Volatile Profile in Greek Grape Marc Spirits with HS-SPME-GC-MS and Chemometrics: Evaluation of Terroir Impact.

Greek grape marc spirits, although being traditional products, have lately attracted increasing attention as a revisited spirit product category. A headspace solid-phase microextraction coupled to the gas chromatography-mass spectrometry method was optimized in order to determine the volatile composition of 39 samples of grape marc spirits from eight major geographical regions of Greece and 4 protected designations of origin (PDOs). This untargeted analysis approach yielded the identification and quantification of 200 volatiles characterizing those products. Principal component analysis and partial least-squares-discriminant analysis (PLS-DA) combined with the volatile matrix were further employed to identify terroir denomination. Initial results showed a clear separation of the PDO products from Tyrnavos from the rest. Variable importance in projection data pretreatment was further adopted, and with 31 volatiles subsequently used with PLS-DA, products were correctly classified according to region of origin at 76.92% and for 3 out of 4 PDO denominations at 100%.

Pulsed electric field: A "green" extraction technology for biomolecular products from glycerol with fermentation of non-Saccharomyces yeasts.

Glycerol is the main organic by-product of the biodiesel industry and it can be a source of carbon for fermentations or a substrate for biotransformations. This work investigates a process that uses pulsed electric field (PEF) to enhance polyol and propanediols extraction from a glycerol/glucose fermentation broth. Three different commercial, non-Saccharomyces strains, Torulaspora delbrueckii Prelude (Hansen), Torulaspora delbrueckii Biodiva 291 (Lallemand) and Metschnikowia pulcherrima (Lallemand) were studied. The results revealed that PEF had a positive impact on the extraction of polyols ranging from 12 to 191%, independently of fermentation conditions. Torulaspora delbrueckii Biodiva 291 (Lallemand) was found to be more efficient at pH 7.1. An optimized chromatography-based method for the qualitative and quantitative determination of the formed products evaluated. The experiments were carried out either in flasks or in a bioreactor.

Olive Oil Produced from Olives Stored under CO2 Atmosphere: Volatile and Physicochemical Characterization.

In this study, an alternative debittering technique for olives, invented and patented by Prof. Vassilis Dourtoglou, was employed. Olive fruits (Olea europaea cv. Megaritiki) were stored under CO2 atmosphere immediately after harvest for a period of 15 days. After the treatment, a sensory evaluation between the olives stored under CO2 and those stored under regular atmospheric conditions (control) was performed. Additionally, the CO2-treated olives were used for the cold press of olive oil production. The volatile profile of the olive oil produced was analyzed using headspace solid-phase microextraction (HS-SPME) and gas chromatography coupled to mass spectrometry (GC-MS). A total of thirty different volatile compounds were detected. The volatile characteristics of olive oil are attributed, among others, to aldehydes, alcohols, esters, hydrocarbons, alkanes, and terpenes. The volatile compounds' analysis showed many differences between the two treatments. In order to compare the volatile profile, commercial olive oil was also used (produced from olives from the same olive grove with a conventional process in an industrial olive mill). The antioxidant activity, the content of bioactive compounds (polyphenols, α-tocopherol, carotenoids, and chlorophylls), and the fatty acids' profile were also determined. The results showed that the oil produced from CO2-treated olives contains different volatile components, which bestow a unique flavor and aroma to the oil. Moreover, this oil was found comparable to extra virgin olive oil, according to its physicochemical characteristics. Finally, the enhanced content in antioxidant compounds (i.e., polyphenols) not only rendered the oil more stable against oxidation but also better for human health. The overall quality of the olive oil was enhanced and, as such, this procedure holds great promise for future developments.

Optimization of Pulsed Electric Field as Standalone "Green" Extraction Procedure for the Recovery of High Value-Added Compounds from Fresh Olive Leaves.

Olive leaves (OLL) are reported as a source of valuable antioxidants and as an agricultural by-product/waste. Thus, a twofold objective with multi-level cost and environmental benefits arises for a "green" standalone extraction technology. This study evaluates the OLL waste valorization through maximizing OLL extracts polyphenol concentration utilizing an emerging "green" non-thermal technology, Pulsed Electric Field (PEF). It also provides further insight into the PEF assistance span for static solid-liquid extraction of OLL by choosing and fine-tuning important PEF parameters such as the extraction chamber geometry, electric field strength, pulse duration, pulse period (and frequency), and extraction duration. The produced extracts were evaluated via comparison amongst them and against extracts obtained without the application of PEF. The Folin-Ciocalteu method, high-performance liquid chromatography, and differential scanning calorimetry were used to determine the extraction efficiency. The optimal PEF contribution on the total polyphenols extractability (38% increase with a 117% increase for specific metabolites) was presented for rectangular extraction chamber, 25% v/v ethanol:water solvent, pulse duration (tpulse) 2 µs, electric field strength (E) 0.85 kV cm-1, 100 µs period (Τ), and 15 min extraction duration (textraction), ascertaining a significant dependence of PEF assisting extraction performance to the parameters chosen.

