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Due to the high nutritional value as well as the appreciated sensory characteristic Citrus crop is one of the most popular fruits over the world. Albedo and flavedo are commonly discarded as waste and could represent an important by-product of food industry. In a circular economy contest we carried out a characterization of albedo and flavedo of Citrus limon var pompia (pompia) followed by the evaluation of antioxidant potential and cytotoxic activity. The chemical analysis showed the presence of several functional ingredients such as the flavanones, naringin, neohesperidin, nereriocitrin and hesperidin while quinic acid resulted the main phenolic acid detected. The flavedo extract exerted a complete inhibition of the cholesterol oxidative process from 25 μg of extract. Pompia extracts showed an inhibition of lipid oxidation in the β-carotene-linoleate system with an activity equivalent to about 0.31% (albedo) and 0.34% (flavedo) of the BHT activity, while the antiradical activity resulted equivalent to that of 62.90% (albedo) and 60.72% (flavedo) of the trolox activity. Finally, all extracts did not show any cytotoxic effect on differentiated Caco-2 cells by the AlamarBlue assay while exhibited, by MTT assay, a significant decrease in colon cancer Caco-2 cell viability.

Broccoli sprouts are considered functional food as they are naturally enriched in glucoraphanin (GR) that is the biological precursor of the anticancer compound sulforaphane (SFN). Due to its health promoting value, also broccoli sprout juice is becoming very popular. The present study aimed to quantitatively assess the conversion of GR to its hydrolysis products, SFN and SFN-nitrile, during the juice preparation process. We demonstrated that SFN plus SFN-nitrile yield from glucoraphanin is quite low (≈25%) and that some SFN is lost during the juice preparation partially due to the spontaneous conversion to sulforaphane-amine or conjugation to GSH and proteins naturally present in the juice. Our results demonstrate that the detection of the sole SFN free form does not provide reliable information about the real concentration of this functional compound in the juice.

Coffee is the second traded food commodity in the world. Beyond roasted seeds, the most part of the original fruit -and in particular pulp- is discarded as waste, with severe environmental and economic consequences in many developing countries. Our research focused on developing an eco-friendly extraction protocol of phytocomplexes from coffee pulp and evaluating their bioactivity and beneficial effects to human health as food supplements. Antioxidant activity assays (Folin-Ciocalteu and DPPH assays) were adopted to select the most effective extraction technique and results show antioxidant activity of coffee pulp extracts. After analysis of cytotoxicity on human epithelial gastric cells, measurements of IL-8 release of treated or pre-treated cells were performed. Results showed that the use of soft technical equipment and sustainable solvents (i.e. maceration process, aqueous extraction) can extract phytocomplexes with antioxidant properties. Moreover, IL-8 measurements showed impairment of this chemokine release at concentrations that may be reached in vivo in the gastrointestinal tract, following consumption of reasonable amount of extract. Pre-treatments analysis demonstrated the ability of coffee pulp extracts to prevent IL-8 release by gastric epithelial cells. Chemical evaluation performed by liquid chromatography mass spectrometry showed that quinic acid derivatives are abundant in coffee pulp extract together with procyanidins derivatives: those compounds might be responsible for the high biological activity. This evidence supports future applications of coffee pulp extracts as food supplement with high added value, starting from a waste that can be valorized through simple yet efficient extraction methods.

Essential oils from 8 Cuminum cyminum local populations collected in Morocco were investigated for their chemical composition and antimicrobial activity. The chemical composition, investigated by gas chromatographic technique, revealed a common fingerprint in all Moroccan samples: twenty-five compounds were identified with γ-terpinen-7-al being the major component in all samples studied. The antimicrobial activity of cumin essential oils were tested against 10 bacterial strains, belonging to 8 different species, and 6 yeast strains, belonging to 4 species. Lactic acid bacteria showed a good resistance to all essential oil tested while overall the cumin essential oils showed a strong antifungal activity that affected both maximum specific growth rate and lag time

