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…

Effect of the commercial inoculum of arbuscular mycorrhiza in the establishment of a commercial vineyard of the cultivar “Manto negro

The favorable effect of symbiosis with arbuscular mycorrhizal fungi (AMF) has been known and studied since the 60s. Nowadays, many companies took the chance to start promoting and selling commercial inoculants of AMF, in order to be used as biofertilizers and encourage sustainable biological agriculture. However, the positive effect of these commercial biofertilizers on plant growth is not always demonstrated, especially under field conditions. In this study, we used a commercial inoculum on newly planted grapevines of a local cultivar grafted on a common rootstock R110. We followed the physiological status of vines, growth and productivity and functional biodiversity of soil bacteria during the first and second years of 20 inoculated with commercial inoculum bases on Rhizophagus irregularis and Funeliformis mosseaeAMF at field planting time and 20 non-inoculated control plants. All the parameters measured showed a neutral to negative effect on plant growth and production. The inoculated plants always presented lower values of photosynthesis, growth and grape production, although in some cases the differences did not reach statistical significance. On the contrary, the inoculation supposed an increase of the bacterial functional diversity, although the differences were not statistically significant either. Several studies show that the effect of inoculation with AMF is context-dependent. The non-favorable effects are probably due to inoculation ineffectiveness under complex field conditions and/or that, under certain conditions, AMF presence may be a parasitic association. This puts into question the effectiveness of its application in the field. Therefore, it is recommended to only resort to this type of biofertilizer when the cultivation conditions require it (e.g., very low previous microbial diversity, foreseeable stress due to drought, salinity, or lack of nutrients) and not as a general fertilization practice.

Evolution of the amino acids content through grape ripening: Effect of foliar application of methyl jasmonate with or without urea

The parameters that determine the grape quality, and therefore the optimal harvest time, suffer variations during berry ripening, related to climate change, with the widely known problem of the gap between technological and phenolic maturities. However, there are few studies about its incidence on grape nitrogen composition. For this reason, the use of an elicitor, methyl jasmonate (MeJ), alone or with urea, is proposed as a tool to reduce climatic decoupling, allowing to establish the harvest time in order to achieve the optimum grape quality. The aim was to study the effect of MeJ and MeJ+Urea foliar applications on the evolution of Tempranillo amino acids content throughout the grape maturation. Three treatments were foliarly applied, at veraison and 7 days later: control (water), MeJ (10 mM) and MeJ+Urea (10 mM+6 kg N/ha). Grape samples were taken at five stages of maturation: day before the first and second applications, 15 days after the second application (pre-harvest), harvest day, and 15 days after harvest (post-harvest). The amino acids analysis of the samples was carried out by HPLC. Results showed that the evolution of amino acids was similar regardless of the treatment; however, foliar applications influenced the nitrogen compounds content, i.e., there was no qualitative effect but quantitative one. Most of the amino acids reached their maximum concentration in pre-harvest, being higher in grapes from the treatments than in the control. In general, no differences in grape amino acids content were observed between MeJ and MeJ+Urea treatments. Foliar applications with MeJ and MeJ+Urea enhanced the grape amino acids content, without affecting their profile, helping to optimize their quality and allowing to establish a more complete grape ripening standard. Therefore, MeJ and MeJ+Urea foliar applications can be a simple agronomic practice, which has shown promising results in order to enhance the grape quality.

Evaluation of climate change impacts at the Portuguese Dão terroir over the last decades: observed effects on bioclimatic indices and grapevine phenology

In the last decades the growers of the Portuguese Dão winegrowing region (center of Portugal) are experiencing changes in climate that are influencing either grape phenology berry health and ripening. Aiming to study the relationships between climate indices (CI), seasonal weather and grapevine phenology, in this work long-term climate and phenological data collected at the experimental vineyard of the Portuguese Dão research centre between 1958 and 2019 (61 years) for the red variety Touriga Nacional, was analyzed. The trends over time for the classical temperature-based indices (Growing Season Temperature – GST -, Growing Degree Days – GDD, Huglin Index – HI and Cool Night Index – CI) presented a significantly positive slope while the Dryness Index (DI) showed a negative trend over the last 61 years. Regarding grapevine phenology, an average advance of 4.5 days per decade in the harvest day was observed throughout the last 61 years. Consequently, the weather conditions during the ripening period have changed, showing an increasing trend over time in the average temperature (higher magnitude in the maximum than in the minimum temperature) and a decrease in the accumulated rainfall. A regression analysis showed that ~50% of harvest date variability over years was explained by the temperature-based indices variability. These observed effects of climate change on bioclimatic indices and corresponding anticipation of harvest date can still be considered advantageous for the Dão terroir as it allows to achieve an optimal berry ripening before the common equinox rains and, therefore, avoid the potential negative impacts of the rainfall on berry health and composition.

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Climate modeling at local scale in the Waipara winegrowing region in the climate change context

In viticulture, a warming climate can have a very significant impact on grapevine development and therefore on the quality and characteristics of wines across different spatial scales, ranging from global to local. In order to adapt wine-growing to climate change, global climate models can be used to define future scenarios, but only at the scale of major wine regions. Despite the huge progress made over the last ten years in terms of the spatial resolution of climate models (now downscaled to a few square kilometres), they are not yet sufficiently precise to account for the local climate variability associated with such parameters as local topography, in spite of these parameters being decisive for vine and wine characteristics. This study describes a method to downscale future climate scenarios to vineyard scale. Networks of data loggers have been used to collect air temperature at canopy level in the Waipara winegrowing region (New Zealand) over five growing seasons. These measurements allow the creation of fine-scale geostatistical models and maps of temperature (at 100 m resolution) for the growing season. In order to model climate change at pilot site scale, these geostatistical models have been combined with regional climate change predictions for the periods 2031-2050 and 2081-2100 based on the RCP8.5 climate change scenario. The integration of local climate variability with regionalized climate change simulations allows assessment of the impacts of climate change at the vineyard scale. The improved knowledge gained using this methodology results from the increased horizontal resolution that better addresses the concerns of winegrowers. The results provide the local winegrowers with information necessary to understand current processes, as well as historical and future viticulture trends at the scale of their site, thereby facilitating decisions about future response strategies.