In Vitro Digestion of Polyphenolic Compounds and the Antioxidant Activity of Acorn Flour and Pasta Enriched with Acorn Flour.

Acorn flour is a rich source of nutrients and is beneficial to human health due to, among other things, its low glycemic index and polyphenol content. In order to obtain more accurate data on the levels and activities of the substances tested after ingestion and digestion, it may be beneficial to use a simulated in vitro digestion method. Therefore, the objective of the present study was to elucidate the content of polyphenols, individual phenolic acids, flavonoids and antiradical properties of acorn flour and pasta enriched with acorn flour before and after simulated in vitro gastrointestinal digestion. The results indicate that the total polyphenol content (TPC), flavonoid content and radical scavenging activity exhibited an increasing trend following the initial digestion stage and a decreasing trend following the second stage. Nevertheless, the levels of phenolic acids demonstrated an increase in both digestion phases. The digestion processes of polyphenols in acorn flour differ significantly from those in pasta. In the case of pasta, total polyphenols, phenolic acids and flavonoids, as well as free radical scavenging properties, demonstrated a decreasing trend following each digestion stage.

Antioxidant and Cytotoxic Potential of Carlina vulgaris Extract and Bioactivity-Guided Isolation of Cytotoxic Components.

Carlina vulgaris is a poorly understood plant in the context of biological activity, despite its widespread application in ethnomedicine in numerous European countries. The aim of this study was to assess the cytotoxic potential of the plant against human colorectal adenocarcinoma (HT29) and to isolate the plant components linked to this effect. Ultra-high performance liquid chromatography with a high-resolution/quadrupole time-of-flight mass spectrometer (UHPLC-HR/QTOF/MS-PDA) was used for the phytochemical characterization of the extract. Liquid-liquid extraction and preparative chromatography were employed for fractionation purposes. Our investigation demonstrated that the ethyl acetate fraction from C. vulgaris showed significant cytotoxicity, and a bioactivity-guided approach led to the isolation of oxylipins, including traumatic acid, pinellic acid, and 9,10-dihydroxy-8-oxsooctadec-12-enic acid. The structures of the compounds were confirmed by nuclear magnetic resonance spectroscopy. Among these compounds, the last one exhibited significant cytotoxicity, though without selectivity, and traumatic acid was characterized by mild cytotoxicity. The cytotoxicity was linked to intracellular reactive oxygen species generation.

Fresh Chokeberry (Aronia melanocarpa) Fruits as Valuable Additive in Extruded Snack Pellets: Selected Nutritional and Physiochemical Properties.

In this paper, the nutritional value and (selected) physiochemical properties of extruded snack pellets enriched with fresh chokeberry (Aronia melanocarpa) fruits were analyzed from the perspective of being a new product for the functional food sector. The purpose of this study was to determine the effect of the addition of fresh chokeberry and variation in content and screw speed on extruded snack pellet basic compositions, fatty acid profiles, antioxidant activity, as well as water absorption and solubility indexes, fat absorption and color profiles. The obtained results revealed a significant increase in antioxidant activity for all samples (above 90% of free radical scavenging) in comparison to potato-based control samples (just over 20% of free radical scavenging). The total phenolic content assay revealed the most valuable results for samples enriched with 30% chokeberry, while Ultra Performance Liquid Chromatography (UPLC) analysis allowed the determination of the most important phenolic acids. Of interest, chokeberry addition decreased the fat absorption index (FAI) after expansion by frying. Moreover, the highest values of crude protein and crude ash were observed in snack pellets supplemented by the application of 30% chokeberry. In such samples, the crude protein content was at the level of 4.75-4.87 g 100 g-1 and crude ash content at 4.88-5.07 g 100 g-1. Moreover, saturated fatty acids (SFA) content was lower in snack pellets with chokeberry addition, and increasing the amount of chokeberry additive from 10% to 30% in extruded snack pellet recipes resulted in more than double an increase in polyunsaturated fatty acids (PUFA) proportion in the total fatty acids.

