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.

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.

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.

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.

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.

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.

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).

Communicating Function and Co-Creating Healthy Food: Designing a Functional Food Product Together with Consumers.

Functional food is lately an interesting topic from the new product development perspective; complex motivation and expectations of consumers regarding it present a challenge when new products are designed. Co-creation is an interesting alternative to the standard practices by the R & D departments since it directly involves consumers in the various stages of the creation process. This work aims to describe experiences of engaging consumers in different development stages of a functional food product within a project realized at a food research institute. Four consecutive studies were conducted: the first study explored current trends in Serbia regarding the way consumers use functional food and are informed about it; the second study described development of a raspberry seeds extract with antioxidant and anti-proliferative activity confirmed in vitro; the third study tested the same extract in a sample of consumers, validating its usability in food products; and the fourth study described a co-creation session with 18 participants, during which a number of activities were realized to stimulate idea generation. Rather than the final product idea itself, this work is valuable because of detailed insights into the various phases of the co-creation process. It is shown that consumers and food researchers can together engage in the new food product development process as long as the communication between them is rich and with mutual understanding.

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.

Bioaccessibility and uptake by Caco-2 cells of carotenoids from cereal-based products enriched with butternut squash (Cucurbita moschata L.).

Enriching cereals-based products with bioactive compounds is a valuable strategy to improve product quality. We studied carotenoid bioaccessibility and intestinal uptake from a pumpkin-enriched porridge, cookies and sponge cakes by using in vitro digestion coupled with Caco-2 cell uptake. Among the carotenoids recovered in different products, α-carotene was the most important abundant one. However, lutein displayed a significantly higher bioaccessibility compared to α-carotene and ß-carotene in baked products (up to 10.28% compared to 1.22% and 0.88%, respectively). α-Carotene was the only carotenoid recovered in Caco-2 cells after micelle incubation. Cookie micelles led to the highest percentage of α-carotene cell uptake (2.33% and 1.38% for cookies with butter and cookies with vegetable oil, respectively) compared to the other baked products, followed by dry pumpkin puree micelles (1.31%). Overall, our data show that both bioaccessiblity and cell uptake of carotenoids from cereal-based products are variable and highly depend on food formulation and structure.

Simultaneous Hydrolysis of Ellagitannins and Extraction of Ellagic Acid from Defatted Raspberry Seeds Using Natural Deep Eutectic Solvents (NADES).

Defatted raspberry seeds were used as an alternative source of antioxidants and ellagic acid (EA) extracted using Natural deep eutectic solvents (NADES). In the preliminary study, the best NADES combination (citric acid-betaine) and the most influential variables (temperature, time, and NADES/plant ratio) were selected for the further optimization process. All samples were analyzed in terms of total polyphenol, EA content, and antioxidant activity. Two sets of optimal conditions were generated by response surface methodology. The first set (Opt1) was designed for higher conversion of ellagitannins to EA while the latter set (Opt2) for higher EA content/100 g extract. Opt1 and Opt2 had higher values for all investigated responses compared to 80% ethanolic extract but had a lower conversion rate of ellagitannins to EA compared to acidified methanol extract. The third set of parameters (Opt3) selected beyond the initial experimental domain was used to obtain a sample with the highest EA content/100 g extract. Due to their nature, NADES extracts are ready to use and could have various technological roles in products since they are antioxidants, acidifiers, and colorants. NADES raspberry extracts exhibited higher anti-proliferative activity compared to ethanolic extracts in terms of EC50 values. However, the main contributor of anti-cancer activity in NADES raspberry extracts were individual NADES compounds and/or their newly formed NADES structure.

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.

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.

European Database of Carotenoid Levels in Foods. Factors Affecting Carotenoid Content.

Many studies indicate that diets including carotenoid-rich foods have positive effects on human health. Some of these compounds are precursors of the essential nutrient vitamin A. The present work is aimed at implementing a database of carotenoid contents of foods available in the European market. Factors affecting carotenoid content were also discussed. Analytical data available in peer-reviewed scientific literature from 1990 to 2018 and obtained by HPLC/UHPLC were considered. The database includes foods classified according to the FoodEx2 system and will benefit compilers, nutritionists and other professionals in areas related to food and human health. The results show the importance of food characterization to ensure its intercomparability, as large variations in carotenoid levels are observed between species and among varieties/cultivars/landraces. This highlights the significance of integrating nutritional criteria into agricultural choices and of promoting biodiversity. The uncertainty quantification associated with the measurements of the carotenoid content was very rarely evaluated in the literature consulted. According to the EuroFIR data quality evaluation system for food composition tables, the total data quality index mean was 24 in 35, reflecting efforts by researchers in the analytical methods, and less resources in the sampling plan documentation.

