A Comparative UHPLC-Q-Trap-MS/MS-Based Metabolomics Analysis to Distinguish Foeniculum vulgare Cultivars' Antioxidant Extracts.

Among the environmental factors, seasonality is the one which most affects the metabolome of a plant. Depending on the harvest season, the plant may have a variable content of certain metabolites and thus may have different biological properties. Foeniculum vulgare is an annual plant whose cultivation creates large amounts of waste rich in bioactive compounds. The present investigation was performed with the aim of determining the amount of biologically active compounds in F. vulgare wastes obtained from varieties of different seasonality. Ten polyphenolic compounds were quantified in the little stems and leaves of Tiziano, Pegaso, and Preludio cultivars by ultra performance liquid chromatography (UPLC) hyphenated to QTRAP mass spectrometry by using the MRM (multiple reaction monitoring) method. The antioxidant activity of hydroalcoholic extracts was then evaluated using TEAC and DPPH spectrophotometric assays, followed by a multivariate statistical analysis to determine the correlation between metabolite expression and antioxidant activity. The Preludio variety, grown in summer, showed a higher content of bioactive compounds, which guarantees it a better antioxidant power; kaempferol 3-O-glucuronide, quercetin 3-O-glucuronide, and quercetin 3-O-glucoside are the polyphenolic compounds that could be mainly responsible for the antioxidant effect of fennel. The PLS chemometric model, which correlated quantitative data obtained by a sensitive and selective LC-ESI-QTrap-MS/MS analysis of antioxidant activity, resulted in a selective tool to detect the compounds responsible for the activity shown by the extracts in chemical tests.

Effect of Very-Low-Calorie Ketogenic Diet on Psoriasis Patients: A Nuclear Magnetic Resonance-Based Metabolomic Study.

Psoriasis is an inflammatory disease of the epidermis based on an immunological mechanism involving Langerhans cells and T lymphocytes that produce pro-inflammatory cytokines. Genetic factors, environmental factors, and improper nutrition are considered triggers of the disease. Numerous studies have reported that in a high number of patients, psoriasis is associated with obesity. Excess adipose tissue, typical of obesity, causes a systemic inflammatory status coming from the inflammatory active adipose tissue; therefore, weight reduction is a strategy to fight this pro-inflammatory state. This study aimed to evaluate how a nutritional regimen based on a ketogenic diet influenced the clinical parameters, metabolic profile, and inflammatory state of psoriasis patients. To this end, 30 psoriasis patients were subjected to a ketogenic nutritional regimen and monitored for 4 weeks by evaluating the clinical data, biochemical and clinical parameters, NMR metabolomic profile, and IL-2, IL-1ß, TNF-α, IFN-γ, and IL-4 concentrations before and after the nutritional regimen. Our data show that a low-calorie ketogenic diet can be considered a successful strategy and therapeutic option to gain an improvement in psoriasis-related dysmetabolism, with significant correction of the full metabolic and inflammatory status.

LC-ESI/LTQ-Orbitrap-MS Based Metabolomics in Evaluation of Bitter Taste of Arbutus unedo Honey.

Strawberry tree honey is a high-value honey from the Mediterranean area and it is characterised by a typical bitter taste. To possibly identify the secondary metabolites responsible for the bitter taste, the honey was fractionated on a C18 column and the individual fractions were subjected to sensory analysis and then analysed by liquid chromatography coupled with high-resolution tandem mass spectrometry in negative ion mode, using a mass spectrometer with an electrospray source coupled to a hybrid high resolution mass analyser (LC-ESI/LTQ-Orbitrap-MS). A chemometric model obtained by preliminary principal component analysis (PCA) of LC-ESI/LTQ-Orbitrap-MS data allowed the identification of the fractions that caused the perception of bitterness. Subsequently, a partial least squares (PLS) regression model was built. The studies carried out with multivariate analysis showed that unedone (2-(1,2-dihydroxypropyl)-4,4,8-trimethyl-1-oxaspiro [2.5] oct-7-en-6-one) can be considered responsible for the bitter taste of strawberry tree honey. Confirmation of the bitter taste of unedone was obtained by sensory evaluation of a pure standard, allowing it to be added to the list of natural compounds responsible for giving the sensation of bitterness to humans.