Polyphenolic Composition of Carlina acaulis L. Extract and Cytotoxic Potential against Colorectal Adenocarcinoma and Cervical Cancer Cells.

Carlina acaulis is highly valued in the traditional medicine of many European countries for its diuretic, cholagogue, anthelmintic, laxative, and emetic properties. Moreover, practitioners of natural medicine indicate that it has anti-cancer potential. However, its phytochemistry is still little known. In the present study, the polyphenolic composition of the plant was investigated using ultra-high-performance liquid chromatography coupled with a high-resolution/quadrupole time-of-flight mass spectrometer (UHPLC-HR/QTOF/MS-PDA). The fractionation of the extract was carried out using liquid-liquid extraction and preparative chromatography techniques. Cytotoxicity was assessed based on neutral red and MTT assays. The obtained data showed that the species is rich in chlorogenic acids and C-glycosides of luteolin and apigenin. The total amount of chlorogenic acids was 12.6 mg/g. Among flavonoids, kaempferol dihexosidipentose and schaftoside were the most abundant, reaching approximately 3 mg/g, followed by isoorientin, vitexin-2-O-rhamnoside, and vicenin II, each with a content of approximately 2 mg/g. Furthermore, the cytotoxic potential of the plant against human colorectal adenocarcinoma (HT29) and human cervical cancer (HeLa) cells was investigated using the normal epithelial colon cell line (CCD 841CoTr) as a reference. It has been demonstrated that the ethyl acetate fraction was the most abundant in polyphenolic compounds and had the most promising anticancer activity. Further fractionation allowed for the obtaining of some subfractions that differed in phytochemical composition. The subfractions containing polyphenolic acids and flavonoids were characterized by low cytotoxicity against cancer and normal cell lines. Meanwhile, the subfraction with fatty acids was active and decreased the viability of HeLa and HT29 with minimal negative effects on CCD 841CoTr. The effect was probably linked to traumatic acid, which was present in the fraction at a concentration of 147 mg/g of dried weight. The research demonstrated the significant potential of C. acaulis as a plant with promising attributes, thus justifying further exploration of its biological activity.

Phytochemical Profiling, Antioxidant Activity, and Protective Effect against H2O2-Induced Oxidative Stress of Carlina vulgaris Extract.

Carlina vulgaris is a little-understood plant with unexplored biological potential, and the papers regarding its chemical composition are scarce. In our study, for the first time, the phytochemical profile of the plant, focusing on polar metabolites, was established using modern chromatographic techniques including LC-HRMS-QTOF-CAD, UHPLC-PDA-MS. Phytochemical analysis revealed that the species is a rich source of polyphenolic components, with the most abundant being chlorogenic acid and C-glycosides of luteolin, including carlinoside, orientin, isoorientin, and C-glycosides of apigenin, schaftoside, isoschaftoside, and vitexin. Furthermore, we assessed the impact of the polyphenolic-rich fraction of C. vulgaris extracts on human skin fibroblasts using the MTT and NR assays. It was found that the extract was non-toxic and exhibited potent antioxidant activity in the cells subjected to induced oxidative stress. Additionally, it effectively protected the cells against H2O2-induced cytotoxicity. Our study contributes to the general trend of searching for new phytotherapeutics with potential applications in pharmacy and medicine. The results indicate that further exploration of C. vulgaris species is worthwhile, as they can serve as valuable plant material for cosmetic use.

The Effect of In Vitro Digestion on Polyphenolic Compounds and Antioxidant Properties of Sorghum (Sorghum bicolor (L.) Moench) and Sorghum-Enriched Pasta.