Pulsed Electric Field and Salvia officinalis L. Leaves: A Successful Combination for the Extraction of High Value Added Compounds.

The present study aimed to evaluate the pulsed electric field (PEF)-assisted extraction of phytochemicals from Salvia officinalis L. leaves. The study parameters included a PEF pulse duration of 10 or 100 µs for 30 min, using different "green" extraction solvents: pure ethanol, pure water, and their mixtures at 25, 50, and 75% v/v concentrations. The resulting extracts were evaluated against reference extracts obtained without PEF. For estimation of the extraction efficiency, the content in total polyphenols, individual polyphenols, and volatile compounds, as well as the resistance to oxidation, were determined. The optimal PEF contribution on the total and individual polyphenols, rosmarinic acid, extractability (up to 73.2% and 403.1% increase, respectively) was obtained by 25% v/v aqueous ethanol solvent using a pulse duration of 100 µs. PEF was proven to also affect the final concentration and composition of volatile compounds of the extracts obtained.

In situ Creation of the Natural Phenolic Aromas of Beer: A Pulsed Electric Field Applied to Wort-Enriched Flax Seeds.

To fine tune the production of phenolic aromas in beer, a pulsed electric field (PEF) was applied to beer wort, which was enriched with flax seeds. The choice of flax seeds as a source of FA is based on its high content of ferulic precursors and their intrinsic nutritional value. PEF was applied to ground flax seeds, with and without beta glycosidase. Fermentation was carried out with Saccharomyces and non-Saccharomyces yeast strains. Moreover, 4-vinylguaiacol (4-VG), a flavor highly active derived from volatile phenol, was produced by decarboxylation of ferulic acid (FA), or its precursor and flavor-inactive (4-hydroxy-3-methoxycinnamic acid). All yeast strains could metabolize FA into 4-VG, using the pure compound in the synthetic medium or in flax seeds, with the best quantity produced by Saccharomyces cerevisiae as a precursor. The method yields 4-VG production efficiencies up to 120% (mgL-1). Experimental treatment conditions were conducted with E= 1 kV/cm, total time treatment 15 min (peak time ti = 1 µs, pause time tp = 1 ms, Total pulses 9003). Treatment efficacy is independent of the fermentation yeast.

Addition of yogurt to wort for the production of spirits: Evaluation of the spirit aroma over a two-year period.

Triggered by the development of lactic acid bacteria (LAB) during the production of Scotch whisky, this study examined the influence of yeast and LAB inoculation on whisky flavor. Four new spirits were produced using the same process. LAB were added as a form of a Greek yogurt's live culture. In each category (barley and rye), one sample was fermented with Greek yogurt while the other was fermented without it. The spirits were matured and analyzed at five different points. Results from gas chromatography-mass spectrometry (GC-MS) analysis showed basic volatile compounds, along with some important extra compounds with yogurt culture. The most obvious differences were observed in the concentration of butanoic acid, a characteristic acid in spirits undergoing lactic acid fermentation: to identify esters such as ethyl butanoate, ethyl isobutanoate, isoamyl butanoate, and 2-phenylethyl butanoate, they are not typical compounds in whisky.

Pulsed Electric Field Extraction of α and ß-Acids From Pellets of Humulus lupulus (Hop).

This paper investigates the process of extracting hop pellets (hops) utilizing the pulsed electric field (PEF) technique and the contrasting effects of the technique between two distinct hop varieties (one bitter and one aromatic). The effect of PEF on the extraction was evaluated by measuring the concentration of α-acids and ß-acids (humulones and lupulones). Regarding the aromatic character, the hop's volatile caryophyllene, humulene and ß-myrcene were analyzed both with and without employing the PEF treatment. In order to analyze the acids and the volatile fraction, the analytical method of UV-vis spectrophotometry was applied followed by gas chromatography coupled with mass spectrometry. For the second technique, the extracts were previously purified through a Graphitized Carbon Black syringe for Solid Phase Extraction. The results revealed that PEF had a positive impact on the alpha acids of bitter hops by increasing the extraction rate of these acids by 20%, while the volatiles demonstrated an increase of 5.6 and 7.4% for humulene and caryophyllene, respectively. Concerning the aromatic variety of hops, the PEF treatment appeared to have no noteworthy effects.

Nano-formulation enhances insecticidal activity of natural pyrethrins against Aphis gossypii (Hemiptera: Aphididae) and retains their harmless effect to non-target predators.

