OENO IVAS 2019 banner
IVES 9 IVES Conference Series 9 OENO IVAS 9 OENO IVAS 2019 9 Analysis and composition of grapes, wines, wine spirits 9 Dispersive liquid-liquid microextraction for the quantification of terpens in wines

Dispersive liquid-liquid microextraction for the quantification of terpens in wines

Abstract

In a highly competitive worldwide market, a current challenge for the beverage sector is to diversify the range of products and to offer wines and spirits with typicity and character. 

During alcoholic fermentation, wine yeasts generate a large variety of volatile metabolites, including acetate esters, ethyl fatty acid esters, higher alcohols, volatile fatty acids and volatile sulfur compounds that contribute to the aroma profile of wine. These molecules, refered as fermentative aromas, are the most abundant volatile compounds synthetized by yeasts and the metabolic pathways involved in their formation have been well characterized. Furthermore, other molecules with a major organoleptic impact may be produced during wine fermentation including terpene derivatives. However, little information is available on the contribution of yeasts to the formation of these molecules, in particular on their ability to synthethise de novo the terpens derivatives or to produce hydrolytic enzymes involved in the release of varietal precursors. 

To study the yeasts ability to produce these molecules, a dispersive liquid-liquid microextraction (DLLME) gas chromatography mass spectrometry was developed for their quantification in white wines, synthetic wine and fermented synthetic medium. A mixture of acetone (dispersive solvent) and dichloromethane (extractive solvent) was added to 5 ml of sample. The proposed method showed no matrix effect, a good linearity in enological range (from 10 to 300 μg/L), good recoveries, inter-day precision and good reproducibility. The developed method was applied to the analysis of the capacities of 41 yeast strains to produce terpene compounds in Chardonnay must and in synthetic meidum. Interestingly, the majority of the studied compound has been detected and quantified in the resulting wines. 

This sample-preparation technique is very interesting for high-throughput studies and for economic and environmental reasons because it is fast, easy to operate with a high enrichment, and consumes low volume of organic solvent.

DOI:

Publication date: June 23, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Guillaume Bergler, Michel Brulfert, Anne Ortiz-Julien, Carole Camarasa, Audrey Bloem

Martell-Mumm-Perrier Jouët, Pernod Ricard, Cognac, France 
Lallemand SAS, Blagnac 
UMR SPO, INRA Montpellier 2 place Pierre Viala, 34060 Montpellier, France 

Contact the author

Keywords

DLLME, Terpens, Alcoholic fermentation, Wine yeast 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

Mechanisms involved in the heating of the environment by the aerodynamic action of a wind machine to protect a vineyard against spring frost

One of the main consequences of global warming is the rise of the mean temperature. Thus, the heat summation by the plants begins sooner in the early spring, and by cumulating growing degree-days, phenological development tends to happen earlier. However, spring frost is still a recurrent phenomenon causing serious damages to buds and therefore, threatening the harvests of the winegrowers. The wind machine is a solution to protect fruit crops against spring frost that is increasingly used. It is composed of a 10-m mast with a blowing fan at its peak. By tapping into the strength of the nocturnal thermal inversion, it sweeps the crop by propelling warm air above to the ground. Thus, stratification is momentarily suppressed. Furthermore, the continuous action of the machine, alone or in synergy, or the addition of a heater allow the bud to be bathed in a warmer environment. Also, the punctual action of the tower’s warm gust reaches the bud directly at each rotation period. All these actions allow the bud to continuously warm up, but with different intensities and over a different period. Although there is evidence of the effectiveness of the wind machines, the thermal transfers involved in those mechanisms raise questions about their true nature. Field measurements based on ultrasonic anemometers and fast responding thermocouples complemented by laboratory measurements on a reduced scale model allow to characterize both the airflow produced by the wind machine and the local temperature in its vicinity. Those experiments were realized in the vineyard of Quincy, in the framework of the SICTAG project. In the future paper, we will detail the aeraulic characterization of the wind machine and the thermal effects resulting from it and we will focus on how the wind machine warms up the local atmosphere and enables to reduce the freezing risk.

Terroir traceability in grapes, musts and wine: results of research on Gewürztraminer and Sauvignon Blanc grape varieties in northern Italy

In the study of terroir, a separate analysis of its many component factors can be of great help in accurately identifying a vineyard’s natural elements that impact wine quality and typicity. This research used a dedicated pluri-disciplinary approach to investigate the ecological characteristics, including geology and geographical features, of 14 vineyards that produce Gewürztraminer and Sauvignon Blanc cultivars in the alpine Alto Adige DOC wine region. Both the geopedological method using Vineyards Geological Identity (VGI) and the new Solar Radiaton Identity (SRI) topoclimatic classification method were used to provide analytical measurements and qualitative/quantitative characterisations. In addition, wide-ranging targeted and untargeted oenological and chemical analyses were carried out on grapes, musts and wines to correlate the soils’ geomineral and physical conditions with the biochemical properties of their fruits and wines. The research identified strong correlations between vineyard geo-identity and wine biofingerprint, confirming a mineral traceability of strontium rubidium ratio and some minerals distinctive to the local geology, such as K, Ca, Ag, Ba and Mn.  The study also discovered that particular geomineral and physical soil conditions of the studied vineyards are related to the different amount of amino acids, primary varietal aromas and polyphenols found in grapes, musts and wines. The research confirmed that winemaking technologies support oenological quality, although in some cases, human practices can overpower certain characteristic elements in wine, erasing the typical imprint left by the vineyards’ natural terroir, which becomes less traceable. Terroir abiotic ecological factors and vineyard identity can be classified in detail using the new VGI and SRI analysis methods to discover interrelationships between geo-pedological and topoclimatic conditions that impact wine quality. These methods are also helpful in identifying which ecological elements are exclusive to a particular vineyard or wine sub-region.

Phenolic composition of Tempranillo Blanco grapes changes after foliar application of urea

Our research aimed to determine the effect and efficiency of foliar application of urea on the phenolic composition of Tempranillo Blanco grapes. The field experiment was carried out in 2019 and 2020 seasons and the plot was located in D.O.Ca Rioja (North of Spain). The vineyard was Vitis vinifera L. Tempranillo Blanco and grafted on Richter-110 rootstock. The treatments were control (C), whose plants were sprayed with water and three doses of urea: plants were sprayed with urea 3 kg N/ha (U3), 6 kg N/ha (U6) and 9 kg N/ha (U9). The applications were performed in two phenological stages, pre-veraison (Pre) and veraison (Ver). Also, each of the treatments was repeated one week later. Control and treatments were performed in triplicate and arranged in a randomised block design. Grapes were harvested at optimum ripening stage. High-performance liquid chromatography was used to analyse the phenolic composition of the grapes. Finally, the results obtained from the analytical determinations – flavonols, flavanols and non-flavonoid (hydroxybenzoic acids, hydroxycinnamic acids and stilbenes) – were studied statistically by analysis of variance. The results showed that, in 2019, U6-Pre and U9-Pre treatments increased the hydroxybenzoic acid content in grapes, and also all foliar treatments applied at Pre enhanced the stilbene concentration. Moreover, U3-Ver was the only treatment that rose flavonol and stilbene contents in the Tempranillo Blanco grapes. In 2020, all treatments applied at Pre enhanced the flavonol concentration in grapes. Furthermore, U3-Pre and U9-Pre treatments increased stilbene content in grapes. Nevertheless, the hydroxybenzoic acid content was improved by U6-Ver and U9-Ver and besides, hydroxycinnamic acid concentration in grapes was increased by all treatments applied at Ver. In conclusion, the lower and highest dose of urea (U3 and U9), applied at pre-veraison, were the best treatments to improve the Tempranillo Blanco grape phenolic composition.

Effects of organic mulches on the soil environment and yield of grapevine

Farming management practices aiming at conserving soil moisture have been developed in arid and semiarid-areas facing water scarcity problems. Organic mulching is an effective method to manipulate the crop-growing microclimate increasing crop yield by controlling soil temperature, and retaining soil moisture by reducing soil evaporation. In this sense, the effectiveness of different organic mulching materials (straw mulch and grapevine pruning debris) applied within the row of a vineyard was evaluated on the soil and on the vine in a Tempranillo vineyard located in La Rioja (Spain). Organic mulches were compared with a traditional bare soil management technique (based on the use of herbicides to avoid weed incidence). Mulching coverages favourably influenced the soil water retention throughout all the grapevine vegetative cycle. However, the soil-moisture variation was not the same under different mulching materials, being the straw mulch (SM) the one that retained more water in comparison with grapevine pruning debris (GPD) based-cover. The changes of soil moisture in the upper surface layer (0–10 cm) were highly dynamic, probably due to water vapour fluxes across the soil-atmospheric interface. However, both, SM and GPD reduced these fluctuations as compared with bare soils. A similar trend occurred with soil temperature. Both organic mulches altered soil temperature in comparison with bare soil by reducing soil temperature in summer and raising it in winter. Moreover, the same buffering effect for the temperature on the covered soil also remains in the deeper layers. To conclude, we could see that organic mulching had a positive impact on soil-moisture storage and soil temperature and the extent of this effect depends on the type of mulching materials. These changes led to higher rates of photosynthesis and stomatal conductivity compared to bare soils, also favouring crop growth and grape yields.

Combining effect of leaf removal and natural shading on grape ripening under two irrigation strategies in Manto negro (Vitis vinifera L.)

The increasingly frequent heat waves during grape ripening pose challenges for high quality wine grape production. Defoliation is a common practice that can improve the control of diseases in bunches, but also it increases the exposure to sunlight. Grapes exposed to solar radiation reach temperatures over the optimum for berry development and maturation. This makes the development of irrigation and canopy management techniques of great importance to maximize yield and grape quality. A field experiment was carried out during 2021 using Manto negro wine grapes to study the effect of applied irrigation and different light exposure levels on grape quality. Two irrigation treatments were imposed based on the frequency and amount of water doses in a four-block experimental vineyard at Bodega Ribas (Mallorca). Three light exposure treatments were randomly applied in each irrigation plot. The light treatments included exposed clusters from pea size, non-exposed clusters, and shaded clusters after softening. Leaf area index and canopy porosity was estimated every 2 weeks. Midday leaf water potential was measured weekly. Additionally, apparent electrical conductivity was measured between rows to estimate the soil water content variability. Light and temperature sensors were installed at the bunch level to quantify the differences in bunch temperature and light intensity among treatments. The effect of irrigation and cluster light exposure on berry weight, TSS, TA, malic acid, tartaric acid, K+, and pH were analysed at 5 moments along grape ripening. During different heat waves, the natural shading technique decreased the maximum bunch temperature around 10 °C respect to the exposed bunches in both irrigation strategies. The combination of defoliation and shading techniques after softening decreased TSS at harvest and affected most of the quality parameters during the last stages of ripening, showing an interesting technique to delay ripening in warm viticulture areas.