OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Analytical developments from grape to wine, spirits : omics, chemometrics approaches… 9 Development of a LC-FTMS method to quantify natural sweeteners in red wines

Development of a LC-FTMS method to quantify natural sweeteners in red wines

Abstract

The quality of a wine is largely related to the balance between its sourness, bitterness and sweetness. Recently, molecules coming from grapes have been showed to notably contribute to sweet taste of dry wines. To study the viticultural and oenological parameters likely to affect their concentration, their quantification appears of high interest and subsequently requires powerful analytical techniques. 

Therefore, a new method using liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS) was developed and validated to quantify epi-DPA-3′-O-β-glucopyranoside acid (epi-DPA-G) and astilbin, sweet molecules identified in wine. Three gradients were tested on five different C18 columns (Hypersil Gold, HSS T3, BEH, Syncronis and Kinetex). 

The finest results were obtained upon using Hypersil Gold C18 and a gradient elution composed of 0.1 % formic acid in water and 0.1 % formic acid in acetonitrile. Satisfactory linearity with correlative coefficient (r2) higher than 0.995 was achieved for both compounds with recoveries higher than 89 %. Good sensitivity (LOD ≤ 7 μg L-1) and repeatability (RSD ≤ 3 %) were obtained. 

The developed method was applied to screen epi-DPA-G and astilbin in red wines coming from several vintages over one century. Both compounds have been detected in all wines, at concentrations varying from 1.4 to 14.7 mg L-1 for epi-DPA-G and from 0.5 to 32.2 mg L-1 for astilbin. These results demonstrate the reliability of the developed method to quantifiy epi-DPA-G and astilbin in wine and suggest their oenological interest. Moreover, the method was used to study the influence of various winemaking parameters on epi-DPA-G and astilbin concentrations. The results opened promising perspectives for a better monitoring of extraction during vatting.

DOI:

Publication date: June 23, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Syntia Fayad, Blandine Cretin, Axel Marchal 

Unité de recherche Oenologie, EA 4577, USC 1366 INRA, ISVV, Université de Bordeaux, Bordeaux INP, F33882 Villenave d’Ornon France 

Contact the author

Keywords

Orbitrap, method validation, wine, sweetness

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

An analytical framework to site-specifically study climate influence on grapevine involving the functional and Bayesian exploration of farm data time series synchronized using an eGDD thermal index

Climate influence on grapevine physiology is prevalent and this influence is only expected to increase with climate change. Although governed by a general determinism, climate influence on grapevine physiology may present variations according to the terroir. In addition, these site-specific differences are likely to be enhanced when climate influence is studied using farm data. Indeed, farm data integrate additional sources of variation such as a varying representativity of the conditions actually experienced in the field. Nevertheless, there is a real challenge in valuing farm data to enable grape growers to understand their own terroir and consequently adapt their practices to the local conditions. In such a context, this article proposes a framework to site-specifically study climate influence on grapevine physiology using farm data. It focuses on improving the analysis of time series of weather data. The analytical framework includes the synchronization of time series using site-specific thermal indices computed with an original method called Extended Growing Degree Days (eGDD). Synchronized time series are then analyzed using a Bayesian functional Linear regression with Sparse Steps functions (BLiSS) in order to detect site-specific periods of strong climate influence on yield development. The article focuses on temperature and rain influence on grape yield development as a case study. It uses data from three commercial vineyards respectively situated in the Bordeaux region (France), California (USA) and Israel. For all vineyards, common periods of climate influence on yield development were found. They corresponded to already known periods, for example around veraison of the year before harvest. However, the periods differed in their precise timing (e.g. before, around or after veraison), duration and correlation direction with yield. Other periods were found for only one or two vineyards and/or were not referred to in literature, for example during the winter before harvest.

