IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Study of the grape glycosidic aroma precursors by crossing SPE-GC/MS, SPME-GC/MS and LC/QTOF methods

Study of the grape glycosidic aroma precursors by crossing SPE-GC/MS, SPME-GC/MS and LC/QTOF methods

Abstract

Depending on the variety, grapes contain several chemical classes of aromatic compounds (i.e., terpenols, norisoprenoids, benzenoids) mainly stored as glycosides in berry skin. These secondary metabolites are the aromatic precursors fraction of grape which is liberated in wine during fermentation. Knowledge of their profile is often required to estimate the aromatic potential transferable to the wine and for chemotaxonomic aims (Nasi et al., 2008; Ferreira and Lopez, 2019).

In general, the methods used to study glycosidic aroma profile involve sample extraction and concentration by passage of large volumes of must or grape extract through a SPE cartridge (the stationary phases commonly used are 1-10 g of C18 or polystyrene-divinylbenzene), then the methanolic fraction eluted containing the glycoside compounds is carried to dryness, resolubilized using a citrate pH 5 buffer, and an enzymatic hydrolysis is carried out overnight to liberate the aglycones which are then analyzed by GC/MS. Main advantage of SPE is until 1000-fold concentration of sample which allows to detect also compounds present at low level but which can play important role in determining the organoleptic characteristics of wine. Usually, the selectivity of SPE towards the compounds studied is low, so performing quantitation by expressing the compounds as mg internal standard/kg grape provides accuracy acceptable for the aim of the study. On the other hand, SPE is laborious, needs long time and is hardly applicable in quality control laboratories. SPME is faster but the selectivity of fiber towards the analytes is often very different and to perform acceptable quantitative analysis it is essential the calculation of calibration curves. Unfortunately, just few standards of the grape aroma compounds are commercially available (Panighel et al., 2014).

In this study SPE-GC/MS and SPME-GC/MS methods are compared by performing analysis of a set of model standard solutions and grape must samples. The use of several internal standards allows to estimate recoveries of the analytes and calculation of corrective coefficients between the two methods. To have also information free of enzymatic artifacts, GC/MS results are crossed with profile of glycosidic aroma precursors determined by LC/QTOF analysis (Flamini et al., 2014).

The study is finalized to develop a quick SPME-GC/MS method which provides exhaustive and reliable qualitative and semi-quantitative information on the grape glycosidic aroma precursors

References

Nasi A., Ferranti P., Amato S., Chianese L. (2008). Food Chem. 110: 762-768
Ferreira V., Lopez R. (2019). Biomolecules 9(12): 818- doi:10.3390/biom9120818
Panighel A., Flamini R. (2014). Molecules 19: 21291-21309 doi:10.3390/molecules191221291
Flamini R., De Rosso M., Panighel A., Dalla Vedova A., De Marchi F., Bavaresco L. (2014). J. Mass Spec. 49(12): 1214-1222 doi:10.1002/jms.34411214

DOI:

Publication date: June 23, 2022

Issue: IVAS 2022

Type: Article

Authors

Panighel Annarita¹, Fugaro Michele², Mazzei Raffaele Antonio², De Rosso Mirko¹, De Marchi Fabiola¹ and Flamini Riccardo¹

¹Council for Agricultural Research and Economics – Viticulture & Oenology (CREA-VE)
²Dipartimento dell’Ispettorato centrale della tutela della qualità e repressione frodi dei prodotti agroalimentari – ICQRF NORD-EST

Contact the author

Keywords

Glycosides, grape, aroma, mass spectrometry

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Teasing apart terroir: the influence of management style on native yeast communities within Oregon wineries and vineyards

Newer sequencing technologies have allowed for the addition of microbes to the story of terroir. The same environmental factors that influence the phenotypic expression of a crop also shape the composition of the microbial communities found on that crop. For fermented goods, such as wine, that microbial community ultimately influences the organoleptic properties of the final product that is delivered to customers. Recent studies have begun to study the biogeography of wine-associated microbes within different growing regions, finding that communities are distinct across landscapes. Despite this new knowledge, there are still many questions about what factors drive these differences. Our goal was to quantify differences in yeast communities due to management style between seven pairs of conventional and biodynamic vineyards (14 in total) throughout Oregon, USA. We wanted to answer the following questions: 1) are yeast communities distinct between biodynamic vineyards and conventional vineyards? 2) are these differences consistent across a large geographic region? 3) can differences in yeast communities be tied to differences in metabolite profiles of the bottled wine? To collect our data we took soil, bark, leaf, and grape samples from within each vineyard from five different vines of pinot noir. We also collected must and a 10º brix sample from each winery. Using these samples, we performed 18S amplicon sequencing to identify the yeast present. We then used metabolomics to characterize the organoleptic compounds present in the bottled wine from the blocks the year that we sampled. We are actively in the process of analysing our data from this study.