The phenol content of sorghum is a unique feature among all cereal grains; hence this fact merits the special attention of scientists. It should be remembered that before polyphenols can be used in the body, they are modified within the digestive tract. In order to obtain more accurate data on the level and activity of tested ingredients after ingestion and digestion in the in vivo digestive tract, in vitro simulated digestion may be used. Thus, the aim of this study was to determine the content of polyphenols, flavonoids, and individual phenolic acids, as well as the antiradical properties, of sorghum and sorghum-enriched pasta before and after in vitro simulated gastrointestinal digestion. We observed that the total content of polyphenols decreased after gastric digestion of sorghum, and slightly increased after duodenal digestion. Moreover, the flavonoid content decreased after the first stage of digestion, while antioxidant properties increased after the first stage of digestion and slightly decreased after the second stage. The digestion of polyphenolics in sorghum is completely different to that in pasta-both in varieties with, and without, the addition of sorghum. For pasta, the content of total polyphenols and flavonoids, and free radical scavenging properties, decrease after each stage of digestion.

The Effect of Fresh Kale (Brassica oleracea var. sabellica) Addition and Processing Conditions on Selected Biological, Physical, and Chemical Properties of Extruded Snack Pellets.

The purpose of this study was to determine the effect of the addition of fresh kale and processing conditions on extruded pellet antioxidant activity and selected physicochemical properties. The results of the applied DPPH, FRAP, and TPC methods indicated that, for both 60 and 100 rpm screw speeds, snack pellet antioxidant activity and phenolic content were strongly linked to the fresh kale content, and these properties increased with the addition of this plant. The amount of fresh kale and the applied processing variables (extruder screw speed and the moisture content of the raw material blends) were also found to significantly affect the water absorption index, water solubility index, fat absorption index, fatty acid profile, and basic chemical composition of the obtained extrudates. The sample with the highest phenolic content (72.8 µg GAE/g d.w.), the most advantageous chemical composition (protein, ash, fat, carbohydrates, and fiber content), and high antioxidant properties was produced at a fresh kale content of 30%, a 36% moisture content, and a 100 rpm screw speed. The following phenolic acids were identified in this sample: protocatechuic, 4-OH-benzoic, vanillic, syringic, salicylic, caffeic, coumaric, ferulic, and sinapic. Sinapic acid was the prevailing phenolic acid.

Effect of the Production Parameters and In Vitro Digestion on the Content of Polyphenolic Compounds, Phenolic Acids, and Antiradical Properties of Innovative Snacks Enriched with Wild Garlic (Allium ursinum L.) Leaves.

A new type of corn snack has been created containing additions of wild garlic (Allium ursinum L.). This medicinal and dietary plant has a long tradition of use in folk medicine. However, studies on wild garlic composition and activity are fairly recent and scarce. This research aimed to investigate the influence of the screw speed and A. ursinum amounts on the antiradical properties as well as the content of polyphenolic compounds and individual phenolic acids of innovative snacks enriched with wild garlic leaves. The highest radical scavenging activity and content of polyphenols and phenolic acids were found in the snacks enriched with 4% wild garlic produced using screw speed 120 rpm. The obtained findings demonstrated that snacks enriched with wild garlic are a rich source of polyphenolic compounds. Since the concentration of such compounds is affected by many factors, e.g., plant material, presence of other compounds, and digestion, the second aim of this study was to determine radical scavenging activity, the content of polyphenols, and individual phenolic acids of snacks after in vitro simulated gastrointestinal digestion. Using an in vitro two-stage model, authors noted a significant difference between the concentration of polyphenolic compounds and the polyphenol content of the plant material before digestion.

Selected Natural Products in Neuroprotective Strategies for Alzheimer's Disease-A Non-Systematic Review.

Neurodegenerative disorders such as Alzheimer's disease (AD) are distinguished by the irreversible degeneration of central nervous system function and structure. AD is characterized by several different neuropathologies-among others, it interferes with neuropsychiatrical controls and cognitive functions. This disease is the number one neurodegenerative disorder; however, its treatment options are few and, unfortunately, ineffective. In the new strategies devised for AD prevention and treatment, the application of plant-based natural products is especially popular due to lesser side effects associated with their taking. Moreover, their neuroprotective activities target different pathological mechanisms. The current review presents the anti-AD properties of several natural plant substances. The paper throws light on products under in vitro and in vivo trials and compiles information on their mechanism of actions. Knowledge of the properties of such plant compounds and their combinations will surely lead to discovering new potent medicines for the treatment of AD with lesser side effects than the currently available pharmacological proceedings.

