Hieracium waldsteinii (Asteraceae) and Onosma stellulata (Boraginaceae) as a Source of Antioxidant and Antimicrobial Agents.

The current study aimed to phytochemically characterize (including a detailed phenolic profile) two endemic Balkan's species (Hieracium waldsteinii and Onosma stellulata) and determine their possible application as a source of natural antioxidant and antimicrobial agents. The main phenolic compound in both species (in all examined parts) was chlorogenic acid. Eriodictyol, genistein and naringenin were quantified only in H. waldsteinii while isorhamnetin-3-O-rutinoside and sinapic acid were characteristic for O. stellulata. The highest antioxidant activity (98 mg AAE/g dry weight for TAC assay) was ascribed to the flower extract of H. waldsteinii while the lowest results (∼4.3 mg AAE/g dry weight for FRP assay) were exhibited by the extracts obtained from the plant's stem. Antimicrobial assays showed moderate antibacterial, i. e., moderate/strong activity against several tested fungi (in particular Trichoderma viride). Correlation analysis revealed strong positive connection between phenolic compounds and reducing power of extracts as well as between total phenolic and flavonoid content and the obtained minimal inhibitory concentration recorded in antibacterial assays.

Establishing the chromatographic fingerprints of flavan-3-ols and proanthocyanidins from rose hip (Rosa sp.) species.

The profile of flavan-3-ols and proanthocyanidins in five different Rosa species (R. canina, R. glutinosa, R. rubiginosa, R. multiflora, and R. spinosissima) was estimated on high performance thin layer chromatography cellulose plates. Differences in flavanol and proanthocyanidin profiles of the extracts were evident, among which Rosa spinosissima stood out with catechin as the only detected flavanol and red zones as indication of anthocyanins. Furthermore, the elution solvent for thin layer chromatography with mass spectrometry analyses of glycosylated flavan-3-ols and proanthocyanidins was optimized, enabling identification of catechin, (epi)catechin hexoside, proanthocyanidin dimer, and proanthocyanidin dimers and trimers hexosides. A total of 15 flavanols and their derivatives were identified using ultra-high-performance liquid chromatography with linear trap quadrupole-Orbitrap mass analyzer and epicatechin, gallocatechin, and proanthocyanidin trimer were identified only using this technique. However, proanthocyanidin trimer trihexoside was identified only by thin-layer chromatography with mass spectrometry. To establish the relationships between the flavanols and proanthocyanidins composition of rose hip and their origin, principal component analysis was performed on the entire set of liquid chromatography/mass spectrometry data. Both principal components' scores plots showed that Rosa spinosissima could be considered as an outlier. Our study demonstrated that flavanol and proanthocyanidin profiles of different rose hips depend on the geographical origin rather than on the cultivar and genotype.

Phytochemical Analysis and Total Antioxidant Capacity of Rhizome, Above-Ground Vegetative Parts and Flower of Three Iris Species.

This study was aimed at investigating the phytochemical composition and antioxidant capacity of rhizomes, above-ground vegetative parts and flowers of three Iris species: Iris humilis Georgi, Iris pumila L. and Iris variegata L. UHPLC-Orbitrap MS analysis was used for determination of phytochemical profile. Total pigments, phenolics, flavonoids, soluble sugars and starch content as well as ABTS antioxidant capacity were also determined. In total, 52 phenolics compounds were identified with 9 compounds (derivatives of iriflophenone, apigenin C-glycosides, luteolin O-glycoside, isoflavones derivatives of iristectorigenin, dichotomitin, nigracin and irilone) never reported before in Iris spp. Differences in phenolic composition profile, pigments, soluble sugar, starch, total phenolics and flavonoids content and total antioxidant capacity were found among Iris species and different part of plants. Significant correlation between total phenolic content and antioxidant capacity was determined. The obtained results are comparable with those obtained for medical plants. These findings could be useful for fingerprinting characterization of Iris species and estimation of possible use in pharmaceutical industries.

Identification of Phenolic Compounds from Seed Coats of Differently Colored European Varieties of Pea (Pisum sativum L.) and Characterization of Their Antioxidant and In Vitro Anticancer Activities.

