Fully automated non-targeted GC-MS data analysis

Abstract

Non-targeted analysis is applied in many different domains of analytical chemistry such as metabolomics, environmental and food analysis. In contrast to targeted analysis, non-targeted approaches take information of known and unknown compounds into account, are inherently more comprehensive and give a more holistic representation of the sample composition. 

Besides chromatographic techniques coupled to high resolution mass spectrometry such as LC-HRMS, gas chromatography with unit resolution mass spectrometry is still regularly utilized for non-targeted profiling or fingerprinting. This is mainly due to high separation power of GC and a wide availability and low costs of quadrupole mass spectrometers. 

Although several non-targeted approaches have been developed, data processing still remains a serious bottleneck. Baseline correction, feature detection, and retention time alignment can be prone to errors and time-consuming manual corrections are often necessary. We therefore developed an automated strategy to non-targeted GC-MS data avoiding feature detection and retention time alignment. The novel automated approach includes segmentation of chromatograms along the retention time axis, multiway decomposition of transformed segments followed by a supervised machine learning pipeline based on gradient boosted tree classification on the decomposed tensor [1, 2]. 

In order to make this novel data analysis strategy available to scientists without programming background, we developed a convenient browser based application. For the here presented interactive browser application the open source Python packages Bokeh and HoloViews were used. The application will be online freely available soon. 

[1] J. Vestner, G. de Revel, S. Krieger-Weber, D. Rauhut, M. du Toit, A. de Villiers, Toward automated chromatographic fingerprinting: A non-alignment approach to gas chromatography mass spectrometry data. Acta Chimica Acta 911 (2016) 42-58 
[2] K. Sirén, U. Fischer, J. Vestner, Automated supervised learning pipeline for non-targeted GC-MS data analysis. Analytica Chimica Acta: X 1 (2019) 100005

DOI:

Publication date: June 19, 2020

Issue: OENO IVAS 2019

Type: Article

Authors

Jochen Vestner, Kimmo Sirén, Pierre Le Brun, Ulrich Fischer

Institute for Viticulture and Oenology, DLR Rheinpfalz, Breitenweg 71, D-67435 Neustadt, Germany
Institut National Supérieur des Sciences Agronomiques de l’Alimentation et de l’ Environnement, Agrosup Dijon, 6 boulevard Docteur Petitjean, 21000 Dijon, France
Department of Chemistry, University of Kaiserslautern, Erwin-Schroedinger-Strasse 52, D-67663 Kaiserslautern

Contact the author

Keywords

metabolomics, non-targeted, GC-MS, exploratory data analysis 

Tags

IVES Conference Series | OENO IVAS 2019

Citation

Related articles…

Green berries on Gewürztraminer (Vitis vinifera L.) in South Tyrol (Italy)

The grape variety Gewürztraminer is known to be affected by two physiological disorders namely berry shrivel and bunch stem necrosis. During the season 2014 we noticed a new symptomatology type of ripening disorder on the variety. The new symptom showed not all berries fallowing the normal maturation stages, but single berries remaining at a soft but green stage till harvest. The broad distribution of these so called “green berries” symptoms in different production sites of our region, caused huge damage due to the difficulty of eliminating single berries per bunch before harvesting. Therefore, the Research Centre Laimburg began to investigate the reasons and origins of this new symptom. This work shows the results of first attempts to find causes for the symptom as well as the resulting approach to mitigate symptoms. Applications of magnesium leaf fertilizer showed first promising results against this putative disorder. To study the causal effect of the green berries 30 symptomatic vineyards in 2014 have been selected for a monitoring during the season 2016. To evaluate the foliar nutrient treatment two vineyards have been selected for application of magnesium sulfate and magnesium chloride. Leaf and berry nutrient analysis, as well as the main quality parameters during ripening have been performed. As soon as “green berries” symptoms appeared, incidence and severity have been evaluated. Most of the symptomatic vineyards of the 2016 monitoring showed light to clear magnesium deficit symptoms on their foliage. Only during the seasons 2020 and 2021 “green berries” symptoms could be found in the leaf fertilizer treatment vineyards. Both seasons showed a significant effect of the magnesium treatments to reduce the incidence and severity of the symptom. It seems that the appearance of the “green berries” symptom on Gewürztraminer is correlated to a disturbed uptake of magnesium of the vines.

How does aromatic composition of red wines, resulting from varieties adapted to climate change, modulate fruity aroma?

