High Voltage Electrical Discharges as an Alternative Extraction Process of Phenolic and Volatile Compounds from Wild Thyme (Thymus serpyllum L.): In Silico and Experimental Approaches for Solubility Assessment.

The objective of this study was to evaluate the potential of green solvents for extractions of bioactive compounds (BACs) and essential oils from wild thyme (Thymus serpyllum L.) using theoretical and experimental procedures. Theoretical prediction was assessed by Hansen solubility parameters (HSPs) and conductor-like screening model for realistic solvents (COSMO-RS), to predict the most suitable solvents for extraction of BACs. An experimental procedure was performed by nonthermal technology high voltage electrical discharge (HVED) and it was compared with modified conventional extraction (CE). Obtained extracts were analyzed for chemical and physical changes during the treatment. Theoretical results for solution of BACs in ethanol and water, as green solvents, were confirmed by experimental results, while more accurate data was given by COSMO-RS assessment than HSPs. Results confirmed high potential of HVED for extraction of BACs and volatile compounds from wild thyme, in average, 2.03 times higher yield of extraction in terms of total phenolic content was found compared to CE. The main phenolic compound found in wild thyme extracts was rosmarinic acid, while the predominant volatile compound was carvacrol. Obtained extracts are considered safe and high-quality source reach in BACs that could be further used in functional food production.

Croatian Wines from Native Grape Varieties Have Higher Distinct Phenolic (Nutraceutic) Profiles than Wines from Non-Native Varieties with the Same Geographic Origin.

Croatian wines made from native (Plavac mali and Teran) and non-native grape varieties (Cabernet Sauvignon and Merlot), all grown in Croatian coastal regions, were investigated. Analyses included measurements of antioxidant activities, total phenolic contents and concentrations of non-colored phenolic compounds, chosen based on their known nutraceutical properties. Plavac mali wines were distinguished by higher antioxidant activity, total phenolic content and catechin concentrations but lower flavonol concentrations. Teran wines had higher hydroxytyrosol, myricetin and resveratrol concentrations. Merlot and Cabernet Sauvignon wines had higher flavonol concentrations (except myricetin). Canonical analysis was successful in discriminating Plavac mali from Teran wines, and both varieties were separated from non-native varieties. The results indicate distinct genetic potentials of studied varieties and enable wine authentication based on the investigated bioactive compounds.

Effect of winemaking on phenolic profile, colour components and antioxidants in Crljenak kastelanski (sin. Zinfandel, Primitivo, Tribidrag) wine.

Crljenak kastelanski or Tribidrag is a Croatian autochthonous grape variety which is a parent of the America's most popular variety Zinfandel or Italian Primitivo. The aim of this study was to investigate the dynamics of extraction of phenolics and changes in antioxidant activity of the samples collected during maceration of C. kastelanski grapes processed using two different macerating enzymes. According to the obtained results it can be concluded that wines from C. kastelanski grapes are great source of bioactive phenolics although differences in phenolic profiles between control and enzyme-treated wines were detected. The highest content of total phenolics was detected in control wine (2691 mg GAE/L). Use of pectolitic enzyme Vinozym Vintage improved anthocyanin extraction, while higher colour parameters were observed for wine samples produced using Sihazym Extro. The statistical analysis confirmed great influence of total phenolics and concentrations of some individual phenolic compounds (e.g. catechin, gallic acid, epicatechin) on reducing and free radical scavenging activity of wine samples while the impact of anthocyanins was not detected. According to the obtained results it can be concluded that use of enzymes has slightly negative effect on total phenolics and wine antioxidant properties, but it increases the extraction yield of wine colour components what enables shorter maceration and prevents colour losses during the aging process.

Geographical Discrimination of Croatian Wines by Stable Isotope Ratios and Multielemental Composition Analysis.