To date little has been done on identification of major phenolic compounds responsible for anticancer and antioxidant properties of pea (Pisum sativum L.) seed coat extracts. In the present study, phenolic profile of the seed coat extracts from 10 differently colored European varieties has been determined using ultrahigh-performance liquid chromatography-linear trap quadrupole orbitrap mass spectrometer technique. Extracts of dark colored varieties with high total phenolic content (up to 46.56 mg GAE/g) exhibited strong antioxidant activities (measured by 2,2-diphenyl-1-picrylhydrazyl or DPPH assay, and ferric ion reducing and ferrous ion chelating capacity assays) which could be attributed to presence of gallic acid, epigallocatechin, naringenin, and apigenin. The aqueous extracts of dark colored varieties exert concentration-dependent cytotoxic effects on all tested malignant cell lines (human colon adenocarcinoma LS174, human breast carcinoma MDA-MB-453, human lung carcinoma A594, and myelogenous leukemia K562). Correlation analysis revealed that intensities of cytotoxic activity of the extracts strongly correlated with contents of epigallocatechin and luteolin. Cell cycle analysis on LS174 cells in the presence of caspase-3 inhibitor points out that extracts may activate other cell death modalities besides caspase-3-dependent apoptosis. The study provides evidence that seed coat extracts of dark colored pea varieties might be used as potential cancer-chemopreventive and complementary agents in cancer therapy.

Chemical profile of major taste- and health-related compounds of Oblacinska sour cherry.

BACKGROUND: Oblacinska sour cherry, an autochthonous cultivar, is the most planted cultivar in Serbian commercial orchards. Owing to its long cultivation under different agro-ecological conditions, it is a mixture of different clones. To obtain comprehensive information on Oblacinska sour cherry fruit, the total carbohydrate composition, total phenolic content, total anthocyanin content, antioxidant activity and polyphenolic profiles of 39 clones were investigated. RESULTS: Fructose was found to be the dominant sugar, followed by glucose, sorbitol and sucrose. Rutin and chlorogenic acid were the most abundant polyphenols. Some clones contained pinobanksin, hesperetin and galangin, which, to best knowledge, is the first time these compounds have been reported in sour cherry fruit. CONCLUSION: The data obtained showed huge variation within the germplasm studied, confirming the fact that Oblacinska sour cherry is not a cultivar but a population.

Monofloral Corn Poppy Bee-Collected Pollen-A Detailed Insight into Its Phytochemical Composition and Antioxidant Properties.

The aim of this study was to compile a detailed phytochemical profile and assess the antioxidant properties of bee-collected pollen (PBP) obtained from corn poppy (Papaver rhoeas L.) plants. To achieve this, a lipid fraction was prepared for quantifying fatty acids using GC-FID. Extractable and alkaline-hydrolysable PBP fractions (obtained from a defatted sample) were used to determine the qualitative and quantitative profiles of phenolic compounds, phenylamides and alkaloids using UHPLC/Q-ToF-MS. Additionally, various spectrophotometric assays (TAC, FRP, CUPRAC, DPPH⦁) were conducted to evaluate the antioxidant properties. Phenolic compounds were more present in the extractable fraction than in the alkaline-hydrolysable fraction. Luteolin was the predominant compound in the extractable fraction, followed by tricetin and various derivatives of kaempferol. This study presents one of the first reports on the quantification of tricetin aglycone outside the Myrtaceae plant family. The alkaline-hydrolysable fraction exhibited a different phenolic profile, with a significantly lower amount of phenolics. Kaempferol/derivatives, specific compounds like ferulic and 5-carboxyvanillic acids, and (epi)catechin 3-O-gallate were the predominant compounds in this fraction. Regarding phenylamides, the extractable fraction demonstrated a diverse range of these bioactive compounds, with a notable abundance of different spermine derivatives. In contrast, the hydrolysable fraction contained six spermine derivatives and one spermidine derivative. The examined fractions also revealed the presence of seventeen different alkaloids, belonging to the benzylisoquinoline, berberine and isoquinoline classes. The fatty-acid profile confirmed the prevalence of unsaturated fatty acids. Furthermore, both fractions exhibited significant antioxidant activity, with the extractable fraction showing particularly high activity. Among the assays conducted, the CUPRAC assay highlighted the exceptional ability of PBP's bioactive compounds to reduce cupric ions.

