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…

Assessment of the impact of actions in the vineyard and its surrounding environment on biodiversity in Rioja Alavesa (Spain)

Traditional viticulture areas have experienced in the last decades an intensification of field practices, linked to an increased use of fertilisers and phytosanitary products, and to a more intensive mechanization and uniformization of the landscape. This change in management has sometimes led to higher rates of soil erosion andloss of soil structure, fertility decline, groundwater contamination, and to an increased pressure of pests and diseases. Additionally, intensification usually leads to a simplification of landscapes, of particular concern in prestigious wine grape regions where the economical revenue encourages the conversion of land use from natural habitats to high value wine grape production. To revert this trend, it is necessary that growers implement actions that promote biodiversity in their vineyards. The aim of this study is to assess the impact of the implementation of cover crops, vegetational corridors, dry stone walls and vineyard biodiversity hotspots estimated through the study of arthropods. The work has been carried out in four vineyards in Rioja Alavesa belonging to Ostatu winery, where these infrastructures were implemented in 2020. The presence and diversity of arthropods was studied by capturing them at different times in the season and at different distances from the infrastructure using pit-fall traps in the soil and yellow, white and blue chromatic traps at the canopy level. This is a preliminary study in which all adult insects were sorted to the taxonomic level of order and Coleoptera were classified to morphospecies. The results obtained show that there is a relationship between the basic characteristics of the vineyard and the arthropods captured, with a positive effect, although also dependent on the vineyard, of the presence of infrastructure.

Characterization of variety-specific changes in bulk stomatal conductance in response to changes in atmospheric demand and drought stress

In wine growing regions around the world, climate change has the potential to affect vine transpiration and overall vineyard water use due to related changes in atmospheric demand and soil water deficits. Grapevines control their transpiration in response to a changing environment by regulating conductance of water through the soil-plant-atmosphere continuum. Most vineyard water use models currently estimate vine transpiration by applying generic crop coefficients to estimates of reference evapotranspiration, but this does not account for changes in vine conductance associated with water stress, nor differences thought to exist between varieties. The response of bulk stomatal conductance to daily weather variability and seasonal drought stress was studied on Cabernet-Sauvignon, Merlot, Tempranillo, Ugni blanc, and Semillon vines in a non-irrigated vineyard in Bordeaux France. Whole vine sap flow, temperature and humidity in the vine canopy, and net radiation absorbed by the vine canopy were measured on 15-minute intervals from early July through mid-September 2020, together with periodic measurement of leaf area, canopy porosity, and predawn leaf water potential. From this data, bulk stomatal conductance was calculated on 15-minute intervals, and multiple regression analysis was performed to identify key variables and their relative effect on conductance. Attention was focused on addressing multicollinearity and time-dependency in the explanatory variables and developing regression models that were readily interpretable. Variability of vapor pressure deficit over the day, and predawn water potential over the season explained much of the variability in conductance, with relative differences in response coefficients observed across the five varieties. By characterizing this conductance response, the dynamics of vine transpiration can be better parameterized in vineyard water use modeling of current and future climate scenarios.

Variations of soil attributes in vineyards influence their reflectance spectra

Knowledge on the reflectance spectrum of soil is potentially useful since it carries information on soil chemical composition that can be used to the planning of agricultural practices. If compared with analytical methods such as conventional chemical analysis, reflectance measurement provides non-destructive, economic, near real-time data. This paper reports results from reflectance measurements performed by spectroradiometry on soils from two vineyards in south Brazil. The vineyards are close to each other, are on different geological formations, but were subjected to the same management. The objective was to detect spectral differences between the two areas, correlating these differences to variations in their chemical composition, to assess the technique’s potential to predict soil attributes from reflectance data.To that end, soil samples were collected from ten selected vine parcels. Chemical analysis yield data on concentration of twenty-one soil attributes, and spectroradiometry was performed on samples. Chemical differences significant to a 95% confidence level between the two studied areas were found for six soil attributes, and the average reflectance spectra were separated by this same level along most of the observed spectral domain. Correlations between soil reflectance and concentrations of soil attributes were looked for, and for ten soil traits it was possible to define wavelength domains were reflectance and concentrations are correlated to confidence levels from 95% to 99%. Partial Least Squares Regression (PLSR) analyses were performed comparing measured and predicted concentrations, and for fifteen out of 21 soil traits we found Pearson correlation coefficients r > 0.8. These preliminary results, which have to be validated, suggest that variations of concentration in the investigated soil attributes induce differences in reflectance that can be detected by spectroradiometry. Applications of these observations include the assessment of the chemical content of soils by spectroradiometry as a fast, low-cost alternative to chemical analytical methods.

