A systematic UHPLC Q-ToF MS approach for the characterization of bioactive compounds from freeze-dried red goji berries (L. barbarum L.) grown in Serbia: Phenolic compounds and phenylamides.

The aim of this study was to identify and characterise different classes of bioactive compounds from freeze-dried red goji berries (RGB) grown in Serbia, using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC Q-ToF MS). In addition, this study aims to demonstrate the importance of applying the advanced UHPLC Q-ToF MS technique in the identification of various biocompounds. The analysis showed the presence of 28 phenolic compounds, 3 organic acids, and 26 phenylamides. The 2-O-ß-d-glucopyranosyl-l-ascorbic acid (AA-2ßG) was identified by UHPLC Q-ToF MS and quantified by standardised UHPLC-DAD method. Most of the compounds detected were derivatives of caffeic acid and ferulic acid, followed by quercetin derivatives. Among the phenylamides, several glucosylated caffeoyl and/or dihydrocaffeoyl derivatives of spermidine and spermine were characterized, confirming their recent characterization. Some glycosylated/non-glycosylated putrescine derivatives and caffeoyl-dihydrocaffeoyl-feruloyl spermidines were identified in goji berriesfor the first time. Their tentative structures and fragmentations were proposed.

Potential application of green extracts rich in phenolics for innovative functional foods: natural deep eutectic solvents as media for isolation of biocompounds from berries.

The health-promoting effects of berries have attracted attention due to the possible application of their extracts as functional ingredients in food products. Natural deep eutectic solvents (NADESs) are a new generation of environmentally friendly solvents for the extraction of natural products, and they are green alternatives to organic solvents, and they can improve the solubility, stability, and bioavailability of isolated biocompounds. In this study, an efficient eco-friendly method was used for the extraction of phenolic compounds from different berries: chokeberries, blueberries, and black goji berries with a range of eutectic solvents consisting of hydrogen bond acceptors (HBAs) such as choline chloride, L-proline, L-glycine, and L-lysine and hydrogen bond donors (HBDs) such as malic, citric, tartaric, lactic and succinic acids, glucose and glycerol. The obtained results indicated the ability of NADESs towards selective extraction of phenolics; the eutectic system choline chloride : malic acid showed selective extraction of anthocyanins, while choline chloride : glycerol and choline chloride : urea showed selectivity towards flavonoids and phenolic acids. The methodology for screening of the NADES extraction performance, which included chromatographic profiling via high-performance thin layer chromatography combined with chemometrics and spectrophotometric essays, allowed effective assessment of optimal eutectic solvents for isolation of different groups of phenolics. Great antioxidant and antimicrobial activities of extracts, along with the green nature of eutectic solvents, enable NADES berry extracts to be used as "green-labelled" functional foods or ingredients.

Broccoli, Amaranth, and Red Beet Microgreen Juices: The Influence of Cold-Pressing on the Phytochemical Composition and the Antioxidant and Sensory Properties.

The aim of this study was to analyze in detail the phytochemical composition of amaranth (AMJ), red beet (RBJ), and broccoli (BCJ) microgreens and cold-pressed juices and to evaluate the antioxidant and sensory properties of the juices. The results showed the presence of various phenolic compounds in all samples, namely betalains in amaranth and red beet microgreens, while glucosinolates were only detected in broccoli microgreens. Phenolic acids and derivatives dominated in amaranth and broccoli microgreens, while apigenin C-glycosides were most abundant in red beet microgreens. Cold-pressing of microgreens into juice significantly altered the profiles of bioactive compounds. Various isothiocyanates were detected in BCJ, while more phenolic acid aglycones and their derivatives with organic acids (quinic acid and malic acid) were identified in all juices. Microgreen juices exhibited good antioxidant properties, especially ABTS•+ scavenging activity and ferric reducing antioxidant power. Microgreen juices had mild acidity, low sugar content, and good sensory acceptability and quality with the typical flavors of the respective microgreen species. Cold-pressed microgreen juices from AMJ, RBJ, and BCJ represent a rich source of bioactive compounds and can be characterized as novel functional products.

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.

Authenticity assessment of cultivated berries via phenolic profiles of seeds.