Bioaccessibility of Phenolic Compounds and Antioxidant Properties of Goat-Milk Powder Fortified with Grape-Pomace-Seed Extract after In Vitro Gastrointestinal Digestion.

This study deals with the evaluation of the bioaccessibility and antioxidant properties of phenolic compounds from heat-treated skim goat-milk powder fortified with grape-pomace-seed extract, after in vitro gastrointestinal digestion. Ultra-high performance liquid chromatography coupled to diode array detection and mass spectrometry (UHPLC-DAD MS/MS) analysis confirmed the abundant presence of phenolic acids and flavan-3-ols in the grape-pomace-seed extract (SE) and heat-treated skim goat-milk/seed-extract powder (TME). After in vitro digestion of TME powder and recovery of total quantified phenolics, flavan-3-ols and phenolic acids were 18.11%, 24.54%, and 1.17%, respectively. Low recovery of grape-pomace-seed phenolics indicated strong milk protein-phenolic interactions. Electrophoretic analysis of a soluble fraction of digested heat-treated skim goat milk (TM) and TME samples showed the absence of bands originating from milk proteins, indicating their hydrolysis during in vitro gastrointestinal digestion. The digested TME sample had better antioxidant properties in comparison to the digested TM sample (except for the ferrous ion-chelating capacity, FCC), due to the presence of bioaccessible phenolics. Taking into account the contribution of the digestive cocktail, digested TME sample had lower values of total phenolic content (TPC), in vitro phosphomolybdenum reducing capacity (TAC) and ferric reducing power (FRP), compared to the undigested TME sample. These results could be attributed to low recovery of phenolic compounds. TME powder could be a good carrier of phenolics to the colon; thus, TME powder could be a promising ingredient in the formulation of functional food.

Phytochemical Profile and Antioxidant Properties of Bee-Collected Artichoke (Cynara scolymus) Pollen.

The current study intended to determine, for the first time, phenolic and fatty acid profile, antioxidant and certain nutritional properties of monofloral bee-collected artichoke (Cynara scolymus) pollen. Based on UHPLC-DAD MS-MS analysis the main phenolics in extractable fraction were different flavonol glycosides (in particular Isorhamnetin-3-O-glucoside, 49.2 mg/kg of dry weight) while ferulic acid was the predominant phenolic compound (39.4 mg/kg of dry weight) in the alkaline hydrolyzable fraction. Among fatty acids (FAs), results of GC-FID analysis revealed prevalence of unsaturated FAs with cis-5,8,11,14,17-eicosapentaenoic acid (EPA) and oleic acid as the main ones- 28.4% and 24.9%, respectively. Based on the FA composition, nutritional analysis proved that artichoke bee-collected pollen had balanced ω-6 and ω-3 FAs content. To determine the antioxidant properties of pollen, five different assays were applied. It was proved that bioactive compounds in artichoke pollen possessed significant ability to quench DPPH radical as well as ABTS radical cation. In addition, in vitro phosphomolybdenum assay confirmed that artichoke pollen is an excellent source of different antioxidants. Pollen extracts exhibited moderate ferric reducing power as well as low ferrous chelating ability. Some further antioxidant studies (preferably in vivo) should be performed to confirm the observed results.

Skimmed Goat's Milk Powder Enriched with Grape Pomace Seed Extract: Phenolics and Protein Characterization and Antioxidant Properties.

The aim of this research was phenolics and protein characterization and antioxidant properties evaluation of skimmed thermally treated goat's milk powder enriched with different concentration of grape pomace seed extract (SE). The dominant phenolics in SE were phenolic acids, flavan-3-ols and procyanidins. Different electrophoretic techniques together with UHPLC-MS/MS analysis revealed the presence of phenolics-protein interactions in the samples, mainly procyanidins with whey protein/caseins complexes. Addition of SE into thermally treated goat's milk significantly improved antioxidant properties of goat's milk such as TAC, FRP, DPPH• and ABTS•+ scavenging activity. Gallic acid, catechin, and procyanidins mostly contributed to these activities. The schematic representation of phenolics-casein micelles interactions in thermally treated goat's milk enriched with SE was given. The addition of SE into thermally treated goat's milk can be a promising strategy in food waste recovery and to enhance the beneficial health effects of goat's milk-based functional foods.

