IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Selective and sensitive quantification of wine biogenic amines using a dispersive solid-phase extraction clean-up/concentration method

Selective and sensitive quantification of wine biogenic amines using a dispersive solid-phase extraction clean-up/concentration method

Abstract

Biogenic amines exist in numerous foods, including wine. They can have aliphatic (putrescine, cadaverine, spermine, and spermidine), aromatic (tyramine and phenylethylamine) and heterocyclic structure (histamine and tryptamine). In wine, the biogenic amines have three possible origins, they can be present in the grape juice, can be formed during alcoholic fermentation by yeasts, or during malolactic fermentation by the action of lactic acid bacteria that can decarboxylate amino acids present in wine. Therefore, the main request for the formation of biogenic amines is the presence of free amino acids, the existence of decarboxylase-positive microorganisms, and environmental conditions that permit bacterial growth and decarboxylase synthesis and activity [1]. In low levels, biogenic amines contribute to physiological functions like regulation of stomach pH, body temperature, or brain activity. Nevertheless, the ingestion of wines comprising high levels of biogenic amines, numerous toxicological effects may happen for example headaches, nausea, and in severe situations intracerebral hemorrhage or even death [2].
Monitoring the existence of these compounds in wine is essential, not only from the toxicological perspective but also as an indicator of wine spoilage [3]. In this work, a simple dispersive solid-phase extraction (dSPE) was developed for sample clean-up and pre-concentration of biogenic amines in wine. The dSPE using a strong cation exchange resin increased the selectivity and sensitivity of the analysis by elimination of interfering compounds and a five-fold enrichment of biogenic amines. The derivatization with benzoyl chloride and then the extraction with diethyl ether steps were optimized. HPLC with diode array detector was used as an analytical technique and this method was validated for twelve biogenic amines – ethylamine, propylamine, butylamine, putrescine, cadaverin, typtamine, b-phenylethylamine, amylamine, spermidine, hexylamine, spermine, and histamine. The method presented an adequate precision and linearity with detection limits ranging from 0.133 to 0.509 mg/L. Recoveries ranging from 72 to 99% prove the accuracy of the method for determining biogenic amines in red, white, and Tawny Port wine samples yielding chromatograms clean from interferents [4]. The method was applied successfully to the analysis of 31 young commercial red wines from the 2016 vintage collected in wineries located in different Portuguese demarcated wine regions. The dSPE method developed is a simple, cheap, quick, and green sample clean-up strategy for biogenic amine analysis. Increasing their selective and sensitive UV detection, the more used detector in liquid chromatography. The results indicated that this method is suitable for the intended purpose with a good recovery, precision, detection, and quantification limits, and with a suitable range for the amounts of biogenic amines existing in wine. 

References

[1]R. E. Anli, M. Bayram, Food Reviews International, 25:1 (2008) 86-102.
[2] A. C. Manetta, L. D. Guiseppe, R., Tofalo, M. Martuscelli, M. Schirone, M. Giammarco, G. Suzzi. Food Control. 2016. 65, 351-356.
[3] L. Beneduce, A. Romano, V. Capozzi, P. Lucas, L. Barnavon, B. Bach, P. Vuchot, F. Grieco, G. Spano. Ann. Microbiol. 2010, 60, 573-578.
[4]J. Milheiro, L. C. Ferreira, L. Filipe-Ribeiro, F. Cosme, F. M. Nunes, Food Chemistry, 274 (2019) 110-117.

DOI:

Publication date: June 24, 2022

Issue: IVAS 2022

Type: Poster

Authors

Cosme Fernanda1, Milheiro Juliana1, Ferreira Leonor C.1, Filipe-Ribeiro Luís1 and Nunes Fernando M.1

1Chemistry Research Centre-Vila Real (CQ-VR), Food and Wine Chemistry Laboratory, University of Trás-os-Montes and Alto Douro, School of Life Sciences and Environment

Contact the author

Keywords

Red wine; Biogenic amines; Dispersive solid phase extraction; Derivatization, Histamine.