Considering the health-benefits of berry fruits consumption and increased market demands for food authenticity as one of the most important quality assurances, phenolic profiling by high-performance thin layer chromatography and ultra-high-performance liquid chromatography hyphenated with mass spectrometry was combined with multivariate analysis for phytochemical characterization and intercultivar discrimination of cultivated berry seeds. The phenolic profiles of 45 berry seeds from nine genuine Serbian cultivated fruit species (strawberry, raspberry, blackberry, black currant, blueberry, gooseberry, cape gooseberry, chokeberry, and goji berry) revealed a good differentiation according to botanical origin. In order to determine biomarkers responsible for the classification, a total of 103 phenolic compounds were identified, including 53 phenolic acids and their derivatives, 26 flavonoids and 24 glycosides. Biomarkers derived from the phenolic profile of berry seeds proved to be a powerful tool in the authentication of botanical origin, and may be useful in detection of frauds in berry-based seed-containing product.

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.

Antibacterial Fractions from Erodium cicutarium Exposed-Clinical Strains of Staphylococcus aureus in Focus.

Followed by a buildup of its phytochemical profile, Erodium cicutarium is being subjected to antimicrobial investigation guided with its ethnobotanical use. The results of performed in vitro screening on Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans strains, show that E. cicutarium has antimicrobial activity, with a particular emphasis on clinical S. aureus strains-both the methicillin sensitive (MSSA) and the methicillin resistant (MRSA) S. aureus. Experimental design consisted of general methods (the serial microdilution broth assay and the agar well diffusion assay), as well as observing bactericidal/bacteriostatic activity through time (the "time-kill" assay), investigating the effect on cell wall integrity and biofilm formation, and modulation of bacterial hemolysis. Observed antibacterial activity from above-described methods led to further activity-guided fractionation of water and methanol extracts using bioautography coupled with UHPLC-LTQ OrbiTrap MS4. It was determined that active fractions are predominantly formed by gallic acid derivatives and flavonol glycosides. Among the most active phytochemicals, galloyl-shikimic acid was identified as the most abundant compound. These results point to a direct connection between galloyl-shikimic acid and the observed E. cicutarium antibacterial activity, and open several new research approaches for future investigation.

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.

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.

Polyphenol bioaccessibility and antioxidant properties of in vitro digested spray-dried thermally-treated skimmed goat milk enriched with pollen.

The aim of research was to determine polyphenols bioaccessibility and antioxidant properties of thermally-treated skimmed goat milk enriched with sunflower bee-collected pollen through in vitro digestion. HPLC analysis confirmed that pollen-enriched milk contained flavonols as the main phenolic fraction (80.7-76.2%) followed by phenolic acids (14.2-17.4%). Among individual compounds quercetin-3-O-glucoside (155.1-197.2 µg/L) and p-coumaric acid (29.5-30.7 µg/L) were the main quantified flavonols and phenolic acids, respectively. After digestion of milk/pollen sample, total polyphenols recovery was 30.71% with higher phenolic acids recovery (40.1%) compared to flavonols (28.3%) indicating strong interactions between caprine milk casein micelles and pollen polyphenols. Applied antioxidant assays (phosphomolybdenum, ABTS•+scavenging activity and ferrous-ion-chelating capacity) have confirmed complexity of prepared product- it had high ability to quench ABTS•+ radicals and to form chelating complexes with Fe2+ ions. Digestion provoked 20% reduction in total antioxidant capacity compared to the initial sample. TTSG milk/pollen powder could be good functional ingredient.

Chemical characterization and in vitro immunomodulatory effects of different extracts of moss Hedwigia ciliata (Hedw.) P. Beauv. from the Vrsacke Planine Mts., Serbia.

Bioactive compounds from natural sources are of great importance because of their potential pharmacological activity and tremendous structural diversity. In this study, the chemical composition of different moss extracts of Hedwigia ciliata P. Beauv. have been examined, as well as their antioxidant, antineurodegenerative/anti-neuroinflammatory, antidiabetic, and antiproliferative potential. The extracts were prepared by Soxhlet extractor using solvents of different polarity. Chemical characterization of the extracts revealed the presence of phenolics and flavonoid compounds, together with triterpenoids as secondary metabolites of high biological activity. Significant antioxidant properties of all the extracts were exhibited using the ß-carotene assay. The highest activities were found for water:ethanol extract (with the highest inhibition rate of 96%), but also significant inhibition was measured for ethanol and ethyl acetate extracts (80% and 70%, respectively). Confirmation of biocompatibility of investigated moss extracts has been performed using normal human fibroblast cell line, MRC-5. The H. ciliata extracts exhibited significant antiproliferative activity (~ 50%) against the MDA-MB-231 (human breast adenocarcinoma cell line), which has not previously been reported elsewhere. The Griess assay confirmed the potential anti-neuroinflammatory activity of the extracts, as significant effects in reducing NO production by LPS-stimulated BV2 (normal murine microglia cell line) was observed. This data is in line with noted antineurodegenerative potential measured by the inhibition of acetylcholinesterase (with the highest inhibition rate of 60% for ethyl acetate extract) and tyrosinase (with the highest inhibition rate of 70% for ethanol extract). Additionally, the H. ciliata extracts exhibited significant antidiabetic effect mediated by α-glucosidase inhibition (with the highest inhibition rate of 80% for ethyl acetate extract). The obtained data suggest the presence of immunomodulatory effects of the moss extracts in vitro, which allows the design of new experiments aimed at detecting and characterizing bioactive compounds of the extracts and additionally elucidate detailed mechanisms of their effects.

