Valorization of Sour Cherry Kernels: Extraction of Polyphenols Using Natural Deep Eutectic Solvents (NADESs).

The objective of this research was to optimize the natural deep eutectic solvent (NADES) extraction process from sour cherry kernels (Prunus cerasus L.). For polyphenol isolation, conventional solid-liquid extraction was employed using different concentrations of ethanol (0, 10, 20, 30, 40, 50, 60, 70, 80, 90, and 96%), as well as the innovative NADES extraction technique. In the initial phase of the research, a screening of 10 different NADESs was conducted, while extraction was carried out under constant parameters (50 °C, 1:20 w/w, 60 min). NADES 4, composed of lactic acid and glucose in a molar ratio of 5:1, exhibited the highest efficiency in the polyphenol isolation. In the subsequent phase of the research, response surface methodology (RSM) was utilized to optimize the extraction process. Three independent variables, namely temperature, extraction time, and solid-liquid (S/L) ratio, were examined at three different levels. The extracted samples were analyzed for total phenol (TP) and antioxidant activity using the DPPH, ABTS, and FRAP assays. ANOVA and descriptive statistics (R2 and CV) were performed to fit the applied model. According to RSM, the optimal extraction conditions were determined as follows: temperature of 70 °C, extraction time of 161 min, and S/L ratio of 1:25 w/w.

A comprehensive review on carotenoids in foods and feeds: status quo, applications, patents, and research needs.

Carotenoids are isoprenoids widely distributed in foods that have been always part of the diet of humans. Unlike the other so-called food bioactives, some carotenoids can be converted into retinoids exhibiting vitamin A activity, which is essential for humans. Furthermore, they are much more versatile as they are relevant in foods not only as sources of vitamin A, but also as natural pigments, antioxidants, and health-promoting compounds. Lately, they are also attracting interest in the context of nutricosmetics, as they have been shown to provide cosmetic benefits when ingested in appropriate amounts. In this work, resulting from the collaborative work of participants of the COST Action European network to advance carotenoid research and applications in agro-food and health (EUROCAROTEN, www.eurocaroten.eu, https://www.cost.eu/actions/CA15136/#tabs|Name:overview) research on carotenoids in foods and feeds is thoroughly reviewed covering aspects such as analysis, carotenoid food sources, carotenoid databases, effect of processing and storage conditions, new trends in carotenoid extraction, daily intakes, use as human, and feed additives are addressed. Furthermore, classical and recent patents regarding the obtaining and formulation of carotenoids for several purposes are pinpointed and briefly discussed. Lastly, emerging research lines as well as research needs are highlighted.

Encapsulation of bioactive compounds extracted from Cucurbita moschata pumpkin waste: the multi-objective optimisation study.

AIM: Artificial neural network (ANN) development to find optimal carriers (pea protein-P, maltodextrin-M, and inulin-I) mixture for encapsulation of pumpkin waste bioactive (ß-carotene and phenolics). METHODS: Freeze-drying encapsulation and encapsulates characterisation in terms of bioactive contents and encapsulation efficiencies, water activity, hygroscopicity, densities, flowability, cohesiveness, particle size (laser diffraction), solubility, colour (CIELab), morphological (SEM), stability and release properties. RESULTS: Optimal encapsulates, OE-T (with highest total bioactive contents; P, M, and I of 53.9, 46.1, and 0%w/w) and OE-EE (with highest bioactive encapsulation efficiencies; P, M, and I of 45.5, 32.0, and 22.5%w/w) had particle diameters of 94.561 ± 1.341 µm and 90.206 ± 0.571 µm, the span of 1.777 ± 0.094 and 1.588 ± 0.089, highest release at pH 7.4 of phenolics of 71.03%w/w after 72 h and 66.22%w/w after 48 h, and ß-carotene of 43.67%w/w after 8 h and 48.62%w/w after 6 h, respectively. CONCLUSION: ANN model for prediction of encapsulates' preparation, showed good anticipation properties (with gained determination coefficients of 1.000).

Composition, texture, sensorial quality, and biological activity after in vitro digestion of durum wheat pasta enriched with carrot waste extract encapsulates.

