Federico J.V. Gomez

Monitoring melatonin and its isomer in Vitis vinifera cv. Malbec by UHPLC-MS/MS from grape to bottle

Abstract:  Several studies have shown the presence of melatonin and related compounds in grapes and wines. The latter provides evidence of the possibility to enhance the nutraceutical properties of premium wines. However, there are many external factors that can influence the levels of this indolamine in grape and wines. In this study, the monitoring of melatonin and its tentatively identified isomer was carried out during the entire winemaking process in Vitis vinifera cv. Malbec by ultra high‐performance liquid chromatography‐tandem mass spectrometry. Laboratory and pilot studies were carried out to elucidate the role of grape, yeasts, and tryptophan in the evolution of the indolamines during the fermentation process. Melatonin was detected in grape extract within the range 120–160 ng/g while its isomer was found in musts and finished wines. Our results demonstrate that Saccaromyces cervisiae plays a decisive role in contributing to the content of melatonin and its isomer in wine.

Analytical tools for elucidating the biological role of melatonin in plants by LC-MS/MS

Melatonin (MT) presence in higher plants was recently discovered and the knowledge of its function in vivo is limited. Several studies have recently shown the occurrence of MT and related compounds in grapes and wines. The analysis of MT in plants and foods represents a highly challenging task due to its wide concentration range, the difficulty in the selection of the extraction solvents because of its amphipathic nature, and the fact that it reacts quickly with other matrix components. Thus, sample processing factors; preparation/cleanup procedures; and chromatographic/detection parameters, such as HILIC and reverse phase (C8 and C18) chromatographic modes, ESI, and atmospheric pressure chemical ionization (APCI) in both negative and positive modes were evaluated. Taken together, we have demonstrated that optimal conditions were quite different for each of the matrices under study. A sonication‐mediated extraction step was necessary for grape skin (100% v/v methanol) and plant tissues (50% v/v methanol), while wine and must required a SPE preconcentration step. HILIC−(+) APCI ionization was better for MT standards, while C8−(+) APCI was the best choice for grape skin and C18−(+ESI) was suitable for wine. On the other hand, C8−(+)ESI was the most appropriate for vegetal tissues of Arabidopsis thaliana. Proposed methods were validated and the LODs were in the low picogram levels range. The optimized approaches were applied to the determination of MT and its isomer in different vegetal/food samples; levels found within the range: 4.9–440 ng/g.

Melatonin in Arabidopsis thaliana acts as plant growth regulator at low concentrations and preserves seed viability at high concentrations

Since the discovery of melatonin in plants, several roles have been described for different species, organs, and developmental stages. Arabidopsis thaliana, being a model plant species, is adequate to contribute to the elucidation of the role of melatonin in plants. In this work, melatonin was monitored daily by UHPLC-MS/MS in leaves, in order to study its diurnal accumulation as well as the effects of natural and artificial light treatments on its concentration. Furthermore, the effects of exogenous application of melatonin to assess its role in seed viability after heat stress and as a regulator of growth and development of vegetative tissues were evaluated. Our results indicate that melatonin contents in Arabidopsis were higher in plants growing under natural radiation when compared to those growing under artificial conditions, and its levels were not diurnally-regulated. Exogenous melatonin applications prolonged seed viability after heat stress conditions. In addition, melatonin applications retarded leaf senescence. Its effects as growth promoter were dose and tissue-dependent; stimulating root growth at low concentrations and decreasing leaf area at high doses.

Analytical characterization of wine and its precursors by capillary electrophoresis.

The accurate determination of marker chemical species in grape, musts, and wines presents a unique analytical challenge with high impact on diverse areas of knowledge such as health, plant physiology, and economy. Capillary electromigration techniques have emerged as a powerful tool, allowing the separation and identification of highly polar compounds that cannot be easily separated by traditional HPLC methods, providing complementary information and permitting the simultaneous analysis of analytes with different nature in a single run. The main advantage of CE over traditional methods for wine analysis is that in most cases samples require no treatment other than filtration. The purpose of this article is to present a revision on capillary electromigration methods applied to the analysis of wine and its precursors over the last decade. The current state of the art of the topic is evaluated, with special emphasis on the natural compounds that have allowed wine to be considered as a functional food. The most representative revised compounds are phenolic compounds, amino acids, proteins, elemental species, mycotoxins, and organic acids. Finally, a discussion on future trends of the role of capillary electrophoresis in the field of analytical characterization of wines for routine analysis, wine classification, as well as multidisciplinary aspects of the so‐called “from soil to glass” chain is presented.