Structure-retention relationship study of arylpiperazines by linear multivariate modeling.

A quantitative structure-retention relationship study has been performed to correlate the retention of 33 newly synthesized arylpiperazines with their molecular characteristics, using thin-layer chromatography. Principal component analysis followed by multiple linear regression (MLR), principal component regression (PCR) and partial least squares (PLS) was performed to identify the most important factors, to quantify their influences, and to select descriptors that best describe the behavior of the compounds investigated. The best statistical performance was achieved by applying PLS regression, leading to the lowest value of the standard error (root mean square errors of calibration of 0.159 and cross-validated value RMSE cross-validation=0.231 units), followed by the PCR (root mean square errors of calibration=0.195 and RMSE cross-validation=0.305) and MLR (R(adj)(2)=0.9499, F=102.017, mean square error=0.052 and predicted residual error sum of squares=2.23). Two factors of the highest influence: surface tension and hydrophilic-lipophilic balance appear as the part of obtained models. In addition, polar surface area and hydrophilic surface area are included by both PLS and PCR models. Moreover, logP has been added to the PLS model. Besides, PCR model includes following descriptors: hydrogen bond acceptor, hydrogen bond donor and LUMO energy, whereas topological descriptors: connectivity indices 0 and 2, and valence index 3 are included in the MLR model.

The Application of Pollen as a Functional Food and Feed Ingredient-The Present and Perspectives.

Pollen is recognized as an excellent dietary supplement for human nutrition, which is why it can be found in different forms on the market (granules, capsules, tablets, pellets, and powders). But, the digestibility of pollen's nutrients is strongly affected by the presence of a pollen shell, which can decrease the bioavailability of nutrients by 50% and more. Since consumers have become more aware of the benefits of a healthy diet and the necessity to improve pollen digestibility, different pollen-based functional food products have been developed and extensive studies were done to estimate the beneficial effects of pollen-based feed on animal growth, health, and rigor mortise stage. Considering the positive effects of pollen nutrients and phytometabolites on human and animal health, the aim of this paper was to give an overview of recent achievements in the application of pollen in the formulation of functional food and animal diets. Special attention was paid to the effects of pollen's addition on the nutritional, functional, techno-functional, and sensory properties of the new formulated food products. Anti-nutritional properties of pollen were also discussed. This review points out the benefits of pollen addition to food and feed and the possible directions in the further development of functional food and feed for the wellbeing of everyone.

Mycotoxins and Mycotoxin Producing Fungi in Pollen: Review.

Due to its divergent chemical composition and good nutritional properties, pollen is not only important as a potential food supplement but also as a good substrate for the development of different microorganisms. Among such microorganisms, toxigenic fungi are extremely dangerous as they can synthesize mycotoxins as a part of their metabolic pathways. Furthermore, favorable conditions that enable the synthesis of mycotoxins (adequate temperature, relative humidity, pH, and aw values) are found frequently during pollen collection and/or production process. Internationally, several different mycotoxins have been identified in pollen samples, with a noted predominance of aflatoxins, ochratoxins, fumonisins, zearalenone, deoxynivalenol, and T-2 toxin. Mycotoxins are, generally speaking, extremely harmful for humans and other mammals. Current EU legislation contains guidelines on the permissible content of this group of compounds, but without information pertaining to the content of mycotoxins in pollen. Currently only aflatoxins have been researched and discussed in the literature in regard to proposed limits. Therefore, the aim of this review is to give information about the presence of different mycotoxins in pollen samples collected all around the world, to propose possible aflatoxin contamination pathways, and to emphasize the importance of a regular mycotoxicological analysis of pollen. Furthermore, a suggestion is made regarding the legal regulation of pollen as a food supplement and the proposed tolerable limits for other mycotoxins.

Application of Polyphenol-Loaded Nanoparticles in Food Industry.