Aim of this research was to evaluate durum-wheat pasta enriched with encapsulated carrot waste extracts in oil, obtained by freeze drying (FDE) or spray drying techniques (SDE). Five pastas (control, enriched with 10% FDE, or 10% SDE, or 20% FDE, or 20% SDE) were evaluated for furosine, carotenoids, tocols, colour, in vitro bioactivities, cooking performance, texture and sensory quality. The encapsulates added to the enriched pastas α-carotene (0.58-1.24 mg/kg), ß-carotene (1.43-3.29 mg/kg), cis-ß-carotene (0.51-1.11 mg/kg) and total tocols (10.9-33.6 mg/kg). The carotenes were stable and the tocols diminished (-13%) during pasta manufacturing; both decreased (2-18% and 4-15%, respectively) during cooking, but they were still more abundant in the enriched pastas. Antioxidant, anti-hyperglycaemic, anti-inflammatory and anti-proliferative activities after in vitro digestion of cooked pastas improved, while sensory acceptability of control and 10% enriched pastas were similar. The encapsulates addition significantly improved the nutritional and technological qualities of durum-wheat pasta.

Natural Deep Eutectic Solvent (NADES) Extraction Improves Polyphenol Yield and Antioxidant Activity of Wild Thyme (Thymus serpyllum L.) Extracts.

Wild thyme (Thymus serpyllum L.) herbal dust has been recognized as a potential underutilized resource for the recovery of antioxidants. The aim of this paper was to optimize natural deep eutectic solvent (NADES) extraction of polyphenols to obtain improved antioxidant activity of extracts determined by selected in vitro assays (DPPH, FRAP, and ABTS). Twenty different NADES systems were investigated in the first step of the screening of the extraction solvent and l-proline (Pro)-glycerine (Gly) based solvents provided the best results. Preliminary experiments organized by 25-1 fractional factorial design narrowed down the number of extraction factors from five (temperature, extraction time, NADES type, water content and L/S ratio) to three and determined their experimental domain for the final step. A face-centered central composite design with temperature (40-55-70 °C), extraction time (60-120-180 min) and L/S ratio (10-20-30 g NADES/g sample) was applied for influence analysis and process optimization. Multi-response optimization suggested a temperature of 65 °C, time of extraction of 180 min and L/S ratio of 28 g NADES/g DW as optimal extraction parameters. Experimental validation confirmed good agreement between experimental and predicted results in the extract obtained at optimal conditions and the interactions in the most suitable NADES (N16; Pro-Gly-H2O; 1:2:1) were confirmed by the 1H-NMR.

Phytochemical characterization and effects on cell proliferation of Pinus nigra Arn. bark.

Pinus nigra Arn. bark extracts from Mokra gora (MG) and Tara mountains were analyzed with regard to their polyphenolic profile and antioxidative and antiproliferative activity. The ethanol extract from MG showed the highest phenolic, flavonoid, tannin, and proanthocyanidin content when compared with the acetone and methanol extracts from both sites. The same extract exhibited the highest ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt) and ferric reducing antioxidant power (FRAP) radical scavenging ability and total antioxidant activity (TAA). On the basis of high-performance liquid chromatography analysis, catechin, caffeic, syringic, p-coumaric, and ferulic acids were predominantly present in the MG extracts. The ethanol extract from MG was rich in syringic acid, epicatechin and its derivatives, and ferulic acid and its derivatives. The bark extracts also exerted a high cytotoxic bioactivity against all evaluated cell lines (HeLa, MCF7, HT-29, and MRC-5). The antiproliferative activity of P. nigra bark is probably related to the ferulic acid content and its synergistic activity to caffeic acid and taxifolin. The antioxidative role of the presented phenols was confirmed through the obtained significant linear correlation between the total phenolic content and DPPH (r = .934) as well as the FRAP% of the extracts (r = .948). Also, the TAA significantly depended on the proanthocyanidins (r = .902) and tannin contents (r = .914). The composition of the presented compounds could be related to promising antioxidant and antiproliferative efficacy of MG bark.