Carotenoids: Considerations for Their Use in Functional Foods, Nutraceuticals, Nutricosmetics, Supplements, Botanicals, and Novel Foods in the Context of Sustainability, Circular Economy, and Climate Change.

Carotenoids are versatile isoprenoids that are important in food quality and health promotion. There is a need to establish recommended dietary intakes/nutritional reference values for carotenoids. Research on carotenoids in agro-food and health is being propelled by the two multidisciplinary international networks, the Ibero-American Network for the Study of Carotenoids as Functional Foods Ingredients (IBERCAROT; http://www.cyted.org) and the European Network to Advance Carotenoid Research and Applications in Agro-Food and Health (EUROCAROTEN; http://www.eurocaroten.eu). In this review, considerations for their safe and sustainable use in products mostly intended for health promotion are provided. Specifically, information about sources, intakes, and factors affecting bioavailability is summarized. Furthermore, their health-promoting actions and importance in public health in relation to the contribution of reducing the risk of diverse ailments are synthesized. Definitions and regulatory and safety information for carotenoid-containing products are provided. Lastly, recent trends in research in the context of sustainable healthy diets are summarized.

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.

A comprehensive study of parameters correlated with honey health benefits.

One hundred honey samples of different floral origin (acacia, sunflower, meadow, and forest) collected from nine European countries (Serbia, Albania, Croatia, Montenegro, Romania, Bulgaria, Bosnia and Herzegovina, North Macedonia and Hungary) were analysed for various physicochemical, sensory, antioxidant and antibacterial parameters. The relative antioxidant capacity index and relative antibacterial index were calculated, integrated and expressed as a new property - Power of Honey, intended to be used to predict the health potential of a honey based on its antioxidant and antibacterial activities. Free acidity and colour coordinates L* and a* were chosen for building an artificial neural network model for the prediction of honey health potential. These were chosen based on the obtained correlations between the investigated parameters and in light of the simplicity of the analysis. This model successfully predicted the Power of Honey with a gained coefficient of determination of 0.856. Forest honey samples exhibited the highest Power of Honey. This novel approach should make it possible for honey producers to predict the honey health potential of a particular honey based on a quick and simple analysis.

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.

Influence of different extraction techniques on the chemical profile and biological properties of Anthemis cotula L.: Multifunctional aspects for potential pharmaceutical applications.

The phytochemical composition of different extracts obtained from stinking chamomile (Anthemis cotula L.) was investigated. Ethanol was used as solvent and accelerated solvent extraction (ASE), microwave assisted extraction (MAE), maceration, soxhlet extraction (SE), and ultrasound assisted extraction (UAE) were applied on plant material. Comparison of the phytochemical contents, antioxidant, and enzyme inhibitory properties were performed. The most abundant sesquiterpene in the extracts was anthecotuloide, while the most present phenolics were caffeoyl quinic acid, quercetin, and kaempferol derivatives. UAE extract was the most efficient in the extraction of sesquiterpenoids and polyphenols. Considering the assays on antioxidant activity and enzyme inhibition, ASE extract showed highest phenolic content (62.92 mg gallic acid equivalent/g extract). Likewise, this extract showed highest radical scavenging (103.44 mg trolox equivalent [TE]/g extract and 155.70 mg TE/g extract, for DPPH and ABTS assays respectively) and reducing power potential (435.32 and 317.89 mg TE/g extract, for CUPRAC and FRAP assays, respectively). The different extracts showed similar results in the enzyme inhibition assays suggesting that the extraction methods used have no effect on observed enzyme activities. Novelty of our findings are the inhibitory action of the ethanol extract of A. cotula aerial parts on key enzymes associated with Alzheimer's disease (acetyl cholinesterase, butyryl cholinesterase), type 2 diabetes (α-amylase, α-glucosidase), and skin hyperpigmentation disorders (tyrosinase). Data collected from the present work further appraises the multiple potential biological properties of stinking chamomile suggesting the need for further investigation on its constituents.