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…

Low-cost sensors as a support tool to monitor soil-plant heat exchanges in a Mediterranean vineyard

Mediterranean viticulture is increasingly exposed to more frequent extreme conditions such as heat waves. These extreme events co-occur with low soil water content, high air vapor pressure deficit and high solar radiant energy fluxes and result in leaf and berry sunburn, lower yield, and berry quality, which is a major constraint for the sustainability of the sector. Grape growers must find ways to proper and effectively manage heat waves and extreme canopy and berry temperatures. Irrigation to keep soil moisture levels and enable adequate plant turgor, and convective and evaporative cooling emerged as a key tool to overcome this major challenge. The effects of irrigation on soil and plant water status are easily quantifiable but the impact of irrigation on soil and canopy temperature and on heat convection from soil to cluster zone remain less characterized. Therefore, a more detailed quantification of vineyard heat fluxes is highly relevant to better understand and implement strategies to limit the effects of extreme weather events on grapevine leaf and berry physiology and vineyards performance. Low-cost sensor technologies emerge as an opportunity to improve monitoring and support decision making in viticulture. However, validation of low-cost sensors is mandatory for practical applicability. A two-year study was carried in a vineyard in Alentejo, south of Portugal, using low-cost thermal cameras (FLIR One, 80×60 pixels and FLIR C5, 160×120 pixels, 8-14 µm, FLIR systems, USA) and pocket thermohygrometers (Extech RHT30, EXTECH instruments, USA) to monitor grapevine and soil temperatures. Preliminary results show that low-cost cameras can detect severe water stress and support the evaluation of vertical canopy temperature variability, providing information on soil surface temperature. All these thermal parameters can be relevant for soil and crop management and be used in decision support systems.

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.

Impact on leaf morphology of Vitis vinifera L. cvs Riesling and Cabernet Sauvignon under Free Air Carbon dioxide Enrichment (FACE)

Atmospheric carbon dioxide (CO2) concentration has continuously increased since pre-industrial times from 280 ppm in 1750, and is predicted to exceed 700 ppm by the end of 21st century. For most of C3 plant species elevated CO2 (eCO2) improve photosynthetic apparatus results in an increased plant biomass production. To investigate the effects of eCO2 on morphological leaf characteristics the two Vitis vinifera L. cultivars, Riesling and Cabernet Sauvignon, grown in the Geisenheim VineyardFACE (Free Air Carbon dioxide Enrichment) system were used. The FACE site is located at Geisenheim University (49° 59′ N, 7° 57′ E, 94 m above sea level), Germany and was implemented in 2014 comparing future atmospheric CO2-concentrations (eCO2, predicted for the mid-21st century) with current ambient CO2-conditions (aCO2). Experiments were conducted under rain-fed conditions for two consecutive years (2015 and 2016). Six leaves per repetition of the CO2 treatment were sampled in the field and immediately fixed in a FAA solution (ethanol, H2O, formaldehyde and glacial acetic acid). After 24 h leaf samples were transferred and stored in an ethanol solution. Subsequently, leaf tissue was dehydrated using ethanol series and embedded in paraffin. By using a rotary microtomesections of 5 µm were prepared and fixed on microscopic slides. Subsequent the samples were stained using consecutive staining and washing solutions. Afterwards pictures of the leaf cross-sections were taken using a light microscope and consecutive measurements were conducted with an open source image software. Differences found in leaf cross-sections of the two CO2 treatments were detected for the palisade parenchyma. Leaf thickness, upper and lower epidermis and spongy parenchyma remained less affected under eCO2 conditions. The observed results within grapevine leaf tissues can provide first insights to seasonal adaptation strategies of grapevines under future elevated CO2 concentrations.

Long-term drought resilience of traditional red grapevine varieties from a semi-arid region

In recent decades, the scarcity of water resources in agriculture in certain areas has been aggravated by climate change, which has caused an increase in temperatures, changes in rainfall patterns, as well as an increase in the frequency of extreme phenomena such as droughts and heat waves. Although the vine is considered a drought-tolerant specie, it has to satisfy important water requirements to complete its cycle, which coincides with the hottest and driest months. Achieving sustainable viticulture in this scenario requires high levels of efficiency in the use of water, a scarce resource whose use is expected to be severely restricted in the near future. In this regard, the use of drought-tolerant varieties that are able to maintain grape yield and quality could be an effective strategy to face this change. During three consecutive seasons (2018-2020) the behavior in rainfed regime of 13 traditional red grapevine varieties of the Spain central region was studied. These varieties were cultivated in a collection at Centro de Investigación de la Vid y el Vino de Castilla-La Mancha (IVICAM-IRIAF) located in Tomelloso (Castilla-La Mancha, Spain). Yield components (yield, mean bunch and berry weight, pruning weight), physicochemical parameters of the musts (brix degree, total acidity, pH) and some physiological parameters related with water stress during ripening period (δ13C, δ18O) were analysed. The application of different statistical techniques to the results showed the existence of significant differences between varieties in their response to stressful conditions. A few varieties highlighted for their high ability to adapt to drought, being able to maintain high yields due to their efficiency in the use of water. In addition, it was possible quantify to what extent climate can be a determinant in the δ18O of musts under severe water stress conditions.

Climate projections over France wine-growing region and its potential impact on phenology

Climate change represents a major challenge for the French wine industry. Climatic conditions in French vineyards have already changed and will continue to evolve. One of the notable effects on grapevine is the advancing growing season. The aim of this study is to characterise the evolution of agroclimatic indicators (Huglin index, number of hot days, mean temperature, cumulative rainfall and number of rainy days during the growing season) at French wine-growing regions scale between 1980 and 2019 using gridded data (8 km resolution, SAFRAN) and for the middle of the 21th century (2046-2065) with 21 GCMs statistically debiased and downscaled at 8 km. A set of three phenological models were used to simulate the budburst (BRIN, Smoothed-Utah), flowering, veraison and theoretical maturity (GFV and GSR) stages for two grape varieties (Chardonnay and Cabernet-Sauvignon) over the whole period studied. All the French wine-growing regions show an increase in both temperatures during the growing season and Huglin index. This increase is accompanied by an advance in the simulated flowering (+3 to +9 days), veraison (+6 to +13 days) and theoretical maturity (+6 to +16 days) stages, which are more noticeable in the north-eastern part of France. The climate projections unanimously show, for all the GCMs considered, a clear increase in the Huglin index (+662 to 771 °C.days compared to the 1980-1999 period) and in the number of hot days (+5.6 to 22.6 days) in all the wine regions studied. Regarding rainfall, the expected evolution remains very uncertain due to the heterogeneity of the climates simulated by the 21 models. Only 4 regions out of 21 have a significant decrease in the number of rainy days during the growing season. The two budburst models show a strong divergence in the evolution of this stage with an average difference of 18 days between the two models on all grapevine regions. The theoretical maturity is the most impacted stage with a potential advance between 40 and 23 days according to wine-growing regions.