Electrospray ionization mass spectrometry characterization of ubiquitous minor lipids and oligosaccharides in milk of the camel (Camelus dromedarius) and their inhibition of oxidative stress in human plasma.

The aim of this study was to characterize minor lipids in methanol fraction extracted from raw camel milk after loading it on a water-preconditioned short C18 open column and fractionating with a gradient of methanol/water. The C18 column showed high fractionation efficiency of minor lipids, such as glycosphingolipids, lipopolysaccharides, or oligosaccharides, when compared with other constituents, in particular polysaccharides, proteins, and free fatty acids. Liquid chromatography electrospray ionization tandem mass spectrometry in negative ion mode was used to identify 21 new glycosphingolipids, lipopolysaccharides, and oligosaccharides. Electrospray ionization tandem mass spectrometry was qualified to provide relevant data for recognizing the molecular mass, glycosylation sequences, and structure of saccharide moieties for the revealed compounds. The sequence of combinations of one selected lipopolysaccharide, which was considered the backbone of the remaining lipopolysaccharides, was confirmed in a density functional theory study. The obtained results showed that the tested fraction is a rich source of glycosphingolipids, lipopolysaccharides, and oligosaccharides with antioxidant activity.

Novel Phenolic Constituents of Pulmonaria officinalis L. LC-MS/MS Comparison of Spring and Autumn Metabolite Profiles.

Lungwort (Pulmonaria officinalis L., Boraginaceae) is considered to possess therapeutic properties and it has been traditionally used as a remedy against various lung disorders in many countries. Nevertheless, very few data concerning its phytochemical composition are available. This research aims to provide a detailed description of specialized metabolites from the aerial parts of lungwort. Nine previously undescribed and 36 known phenolic compounds were detected in the 50% methanolic extract. Following multistep preparative procedures, structures of newly discovered compounds were determined using one- and two-dimensional techniques of NMR spectroscopy. Among the identified compounds were caffeic acid esters with aliphatic hydroxycarboxylic acids, conjugates of dicaffeic acid with rosmarinic acid, and previously unknown isomers of isosalvianolic acid A and yunnaneic acid E, as well as other lignans. Concentrations of all identified phenolic derivatives in the investigated herbal material were estimated using a method based on liquid chromatography with high-resolution mass spectrometry detection. Seasonal changes in the concentration of metabolites were also investigated using targeted and untargeted metabolomics techniques.

γ-Pyrone compounds: flavonoids and maltol glucoside derivatives from Herniaria glabra L. collected in the Ternopil region of the Ukraine.

The phytochemical investigation of the whole plant extracts of Herniaria glabra L. (Caryophyllaceae) led to the identification and isolation of four known flavonoids, one known and three undescribed maltol derivatives, and benzyl ß-gentiobioside. The structures were established by extensive 1D and 2D NMR spectroscopic analyses, as well as HRESIMS data. For the first time in Herniaria genus, as well as in Caryophylaceae family the presence of apiorutin {quercetin 3-O-[(D-apio-ß-d-furanosyl-(1 → 2)-O-[-α-l-rhamnopyranosyl-(1 → 6)]-ß-d-glucopyranoside]} and licoagroside B {maltol 3-O-[6-O-(3-hydroxy-3-methylglutaroyl)]-ß-d-glucopyranoside} were revealed. Additionally, antioxidant actions of apiorutin, rutin, narcissin (isorhamentin 3-O-ß-d-rutinoside) and licoagroside B were assessed in human blood plasma, exposed to the peroxynitrite-induced oxidative stress in vitro. The isolates partly reduced oxidative (oxidation of thiol groups) and nitrative (tyrosine nitration) damage to blood plasma proteins, decreased plasma lipid peroxidation as well as enhanced the non-enzymatic antioxidant capacity of blood plasma. No cytotoxicity of the examined substances towards peripheral blood mononuclear cells was found.