The perspectives of natural deep eutectic solvents in agri-food sector.

The principles of 'green chemistry' are gaining importance in agri-food sector due to the need to reduce pollution from toxic chemicals, make industrial processes safer and more sustainable, and to offer 'clean-labeled products' required by the consumers. The application of natural deep eutectic solvents (NADES) - natural product-based green liquids is considered the promising alternative to conventional organic solvents. This review is intended to summarize and discuss recent advances related to physicochemical properties of NADES, their applications, compatibility with analytic techniques and toxicological profile, pointing out the challenges and necessary improvements for their wider utilization in agri-food sector. NADES allow extraction of wide range of food compounds and they are proven to be convenient for food-related applications. However, their potential for industrial scale-up utilization is not completely investigated. Examined NADES are readily biodegradable, but only preliminary studies on NADES toxicity which include limited number of NADES molecules are available. Apart from fundamental research dealing with NADES formation and the nature of the interactions and structure underpinning the liquid phase formation, the question of purity of NADES obtained by different synthetic methodologies need to be addressed in the future. Data on physicochemical properties of synthetized NADES are still needed as they are relevant for industrial applications.

QSAR characterization of new synthesized hydantoins with antiproliferative activity.

Hydantois have been identified as constituents of a number of pharmacologically active molecules. In the present study, we have examined in vitro antiproliferative activity against human colon cancer cell lines HCT-116 of three series of 3-(4-substituted benzyl)-hydantoins with various substituent attached in position 5 of the hydantoin ring. Since the investigated compounds have recently been synthesized and show antiproliferative activity, a good understanding of the properties of the potential drug responsible for their pharmacokinetics is an important goal for their further development. One of the important properties is lipophilicity. Lipophilicity has been assessed by reversed-phase liquid chromatography (high-performance thin-layer chromatography and high-pressure liquid chromatography) by means of direct and indirect (using calibration curve) methods. Chromatographic lipophilicity indices in addition to calculated logP values were compared by hierarchical cluster analysis. The linear solvation energy relationship approach was used to understand and compare the types and relative strength of the molecular interactions that occur in the chromatographic as well as in the n-octanol-water partitioning systems. Finally, correlation between in silico pharmacokinetic predictors and antiproliferative activity was examined. Preliminary quantitative structure-activity relationship modeling indicates that pharmacokinetic predictors capture only one-quarter of all chemical features that are important for antiproliferative activity itself. Among selected descriptors are chromatographic lipophilicity indices.

Meripilus giganteus ethanolic extract exhibits pro-apoptotic and anti-proliferative effects in leukemic cell lines.

BACKGROUND: The interest towards botanicals and plant extracts has strongly risen due to their numerous biological effects and ability to counteract chronic diseases development. Among these effects, chemoprevention which represents the possibility to counteract the cancerogenetic process is one of the most studied. The extracts of mushroom Meripilus giganteus (MG) (Phylum of Basidiomycota) showed to exert antimicrobic, antioxidant and antiproliferative effects. Therefore, since its effect in leukemic cell lines has not been previously evaluated, we studied its potential chemopreventive effect in Jurkat and HL-60 cell lines. METHODS: MG ethanolic extract was characterized for its antioxidant activity and scavenging effect against different radical species. Moreover, its phenolic profile was evaluated by HPLC-MS-MS analyses. Flow cytometry (FCM) analyses of Jurkat and HL-60 cells treated with MG extract (0-750 µg/mL) for 24-72 h- allowed to evaluate its cytotoxicity, pro-apoptotic and anti-proliferative effect. To better characterize MG pro-apoptotic mechanism ROS intracellular level and the gene expression level of FAS, BAX and BCL2 were also evaluated. Moreover, to assess MG extract selectivity towards cancer cells, its cytotoxicity was also evaluated in human peripheral blood lymphocytes (PBL). RESULTS: MG extract induced apoptosis in Jurkat and HL-60 cells in a dose- and time- dependent manner by increasing BAX/BCL2 ratio, reducing ROS intracellular level and inducing FAS gene expression level. In fact, reduced ROS level is known to be related to the activation of apoptosis in leukemic cells by the involvement of death receptors. MG extract also induced cell-cycle arrest in HL-60 cells. Moreover, IC50 at 24 h treatment resulted 2 times higher in PBL than in leukemic cell lines. CONCLUSIONS: Our data suggest that MG extract might be considered a promising and partially selective chemopreventive agent since it is able to modulate different mechanisms in transformed cells at concentrations lower than in non-transformed ones.

