IVAS 2022 banner

Development, validation and application of a fast UHPLC-HRMS method for the analysis of amino acids and biogenic amines in wines and musts.

Abstract

The amino acids in grape juice are an important nitrogen source for yeast during alcoholic fermentation. Additionally, certain AAs are precursors to some of the volatile compounds found in wine and overall, they have an important role in the aromatic complexity of wines. Biogenic amines are produced during the fermentation process by microbial decarboxylation of the corresponding amino acid precursors. Yet, their fate is not only determined by the presence of microorganisms as they are also produced by the grape berries in response to abiotic factors. The presence of biogenic amines affect the sensory attributes of wines by reducing the varietal character and giving rise to meaty and metallic aromas in wines having higher pH values. Moreover, they also have a detrimental impact on consumer health. Due
to the importance of those compounds, several detection and quantification methods have been designed and published. However, to the best of our knowledge, none of them entailed the use of ultra-high performance liquid chromatography (UHPLC) coupled to a high-resolution mass spectrometry (HRMS). In this study, an innovative UHPLC-HRMS method useful for fast quantification of a broad range of amino acids and amines was developed. Twenty-five amino acids, twelve biogenic amines as well as glutathione and S-methylmethionine were identified and quantified in a single chromatographic run taking only 12 minutes. Additionally, a second run of the same length involving the use of o-phthalaldehyde derivatisation reagent was developed to quantify two more amines and ammonium. Validation of the method was performed in relation to the limit of detection, limit of quantification, linearity range, repeatability, reproducibility, and recovery. Once validated, the method was successfully tested on commercial oenological samples and grape musts, demonstrating its applicability to fast routine analysis of musts and wines

DOI:

Publication date: June 24, 2022

Issue: IVAS 2022

Type: Poster

Authors

Delaiti Simone¹, Nardin Tiziana¹and Larcher Roberto

¹Edmund Mach Foundation (FEM

Contact the author

Keywords

amino acids, amines, UHPLC-HRMS, wine, must

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

Downscaling of remote sensing time series: thermal zone classification approach in Gironde region

In viticulture, the challenges of local climate modelling are multiple: taking into account the local environment, fine temporal and spatial scales, reliable time series of climate data, ease of implementation and reproducibility of the method. At the local scale, recent studies have demonstrated the contribution of spatialization methods for ground-based climate observation data considering topographic factors such as altitude, slope, aspect, and geographic coordinates (Le Roux et al, 2017; De Rességuier et al, 2020). However, these studies have shown questions in terms of the reproducibility and sustainability of this type of climate study. In this context, we evaluated the potential of MODIS thermal satellite images validated with ground-based climate data (Morin et al, 2020). Previous studies have been encouraging, but questions remain to be explored at the regional scale, particularly in the dynamics of the massive use of bioclimatic indices to classify the climate of wine regions. The results at the local scale were encouraging, but this approach was tested in the current study at the regional scale. Several objectives were set: 1) to evaluate the downscaling method for land surface temperature time series, 2) to identify regional thermal structure variations. We used weekly minimum and maximum surface temperature time series acquired by MODIS satellites at a spatial resolution of 1000 m and downscaled at 500 m using topographical variables. Two types of analyses were performed:

Under-vine management effects on grapevine production, soil properties and plant communities in South Australia

Under-vine (UV) management has traditionally consisted of synthetic herbicide use to limit competition between weeds and grapevines. With growing global interest towards non-synthetic chemical use, this study aimed to capture the effects of alternative UV management at two commercial Shiraz vineyards in South Australia, where the sole management variables were UV management since 2016. In adjacent treatment blocks, cultivation (CU) was compared to spontaneous vegetation (SV) in McLaren Vale (MV), and herbicide was compared to SV in Eden Valley (EV). Soil water infiltration rates were slower and grapevine stem water potential was lower in CU compared to SV in MV, with the latter having a plant community dominated by soursob (Oxalis pes-caprae) during winter; while in EV, there was little separation between the treatments. Yields were affected at both sites, with SV being higher in MV and HE being higher in EV. In MV, the only effect on grape must was a lower 13C:12C isotope ratio in CU, indicating greater grapevine water stress. In the grape must at EV, SV had higher total soluble solids, total phenolics, anthocyanins, and yeast available nitrogen; and lower pH and titratable acidity. Pruning weights were not affected by the treatments in MV, while they were higher in HE at EV. Assessments revealed that the differing soil types at the two sites were likely the main determinants of the opposing production outcomes associated with UV management. In the silty loam soil of MV, the higher yields in SV were likely due to more plant-available water, as a potential result of the continuous soil bio-pores formed by winter UV vegetation. Conversely, in the loamy sand soils of EV with a lower cation exchange capacity, the lower yields and pruning weights in SV suggest the UV vegetation competed significantly with the grapevines for available water and nutrients.