Comparative Phytochemical Characterization, Genetic Profile, and Antiproliferative Activity of Polyphenol-Rich Extracts from Pigmented Tubers of Different Solanum tuberosum Varieties.

Plants produce a vast array of biomolecules with beneficial effects for human health. In this study, polyphenol and anthocyanin-rich extracts (PAE) from pigmented tubers of Solanum tuberosum L. varieties "Blue Star", "Magenta Love", and "Double Fun" in comparison with the more extensively studied "Vitelotte" were evaluated and compared for antiproliferative effects in human leukemia cells, and their phytochemical and genetic profiles were determined. In U937 cells, upon treatment with PAE, it was possible to reveal the expression of specific apoptotic players, such as caspase 8, 9, 3, and poly (ADP-ribose) polymerase (PARP), as well as the induction of monocyte and granulocyte differentiation. A liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) investigation revealed the presence of polyphenolic compounds in all the varieties of potatoes analyzed, among which caffeoyl and feruloyl quinic acid derivatives were the most abundant, as well as several acylated anthocyanins. Each pigmented variety was genotyped by DNA-based molecular markers, and flavonoid-related transcription factors were profiled in tubers in order to better characterize these outstanding resources and contribute to their exploitation in breeding. Interesting biological activities were observed for "Blue Star" and "Vitelotte" varieties with respect to the minor or no effect of the "Double Fun" variety.

Evaluation of bioactive compounds and antioxidant capacity of edible feijoa (Acca sellowiana (O. Berg) Burret) flower extracts.

ABSTRACT: The research aimed to evaluate the polyphenolic composition and the antioxidant capacity of edible extracts of feijoa (Acca sellowiana (O. Berg) Burret) flowers. Phenolic compounds of whole feijoa flower (FM), feijoa petals (PM) and feijoa petals juice (PJ) were identified by high-performance liquid chromatography coupled with electrospray mass spectrometry and quantified by liquid chromatography coupled with ultraviolet/visible detection. Moreover, the total polyphenol (TP) content was measured spectrophotometrically and the antioxidant capacities of the extracts were evaluated by FRAP, CUPRAC, DPPH·, and ABTS·+ assays. The FM showed TP content (395.14 ± 7.91 mg GAE/L) higher than PM and PJ, and exhibited better antioxidant capacities. FM extracts were characterized by the high content of anthocyanins (115.3 ± 3.6 mg/L), flavonols (42.9 ± 3.3 mg/L) and the presence of ellagic acid (7.9 ± 0.2 mg/L) and other galloyltannins. In addition, cyanidin-3-O-glucoside and apigenin were detected in all the three extracts. The present study provided an overview on particular bioactive compounds that characterise different parts of edible feijoa flowers. Among the latter, FM proved to be the most suitable for exploitation in the food and health manufactory.

Plant Specialized Metabolites in Hazelnut (Corylus avellana) Kernel and Byproducts: An Update on Chemistry, Biological Activity, and Analytical Aspects.

Corylus avellana (hazelnut) is one of the most popular tree nuts on a worldwide basis. The main products of C. avellana are kernels, a nutritious food, with a high content of healthy lipids, contained in a hard shell. In recent years, along with the ongoing research carried out on hazelnut kernels, a growing interest has been addressed to the hazelnut byproducts including hazelnut skin, hazelnut hard shell, and hazelnut green leafy cover as well as hazelnut tree leaf. These byproducts deriving from the roasting, cracking, shelling/hulling, and harvesting processes have been found as a source of "phytochemicals" with biological activity. The aim of this review is to provide a comprehensive and critical update on the chemistry and biological activity of specialized metabolites occurring in hazelnut kernels and byproducts. Phenolics are the most abundant phytochemicals not only in the kernels, but also in other processing byproducts. Attention has been also devoted to taxane derivatives isolated from C. avellana leaves. An overview on the biological activity, mainly antioxidant, antiproliferative, and antimicrobial along with less common biological effects, has been provided, contributing to highlight C. avellana as a source of bioactive phytochemicals with the potential to exert beneficial effects on human health. Finally, analytical techniques for the quali-quantitative analysis of specialized metabolites occurring in the different parts of C. avellana have been reviewed.