Valorisation of raspberry and blueberry pomace through the formulation of value-added gluten-free cookies.

Successful valorisation of raspberry and blueberry pomace was achieved through their application, as dried and ground powders, in the formulation of value-added gluten-free cookies. Simplex-lattice mixture design approach was applied to obtain the product with the best sensory properties, nutritional profile and antioxidant activity. The highest desirability of 90.0 % was accomplished with the substitution of gluten-free flour mixture with 28.2 % of blueberry and 1.8 % of raspberry pomace. The model was verified. Optimized cookies had similar protein (3.72 %) and carbohydrate (66.7 %) contents as gluten-containing counterparts used for comparison, but significantly lower fat content (10.97 %). Daily portion of the optimized cookies meets: 5.00 % and 7.73 % of dietary reference intakes (DRIs) for linoleic acid, 23.6 % and 34.3 % of DRIs for α-linolenic acid and 10.3 % and 15.6 % of DRIs for dietary fibers, for male and female adults, respectively. The nutritional profile of the optimized formulation makes it comparable with added-value gluten-containing counterparts.

Antioxidant Capacity, Mineral Content and Sensory Properties of Gluten-Free Rice and Buckwheat Cookies.

Light buckwheat flour was used to substitute rice flour at the level of 10, 20 and 30% to produce gluten-free cookies. The substitution of gluten-free cookie formulation with light buckwheat flour contributed to the significantly higher mineral content, especially magnesium, potassium, iron and copper, in comparison with the control rice cookies (p<0.05). Gluten-free cookies made with rice flour and buckwheat flour exhibited significantly higher total phenolic and rutin content, scavenging activity against 1,1-diphenyl-2-picrylhydrazyl radicals (DPPH•), antioxidant activity and reducing power than the control cookies (p<0.05). Comparing all evaluated sensory properties, cookies containing 20% of light buckwheat flour had the most acceptable sensory properties. The obtained results of principal component analysis showed that the cookies with 20 and 30% buckwheat flour had better antioxidant and sensory properties in comparison with other two cookie samples.

PhInd database - Polyphenol content in Agri-food by-products and trends in extraction technologies: A critical review.

Sustainable Development Goal 12 and target 12.3 set by the United Nations aims to reduce"food waste" per capita global for 50% losses by 2030. Databases such as the PhInd could help us to achieve set goals via mapping the potential ways for valorization of polyphenols from the agri-food by-products and waste. Fruit by-products (73.2% of the PhInd entries) are the most studied sources of polyphenols and future studies might be more focused on vegetables. More than half (55.8%) of entries were evaluated polyphenols in samples created in laboratory. These samples could have significantly different composition from industrial samples. Solid-liquid extraction (53.5%) and solvents like water, ethanol and aqueous ethanol (51.5%) were the most often used for extraction of polyphenols. Green solvents as NADES (0.4%) are rarely used in studies and should be more explored.

PhInd-Database on Polyphenol Content in Agri-Food By-Products and Waste: Features of the Database.

Timely access to topic-relevant datasets is of paramount importance for the development of any successful strategy (food waste reduction strategy), since datasets illuminate opportunities, challenges and development paths. PhInd is the first comprehensive database on polyphenol content in plant-based by-products from the agri-food sector or the wastewater sector and was developed using peer-reviewed papers published in the period of 2015-2021. In total, >450 scientific manuscripts and >6000 compound entries were included. Database inclusion criteria were polyphenol contents = determined using HPLC/UHPLC quantitative methods. PhInd can be explored through several criteria which are either 'open' or checkboxes. Criteria are given in subsections: (a) plant source; (b) by-product industrial processing; (c) pre-treatment of by-products before the isolation of polyphenols; and (d) the extraction step of polyphenols. Database search results could be explored on the website directly or by downloading Excel files and graphs. This unique database content is beneficial to stakeholders-the food industry, academia, government and citizens.