This study aims to identify and quantify the glucosinolates from different parts of wild radish R. raphanistrum (leaves, flowers, fruits, roots) using liquid chromatography-tandem mass spectrometry. Glucoraphenin is the predominant compound, accounting for about 87% (w/w) of total glucosinolate content, followed by glucobrassicin, glucoraphasatin and glucoraphanin (153 mg 100 g−1, 149 mg 100 g−1 and 141 mg 100 g−1 FW, respectively) in fruits; followed by glucoraphasatin (3 mg 100 g−1 FW) in flowers and by glucobrassicin, 4-hydroxyglucobrassicin and glucoraphasatin (145 mg 100 g−1, 27 mg 100 g−1 and 24 mg 100 g−1 FW, respectively) in leaves. In roots the major glucosinolate is glucoraphasatin (56 mg 100 g−1 FW) followed by the glucoraphenin and methoxyglucobrassicin (16 mg 100 g−1 and 7 mg 100 g−1 FW, respectively). Principal component analysis allowed the discrimination of fruit samples from other parts of the plant for the majority of glucosinolates and the fruits are highlighted as sources of glucosinolates. The results are interesting given that wild radish is one of the most important weeds of crops in the Mediterranean region and is popular for home vegetable production and for its employment in human nutrition both as a food as well as for medicinal purposes.

The phytochemical content of Fragaria vesca L. (wild strawberry) grown in Campania Region (South Italy) was investigated. Wild and cultivated fruit were collected from different geographic areas and from bothautochthonous and non-autochthonous germplasm. Extracts were submitted to untargeted liquid chromatography coupled to high resolution mass spectrometry (LC–ESI-Orbitrap–MS) for metabolite profiling. 39 different phenolic compounds have been detected and identified in the methanolic extract of spontaneous and cultivated strawberries from different origins. Targeted quantitative analyses of selected known metabolites were performed using UHPLC interfaced to Q-Trap mass spectrometer in Multiple Reaction Monitoring mode. Principal Component Analysis (PCA) was applied to both untargeted and targeted results. PCA was applied to the untargeted profiling data indicating that fruits from locations with different pedoclimatic conditions can be discerned. Results from quantitative analysis indicate that autochthonous strawberries display significantly higher levels of anthocyanins, especially cyanidin 3-O-glucoside, and other phenolic compounds, like gallic acid and procyanidin B1, compared to the one from non-autochthonous germplasm. The targeted and untargeted metabolite profiling approach used in this work is proposed for the assessment of geographical origin of the berries. © 2016 Elsevier Ltd

Capparis spinosa L. (Capparidaceae), also known as caper, is widely known for its very aromatic flower buds (capers),that are largely employed as a flavouring in cooking. Capparis species are regarded as a potential source of important bioactive compounds, in fact, due to their botanical relationship with Brassica species; they contain glucosinolates, secondary plant metabolites, that have been studied for their potential anticarcinogenic properties. In addition, the presence of other numerous beneficial compounds such as polyphenols, alkaloids, lipids, vitamins and minerals have been reported. The aim of this study was to individuate and determinate the principal bioactive compounds occurring in different part (leaves, buds and flowers) of wild and cultivated C. spinosa collected from different area of Sardinia (Italy). Ultra-high performance liquid chromatography-triple quadrupole/linear ion trap tandem mass spectrometry methods were used for identification and simultaneous determination of 27 bioactive molecules. Analysis of different samples revealed qualitative and quantitative differences in the content of flavonoids, glucosinolates, anthocyanins and phenolic acids. In particular, glucocapparin resulted the most abundant with values ranging from 112 to 364 mg/100 g Fresh Weight (FW); followed by rutin with highest value of 126 mg/100 g FW, 4-hydroxyglucobrassicin with highest value of 42 mg/100 g FW and isorhamnetin 3-O-rutinoside with highest value of 24 mg/100 g FW. Based on this metabolomic targeted approach, quantitative results were treated by principal component analysis to explore and visualise correlation and discrimination among collections of C. spinosa samples. Copyright © 2016 John Wiley & Sons, Ltd.

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.

Epidemiological studies show an inverse association between Brassica consumption and chronic diseases. Phytochemicals are thought to be beneficial for human health and therefore responsible for this protective effect. Increasing their levels into Brassica food is considered an expedient nutritional strategy that can be achieved through the manipulation of growth conditions by elicitors. In this work we systematically evaluated the influence of treatment with different elicitors (sucrose, mannitol, NaCl, 1-aminocyclopropane-L-carboxylic acid, salicylic acid and methyl jasmonate) on the phytochemical composition of broccoli sprouts. The content of total and single glucosinolates, total phenolic compounds, total flavonoids, total anthocyanins, vitamin C and E and β-carotene was assessed. The exposure to different elicitors produced concentration- and elicitor-dependent specific changes in the content of all the phytochemicals considered. Sucrose, identified as the most effective elicitor by principal component analysis, induced a significant increase of total and specific glucosinolates, vitamin C, total anthocyanins and polyphenols. Sucrose is likely to represent an effective tool to increase the nutritional value of broccoli sprouts.