The insecticidal activity of a new nano-formulated natural pyrethrin was examined on the cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), and the predators Coccinella septempunctata L. (Coleoptera: Coccinellidae) and Macrolophus pygmaeus Rambur (Hemiptera: Miridae), in respect with the nano-scale potential to create more effective and environmentally responsible pesticides. Pyrethrin was nano-formulated in two water-in-oil micro-emulsions based on safe biocompatible materials, i.e., lemon oil terpenes as dispersant, polysorbates as stabilizers, and mixtures of water with glycerol as the dispersed aqueous phase. Laboratory bioassays showed a superior insecticidal effect of the pyrethrin micro-emulsions compared to two commercial suspension concentrates of natural pyrethrins against the aphid. The nano-formulated pyrethrins were harmless, in terms of caused mortality and survival time, to L3 larvae and four-instar nymphs of the predators C. septempunctata and M. pygmaeus, respectively. We expect that these results can contribute to the application of nano-technology in optimization of pesticide formulation, with further opportunities in the development of effective plant protection products compatible with integrated pest management practices.

Biocompatible colloidal dispersions as potential formulations of natural pyrethrins: a structural and efficacy study.

Biocompatible colloidal dispersions of the micro- and nanoemulsion type based on lemon oil terpenes, polysorbates, water, and glycerol were used for the formulation of pyrethrins, botanical insecticides derived from the white pyrethrum daisy, Tanacetum cinerariifolium. The proposed formulation is based on pyrethrin-containing water-in-oil (W/O) microemulsions that could be diluted in one step with an aqueous phase to obtain kinetically stable oil-in-water (O/W) nanoemulsions. Structural characteristics of the micro- and nanoemulsions were evaluated by electron paramagnetic resonance (EPR) spectroscopy, dynamic light scattering (DLS), small angle X-ray scattering (SAXS), and electrical conductivity. Dynamic properties of the surfactant monolayer as evidenced by EPR measurements were affected by the water content, the surfactant, and also the presence of the biocide. DLS and SAXS experiments of the nanoemulsions indicated the existence of two populations of oil droplets dispersed in the aqueous phase, globular droplets of 36-37 nm in diameter, and also larger droplets with diameters >150 nm. All of the applied techniques for structural determination revealed the participation of the biocide in the nanostructure. The insecticidal effect of the encapsulated natural pyrethrin was evaluated in laboratory bioassays upon a target-insect pest, the cotton aphid Aphis gossypii Glover (Hemiptera: Aphididae) in eggplant, and was found to be increased compared to the commercial pyrethrin formulation.

Discrimination of varietal wines according to their volatiles.

A method is being proposed in order to discriminate bottled wines of different varieties when no other information is known. The advantages of the method consist in the fact that anyone who wants to certify the variety, which is written on the label or the area of origin, can use such a technique to achieve the conformity. Additionally, the method can be easily applied by laboratories equipped with a GC. The differentiation has been achieved by using only seven of the total extracted volatiles, mainly higher alcohols and higher alcohol esters, namely 3-methyl-1-butanol, 2,3-butanediol, ethyl lactate, 3-methyl-1-butyl acetate, 2-phenylethanol, phenyl ethyl acetate and p-hydroxy phenyl ethanol. These key compounds are not relevant to a single variety. The proposed method does not take into account variables such as the year of vintage and fermentation procedures (agitation, temperature).

Storage of olives (Olea europaea L.) under CO2 atmosphere: liquid chromatography-mass spectrometry characterization of indices related to changes in polyphenolic metabolism.

Olives (Olea europaea cv. Chondrolia) were stored under a CO2 atmosphere immediately after harvesting for a period of 12 days. Samples obtained at 24-h intervals were analyzed by HPLC to identify components that may reflect changes in the biochemical behavior of the tissue. Four substances were shown to undergo significant fluctuations during storage, while their evolution was found to be different in olives stored under CO2 from those stored under regular atmospheric conditions (control). On the basis of data provided by liquid chromatography-electrospray ionization mass spectrometry, these substances were tentatively identified as hydroxytyrosol glucoside, demethylated ligstroside aglycone, ibotalactone A methyl ester, and verbascoside. The data are discussed in relation to the effect of postharvest treatments of olives for purposes of manipulating their polyphenolic content and plausible development of novel debittering processes.

The simultaneous determination of 1-aminocyclopropane-1-carboxylic acid and cyclopropane-1,1-dicarboxylic acid in Lycopersicum esculentum by high-performance liquid chromatography--electrospray tandem mass spectrometry.

Varying concentrations of cyclopropane-1,1-dicarboxylic acid (CDA), an inhibitor of 1-aminocyclopropane-1-carboxylic acid oxidase, added to the solid culture medium of tomato nodal shoot segments resulted in a reduction in the level of endogenous ethylene according to the concentration of inhibitor applied. Following treatment with inhibitor, plants were homogenised and the concentrations of CDA and of 1-aminocyclopropane-1-carboxylic acid (ACC) were measured simultaneously in the resulting juice using an HPLC-ESI/MS-MS method. The levels of CDA and ACC measured in the plant tissues were associated with the concentration of inhibitor added to the solid medium. The HPLC-ESI/MS-MS method described produced limits of detection of 0.8 pmol for ACC and of 4 pmol for CDA.