Novel approaches mediated by tailor-made green solvents for the extraction of phenolic compounds from agro-food industrial by-products

An environmentally friendly method for the phenolic compound extraction from agro-food industrial by-products was developed in order to contribute with their sustainable valorization. A Natural Deep Eutectic Solvent was chemometrically-designed for the first time and compared with traditional solvents in terms of analyte stabilization. The combination of lactic acid, glucose and 15% water (LGH-15) was selected as optimal. A high-efficiency ultrasound-assisted extraction mediated by LGH-15 prior to HPLC-DAD allows the determination of 14 phenols in onion, olive, tomato and pear industrial by-products. NADES synthesis as well as the extraction procedures were optimized by Response Surface Methodology. Thus, phenolic determination in these complex samples was achieved by a simple, non-expensive, eco-friendly and robust system. The application to different matrices demonstrated the versatility of the proposed method. NADES opens interesting perspectives for their potential use as vehicles of bioactive compounds as food additives or pharmaceuticals.

Pencil graphite electrodes for improved electrochemical detection of oleuropein by the combination of Natural Deep Eutectic Solvents and graphene oxide

A novel methodology is presented for the enhanced electrochemical detection of oleuropein in complex plant matrices by Graphene Oxide Pencil Grahite Electrode (GOPGE) in combination with a buffer modified with a Natural Deep Eutectic Solvent, containing 10% (v/v) of Lactic acid, Glucose and H2O (LGH). The electrochemical behavior of oleuropein in the modified‐working buffer was examined using differential pulse voltammetry. The combination of both modifications, NADES modified buffer and nanomaterial modified electrode, LGH‐GOPGE, resulted on a signal enhancement of 5.3 times higher than the bare electrode with unmodified buffer. A calibration curve of oleuropein was performed between 0.10 to 37 μM and a good linearity was obtained with a correlation coefficient of 0.989. Detection and quantification limits of the method were obtained as 30 and 102 nM, respectively. In addition, precision studies indicated that the voltammetric method was sufficiently repeatable, %RSD 0.01 and 3.16 (n = 5) for potential and intensity, respectively. Finally, the proposed electrochemical sensor was successfully applied to the determination of oleuropein in an olive leaf extract prepared by ultrasound‐assisted extraction. The results obtained with the proposed electrochemical sensor were compared with Capillary Zone Electrophoresis analysis with satisfactory results.

Carbon tape as a convenient electrode material for electrochemical paper-based microfluidic devices (ePADs)

Electrochemical paper-based analytical devices represent an innovative and versatile platform for fluid handling and analysis. Nevertheless, the intrinsic structure of the paper can impose limitations to both the selection of the electrode material and the method selected to attach the electrodes to the device, potentially affecting the analytical performance of the device. To address these limitations, we herein propose carbon tape as a simple and low cost alternative to develop ePADs. The proposed material (in the form of tape or tabs) was first characterized using a combination of contact angle analysis, resistivity, Raman spectroscopy, cyclic voltammetry, and electrochemical impedance spectroscopy. Upon this initial assessment, carbon tape was selected and modified with carbon nanotubes, to provide not only a better surface for proteins to adhere to, but also an enhanced electroactive surface. The analytical performance of the resulting device was assessed by integrating three enzymes that facilitate the oxidation of ethanol, glucose, and phenol, and by performing the detection of these analytes in beer samples. The resulting device, for which materials cost less than a dollar, represents a simple alternative material for ePADs, applied in this case to monitor three of the most important parameters during the production of beers.

Natural deep eutectic solvents-mediated extractions: The way forward for sustainable analytical developments

The concept of sustainable development has impacted in analytical chemistry changing the way of thinking processes and methods. It is important for analytical chemists to consider how sample preparation can integrate the basic concepts of Green Chemistry. In this sense, the replacement of traditional organic solvents is of utmost importance. Natural Deep Eutectic Solvents (NADES) have come to light as a green alternative. In the last few years, a growing number of contributions have applied these natural solvents proving their efficiency in terms of extraction ability, analyte stabilization capacity and detection compatibility. However, the arising question that has to be answered is: the use of NADES is enough to green an extraction process? This review presents an overview of knowledge regarding sustainability of NADES-based extraction procedures, focused on reported literature within the timeframe spanning from 2011 up to date. The contributions were analyzed from a green perspective in terms of energy, time, sample and solvent consumption. Moreover, we include a critical analysis to clarify whether the use of NADES as extraction media is enough for greening an analytical methodology; strategies to make them even greener are also presented. Finally, recent trends and future perspectives on how NADES-based extraction approaches in combination with computational methodologies can contribute are discussed.