HR-LC-ESI-Orbitrap-MS based metabolite profiling of Prunus dulcis Mill. (Italian cultivars Toritto and Avola) husks and evaluation of antioxidant activity.

INTRODUCTION: Prunus dulcis Mill. is a plant cultivated for the production of its edible seeds, known as sweet almonds. In the last decades almond demand has highly increased due to their wide use in the industrial manufacturing of almond-based foods; this has led to a huge production of waste materials, mainly corresponding to shells and husks. OBJECTIVE: To achieve a deeper understanding of the chemical composition of almond husks with the aim of exploring these byproducts as a source of bioactives to be used in nutraceutical and cosmetic formulations. METHODS: Methanol, ethanol and ethanol-water extracts of the almond husks were analysed by high-resolution liquid chromatography electrospray ionisation Orbitrap mass spectrometry (HR-LC-ESI-Orbitrap-MS) in negative ion mode. Tandem mass spectrometry (MS/MS) data were acquired by using the Data-Dependent Scan experiment, allowing the precursor ion to be selected as the most intense peak during LC-MS analyses. Nuclear magnetic resonance (NMR) experiments were performed on a Bruker DRX-600 spectrometer. Folin-Ciocalteu, DPPH• (2,2-diphenyl-1-picrylhydrazyl) and TEAC (Trolox equivalent antioxidant capacity) assays were employed to determine the total phenolic content and the radical scavenging activity of the extracts. RESULTS: The LC-MS/MS analysis of the methanol extract guided the isolation of phloroglucinol derivatives, flavonoids and terpenes. Eco-friendly extraction methods showed to be selective in extracting flavonoids while the comparison of the LC-MS profiles of the Italian cultivars Toritto and Avola showed significant differences, confirming how different growing conditions may influence the metabolome of a plant species. CONCLUSION: This study led to a deeper insight into the chemical constituents of almond husks and showed how the eco-friendly extraction resulted in an effective method to obtain extracts rich in antioxidant sources.

Profiling and Simultaneous Quantitative Determination of Anthocyanins in Wild Myrtus communis L. Berries from Different Geographical Areas in Sardinia and their Comparative Evaluation.

INTRODUCTION: Myrtus communis L. (Myrtaceae) is a self-seeded shrub, widespread in Sardinia, with anti-inflammatory, antiseptic, antimicrobial, hypoglycemic and balsamic properties. Its berries, employed for the production of sweet myrtle liqueur, are characterised by a high content of bioactive polyphenols, mainly anthocyanins. Anthocyanin composition is quite specific for vegetables/fruits and can be used as a fingerprint to determine the authenticity, geographical origin and quality of raw materials, products and extracts. OBJECTIVE: To rapidly analyse and determine anthocyanins in 17 samples of Myrtus communis berries by developing a platform based on the integration of UHPLC-MS/MS quantitative data and multivariate analysis with the aim of extracting the most information possible from the data. METHODOLOGY: UHPLC-ESI-MS/MS methods, working in positive ion mode, were performed for the detection and determination of target compounds in multiple reaction monitoring (MRM) mode. Optimal chromatographic conditions were achieved using an XSelect HSS T3 column and a gradient elution with 0.1% formic acid in water and 0.1% formic acid in acetonitrile. Principal component analysis (PCA) was applied to the quantitative data to correlate and discriminate 17 geographical collections of Myrtus communis. RESULTS: The developed quantitative method was reliable, sensitive and specific and was successfully applied to the quantification of 17 anthocyanins. Peonidin-3-O-glucoside was the most abundant compound in all the extracts investigated. CONCLUSION: The developed methodology allows the identification of quali-quantitative differences among M. communis samples and thus defines the quality and value of this raw material for marketed products. Moreover, the reported data have an immediate commercial value due to the current interest in developing antioxidant nutraceuticals from Mediterranean plants, including Sardinian Myrtus communis. Copyright © 2016 John Wiley & Sons, Ltd.

Light Flux Density and Photoperiod Affect Growth and Secondary Metabolism in Fully Expanded Basil Plants.