Silica Modified with Polyaniline as a Potential Sorbent for Matrix Solid Phase Dispersion (MSPD) and Dispersive Solid Phase Extraction (d-SPE) of Plant Samples.

Polyaniline (PANI) is one of the best known conductive polymers with multiple applications. Recently, it was also used in separation techniques, mostly as a component of composites for solid-phase microextraction (SPME). In the present paper, sorbent obtained by in situ polymerization of aniline directly on silica gel particles (Si-PANI) was used for dispersive solid phase extraction (d-SPE) and matrix solid-phase extraction (MSPD). The efficiency of both techniques was evaluated with the use of high performance liquid chromatography with diode array detection (HPLC-DAD) quantitative analysis. The quality of the sorbent was verified by Raman spectroscopy and microscopy combined with automated procedure using computer image analysis. For extraction experiments, triterpenes were chosen as model compounds. The optimal conditions were as follows: protonated Si-PANI impregnated with water, 160/1 sorbent/analyte ratio, 3 min of extraction time, 4 min of desorption time and methanolic solution of ammonia for elution of analytes. The proposed procedure was successfully used for pretreatment of plant samples.

Tentative Characterization of Polyphenolic Compounds in the Male Flowers of Phoenix dactylifera by Liquid Chromatography Coupled with Mass Spectrometry and DFT.

Phoenix dacylifera is an ancient palm species rich in (poly)phenols. These phenolic compounds were tentatively identified by using liquid chromatography coupled with ion spray mass spectrometry in tandem mode (LC/MS/MS) with negative ion detection. Negative identification of the compounds was based on their retention times and mass spectra in full scan mode (MS), and in different MS/MS modes. For the first time, complete hypothesis, and routs for both p-coumaroylshikimic acids (CoSA) and caffeoylshikimic acids (CSA) were suggested and confirmed by Density Fonctional Theory (DFT) study. Notably, of the 53 compounds characterized, 19 hydroxycinnamates derivatives were tentativelycharacterized in male flowers of date palm and 15 of them were recorded for the first time. In addition, five organic acids, six B-type proanthocyanidins, two anthocyanidin and 21 flavonoid derivatives have been tentatively characterized. Identification of B-type proanthocyanidins were based on the diagnostic ions resulting from heterocyclic ring fission (HRF) and retro-Diels-Alder (RDA) reaction of flavan-3-ol provided information on the hydroxylation pattern and the type of inter-flavan bond proanthocyanidins. The sequence of proanthocyanidins was detected through ions extracted from quinone methide (QM) cleavage of the inter-flavan bond.

Fingerprinting of strong spermatogenesis steroidal saponins in male flowers of Phoenix dactylifera (Date Palm) by LC-ESI-MS.

A fingerprint of steroid saponins, the major constituent in 80% methanolic fraction from the male flowers of Phoenix dactylifera has been established. Under ESI-MS/MS conditions, the fragmentation patterns of [M - H]- ions exclusively displayed signals corresponding to the cleavage of the glycosidic bonds, thus allowing a rapid identification of 21 steroidal saponins. Moreover, two unique among them conjugated with histidine were detected by LC-ESI (-)-MS and DFT and were given tentative names of 3-o-histidine-26-o-hexosyl-dioscin and 3-o-histidine-26-o-dihexosyl-hydroxydioscin. Their steroidal saponins exhibited a significant improvement of the sperm cells count, motility and viability in male rats. These effects could be attributed to enhancing the levels of sex hormones.