Essential oils (EOs) from several individuals of Myrtus communis L. (M. communis) growing in different habitats in Sardinia have been studied. The analyses were focused on four groups of samples, namely cultivated and wild M. communis var. melanocarpa DC, characterized by red/purple berries, and cultivated and wild M. communis var. leucocarpa DC, characterized by white berries. Qualitative and quantitative analyses demonstrated different EO fingerprints among the studied samples: cultivated and wild leucocarpa variety differs mainly from the melanocarpa variety by a high amount of myrtenyl acetate (>200 mg/mL and 0.4 mg/mL in leucocarpa and melanocarpa varieties respectively). Conversely, the wild group is characterized by a higher amount, compared with the cultivated species, of linalool (about 110 mg/mL and 20 mg/mL respectively), linalyl acetate (about 24 mg/mL and about 6 mg/mL respectively) whereas EOs of the cultivated plants were rich in pinocarveol-cis compared with wild plants (about 2 mg/mL and about 0.5 mg/mL respectively). Principal component analysis applied to the chromatographic data confirm a differentiation and classification of EOs from the four groups of M. communis plants. Finally, antioxidant activity of the studied EOs shows differences between the various categories of samples.

Helichrysum italicum (Asteraceae) is a small shrub of the Mediterranean area which colonizes a wide range of habitats. In Sardinia, Italy, the species is widely dispersed in different environments ranging from sandy beaches to holm hoak forests 1250 m above sea level (a. s. l.). Recently, the species has been attributed an increasing economical value mainly for its pharmacological, aromatic proprieties and ornamental uses. In this study, the chemical profiles of 146 H. italicum ssp. microphyllum genotypes from two contrasting habitats (seaside and mountains) in Sardinia were explored. Significant differences in volatile fractions between the two groups were found, displaying a correlation between secondary metabolite production and habitat type. Multivariate analysis of variance revealed that monoterpenes, sesquiterpenes (oxygenated/not oxygenated), alcohols, ester were quantitatively differentially produced in the plants growing under different environmental conditions (P Considering the importance of climatic conditions in plant development and growth, the relationship between meteorological variables and chemical compounds of the volatile fraction was explored by principal component analysis. Nerolidol is mostly positively correlated to mean winter temperature, while italicene, bergamotene, nerol and curcumene are positively influenced by spring and summer precipitation. Finally, Mantel test revealed a significant correlation (Spearman, P = 0.002) between chemical and meteorological matrices of Euclidean distances, suggesting that the most diverse chemical profiles belonged to plants from the farther climatic locations.

Extra virgin olive oils (EVOOs) from the fruits of Italian autochthonous varietiesTonda di Villacidro,Tonda di Cagliari,Semidana, andBosanawere investigated to promote their quality aspects. All the analyzed EVOOs showed low values of acidity (≤0.45%) and of peroxide value (≤6.22 mEq O2/kg). There were no relevant differences in fatty acids and triacylglycerols composition among the four EVOOs. Tocopherols determined by HPLC-FL revealed thatBosanaoil was characterized by the highestα-tocopherol level (213.3 ± 55.4 mg/kg). Chlorophylls, carotenoids, and total phenol (TP) contents as well as antioxidant activity (FRAP, DPPH∙, and ABTS∙+assays) of the oils hydrophilic fractions (HFs) were assessed by spectrophotometric methods. Some differences concerning the antioxidant activity and the TP content were observed:Bosanaoil HF activity was the most pronounced (1.17 ± 0.37 mmol TEAC/kg) and it contained the highest TPs amount (335.20 ± 121.34 mg/kg). HFs phenolic composition was determined by HPLC-DAD. The main identified phenols were secoiridoids, dominating inBosanaoil, such as decarboxymethyl ligstroside aglycone (p-HPEA-EDA, 35.8 ± 19.9 mg/kg) and oleuropein aglycone (3,4-HPEA-EA up to 84.7 mg/kg). In summary, all the four varieties showed good characteristics for the use as quality EVOO.

