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…

1H-NMR-based Metabolomics to assess the impact of soil type on the chemical composition of Mediterranean red wines

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.

Impact of climate variability and change on grape yield in Italy

Viticulture is entangled with weather and climate. Therefore, areas currently suitable for grape production can be challenged by climate change. Winegrowers in Italy already experiences the effect of climate change, especially in the form of warmer growing season, more frequent drought periods, and increased frequency of weather extremes.
The aim of this study is to investigate the impact of climate variability and change on grape yield in Italy to provide winegrowers the information needed to make their business more sustainable and resilient to climate change. We computed a specific range of bioclimatic indices, selected by the International Organisation of Vine and Wine (OIV), and correlated them to grape yield data. We have worked in collaboration with some wine consortiums in northern and central Italy, which provided grape yield data for our analysis.
Using climate variables from the E-OBS dataset we investigate how the bioclimatic indices changed in the past, and the impact of this change on grape productivity in the study areas. The climate impact on productivity is also investigated by using high-resolution convection-permitting models (CPMs – 2.2 horizontal resolution), with the purpose of estimating productivity in future emission scenarios. The CPMs are likely the best available option for this kind of impact studies since they allow a better representation of small-scale processes and features, explicitly resolve deep convection, and show an improved representation of extremes. In our study, we also compare CPMs with regional climate models (RCMs – 12 km horizontal resolution) to assess the added value of high-resolution models for impact studies. Further development of our study will lead to assessing the future suitability for vine cultivation and could lead to the construction of a statistical model for future projection of grape yield.

Grape must quality and mesoclimatic variability in Fruška Gora wine-growing region, Serbia

The Fruška Gora mountain is a traditional wine-growing region in Serbia situated in the Pannonian Basin. Due to such a position, the vicinity of the Danube River and the presence of concave configuration, it is suitable for grape production. This paper provides analyses of spatial variations in meteorological parameters and grape juice quality within Fruška Gora wine region over three consecutive vintages (2018-2020). The examined period can be defined as warm with cool nights during September (AVG 18,9°C; GDD 1918°C; CI 12°CF) and with the presence of mesoclimatic variability. The East part of the study area was somewhat drier and hotter compared to other parts of the region. The analyses of grape must samples (190 in total) of five cultivars (Cabernet-Sauvignon, Merlot, Chardonnay, Sauvignon blanc and Grašac (Welschriesling)) commonly grown across the region (19 sites), were performed using Fourier Transform Infrared Technology (FTIR). Among all cultivars, Sauvignon blanc was harvested first in the East area (DOY=246±5, GDD at harvest=1552±74, 22.2±0.7 °Brix), while the latest harvest was recorded for Cabernet-Sauvignon in the West (DOY=283±5, GDD at harvest=1936±187, 23.4±1.0 °Brix ). Both the red and white cultivars had higher acidity and YAN in the grape must if the vines were grown in the North and East compared to South and West areas. According to PCA analysis, Grašac showed the lowest variation in grape must chemical composition. Thus, the results confirm that Grašac is the most stable cultivar in Fruška Gora. All monitored cultivars reached technological fruit ripeness by the end of the growing season. However, it was difficult to reach full ripeness of red cultivars, mostly beacuse of uncoupling of technolocical and phenolic ripeness. Thus, Cabernet-Sauvignon had higher variations in GDD sums at harvest compared to other cultivars, which probably increased variations in grape must quality.

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.

Sustainable fertilisation of the vineyard in Galicia (Spain)

Excessive fertilization of the vineyard leads to low quality grapes, increased costs and a negative impact on the environment. In order to establish an integrated management system aimed at a sustainable fertilization of the vineyards, nutritional reference levels were established. For this purpose, 30 representative vineyards of the Albariño variety were studied, in which soil and petiole analyses were carried out for two years and grape yield and quality at harvest were measured. In both years of study, soil pH, calcium, sodium and cation exchange capacity were positively correlated with calcium content and negatively correlated with manganese in grapes. Irrigated vineyards had higher levels of aluminium in soil and lower levels of calcium in petiole. Climatic conditions were very different in the years of the study. The year 2019 was colder than usual, in 2020 there was a marked water stress with high summer temperatures. This resulted in medium-high acidity in grapes in 2019 and low acidity in 2020, with sugar levels being similar both years. A very marked decrease in must amino nitrogen was observed in 2020, with ammonia nitrogen remaining stable. The correlation of acidity and sugar values in grapes with soil and petiole analysis data made it possible to establish reference levels for the nutritional diagnosis of the Albariño variety in this region. Based on these results, an easy-to-use TIC application is currently being created for grapegrowers, aimed at improving the sustainability of the vineyard through reasoned fertilization. This study has now been extended to other Galician vine varieties.