Indoor production of basil (Ocimum basilicum L.) is influenced by light spectrum, photosynthetic photon flux density (PPFD), and the photoperiod. To investigate the effects of different lighting on growth, chlorophyll content, and secondary metabolism, basil plants were grown from seedlings to fully expanded plants in microcosm devices under different light conditions: (a) white light at 250 and 380 µmol·m-2·s-1 under 16/8 h light/dark and (b) white light at 380 µmol·m-2·s-1 under 16/8 and 24/0 h light/dark. A higher yield was recorded under 380 µmol·m-2·s-1 compared to 250 µmol·m-2·s-1 (fresh and dry biomasses 260.6 ± 11.3 g vs. 144.9 ± 14.6 g and 34.1 ± 2.6 g vs. 13.2 ± 1.4 g, respectively), but not under longer photoperiods. No differences in plant height and chlorophyll content index were recorded, regardless of the PPFD level and photoperiod length. Almost the same volatile organic compounds (VOCs) were detected under the different lighting treatments, belonging to terpenes, aldehydes, alcohols, esters, and ketones. Linalool, eucalyptol, and eugenol were the main VOCs regardless of the lighting conditions. The multivariate data analysis showed a sharp separation of non-volatile metabolites in apical and middle leaves, but this was not related to different PPFD levels. Higher levels of sesquiterpenes and monoterpenes were detected in plants grown under 250 µmol·m-2·s-1 and 380 µmol·m-2·s-1, respectively. A low separation of non-volatile metabolites based on the photoperiod length and VOC overexpression under longer photoperiods were also highlighted.

Metabolite Profiling for Typization of "Rucola della Piana del Sele" (PGI), Eruca sativa, through UHPLC-Q-Exactive-Orbitrap-MS/MS Analysis.

In August 2020, the Eruca sativa cultivar "Rucola della Piana del Sele" obtained from the European Union the prestigious PGI (protected geographical indication) label, which certifies the uniqueness of its characteristics and increases its prestige both nationally and, above all, internationally. This plant is recognized as a product of excellence, with a unique flavor and unmistakable aroma. Therefore, since there are no methods to characterize the PGI product, a metabolomic approach was applied to characterize E. sativa grown in the Piana del Sele and different geographical areas. As E. sativa has very wide cultivation, this study sought to compare the metabolite profiles of rocket grown in Piana del Sele, Bergamo, and Brescia, as well as in Switzerland, making a comparison also with the metabolite profile of E. sativa grown spontaneously. To determine the best procedure to distinguish "Rucola della Piana del Sele" from the others, different extraction procedures were carried out using different solvents and fresh or freeze-dried plant matrices. The different extracts were analyzed by liquid chromatography coupled with high-resolution mass spectrometry experiments, using chemometric analyses to identify biomarker metabolites that characterize the PGI product. The LC-ESI-Q-Exactive-MS/MS profiles of methanol and hydroalcoholic extracts of different cultivars of E. sativa were found to be rich in bioactive compounds such as glucosinolates, glycosylated flavonoids, fatty acids, and lipids. The LCMS data were analyzed by principal component analysis (PCA); the score scatter plot shows significant separation among Eruca samples grown in different geographical areas. In detail, loading the scatter plot revealed Eruca grown in Piana del Sele to be richer than other cultivars in glycosylated quercetin 3,3',4'-O-triglucoside (7), quercetin-3,4'-O-diglucoside-3'-O-(6-sinapoyl-glucoside) (10), and quercetin diglucoside (30). Finally, considering the biological interest in erucin, the myrosinase product of glucoerucin, the latter was quantified in the extracts by LC-ESI/QTrap/MS/MS using the multiple reaction monitoring (MRM) method; E. sativa from Piana del Sele showed the highest content of glucoerucin.

Sustainable Extraction Methods Affect Metabolomics and Oxidative Stability of Myrtle Seed Oils Obtained from Myrtle Liqueur By-Products: An Electron Paramagnetic Resonance and Mass Spectrometry Approach.