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

Updating the Winkler index: An analysis of Cabernet sauvignon in Napa Valley’s varied and changing climate

This study aims to create an updated, agile viticultural climate index (similar to the Winkler Index) by performing in-depth analyses of current and historical data from industry partners in several major winegrowing regions. The Winkler Index was developed in the early twentieth century based on analysis of various grape-growing regions in California. The index uses heat accumulation (i.e. Growing Degree Days) throughout the growing season to determine which grape varieties are best suited to each region. As viticultural regions are increasingly subject to the complexity and uncertainty of a changing climate, a more rigorous, agile model is needed to aid grape growers in determining which cultivars to plant where. For the first phase of this study, 21 industry partners throughout Napa Valley shared historical phenology, harvest, viticultural practice, and weather data related to their Cabernet sauvignon vineyard blocks. To complement this data, berry samples were collected throughout the 2021 growing season from 50 vineyard blocks located throughout 16 American Viticultural Areas that were then analyzed for basic berry chemistry and phenolics. These blocks have been mapped using a Geographic Information System (GIS), enabling analysis of altitude, vineyard row orientation, slope, and remotely sensed climate data. Sampling sites were also chosen based on their proximity to a weather station. By analyzing historical data from industry partners and data specifically collected for this study, it is possible to identify key parameters for further analysis. Initial results indicate extreme variability at a high spatial resolution not currently accounted for in modern viticultural climate indices and suggest that viticultural practices play a major role. Using the structure of data collection and analyses developed for the first phase, this project will soon be expanded to other wine regions globally, while continuing data collection in Napa Valley.

Permanent cover cropping with reduced tillage increased resiliency of wine grape vineyards to climate change

Majority of California’s vineyards rely on supplemental irrigation to overcome abiotic stressors. In the context of climate change, increases in growing season temperatures and crop evapotranspiration pose a risk to adaptation of viticulture to climate change. Vineyard cover crops may mitigate soil erosion and preserve water resources; but there is a lack of information on how they contribute to vineyard resiliency under tillage systems. The aim of this study was to identify the optimum combination of cover crop sand tillage without adversely affecting productivity while preserving plant water status. Two experiments in two contrasting climatic regions were conducted with two cover crops, including a permanent short stature grass (P. bulbosa hybrid), barley (Hordeum spp), and resident vegetation under till vs. no-till systems in a Ruby Cabernet (V. vinifera spp.) (Fresno) and a Cabernet Sauvingon (Napa) vineyard. Results indicated that permanent grass under no-till preserved plant available water until E-L stage 17. Consequently, net carbon assimilation of the permanent grass under no-till system was enhanced compared to those with barley and resident vegetation. On the other hand, the barley under no-till system reduced grapevine net carbon assimilation during berry ripening that led to lower content of nonstructural carbohydrates in shoots at dormancy. Components of yield and berry composition including flavonoid profile at either site were not adversely affected by factors studied. Switching to a permanent cover crop under a no-till system also provided a 9% and 3% benefit in cultural practices costs in Fresno and Napa, respectively. The results of this work provides fundamental information to growers in preserving resiliency of vineyard systems in hot and warm climate regions under context of climate change.

Estimating bulk stomatal conductance of grapevine canopies

In response to changes in their environment, grapevines regulate transpiration using various physiological mechanisms that alter conductance of water through the soil-plant-atmosphere continuum. Expressed as bulk stomatal conductance at the canopy scale, it varies diurnally in response to changes in vapor pressure deficit and net radiation, and over the season to changes in soil water deficits and hydraulic conductivity of both soil and plant. It is necessary to characterize the response of conductance to these variables to better model how vine transpiration also responds to these variables. Furthermore, to be relevant for vineyard-scale modeling, conductance is best characterized using data collected in a vineyard setting. Applying a crop canopy energy flux model developed by Shuttleworth and Wallace, bulk stomatal conductance was estimated using measurements of individual vine sap flow, temperature and humidity within the vine canopy, and estimates of net radiation absorbed by the vine canopy. These measurements were taken on several vines in a non-irrigated vineyard in Bordeaux France, using equipment that did not interfere with ongoing vineyard operations. An inverted Penman-Monteith equation was then used to calculate bulk stomatal conductance on 15-minute intervals from July to mid-September 2020. Time-series plots show significant diurnal variation and seasonal decreases in conductance, with overall values similar to those in the literature. Global sensitivity analysis using non-parametric regression found transpiration flux and vapor pressure deficit to be the most important input variables to the calculation of bulk stomatal conductance, with absorbed net radiation and bulk boundary layer conductance being much less important. Conversely, bulk stomatal conductance was one of the most important inputs when calculating vine transpiration, further emphasizing the need for characterizing its response to environmental changes for use in vineyard water use modeling.