Eco-Friendly Extraction: A green approach to maximizing bioactive extraction from pumpkin (Curcubita moschata L.).

The research focused on optimizing the accelerated solvent extraction (ASE) of carotenoids and polyphenols from pumpkin powder. The study optimized accelerated solvent extraction (ASE) of carotenoids and polyphenols from pumpkin powder. Using a mix of standard score (SS) and artificial neural network (ANN) methods, the extraction process was fine-tuned. The ANN model assessed extraction parameters' significance, achieving high predictability for total carotenoid content (TCC), total phenolic content (TPC), and free radical scavenging capacity (DPPH and ABTS methods). The analysis highlighted the most effective extraction at 50 % concentration, 120 °C temperature, 5 min duration, and 2 cycles, yielding high carotenoid and phenolic content (TCC 571.49 µg/g, TPC 7.85 mg GAE/g). HPLC-DAD profiles of the optimized ASE extract confirmed major carotenoids and phenolic compounds. Strong correlations were found between bioactive compounds and antioxidant activity, emphasizing potential health benefits.

Role of Arthrospira Platensis in Preventing and Treating High-Fat Diet-Induced Hypercholesterolemia in Adult Rats.

Hyperlipidaemia is a recognised risk factor for cardiovascular disease. In this study, the antihyperlipidaemic properties of spirulina (Arthrospira platensis, strain S2 from Serbia) were tested in adult Wistar rats before and after induction of hypercholesterolaemia by a high-fat diet (HFD) to compare the preventive with the curative effect. Total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), alanine transaminase (ALT) and aspartate transaminase (AST) levels were measured in the blood samples. The chemical composition (lipids, proteins and cholesterol) and the content of bile acids in the faeces of the animals were also analysed. Feeding rats with an atherogenic diet for 10 weeks led to the successful development of hyperlipidaemia, as serum TC and LDL-C levels as well as lipids, cholesterol and bile acids in the animals' faeces were significantly increased. Pre- and post-treatment with spirulina led to a reduction in serum LDL, TC and ALT levels. Administration of spirulina resulted in both a significant increase in primary bile acids excretion and a decrease in bile acids metabolism, with pre-treatment being more effective than post-treatment in some cases. These results suggest that increased excretion of bile acids as well as an effect on the gut microbiota may be the mechanism responsible for the anti-hyperlipidaemic activity of the tested spirulina strain.

Enhancement of antioxidant and isoflavones concentration in gamma irradiated soybean.

Serbian soybean genotype Ana was gamma irradiated at doses of 1, 2, 4, and 10 kGy in order to evaluate the influence of gamma irradiation on isoflavone (genistein, daidzein, and their glycosides genistin and daidzin) contents and hydroxyl radical scavenging effect (HRSE). The increase in genistin and daidzin contents as well as antioxidant activities was observed especially at doses of 4 and 10 kGy. Results were also compared with our previous results relating to total phenol content (TPC), DPPH radical scavenger capacity (DPPH RSC), and ferric reducing antioxidant power (FRAP). Our results indicated that doses up to 10 kGy improve the antioxidant activities of soybean and also nutritional quality with respect to isoflavone content. All results were analyzed by multivariate techniques (correlation matrix calculation and autoscaling transformation of data). Significant positive correlations were observed between genistin, daidzin, DPPH RSC, and HRSE.

Moniliformin Occurrence in Serbian Maize over Four Years: Understanding Weather-Dependent Variability.