Myrtle liqueur production generates high amounts of by-products that can be employed for the extraction of bioactive compounds. Bio-based, non-toxic and biodegradable solvents (ethyl acetate and 2-methyltetrahydrofuran), and a mechanical extraction were applied to myrtle seeds, by-products of the liqueur production, to extract oils rich in phenolic compounds. The oils obtained were characterized for yield, peroxide value (PV), lipid composition, and total phenolic concentration (TPC). The phenolic profile of the oils, determined by LC-MS, the antioxidant activity, and the oxidative stability were also analyzed. A validated UHPLC-ESI-QTRAP-MS/MS analytical method in multiple reaction monitoring (MRM) mode was applied to quantify myricetin and its main derivatives in myrtle oils. The results pointed out clear differences among extraction methods on myricetin concentration. The oxidative stability of myrtle oils was studied with electron paramagnetic resonance (EPR) spectroscopy highlighting the effect of the extraction method on the oxidation status of the oils and the role of phenolic compounds in the evolution of radical species over time. A principal component analysis applied to LC-MS data highlighted strong differences among phenolic profiles of the oils and highlighted the role of myricetin in the oxidative stability of myrtle oils. Myrtle oil, obtained from the by-products of myrtle liqueur processing industry, extracted with sustainable and green methods might have potential application in food or cosmetic industries.

Metabolite Profiling, through LC-ESI/LTQOrbitrap/MS Analysis, of Antioxidant Extracts from Physalis alkekengi L.

Due to the increasing use of Physalis alkekengi L. as a food supplement and starting material for tea preparation, a comprehensive analysis of green extracts was performed. Two different extraction methods were applied to yellow Physalis alkekengi L. fruit and calyx and compared: hydroalcoholic extraction and decoction. Characterization of the metabolome of the calyx and fruit of yellow Physalis alkekengi L. was performed by LC-ESI/LTQOrbitrap/MS followed by LC-ESI/LTQOrbitrap/MS/MS to identify 58 phytocompounds using the two different extraction techniques. Subsequently, through preliminary spectrophotometric assays followed by cell studies, the antioxidant activity of the different Physalis alkekengi L. extracts were evaluated. It was found that Physalis alkekengi L. extracts are a good source of metabolites such as flavonoids, organic acids, phenylpropanoids, physalins and carotenoids, with various biological activities, in particular, antioxidant activity capable of reducing the production of free radicals in intestinal Caco-2 cells. For the first time, an integrated approach (metabolomics approach and antioxidant evaluation) was applied to the study of Physalis alkekengi green extracts and decoctions, the green extraction method mostly used in herbal preparations. An interesting finding was the high antioxidant activity shown by these extracts.

A validated LC-MS/MS method for quantitative determination of L-dopa in Fagioli di Sarconi beans (Phaseolus vulgaris L.).

An analytical method based on ultrasound assisted extraction (UAE) and liquid chromatography coupled to electrospray tandem mass spectrometry (LC-ESI/MS/MS) was validated and applied for determining L-dopa in four ecotypes of Fagioli di Sarconi beans (Phaseolus vulgaris L.), marked with the European label PGI (Protected Geographical Indication). The selectivity of the proposed method was ensured by the specific fragmentation of the analyte. Simple isocratic chromatographic conditions and mass spectrometric detection in multiple reaction monitoring (MRM) acquisition mode were used for sensitive quantification. The LC-ESI/MS/MS method was validated within a linear range of 0.001-5.000 µg/mL. Values of 0.4 and 1.1 ng/mL were obtained for the limits of detection and quantification, respectively. The repeatability, inter-day precision, and recovery values ranges were 0.6%-4.5%, 5.4%-9.9%, and 83%-93%, respectively. Fresh and dried beans, as well as pods, cultivated exclusively with organic methods avoiding any synthetic fertilizers and pesticides were analyzed showing an L-dopa content ranging from 0.020 ± 0.005 to 2.34 ± 0.05 µg/g dry weight.

Effects of White and Blue-Red Light on Growth and Metabolism of Basil Grown under Microcosm Conditions.

Indoor farming of basil (Ocimum basilicum L.) under artificial lighting to support year-round produce demand is an area of increasing interest. Literature data indicate that diverse light regimes differently affect downstream metabolic pathways which influence basil growth, development and metabolism. In this study, basil was grown from seedlings to fully developed plants in a microcosm, an innovative device aimed at growing plants indoor as in natural conditions. Specifically, the effects of white (W) and blue-red (BR) light under a photosynthetic photon flux density of 255 µmol m-2 s-1 on plant growth, photochemistry, soluble nutrient concentration and secondary metabolism were investigated. Plants grew taller (41.8 ± 5.0 vs. 28.4 ± 2.5 cm) and produced greater biomass (150.3 ± 24.2/14.7 ± 2.0 g vs. 116.2 ± 28.3/12.3 ± 2.5 g fresh/dry biomass) under W light compared to BR light. The two lighting conditions differently influenced the soluble nutrient concentration and the translocation rate. No photosynthetic stress was observed under the two lighting regimes, but leaves grown under W light displayed higher levels of maximum quantum yield of PSII and electron transport rate. Sharp differences in metabolic patterns under the two lighting regimes were detected with higher concentrations of phenolic compounds under the BR light.