One of the major issues for the wine sector is the impact of climate change linked to the increasing temperatures which affects physicochemical parameters of the grape varieties planted in Bordeaux vineyard and consequently, the quality of wine. In some varietals, the attenuation of their fresh fruity character is accompanied by the accentuation of dried-fruit notes [1]. As a new adaptive strategy on climate change, some winegrowers have initiated changes in the Bordeaux blend of vine varieties [2]. This study intends to explore the fruitiness in wines produced from grape varieties adapted to the future climate of Bordeaux. 10 commercial single–varietal wines from 2018 vintage made from the main grape varieties in the Bordeaux region (Cabernet franc, Cabernet-Sauvignon and Merlot) as well as from indigenous grape varieties from the Mediterranean basin, such as Cyprus (Yiannoudin), France (Syrah), Greece (Agiorgitiko and Xinomavro), Portugal (Touriga Nacional) and Spain (Garnacha and Tempranillo), were selected among 19 samples using sensory descriptive analyses. Both sensory and instrumental analyses were coupled, to investigate their fruity aroma expression. For sensory analysis, samples were prepared from wine, using a semi preparative HPLC method which preserves wine aroma and isolates fruity characteristics in 25 specific fractions [3,4]. Fractions of interest with intense fruity aromas were sensorially selected for each wine by a trained panel and mixed with ethanol and microfiltered water to obtain fruity aromatic reconstitutions (FAR) [5]. A free sorting task was applied to categorize FAR according to their similarities or dissimilarities, and different clusters were highlighted. Instrumental analysis of the different FAR and wines demonstrated variations in their molecular composition. Results obtained from sensory and gas chromatography analysis enrich the knowledge of the fruity expression of red wines from “new” grape varieties opening up new perspectives in wine technology, including blending, thus providing new tools for producers.

Late frost protection in Champagne

Probably one of the most counterintuitive impacts of climate change on vine is the increased frequency of late frost. Champagne, due to its septentrional position is historically and regularly affected by this meteorological hazard. Champagne has therefore developed a strong experience in frost protection with first experiments dating from the end of 19th century. Frost protection can be divided in two parts: passive and active. Passive protection includes all the methods that do not seek to modify the vine’s environment or resistance at the time of frost. The most iconic passive protection in Champagne is the establishment of the individual reserve. This reserve allows to stock a certain quantity of clear wine during a surplus year to compensate a meteorological hazard like frost during the following years. Other common passive methods are the control of planting area (walls, bushes, topography), the choice of grape variety, late pruning, or the impact of grass cover and tillage. Active frost protection is also divided in two parts. Most of the existing techniques tend to modify vine’s environment. Most of the time they provide warmth (candles, heaters, windmills, heating cables…), or stabilise bud’s temperature above a lethal threshold (water sprinkling). The other way to actively fight is to enhance the resistance of buds to frost (elicitors). The Comité Champagne evaluates frost protection methods following three main axes: the efficiency, the profitability, and the environmental impact through a lifecycle assessment. This study will present the results on both passive and active protection following these three axes.

VINIoT – Precision viticulture service

The project VINIoT pursues the creation of a new technological vineyard monitoring service, which will allow companies in the wine sector in the SUDOE space to monitor plantations in real time and remotely at various levels of precision. The system is based on spectral images and an IoT architecture that allows assessing parameters of interest viticulture and the collection of data at a precise scale (level of grape, plant, plot or vineyard) will be designed. In France, three subjects were specifically developed: evaluation of maturity, of water stress, and detection of flavescence dorée. For the evaluation of maturity, it has been decided first to work at the berry scale in the laboratory, then at the bunch scale and finally in the vineyard. The acquisition of the spectral hyperstal image as well as the reference analyzes to measure the maturity, were carried out in the laboratory after harvesting the berries in a maturity monitoring context. This work focuses on a case study to predict sugar content of three different grape varieties: Syrah, Fer Servadou and Mauzac. A robust method called Roboost-PLSR, developed in the framework of this work (Courand et al., 2022), to improve prediction model performance was applied on spectra after the acquirement of hyperspectral images. Regarding the evaluation of water stress, to work with a significant variability in terms of water status, it has been worked first with potted plants under 2 different water regimes. The facilities have allowed the supervision of irrigation and micro-climatic conditions. The regression models on agronomic variables (stomatal conductance, water potential, …) are studied. To detect flavescence dorée, the experimental plan has consisted of work at leaf scale in the laboratory first, and then in the field. To detect the disease from hyper-spectral imaging, a combination of multivariate curve resolution-alternating least squares (MCR-ALS) and factorial discriminant analysis (FDA) was proposed. This strategy proved the potential towards the discrimination of healthy and infected leaves by flavescence dorée based on the use of hyperspectral images (Mas Garcia et al., 2021).

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: