WAC 2022 banner
IVES 9 IVES Conference Series 9 WAC 9 WAC 2022 9 3 - WAC - Oral 9 Accurate Quantification of Quality Compounds and Varietal Classification from Grape Extracts using the Absorbance-Transmittance Fluorescence Excitation Emission Matrix (A-TEEM) Method and Machine Learning

Accurate Quantification of Quality Compounds and Varietal Classification from Grape Extracts using the Absorbance-Transmittance Fluorescence Excitation Emission Matrix (A-TEEM) Method and Machine Learning

Abstract

Rapid and accurate quantification of grape berry phenolics, anthocyanins and tannins, and identification of grape varieties are both important for effective quality control of harvesting and initial processing for wine making. Current reference technologies including High Performance Liquid Chromatography (HPLC) can be rate limiting and too complex and expensive for effective field operations. Secondary calibrated techniques including UV-VIS and Near and Mid Infrared spectroscopy are insensitive to specific quality compounds and unable to make accurate varietal assignments. In this paper we analyze robotically prepared grape extracts from several key varieties (n=Calibration/p=Prediction samples) including Cabernet sauvignon (64/10), Grenache (16/4), Malbec (14/4), Merlot (56/10), Petit syrah (52/10), Pinot noir (54/8), Syrah (20/2), Terlodego (14/2) and Zinfandel (62/12). Key phenolic and anthocyanin parameters measured by HPLC included Catechin, Epicatechin, Quercetin Glycosides, Malvidin 3-glucoside, Total Anthocyanins and Polymeric Tannins. Separate samples diluted 150 fold in 50% EtOH pH 2 were analyzed in parallel using the A-TEEM method following Multiblock Data Fusion of the absorbance and unfolded EEM data. A-TEEM chemical data were calibrated (n=390) using Extreme Gradient Boost (XGB) Regression and evaluated based on the Root Mean Square Error of the Prediction (RMSEP), the Relative Error of Prediction (REP%) and Coefficient of Variation (R2P) of the Prediction data (n=62). The regression results yielded an average REP% value of 6.0±2.4% and R2P of 0.941±0.024. While we consider the REP% values to be in the acceptable range at <10% we acknowledge that both the grape extraction method repeatability and HPLC reference method repeatability likely contributed the major sources of variation; e.g., A-TEEM sample REP%=1.31 for Polymeric Tannins. Varietal classification was analyzed using XGB discrimination analysis of the Multiblock data and evaluated based on the Prediction data. The classification results yielded 100% True Positive and True negative results for the Prediction Data for all varieties. We conclude that the A-TEEM method requires a minimum of sample preparation and rapid acquisition times (<1 min) and can serve as an accurate secondary method for both grape composition and varietal identification. Importantly, the application of the regression and classification models can be effectively automated for operators.

DOI:

Publication date: June 13, 2022

Issue: WAC 2022

Type: Article

Authors

Adam, Gilmore, Qiang, Sui

Presenting author

Adam, Gilmore – HORIBA Instruments Inc.

E & J Gallo Wines

Contact the author

Keywords

Extreme Gradient Boost – Phenolics – Anthocyanins- Tannins-Grape Variety

Tags

IVES Conference Series | WAC 2022

Citation

Related articles…

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

Analysis of Cabernet Sauvignon and Aglianico winegrape (V. vinifera L.) responses to different pedo-climatic environments in southern Italy

Water deficit is one of the most important effects of climate change able to affect agricultural sectors. In general, it determines a reduction in biomass production, and for some plants, as in the case of grapevine, it can endorse fruit quality. The monitoring and management of plant water stress in the vineyard