UPLC-ESI-QTRAP-MS/MS Analysis to Quantify Bioactive Compounds in Fennel (Foeniculum vulgare Mill.) Waste with Potential Anti-Inflammatory Activity.

Foeniculum vulgare is a perennial aromatic plant whose cultivation produces large amounts of waste rich in bioactive compounds with promising anti-inflammatory activities. Nine selected metabolites were quantified through Ultra Performance Liquid Chromatography (UPLC) hyphenated to QTRAP mass spectrometry by using MRM (multiple reaction monitoring) was performed on four parts of fennel: bulb, stem, little stem, and leaf. Analysis revealed significant differences in the amount of quantified metabolites, suggesting that little stem and leaf are the most valuable parts of the waste. Phenolic acids and glycosylated flavonoids were quantified for their known possible anti-inflammatory activities; in fact, due to this reason their ability to inhibit COX-1 and COX-2 isoforms was evaluated through a fluorometric assay, resulting in specific inhibitors of COX-2 at certain concentrations. In conclusion, as the leaf of fennel may be beneficial to human health, clinical studies should include it in nutraceuticals or functional foods for human consumption.

Effect of Apple Juice Enrichment with Selected Plant Materials: Focus on Bioactive Compounds and Antioxidant Activity.

Using a multi-analytical approach, this paper aimed to investigate the effect of apple juice enrichment with Arbutus unedo and Diospyros kaki fruits, Myrtus communis berry extract, Acca sellowiana, or Crocus sativus flower by-products on both bioactive compounds content and antioxidant activity. Physico-chemical parameters, vitamin C, sugars, organic acids, total polyphenol content, antioxidant activity, and sensory attributes were evaluated. An LC-PDA/MS QTof analysis allowed for the identification of 80 different phenolic compounds. The highest polyphenol content (179.84 and 194.06 mg of GAE/100 g fw) and antioxidant activity (CUPRAC, 6.01 and 7.04 mmol of Fe2+/100 g fw) were observed in products with added A. sellowiana and D. kaki, respectively. Furthermore, the study showed a positive correlation between polymeric procyanidins and antioxidant activity (0.7646-0.8539). The addition of A. unedo fruits had a positively significant influence on the increment of vitamin C (23.68 ± 0.23 mg/100 g fw). The obtained products were attractive to consumers, especially those with 0.1% C. sativus flower juice, M. communis berry extract, and persimmon D. kaki fruits. The synergy among the different analytical techniques allowed us to obtain a complete set of information, demonstrating that the new apple smoothies were enriched in both different beneficial molecules for human health and in antioxidant activity.

Metabolomics Approach to Characterize Green Olive Leaf Extracts Classified Based on Variety and Season.

The huge interest in the health-related properties of plant polyphenols to be applied in food and health-related sectors has brought about the development of sensitive analytical methods for metabolomic characterization. Olive leaves constitute a valuable waste rich in polyphenols with functional properties. A (HR)LC-ESI-ORBITRAP-MS analysis with a multivariate statistical analysis approach using PCA and/or PLS-DA projection methods were applied to identify polyphenols in olive leaf extracts of five varieties from the Apulia region (Italy) in two different seasonal times. A total of 26 metabolites were identified, further finding that although metabolites are common among the different cultivars, they differ in the relative intensity of each peak and within each cultivar in the two seasonal periods taken into consideration. The results of the total phenol contents showed the highest content in November for Bambina and Cima di Mola varieties (1816 and 1788 mg/100 g, respectively), followed by Coratina, Leccino, and Cima di Melfi; a similar trend was found for the antioxidant activity and RapidOxy evaluations by reaching in Bambina values of 45 mmol TE/100 g and 85 min of induction time.