The δ18O and δ13C (analyzed by isotope ratio mass spectrometry, IRMS) and concentration of 22 selected elements (analyzed by inductively coupled plasma-optical emission spectrometry, ICP-OES) in 190 Croatian microvinified and commercial wine samples from continental and coastal winegrowing areas and from three viticultural zones (B, CI, and CII) were measured to investigate whether multivariate statistical methods could provide the fingerprint for geographical origin determination. The highest power for discrimination of wines produced in Croatian winegrowing areas was achieved by general discriminant analysis (GDA) showing correct classification of 97.9% of all investigated samples, 100.0% of microvinified samples and 84.8% of commercial samples in the cross-validation matrix. The most significant markers for discrimination of coastal and continental areas found by GDA were δ18O and Co, followed by K, Rb, Sn, Li, and δ13C in descending order. GDA showed higher levels of correctly classified samples from three viticultural zones in Croatia if only microvinified samples were employed in the analysis (94.9%) than for all samples together (86.3%) or for commercial samples (66.1%) in the cross-validation matrix. The discrimination of viticultural zones B, CI, and CII in Croatia was achieved by δ18O, Co, Rb, Li, K, and Sn. The results obtained showed that the relationships between the isotopic ratios and concentrations of different considered elements combined with appropriate statistical model represent a powerful tool in discrimination of wines produced in different Croatian winegrowing areas.

Occurence of Ochratoxin A and Biogenic Amines in Croatian Commercial Red Wines.

Food safety is one of the main concerns in the world and in wine it depends mostly on metabolites of microbial origin. The aim of this study was to investigate the occurrence of natural contaminants, ochratoxin A and biogenic amines (cadaverine, histamine, putrescine and tyramine), in Croatian commercial red wines originating from different Croatian wine-making regions. Ochratoxin A was detected in 92.8% of samples, however its concentrations in all samples were more than 10-fold lower than the limit set by the European Union (2 µg/kg), marking these wines as safe for consumption. The frequency of occurrence and measured concentrations of ochratoxin A were higher in wines produced in southern regions with highest values obtained in wines from southern Dalmatian islands. All samples were contaminated with cadaverine and putrescine, while 88.2% and 82.7% were contaminated with histamine and tyramine, respectively. Histamine concentrations ranged from below the limit of detection to 8.5 mg/L. Higher histamine concentrations were measured in wines with higher pH values which coincided with southern geographic origin. These results reinforce the need for routine detection and quantification of biogenic amines in Croatian wines to achieve better control of vinification and minimize their formation.

Determination of volatile compounds in grape distillates by solid-phase extraction and gas chromatography.

Solid-phase extraction (SPE) procedure on octadecylsilica (C18) was developed for accumulation of volatile compounds from grape distillates. The procedure was optimised for final analysis by capillary gas chromatography. At mass concentrations in model solutions ranging from 0.1 to 50 mg/l solid-phase extraction recoveries of all analytes ranged from 69% for 2-phenylethanol to 102% for capric acid, with RSD values from 2 to 9%. SPE recoveries of internal standards to be added in the sample solution prior to extraction, higher alcohols 2-ethyl-1-hexanol and 1-undecanol, were 97 and 93%, respectively, with RSD values of 3%. Detection limits of analyzed compounds in model solutions ranged from 0.011 mg/l for isoamyl acetate to 0.037 mg/l for caproic acid. Method efficiency was tested in relation to acetic acid content, volume fraction of ethanol and possible matrix effects. A significant influence of matrix on SPE efficiency for geraniol, cis-2-hexen-1-ol and cis-3-hexen-1-ol was detected. For the same reason, 2-phenylethanol could not be determined by developed SPE method in samples of grape distillates. The developed solid-phase extraction method was successfully applied to determine the differences in volatile compound content in different grape distillates produced by the distillation of crushed, pressed and fermented grapes.

Characterization and differentiation of monovarietal grape marc distillates on the basis of varietal aroma compound composition.

To investigate the varietal aroma compound composition of monovarietal grape marc distillates made from six different varieties (Vitis vinifera L.) in the region of Istria (Croatia), 30 samples were subjected to GC/MS and GC/FID analysis. A total of 73 compounds were identified: 45 monoterpenes, 20 sesquiterpenes, 3 diterpenes, and 5 C(13)-norisoprenoids. The largest number and the highest concentration of monoterpenes were found in Muscat Blanc, followed by Rose Muscat of Porec (Muskat ruza porecki) distillates, which were both characterized as highly aromatic. Lower, but still significant monoterpenol content was determined in distillates made from Istrian Malvasia (Malvazija istarska) grape marc. Chardonnay, Cabernet Sauvignon, and Teran distillates exhibited poorer monoterpene profiles, while Teran distillates contained elevated sesquiterpene concentrations. It was concluded that investigated monovarietal grape marc distillates significantly differ in varietal aroma compound composition. Stepwise linear discriminant analysis provided efficient discrimination models, and extracted various monoterpenes, sesquiterpenes and C(13)-norisoprenoids as important differentiators of distillates according to varietal origin.