Moniliformin (MON) represents one of the most widespread emerging mycotoxins, whose presence in food and feed can potentially cause harmful effects on the health of both the public and animals. In order to investigate MON occurrence, a total of four hundred (n = 400) samples of unprocessed maize were sampled from different regions (Backa, Banat, and Srem) of Serbia during a period of four years (2018-2021) and were analyzed using a validated liquid chromatography with tandem mass spectrometry (LC-MS/MS) method. The influence of regional differences and variations from year to year in terms of weather conditions on the occurrence of MON was also assessed. The obtained validation parameters indicated that the LC-MS/MS method is applicable to the determination of MON in maize samples. It can be observed from the acquired findings that all samples were contaminated with MON, while concentration levels varied between the samples, especially from different sampling years. The maize samples showed the greatest levels of MON concentration during the dry and hot climatic conditions experienced in 2021. In maize samples harvested in the period 2018-2020, the MON concentration levels detected were about two to three times lower compared to the year 2021. Additionally, a comprehensive investigation into the correlation between weather conditions and the occurrence of MON in maize grown in Serbia was undertaken by reviewing the outcomes of research undertaken in the past decade.

Aflatoxins in Maize from Serbia and Croatia: Implications of Climate Change.

Aflatoxins (AFs) represent the most important mycotoxin group, whose presence in food and feed poses significant global health and economic issues. The occurrence of AFs in maize is a burning problem worldwide, mainly attributed to droughts. In recent years, Serbia and Croatia faced climate changes followed by a warming trend. Therefore, the main aim of this study was to estimate the influence of weather on AFs occurrence in maize from Serbia and Croatia in the 2018-2021 period. The results indicate that hot and dry weather witnessed in the year 2021 resulted in the highest prevalence of AFs in maize samples in both Serbia (84%) and Croatia (40%). In maize harvested in 2018-2020, AFs occurred in less than, or around, 10% of Serbian and 20% of Croatian samples. In order to conduct a comprehensive study on the implications of climate change for the occurrence of AFs in maize grown in these two countries, the results of available studies performed in the last thirteen years were searched for and discussed.

Health-promoting phytochemicals of Italian common wheat varieties grown under low-input agricultural management.

BACKGROUND: The increasing interest in organic food products and environmental friendly practices has emphasised the importance of selecting crop varieties suitable for the low-input sector. Moreover, in recent years the relationship between diet and human health has gained much attention among consumers. The aim of this study was to comparatively evaluate the agronomic performance and the nutrient and phytochemical composition of old and modern Italian wheat genotypes grown under low-input management. RESULTS: Research highlighted that several old wheat genotypes were comparable to the modern ones in terms of agronomic performance and nutrient content. Genotype and environmental conditions (growing season), as well as their interaction, significantly affected the phytochemical composition of wheat grains for most of the analysed bioactive compounds. High variability was observed among the wheat genotypes for dietary fibre (154.7-183.3 g kg⁻¹), polyphenol (1.94-2.77 mg g⁻¹), tocopherol (9.1-21.2 mg kg⁻¹) and carotenoid (701.4-3243 µg kg⁻¹) content. CONCLUSION: The comparative study of old and modern wheat varieties highlighted that, under low-input conditions, ancient genotypes may equal modern ones in terms of agronomic traits and additionally provide nutraceutical value-added wheat grains. The most promising ancient varieties for the unique phytochemical profiles are Gentil rosso, Marzuolo d'aqui and Verna.

Potential health benefits of blueberry and raspberry pomace as functional food ingredients: Dietetic intervention study on healthy women volunteers.

The fruit juice industry generates pomace as a valuable by-product especially rich in polyphenols, dietary fibers, vitamins, minerals, and unsaturated fatty acids. In the cookies used in this study, 30% of the gluten-free flour was replaced with dried and ground blueberry and raspberry pomace, rich source of polyphenols, dietary fibers, linoleic and alpha-linolenic acid. In order to examine whether the addition of blueberry and raspberry pomace in cookie formulation can have beneficial effects on certain blood parameters and anthropometric measurements, the designed cookies were tested in 20 healthy, normally fed female subjects, aged 30-50 years (41.35 ± 8.58 years) over four-week dietetic intervention study. Significant changes in the composition of fatty acids serum phospholipids, decrease in LDL-cholesterol level (20.16%), increase in adiponectin level (25.52%) and decrease in ALT and AST values were observed, thus indicating that inclusion of cookies containing blueberry and raspberry dried and ground pomace to usual diet might have positive effects on certain cardiovascular risk factors and liver function indicators.