Nanotechnology is an emerging field of science, and nanotechnological concepts have been intensively studied for potential applications in the food industry. Nanoparticles (with dimensions ranging from one to several hundred nanometers) have specific characteristics and better functionality, thanks to their size and other physicochemical properties. Polyphenols are recognized as active compounds that have several putative beneficial properties, including antioxidant, antimicrobial, and anticancer activity. However, the use of polyphenols as functional food ingredients faces numerous challenges, such as their poor stability, solubility, and bioavailability. These difficulties could be solved relatively easily by the application of encapsulation. The objective of this review is to present the most recent accomplishments in the usage of polyphenol-loaded nanoparticles in food science. Nanoparticles loaded with polyphenols and their applications as active ingredients for improving physicochemical and functional properties of food, or as components of active packaging materials, were critically reviewed. Potential adverse effects of polyphenol-loaded nanomaterials are also discussed.

In vitro digestion of meat- and cereal-based food matrix enriched with grape extracts: How are polyphenol composition, bioaccessibility and antioxidant activity affected?

The aim of this study was to evaluate the effect of enriching a complex food matrix (FM) with grape extracts on polyphenol content, composition, bioaccessibility and antioxidant activity during digestion. The grape extracts and FM were separately tested under the same conditions as controls. The FM by itself contains a significant amount of phenolic acids and flavonols, influencing the final recovery of polyphenols from grape extracts. The FM significantly increased the total recovery of polyphenols after digestion of grape seed extracts compared to those digested without the FM; however, a low recovery of proantocyanidins and total flavonoids was observed. Digestive fluids and FM compounds significantly increased the total polyphenol content of grape digests and significantly contributed to their ABTS+ scavenging activity and ferrous-ion-chelating capacity. The present study suggested that enrichment of meat- and cereal-based products with grape polyphenol extracts could be a good strategy to formulate a healthier diet.

Physicochemical Parameters as a Tool for the Assessment of Origin of Honey.

Honey is a complex mixture of various substances, and its composition depends on both botanical and geographical origin, as well as anthropogenic factors. The accurate identification of honey origin guarantees the satisfaction of consumers' needs and has an impact on the honey market value. Physicochemical parameters, some of which are used in routine analysis of honey quality, could be useful for the assessment of its origin. In this review, special attention is paid to those studies that assessed the sugar and mineral composition of honey, whether they were investigated in terms of botanical or geographical origin, or for the characterization of honey type. The oligosaccharides present in honey and the electrical conductivity of honey correlate strongly with its botanical origin. Mineral content could be indicative for distinguishing honeys according to their botanical and geographical origins because it depends on both the soil composition and the floral type of melliferous plants. This review provides insight into the results obtained by various studies from approximately the last 10 years concerning the sugar profile and the mineral and trace element content of different types of honey. An attempt was made to statistically analyze the results regarding mineral and trace element content in order to identify indicators that could distinguish honey by origin.

Polyphenols as Possible Markers of Botanical Origin of Honey.

In recent years, the botanical and geographical origin of food has become an important topic in the context of food quality and safety, as well as consumer protection, in accordance with international standards. Finding chemical markers, especially phytochemicals, characteristic for some kind of food is the subject of interest of a significant number of researchers in the world. This paper is focused on the use of polyphenols as potential markers for the determination of botanical origin of honey. It includes a review of the polyphenols present in various honey samples and the methods for their separation and identification. Special emphasis in this paper is placed on the identification of honey polyphenols using advanced LC-MS techniques in order to find specific markers of botanical origin of honey. In this regard, this study gives an overview of the literature that describes the use of LC-MS techniques for the isolation and determination of honey polyphenols. This review focuses on the research performed in the past two decades.

The polyphenolics and carbohydrates as indicators of botanical and geographical origin of Serbian autochthonous clones of red spice paprika.