LC-ESI-MS/MS profile of phenolic and glucosinolate compounds in samh flour (Mesembryanthemum forsskalei Hochst. ex Boiss) and the inhibition of oxidative stress by these compounds in human plasma.

Samh flour (Mesembryanthemum forsskalei) is a foodstuff with high protein content, which can be used as a replacement for wheat flour. It is often consumed by Bedouin tribes of northern Saudi Arabia. Very little is known about bioactive molecules present in samh flour, therefore we analyzed its extracts to evaluate the contents of secondary metabolites. A total of 43 secondary metabolites present in 60% MeOH extract of samh flour were tentatively identified using LC-ESI-MS/MS. These compounds represented five major categories: glucosinolates, sinapic acid and sinapoylglycosides, acylated flavonoids, flavonoids, and amide derivatives. Their effect on oxidative damage of proteins and lipids was determined in vitro by assessing levels of protein thiol groups and concentrations of thiobarbituric acid reactive species (TBARS) in human plasma. Obtained results indicated that samh flour is a rich source of compounds with antioxidant activity.

Profiles analysis of proanthocyanidins in the argun nut (Medemia argun--an ancient Egyptian palm) by LC-ESI-MS/MS.

Medemia argun is an ancient palm rich in proanthocyanidins (PACs). These polyphenolic compounds are widely distributed in plants and are an integral part of the human diet. A sensitive high-performance liquid chromatography and electrospray ionization mass spectrometry (HPLC-ESI-MS) method in the negative ion mode for sequencing these ubiquitous and highly beneficial antioxidants is described in order to profile different PACs in M. argun nuts. The analytical protocol based on tandem mass spectrometry was used to sequence dimers, trimers, tetramers and pentamers with different A-type, B-type and A/B-type linkages. Diagnostic ions resulting from heterocyclic ring fission and retro-Diels-Alder reaction of flavan-3-ol provided information on the hydroxylation pattern and the type of interflavan bond. The sequences were discovered through ions derived from quinone methide cleavage of the interflavan bond. The identification of PACs linkages through LC-MS(n) eliminates a number of tedious separation steps. The method was successfully applied to give a view of PAC profile in M. argun nuts. M. argun nuts contained 636.88 mg/g PACs (as equivalent of (þ)-catechin). The data obtained in our research show that M. argun is a rich source of hydrolyzable PACs.

Amides and esters of phenylpropenoic acids from the aerial parts of Trifolium pallidum.

Two new derivatives of phenylpropenoic acids, N-trans-feruloyl-L-DOPA and O-trans-caffeoyl-malic acid dimethyl ester, along with four known N-trans-caffeoyl-L-DOPA (clovamide), N-trans-caffeoyl-L-DOPA-methyl ester, O-trans-caffeoyl-malic acid, O-trans-feruloyl-malic acid and quercetin 3-O-beta-D-glucopyranoside were isolated from the aerial parts of Trifolium pallidum. Their structures were elucidated by extensive spectroscopic methods including 1D- (1H, 13C) and 2D-NMR (DQF-COSY, HSQC, HMBC) experiments as well as mass spectrometry analysis.

Phenolic constituents of Knautia arvensis aerial parts.

A new C-6 flavone glycoside (6), together with seven known compounds, cryptochlorogenic acid (1), chlorogenic acid (2), 2-O-trans-caffeoylhydrocitric acid (3), isovitexin 7-beta-D-glucopyranoside (4), 7,4'-dihydroxy-5-methoxyflavone-6-C-beta-D-glucopyranoside (5), 3,5-O-dicaffeoylquinic acid (7) and 4,5-O-dicaffeoylquinic acid (8), were isolated from the aerial parts of Knautia arvensis. Their structures were elucidated by extensive spectroscopic methods including 1D- (1H, 13C and TOCSY) and 2D-NMR (DQF-COSY, HSQC, HMBC) experiments, as well as ESIMS analysis. Compounds 1, 3-5 and 8 are reported for the first time in Knautia arvensis.