HR-LC-ESI-Orbitrap-MS-Based Metabolic Profiling Coupled with Chemometrics for the Discrimination of Different Echinops spinosus Organs and Evaluation of Their Antioxidant Activity.

This study aimed to assess and correlate the phenolic content and the antioxidant activity of the methanol extracts of the stems, roots, flowers, and leaves of Echinops spinosus L. from north-eastern Algeria. Qualitative analysis was performed by high-resolution mass spectrometry (HR) LC-ESI-Orbitrap-MS and (HR) LC-ESI-Orbitrap-MS/MS). Forty-five compounds were identified in the methanol extracts; some are described for the first time in E. spinosus. Targeted phenolic compounds were quantified by HPLC-DAD and it was shown that caffeoyl quinic derivatives were the most abundant compounds. Chemometric analysis was performed using principal component analysis (PCA) and hierarchical cluster analysis (HCA) based on the qualitative and quantitative LC data. The score plot discriminates different Echinopsis spinosus organs into three distinct clusters, with the stems and flowers allocated in the same cluster, reflecting their resemblance in their secondary metabolites. The antioxidant activities of the methanol extracts were assessed using cupric reducing antioxidant capacity (CUPRAC), ferric reducing antioxidant assay (FRAP), diphenyl picryl hydrazyl radical-scavenging capacity assay (DPPH●), and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS●+). The root extract exhibited the highest antioxidant activity, evidenced by 3.26 and 1.61 mmol Fe2+/g dried residue for CUPRAC and FRAP, respectively, and great free radical-scavenging activities estimated by 0.53 and 0.82 mmol TEAC/g dried residue for DPPH● and ABTS●+, respectively. The methanol extract of the roots demonstrated a significant level of total phenolics (TP: 125.16 mg GAE/g dried residue) and flavonoids (TFI: 25.40 QE/g dried residue TFII: 140 CE/g dried residue). Molecular docking revealed that tricaffeoyl-altraric acid and dicaffeoyl-altraric acid exhibited the best fit within the active sites of NADPH oxidase (NO) and myeloperoxidase (MP). From ADME/TOPAKT analyses, it can be concluded that tricaffeoyl-altraric acid and dicaffeoyl-altraric acid also revealed reasonable pharmacokinetic and pharmacodynamic characteristics with a significant safety profile.

Profiling of phenolics from Tephrosia cinerea.

An HPLC-ESIMS(n) method, based on high-performance liquid chromatography coupled to electrospray positive ionization multistage ion trap mass spectrometry, has been used to rapidly identify and guide the isolation of phenolic constituents from the methanol extract of the aerial parts of T. cinerea. On the basis of the results of the online screening by HPLC-ESIMS (n), 17 phenolic constituents, including two new compounds, namely demethylapollinin 7- O- ß-D-glucopyranoside (1) and cineroside A (17), were isolated, and their structures were unambiguously assigned by the extensive use of 1D- and 2D-NMR experiments.

Antioxidant bibenzyl derivatives from Notholaena nivea Desv.

Four new bibenzyl derivatives were isolated, together with other known bibenzyls, by bioassay-guided fractionation of a CHCl3-MeOH extract of Notholaena nivea Desv. (Pteridaceae) aerial parts. The structures were elucidated by NMR, ESIMS and other spectral analyses. Their antioxidative effects towards superoxide, lipidic peroxidation and the 2,2'-azino-bis-3-ethilbenzothiazoline-6-sulfonic acid (ABTS) radical were assayed. Results showed that the compound 3,12-dihydroxy-5-methoxybibenzyl (6) is the most active compound in the ABTS free-radical scavenging test, while in the coupled oxidation of ß-carotene and linoleic acid assay the compound 5,12-dihydroxy-3-methoxydibenzyl-6-carboxylic acid (1) exerted the highest activity after 1h. A superoxide anion enzymatic test was also carried out and the results were confirmed by an inhibition of xanthine oxidase activity assay. The putative protective role played by compounds 1 and 6 on the injurious effects of reactive oxygen metabolites on the intestinal epithelium, using a Caco-2 human cell line, was investigated. H2O2-induced alterations were prevented by preincubating the cells with compounds 1 and 6.