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

How does aromatic composition of red wines, resulting from varieties adapted to climate change, modulate fruity aroma?

One of the major issues for the wine sector is the impact of climate change linked to the increasing temperatures which affects physicochemical parameters of the grape varieties planted in Bordeaux vineyard and consequently, the quality of wine. In some varietals, the attenuation of their fresh fruity character is accompanied by the accentuation of dried-fruit notes [1]. As a new adaptive strategy on climate change, some winegrowers have initiated changes in the Bordeaux blend of vine varieties [2]. This study intends to explore the fruitiness in wines produced from grape varieties adapted to the future climate of Bordeaux. 10 commercial single–varietal wines from 2018 vintage made from the main grape varieties in the Bordeaux region (Cabernet franc, Cabernet-Sauvignon and Merlot) as well as from indigenous grape varieties from the Mediterranean basin, such as Cyprus (Yiannoudin), France (Syrah), Greece (Agiorgitiko and Xinomavro), Portugal (Touriga Nacional) and Spain (Garnacha and Tempranillo), were selected among 19 samples using sensory descriptive analyses. Both sensory and instrumental analyses were coupled, to investigate their fruity aroma expression. For sensory analysis, samples were prepared from wine, using a semi preparative HPLC method which preserves wine aroma and isolates fruity characteristics in 25 specific fractions [3,4]. Fractions of interest with intense fruity aromas were sensorially selected for each wine by a trained panel and mixed with ethanol and microfiltered water to obtain fruity aromatic reconstitutions (FAR) [5]. A free sorting task was applied to categorize FAR according to their similarities or dissimilarities, and different clusters were highlighted. Instrumental analysis of the different FAR and wines demonstrated variations in their molecular composition. Results obtained from sensory and gas chromatography analysis enrich the knowledge of the fruity expression of red wines from “new” grape varieties opening up new perspectives in wine technology, including blending, thus providing new tools for producers.

Impact of long term agroecological and conventional practices on subsurface soil microbiota in Macabeu and Xarel·lo vineyards

There is a growing trend on the transition from conventional to agroecological management of vineyards. However, the impact of practices, such as reduced-tillage, organic fertilization and cover crops, is not well-understood regarding the soil microbial diversity, and its relationship with the soil physicochemical properties in the subsurface depth near the rooting zone. Soil bacterial diversity is an important contributor towards plant health, productivity and response to environmental stresses. A field experiment was conducted by sampling subsurface soil bacterial community (NGS and qPCR) near to the root zone of Macabeu and Xarel·lo vineyards, located at the Penedes. 3 organic (ECO) and 3 conventional (CON) vineyards, with more than 10 years of respective management were sampled (n=5 each plot). ECO practices did not affect bacterial and fungal abundance but increased significantly the ammonium oxidizing bacteria and alpha-diversity (Inv.Simpson). Interestingly beta-diversity was significantly affected by the management strategy. ANOSIM-tests revealed a significative effect of the management (ecological vs conventional) and plot, on the soil microbial structure (ASV abundance). Main phyla depicted were Proteobacteria, Actinobacteria and Acidobacteria, whose relative abundances were not affected by the management. EdgeR assay revealed a significant increase of Cyanobacteria and decrease of Gemmatimonadetes and Firmicutes phyla in ECO. Interestingly, the grapevine variety was not correlated with the soil microbial community structure. Mantel-test revealed an important correlation (Spearman) of some physicochemical parameters with the soil microbiota structure, in order of importance: texture, EC, pH Ca/Mg, Mg/P, K+, Mg2+, Ca2+, SO42-, and OM. N-NH4 and NTK, which were higher in the ECO managed soils, did not correlated significantly with the soil microbiome population. The results revealed the importance of combining a deep physicochemical characterization of each replicate with the microbial diversity assessment to gain better insights on the relationship between soil microbiome and vineyard management.

Phenological characterization of a wide range of Vitis Vinifera varieties

In order to study the impact of climate change on Bordeaux grape varieties and to assess the adaptation capacities of candidates to the grape varieties of this wine region to the new climatic conditions, an experimental block design composed of 52 grape varieties was set up in 2009 at the INRAE Bordeaux Aquitaine center. Among the many parameters studied, the three main phenological stages of the vine (budburst, flowering and veraison) have been closely monitored since 2012. Observations for each year, stage and variety were carried out on four independent replicates. Precocity indices have been calculated from the data obtained over the 2012-2021 period (Barbeau et al. 1998). This work allowed to group the phenological behaviour of the grapevine varieties, not only based on the timing of the subsequent developmental stages, but also on the overall precocity of the cycle and the total length of the cycle between budburst and veraison. Results regarding the variability observed among the different grape varieties for these phenological stages are presented as heat maps.