared to previous results of the early adaptative phase of the vines [1], our results show little effect of eCO2 treatment on primary metabolites composition in berries. However, total anthocyanins concentration in berry skin was lower for eCO2 treatment in 2020, although the ratio between anthocyanins derivatives did not differ.
[1] Wohlfahrt Y., Tittmann S., Schmidt D., Rauhut D., Honermeier B., Stoll M. (2020) The effect of elevated CO2 on berry development and bunch structure of Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon. Applied Science Basel 10: 2486

20-Year-Old data set: scion x rootstock x climate, relationships. Effects on phenology and sugar dynamics

Global warming is one of the biggest environmental, social, and economic threats. In the Douro Valley, change to the climate are expected in the coming years, namely an increase in average temperature and a decrease in annual precipitation. Since vine cultivation is extremely vulnerable and influenced by the climate, these changes are likely to have negative effects on the production and quality of wine.
Adaptation is a major challenge facing the viticulture sector where the choice of plant material plays an important role, particularly the rootstock as it is a driver for adaptation with a wide range of effects, the most important being phylloxera, nematode and salt, tolerance to drought and a complex set of interactions in the grafted plant.
In an experimental vineyard, established in the Douro Region in 1997, with four randomized blocs, with five varieties, Touriga Nacional, Tinta Barroca, Touriga Franca and Tinta Roriz, grafted in four rootstocks, Rupestris du Lot, R110, 196-17C, R99 and 1103P, data was collected consecutively over 20 years (2001-2020). Phenological observations were made two to three times a week, following established criteria, to determine the average dates of budbreak, flowering and veraison. During maturation, weekly berry samples were taken to study the dynamics of sugar accumulation, amongst other parameters. Climate data was collected from a weather station located near the vineyard parcel, with data classified through several climatic indices.
The results achieved show a very low coefficient of variations in the average date of the phenophases and an important contribution from the rootstock in the dynamic of the phenology, allowing a delay in the cycle of up to10-12 days for the different combinations. The Principal Component Analysis performed, evaluating trends in the physical-chemical parameters, highlighted the effect of the climate and rootstock on fruit quality by grape varieties.

Impact of climate change on the viticultural climate of the Protected Designation of Origin “Jumilla” (SE Spain)

Protected Designation of Origin “Jumilla” (PDO Jumilla) is located in the Spanish provinces of Albacete and Murcia, in the South-eastern part of the Iberian Peninsula, where most of the models predict a severe impact of climate change in next decades. PDO Jumilla covers an area of 247,054 hectares, of which more than 22,000 hectares

A spatial explicit inventory of EU wine protected designation of origin to support decision making in a changing climate

Winemaking areas recognized as protected designations of origin (PDOs) shape important economic, environmental and cultural values that are tied to closely defined geographic locations. To preserve wine products and wine-growing practices adopted in different PDOs these areas are strictly regulated by legal specifications. However, quality viticulture is increasingly under pressure from climate change, which is altering the local conditions of many winegrowing areas. Therefore, maintaining traditional wine products will require the adoption of tailored adaptation strategies, including possible changes in the legal regulation of protected wines. To this end, it is necessary to have a comprehensive knowledge on PDOs including their extension, products and allowed practices. While there have been efforts to build databases that summarize the characteristics for individual wine PDO areas and to quantify the related effects of climate change, much information is still included only in the official documentation of the EU geographical indication register and has never been collected in a comprehensive manner. With this study we aim at filling this gap by building a spatial inventory of European wine PDOs that supports decision making in viticulture in the context of climate change. To map and characterize European wine PDOs, we analysed their legal documents and extracted relevant information useful for climate change adaptation. The output consists of a comprehensive geographical dataset that identifies the boundaries of all 1200 European wine PDOs at unprecedented spatial resolution and includes a set of legally binding regulations, such as authorized vine varieties, maximum yields and planting density. The inventory will allow researchers to analyse the impacts of climate change on European wine PDOs and support decision makers in developing tailored adaptation strategies. This includes, among others, the evaluation of new vineyard site selection, the expansion of cultivated varieties or the authorization of irrigation in vineyards.