Spice peppers (Capsicum annuum L.) var. Lemeska and Lakosnicka paprika were investigated to evaluate their polyphenolic and carbohydrate profiles and antioxidant activity. A total of forty-nine polyphenolics were identified using ultrahigh-performance liquid chromatography (UHPLC) coupled to LTQ OrbiTrap mass analyzer. Twenty-five of them were quantified using available standards, while the other compounds were confirmed by exact mass search of their deprotonated molecule [M-H](-) and its MS(4) fragmentation. Thirteen carbohydrates were quantified using high-performance anion exchange chromatography (HPAEC) with pulsed amperometric detection (PAD). Radical scavenging activity (RSA) ranged from 17.32 to 48.34mmol TE (Trolox equivalent)/kg DW (dry weight) and total phenolics content (TPC) was ranged between 7.03 and 14.92g GAE (gallic acid equivalents)/kg DW. To our best knowledge, five polyphenolic compounds were for the first time tentatively identified in paprika: 5-O-p-coumaroylquinic acid, luteolin 7-O-(2″-O-pentosyl-4″-O-hexosyl)hexoside, quercetin 3-O-(2″-O-hexosyl)rhamnoside, isorhamnetin 3-O-[6″-O-(5-hydroxyferuloyl)hexoside]-7-O-rhamnoside, and luteolin 7-O-[2″-O-(5'″-O-sinapoyl)pentosyl-6″-O-malonyl]hexoside.

Identification and quantification of phenolic compounds in berry skin, pulp, and seeds in 13 grapevine varieties grown in Serbia.

Phenolic profile of 13 grapevine varieties was determined, with respect to three different parts of berries: skin, seed and pulp. Identification and quantification of the phenols was done using ultra-high performance liquid chromatography (UHPLC) coupled with a diode array detector and a triple-quadrupole mass spectrometer. The most abundant phenols in grape seeds were found to be flavan-3-ols, most of which are gallocatechin gallate and catechin. The skins were characterized mostly by flavonols, i.e. quercetin and myricetin. Characterization of anthocyanins in the berry skin by UHPLC coupled with Linear Trap Quadrupole and OrbiTrap mass analyzer revealed a total of twenty derivatives of malvidin, delphinidin, petunidin, cyanidin, and peonidin. To the extent of our knowledge this is the first work that shows the presence of malvidin 3,5-O-dihexoside in the berry skin of 'Merlot', 'Cabernet Franc', 'Shiraz' 'Sangiovese', 'Pinot Noir' and 'Prokupac', untypical for Vitis vinifera Linneo species.

Analysis and characterisation of phytochemicals in mulberry (Morus alba L.) fruits grown in Vojvodina, North Serbia.

In this study, the polyphenolic profile of 11 Morus alba fruits grown in the Vojvodina region was investigated. Ultra high performance liquid chromatography (UHPLC) coupled with Linear Trap Quadrupole and OrbiTrap mass analyzer, and UHPLC coupled with a diode array detector and a triple-quadrupole mass spectrometer were used for the identification and quantification of the polyphenols, respectively. A total of 14 hydroxycinnamic acid esters, 13 flavonol glycosides, and 14 anthocyanins were identified in the extracts with different distributions and contents according to the sampling. The total phenolic content ranged from 43.84 to 326.29 mg GAE/100g frozen fruit. The radical scavenging capacity (50.18-86.79%), metal chelating ability (0.21-8.15%), ferric ion reducing power (0.03-38.45 µM ascorbic acid) and superoxide anion radical scavenging activity (16.53-62.83%) were assessed. The findings indicated that mulberry polyphenolics may act as potent superoxide anion radical scavengers and reducing agents.

Polyphenolic Profile of Pear Leaves with Different Resistance to Pear Psylla (Cacopsylla pyri).

The European pear psylla, Cacopsylla pyri L. (Hemiptera: Psyllidae), is one of the most serious arthropod pests of pear. Since proper control of this pest is essential, better understanding of the complex plant-pest relationship is mandatory. This research deals with constitutive polyphenolic profiles in leaves of 22 pear cultivars of diverse origin (P. communis, P. pyrifolia, and P. pyrifolia × P. communis) and different resistance to psylla. The study was designed to show which differences in the polyphenolic profile of leaves from resistant and susceptible pear cultivars could be utilized as information in subsequent breeding programs. The results demonstrated that the leaves of Oriental pear cultivars contained much higher amounts of p-hydroxybenzoic acid, ferulic acid, aesculin, and naringin, that, together with detected 3-O-(6″-O-p-coumaroyl)-hexoside, apigenin, apigenin 7-O-rutinoside, and hispidulin, indicated a clear difference between the species and might represent phenolics responsible for psylla resistance.