Making sense of available information for climate change adaptation and building resilience into wine production systems across the world

Effects of climate change on viticulture systems and winemaking processes are being felt across the world. The IPCC 6thAssessment Report concluded widespread and rapid changes have occurred, the scale of recent changes being unprecedented over many centuries to many thousands of years. These changes will continue under all emission scenarios considered, including increases in frequency and intensity of hot extremes, heatwaves, heavy precipitation and droughts. Wine companies need tools and models allowing to peer into the future and identify the moment for intervention and measures for mitigation and/or avoidance. Previously, we presented conceptual guidelines for a 5-stage framework for defining adaptation strategies for wine businesses. That framework allows for direct comparison of different solutions to mitigate perceived climate change risks. Recent global climatic evolution and multiple reports of severe events since then (smoke taint, heatwave and droughts, frost, hail and floods, rising sea levels) imply urgency in providing effective tools to tackle the multiple perceived risks. A coordinated drive towards a higher level of resilience is therefore required. Recent publications such as the Australian Wine Future Climate Atlas and results from projects such as H2020 MED-GOLD inform on expected climate change impacts to the wine sector, foreseeing the climate to expect at regional and vineyard scale in coming decades. We present examples of practical application of the Climate Change Adaptation Framework (CCAF) to impacts affecting wine production in two wine regions: Barossa (Australia) and Douro (Portugal). We demonstrate feasibility of the framework for climate adaptation from available data and tools to estimate historical climate-induced profitability loss, to project it in the future and to identify critical moments when disruptions may occur if timely measures are not implemented. Finally, we discuss adaptation measures and respective timeframes for successful mitigation of disruptive risk while enhancing resilience of wine systems.

Amino nitrogen content in grapes: the impact of crop limitation

As an essential element for grapevine development and yield, nitrogen is also involved in the winemaking process and largely affects wine composition. Grape must amino nitrogen deficiency affects the alcoholic fermentation kinetics and alters the development of wine aroma precursors. It is therefore essential to control and optimize nitrogen use efficiency by the plant to guarantee suitable grape nitrogen composition at harvest. Understanding the impact of environmental conditions and cultural practices on the plant nitrogen metabolism would allow us to better orientate our technical choices with the objective of quality and sustainability (less inputs, higher efficiency). This trial focuses on the impact of crop limitation – that is a common practice in European viticulture – on nitrogen distribution in the plant and particularly on grape nitrogen composition. A wide gradient of crop load was set up in a homogeneous plot of Chasselas (Vitis vinifera) in the experimental vineyard of Agroscope, Switzerland. Dry weight and nitrogen dynamics were monitored in the roots, trunk, canopy and grapes, during two consecutive years, using a 15N-labeling method. Grape amino nitrogen content was assessed in both years, at veraison and at harvest. The close relationship between fruits and roots in the maintenance of plant nitrogen balance was highlighted. Interestingly, grape nitrogen concentration remained unchanged regardless of crop load to the detriment of the growth and nitrogen content of the roots. Meanwhile, the size and the nitrogen concentration of the canopy were not affected. Leaf gas exchange rates were reduced in response to lower yield conditions, reducing carbon and nitrogen assimilation and increasing intrinsic water use efficiency. The must amino nitrogen profiles could be discriminated as a function of crop load. These findings demonstrate the impact of plant balance on grape nitrogen composition and contribute to the improvement of predictive models and sustainable cultural practices in perennial crops.

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.