Mobile device to induce heat-stress on grapevine berries

Studying heat stress response of grapevine berries in the field often relies on weather conditions during the growing season. We constructed a mobile heating device, able to induce controlled heat stress on grapes in vineyards. The heater consisted of six 150 W infrared lamps mounted in a profile frame. Heating power of the lamps could be controlled individually by a control unit consisting of a single board computer and six temperature sensors to reach a pre-set temperature. The heat energy applied to individual berries within a cluster decreases by the squared distance to the heat source, enabling the establishment of temperature profiles within individual clusters. These profiles can be measured by infrared thermography once a steady state has been reached. Radiant flux density received by a berry depending on the distance was calculated based on a view factor and measured lamp surface temperature and resulted to 665 Wm-2 at 7cm. Infrared thermography of the fruit surface was in good agreement with measurements conducted with a thermocouple inserted at epidermis level. In combination with infrared thermography, the presented device offers possibilities for a wide range of applications like phenotyping for heat tolerance in the field to proceed in the understanding of the complex response of plants to heat stress. Sunburn necrosis symptoms were artificially induced with the aid of the device for cv. Bacchus and cv. Sylvaner in the 2020 and 2021 growing season. Threshold temperatures for sunburn induction (LT5030min) were derived from temperature data of single berries and visual sunburn assessment, applying logistic regression. A comparison of threshold temperatures for the occurrence of sunburn necrosis confirmed the higher susceptibility of cv. Bacchus. The lower susceptibility of cv. Sylvaner did not seem to be related to its phenolic composition, rendering a thermoprotective role of berry phenolic compounds unlikely.

Photoselective shade films affect grapevine berry secondary metabolism and wine composition

Grapevine physiology and production are challenged by forecasted increases in temperature and water deficits. Within this scenario, photoselective overhead shade films are promising tools in warm viticulture areas to overcome climate change related factors. The aim of this study was to evaluate the vulnerability of ‘Cabernet Sauvignon’ grape berry to solar radiation overexposure and optimize shade film use for berry integrity. A randomized complete block design field study was conducted across two years (2020-2021) in Oakville, Napa Valley, CA, with four shade films (D1, D3, D4, D5) differing in the percent of radiation spectra transmitted and compared to an uncovered control (C0). Integrals for gas exchange parameters and mid-day stem water potential were unaffected by the shade films in 2020 and 2021. By harvest, berries from uncovered and shaded vines did not differ in their size or primary metabolism in either year. Despite precipitation exclusion during the dormant season in the shaded treatments, yield did not differ between them and the control in either season. In 2020, total skin anthocyanins (mg/g fresh mass) in the shaded treatments was greater than C0 during berry ripening and at harvest. Conversely, flavonol concentrations in 2020 were reduced in shaded vines compared to C0. The 2020 growing season highlighted the impact of heat degradation on flavonoids. Flavonoid concentrations in 2021 increased until harvest while flavonoid degradation was apparent from veraison to harvest in 2020 across shaded and control vines. Wine analyses highlighted the importance of light spectra to modify wine composition. Wine color intensity, tonality and anthocyanin values were enhanced in D4 whereas antioxidant properties were enhanced in C0 and D5 wines. Altogether, our results highlighted the need of new approaches in warm viticulture areas given the impact that composition of light has on berry and wine quality.

Drought effect on aromatic and phenolic potential of seven recovered grapevine varieties in Castilla-La Mancha region (Spain)

The effects of climate change are seriously affecting the quality of wine grapes. High temperatures and drought cause imbalances in the chemical composition of grapes. The result is overripe grapes with low acidity and high sugar content, which produce wines with excessive alcohol content, lacking in freshness and not very aromatic. As a consequence, the search of varieties with capacity of produce quality grapes in adverse climate conditions is a good alternative to preserve the sustainability of vineyards. In this work, quality parameters of seven Vitis vinifera L. cultivars (five whites and two reds) recently recovered from extinction and grown under two different hydric regimes (rainfed and irrigated) were analyzed during the 2020 vintage. At harvest time, weight of 100 berries, must physicochemical parameters (brix degree, total acidity, malic acid, pH), and carbon and oxygen isotope ratios (δ13C, δ18O) were determined. Subsequently, varietal aroma potential index (IPAv) and total polyphenol index (TPI) were analyzed. Quality parameters, IPAv and TPI, showed significant differences between varieties and water regimes. Both red varieties, Moribel and Tinto Fragoso, stood out for their high aromatic and phenolic potential, which was higher under rainfed regime. Regarding to white varieties, Montonera del Casar and Jarrosuelto stood out in terms of varietal aroma potential. Montonera del Casar high acidity in its musts and Jarrosuelto showed the highest berry weights.