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Influence of agronomic practices in soil water content in mid-mountain vineyards

In the context of LIFE project MIDMACC (LIFE18 CCA/ES/001099), several pilots have been installed in vineyards in mid mountain areas of Catalonia (NE Spain) to test well stablished agronomic practices to increase the adaptation of Mediterranean mid mountain to climate change. Soil water content (SWC) at three different depths (15, 30 and 45cm) was measured in continuum from August 2020. One pilot (WC) included a well-established green cover (GC), a new GC (NC) and a conventional soil management (CM, tilling+herbicides). NC presented an intermediate state between WC and CM, responding similarly to CM in autumn but quickly reaching similar SWC to WC, then following the same evolution till next spring, with CM presenting lower values along autumn and winter. Then vegetation activation decreased SWC in all plots, (much slower in CM, lacking GC). Sensibility to spring rains is again intermediate for NC, which joins SWC evolution of CM by the end of spring till next autumn. It is expected that NC will resemble WC more and more as its GC develops. In the pilot combining vine training (VSP vs Gobelet) and hillside management (slope vs terrace), no clear pattern could be related with these conditions. However, both terraces seem to be more sensitive to spring rains. A third pilot included new vineyards (7 and 1 year old). In the new vineyard (N), higher canopy development, a spontaneous green cover and row straw resulted in a slower SWC dynamic, not so sensitive to rains but conserving more soil water in spring and most of summer, even with presumably a higher water extraction by vines. In the newest vineyard (VN) the deepest sensor is still sensitive to rain events all over the year and SWC is always highest at this depth, revealing small water capture by vines.

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.

Effect of one-year cover crop and arbuscular mycorrhiza inocululation in the microbial soil community of a vineyard

The microbial composition of the soil is an important factor to consider in viticulture, since its influence on the “terroir” and on the organoleptic properties of the wine have been demonstrated. Different agronomic techniques have the potential to modify the composition and functionality of the soil microbial community. Maintaining green covers is known to increase soil microbial diversity. The direct application of inoculum of beneficial microorganisms to the soil has also been used to increase their abundance. However, the environmental conditions of each site seem to have a determining weight in the result of these practices. In this study, we compared the effect on the microbial community of a cover crop with legumes in autumn and the inoculation of grapevines with commercial inoculum bases on Rhizophagus irregularis and Funeliformis mosseae in the previous spring. The study has been carried out in a vineyard in Binissalem, Mallorca, Spain. After applying the treatments, we will analyze the soil microbial communities using the data obtained from Illumina amplification of soil DNA from the 16S and ITS regions to analyze bacteria and fungi community, respectively. In addition, we will record the physicochemical characteristics of the soil at each sampling point. The result showed that agronomic management, in the short term, has less influence than soil characteristics on the composition of the soil microbiome. With these results, we can conclude that in a vineyard, agricultural techniques should focus on improving the characteristics of the soil to improve the biodiversity of the soil microbiota.

Adaptation to soil and climate through the choice of plant material

Choosing the rootstock, the scion variety and the training system best suited to the local soil and climate are the key elements for an economically sustainable production of wine. The choice of the rootstock/scion variety best adapted to the characteristics of the soil is essential but, by changing climatic conditions, ongoing climate change disrupts the fine-tuned local equilibrium. Higher temperatures induce shifts in developmental stages, with on the one hand increasing fears of spring frost damages and, on the other hand, ripening during the warmest periods in summer. Expected higher water demand and longer and more frequent drought events are also major concerns. The genetic control of the phenotypes, by genomic information but also by the epigenetic control of gene expression, offers a lot of opportunities for adapting the plant material to the future. For complex traits, genomic selection is also a promising method for predicting phenotypes. However, ecophysiological modelling is necessary to better anticipate the phenotypes in unexplored climatic conditions Genetic approaches applied on parameters of ecophysiological models rather than raw observed data are more than ever the basis for finding, or building, the ideal varieties of the future.

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.