The origin and taxonomy of Citrus monstruosa (Rutaceae) are currently unknown and thus a nontargeted metabolomic analysis on the flavedo volatiles was performed. Headspace solid‐phase micro extraction coupled with gas chromatography analysis was applied to eight Citrus spp. including Citrus monstruosa, and the chemical composition of the volatile organic compounds (VOC) was analyzed to give a chemical comparison between different Citrus species. Forty‐four metabolites were detected across the species: several monoterpenes were found in all Citrus spp. and among them limonene comes first in all Citrus spp. ranging from 96.7 to 66.7%. Conversely, some molecules were found to be specific for only a few of the samples. The classical univariate evaluation of the VOC data is unlikely to give objective data for the cultivar classification; a multivariate approach, principal component analysis, is an easy to use, powerful tool. Therefore, a chemometric comparison between chromatographic data of all Citrus spp. was performed with the aim of understanding the role of VOC in the discrimination of the eight studied cultivars.

The consumption of vegetables belonging to the family Brassicaceae (e.g., broccoli and cauliflower) is linked to a reduced incidence of cancer and cardiovascular diseases. The molecular composition of such plants is strongly affected by growing conditions. Here we developed an unbiased metabolomics approach to investigate the effect of light and dark exposure on the metabolome of broccoli sprouts and we applied such an approach to provide a bird’s-eye view of the overall metabolic response after light exposure. Broccoli seeds were germinated and grown hydroponically for five days in total darkness or with a light/dark photoperiod (16 h light/8 h dark cycle). We used an ultra-performance liquid-chromatography system coupled to an ion-mobility, time-of-flight mass spectrometer to profile the large array of metabolites present in the sprouts. Differences at the metabolite level between groups were analyzed using multivariate statistical analyses, including principal component analysis and correlation analysis. Altered metabolites were identified by searching publicly available and in-house databases. Metabolite pathway analyses were used to support the identification of subtle but significant changes among groups of related metabolites that may have gone unnoticed with conventional approaches. Besides the chlorophyll pathway, light exposure activated the biosynthesis and metabolism of sterol lipids, prenol lipids, and polyunsaturated lipids, which are essential for the photosynthetic machinery. Our results also revealed that light exposure increased the levels of polyketides, including flavonoids, and oxylipins, which play essential roles in the plant’s developmental processes and defense mechanism against herbivores. This study highlights the significant contribution of light exposure to the ultimate metabolic phenotype, which might affect the cellular physiology and nutritional value of broccoli sprouts. Furthermore, this study highlights the potential of an unbiased omics approach for the comprehensive study of the metabolism.

Lipidomics and metabolomics have emerged as important tools for the characterization of specific physiological and pathological traits. The selection of the analytical approaches and the choice of a targeted rather than an untargeted strategy in metabolomics find their reasons in the driving hypothesis of the study, sample features and instrumental availability. Moreover, in the last years, -omics approaches have found their application in the study of sex-related dimorphism. In this review, lipidomic and metabolomic analyses are presented in a biomedical perspective. Here, we provide an updated overview covering recent applications of metabolomics and lipidomics in the area of sex-related differences in human and preclinical models. Experimental evidence underlines that sex is one of the most relevant biological variables significantly influencing metabolomic and lipidomic profiles. This knowledge can be exploited for the identification of novel sex-specific biomarkers and innovative targets relevant for gender medicine.

Quality and composition of virgin olive oil (VOO) are strictly dependent on complex processes that take place during the olive fruit crushing and malaxation of the olive paste. In this work, modulation of O2 levels within malaxation chambers (R1: unmodified atmosphere; R2: oxygen: 12.73–4.64 kPa from the beginning to the end of malaxation; R3: 10.46–2.27 kPa; R4: 9.87–0.69 kPa) in two continuous “two-phase” and “three-phase” oil extraction plants was performed. Combined effects on the biosynthesis of nutritionally bioactive molecules and aroma volatiles and on the resulting sensory properties of the produced oils were investigated. Results showed that the type of oil extraction plant markedly affected the level of the phenolic compounds in the oil (and the related sensory attributes of bitter, pungency, astringency and bitter and pungency persistence). Reduction of O2 concentration in the malaxing chamber, while having a minor impact on the presence of phenolic compounds, significantly affected the formation of all the examined volatiles. Particularly, lowered levels of oxygen hindered the formation of lipoxygenase derived volatiles weakening odours and flavours of artichoke, fresh fruity, and fresh cut grass.