Adaptability of grapevines to climate change: characterization of phenology and sugar accumulation of 50 varieties, under hot climate conditions

Climate is the major factor influencing the dynamics of the vegetative cycle and can determine the timing of phenological periods. Knowledge of the phenology of varieties, their chronological duration, and thermal requirements, allows not only for the better management of interventions in the vineyard, but also to predict the varieties’ behaviour in a scenario of climate change, giving the wine producer the possibility of selecting the grape varieties that are best adapted to the climatic conditions of a certain terroir. In 2014, Symington Family Estates, Vinhos, established two grape variety libraries in two different places with distinctive climate conditions (Douro Superior, and Cima Corgo), with the commitment of contributing to a deeper agronomic and oenological understanding of some grape varieties, in hot climate conditions. In these research vineyards are represented local varieties that are important in the regional and national viticulture, but also others that have over time been forgotten — as well as five international reference cultivars. From 2017 to 2021, phenological observations have been made three times a week, following a defined protocol, to determine the average dates of budbreak, flowering and veraison. With the climate data of each location, the thermal requirements of each variety and the chronological duration of each phase have been calculated. During maturation, berry samples have been gathered weekly to study the dynamics of sugar accumulation, between other parameters. The data was analysed applying phenological and sugar accumulation models available in literature. The results obtained show significant differences between the varieties over several parameters, from the chronological duration and thermal requirements to complete the various stages of development, to the differences between the two locations, confirming the influence of the climate on phenology and the stages of maturation, in these specific conditions.

Effect of multi-level and multi-scale spectral data source on vineyard state assessment

Currently, the main goal of agriculture is to promote the resilience of agricultural systems in a sustainable way through the improvement of use efficiency of farm resources, increasing crop yield and quality under climate change conditions. This last is expected to drastically modify plant growth, with possible negative effects, especially in arid and semi-arid regions of Europe on the viticultural sector. In this context, the monitoring of spatial behavior of grapevine during the growing season represents an opportunity to improve the plant management, winegrowers’ incomes, and to preserve the environmental health, but it has additional costs for the farmer. Nowadays, UAS equipped with a VIS-NIR multispectral camera (blue, green, red, red-edge, and NIR) represents a good and relatively cheap solution to assess plant status spatial information (by means of a limited set of spectral vegetation indices), representing important support in precision agriculture management during the growing season. While differences between UAS-based multispectral imagery and point-based spectroscopy are well discussed in the literature, their impact on plant status estimation by vegetation indices is not completely investigated in depth. The aim of this study was to assess the performance level of UAS-based multispectral (5 bands across 450-800nm spectral region with a spatial resolution of 5cm) imagery, reconstructed high-resolution satellite (Sentinel-2A) multispectral imagery (13 bands across 400-2500 nm with spatial resolution of <2 m) through Convolutional Neural Network (CNN) approach, and point-based field spectroscopy (collecting 600 wavelengths across 400-1000 nm spectral region with a surface footprint of 1-2 cm) in a plant status estimation application, and then, using Bayesian regularization artificial neural network for leaf chlorophyll content (LCC) and plant water status (LWP) prediction. The test site is a Greco vineyard of southern Italy, where detailed and precise records on soil and atmosphere systems, in-vivo plant monitoring of eco-physiological parameters have been conducted.