Extracts Characterization and In Vitro Evaluation of Potential Immunomodulatory Activities of the Moss Hypnum cupressiforme Hedw.

Recently, there has been an increasing interest in the chemistry and biological potential of mosses, since a large number of biologically active compounds have been found within these species. This study aimed at examining the chemical composition and immunomodulatory potential (antioxidant, antidiabetic, anti-neuroinflammatory/antineurodegenerative, and antitumor activities) of moss Hypnum cupressiforme Hedw. extracts. Corresponding extracts have been obtained applying Soxhlet extractor. The chemical characterization was performed using spectrophotometric assays and liquid chromatography-mass spectrometry (LC-MS). The extracts were analyzed for antioxidant activity and for inhibitory activities on α-glucosidase, α-amylase, acetylcholinesterase, and tyrosinase. Additionally, extracts were tested against four cell lines-MRC-5, BV2, HCT-116, and MDA-MB-231-for antitumor and anti-inflammatory activities. Chemical analysis of extracts revealed the presence of flavonoids, phenolic acids, and triterpenoids. Major compounds identified by LC-MS in H. cupressiforme were kaempferol and five phenolic acids: p-hydroxybenzoic, protocatechuic, p-coumaric, gallic, and caffeic acid. According to biochemical assays the investigated extracts exhibited significant immunomodulatory potential. Significant antiproliferative potential against MDA-MB-231 cells has been observed together with the promising anti-neuroinflammatory application. The obtained data suggest that moss H. cupressiforme is a valuable natural source of biologically active compounds with potential application in the pharmaceutical industry.

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.

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.

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.

Characterization of Sugar and Polyphenolic Diversity in Floral Nectar of Different 'Oblacinska' Sour Cherry Clones.

'Oblacinska' sour cherry, an autochthonous cultivar, is the most planted cultivar in Serbian orchards. Since fruit trees in temperate zone reward insects by producing nectar which 'quality' affects the efficiency of insect pollination, the aim of this study was analyzing of sugars and polyphenolics in floral nectar of 16 'Oblacinska' sour cherry clones with different yielding potential. The contents of sugars and sugar alcohols were analyzed by ion chromatography, while polyphenolic profile was established using liquid chromatography/mass spectrometry technique. Fourteen sugars and six sugar alcohols were detected in nectar samples and the most abundant were fructose, glucose, and sucrose. Eleven polyphenols were quantified using available standards, while another 17 were identified according to their exact masses and characteristic fragmentations. Among quantified polyphenols, rutin, naringenin, and chrysin were the most abundant in nectar. Principal component analysis showed that some polyphenol components (naringin, naringenin, and rutin) together with sugars had high impact of spatial distribution of nectar samples on score plot.

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.

Fatty acid and phenolic profiles of almond grown in Serbia.

Almond production is not typical for Serbia however the existence of natural populations and unexpectedly suitable agro-climatic conditions initiated this kind of study. Total oil content and concentrations of the fatty acids, total phenolic content and radical-scavenging activity were determined in the kernel oil of 20 local almond selections originating from North Serbia and cultivars 'Marcona', 'Texas' and 'Troito'. Sixteen fatty acids were identified and quantified, with the most abundant being oleic acid and linoleic acid. Nine phenolic acids and nineteen flavonoids were quantified using UHPLC-DAD MS/MS. The predominant polyphenol was catechin, followed by chlorogenic acid and naringenin. Based on oleic acid/linoleic acid ratio, levels of unsaturated fatty acids and specific polyphenols, some selections were chosen for growing and could also be recommended for breeding programs. Our investigation demonstrated that this region could be a suitable for growing almonds with chemical compositions competitive with standard cultivars.

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.