Moringa oleifera is a medicinal plant and an excellent dietary source of micronutrients (vitamins and minerals) and health-promoting phytochemicals (phenolic compounds, glucosinolates and isothiocyanates). Glucosinolates and isothiocyanates are known to possess anti-carcinogenic and antioxidant effects and have attracted great interest from both toxicological and pharmacological points of view, as they are able to induce phase 2 detoxification enzymes and to inhibit phase 1 activation enzymes. Phenolic compounds possess antioxidant properties and may exert a preventative effect in regards to the development of chronic degenerative diseases. The aim of this work was to assess the profile and the level of bioactive compounds in all parts of M. oleifera seedlings, by using different MS approaches. First, flow injection electrospray ionization mass spectrometry (FI-ESI-MS) fingerprinting techniques and chemometrics (PCA) were used to achieve the characterization of the different plant's organs in terms of profile of phenolic compounds and glucosinolates. Second, LC-MS and LC-MS/MS qualitative and quantitative methods were used for the identification and/or determination of phenolics and glucosinolates in M. oleifera.

The consumption of wine and spirits, traditionally aged in oak barrels, exposes humans to roburin ingestion. These molecules belong to a class of ellagitannins (ETs), and their only known source is oak wood. Very little is currently known about roburin bioavailability and biological activity. We reported for the first time human absorption of roburins from a French oak wood (Quercus robur) water extract (Robuvit) by measuring the increase of total phenols (from 0.63 ± 0.06 to 1.26 ± 0.18 μg GAE equiv/mL plasma) and the appearance of roburin metabolites (three different glucoronidate urolithins and ellagic acid), in plasma, after 5 days of supplementation. Robuvit supplementation induced also the increase of plasma antioxidant capacity from 1.8 ± 0.05 to 1.9 ± 0.01 nmol Trolox equiv/mL plasma. Moreover, utilizing a combined ex vivo cell culture approach, we assessed the effect of Q. robur metabolites (present in human serum after supplementation) on gene expression modulation, utilizing an Affymetrix array matrix, in endothelial, neuronal, and keratinocyte cell lines. The functional analysis reveals that Robuvit metabolites affect ribosome, cell cycle, and spliceosome pathways.

Liquid chromatography electro-spray tandem mass spectrometry (LC-ESI/MS/MS) was applied to the determination of sweet glycosides (steviol glycosides), and toxic aglycon steviol in 24 samples of Stevia rebaudiana (Bertoni) aerial parts, which had been experimentally cultivated in Italy, although derived from seeds of different geographical origin. On the basis of the specific fragmentation of these compounds, an LC-MS/MS method was developed with the aim of quantifying analytes in plant material. Although toxic steviol was not detectable in all the samples, the samples with the highest levels of steviol glycosides were identified. Analysis of the different samples revealed that they were good quality samples, quality being directly linked to the presence of sweet glycosides in the plants cultivated in Italy, although there were differences in the content of these compounds according to the origin of the seeds, and in particular, a major concentration of compounds with major sweetness activity and minor toxicity was found in the population coming from Brazil (for example: sample 10, stevioside content 15.74±2.0% p/p and rebaudioside A content 3.09±0.39% p/p of dried plant). Finally, based on this metabolomic targeted approach, the results obtained for the samples were treated by Principal Component Analysis, identifying specific genotypic differences based on the geographic origin of the seeds.

IntroductionArgania spinosa kernel cakes, obtained from argan oil extraction process, are known to contain large amounts of saponins. Only a few have been characterised previously, due to the use of pure ethanol as extracting solvent. The use of aqueous 50% ethanol improved the extraction of more polar saponins. ObjectiveIdentification of polar saponins in kernel cakes of Argania spinosa by liquid chromatography-mass spectrometry and NMR techniques. MethodsDefatted kernel cakes were first extracted with ethanol and then twice with 50% aqueous ethanol. Individual crude extracts were analysed with an ion-trap mass spectrometer in negative mode electrospray MS and MS/MS modes. NMR experiments were run under standard conditions at 300K on a Bruker DRX-600 spectrometer. ResultsThe LC-MS base peak chromatogram of saponins from pure ethanol extract was dominated by 11 large and several small peaks but the UV chromatogram showed only two peaks, corresponding to the main neutral saponins found previously in Argania: arganine A and B. In 50% aqueous ethanol extracts, numerous other saponins were detected. Many of them were glucuronide oleanane-type triterpene carboxylic acid 3,28-O-bidesmosides (GOTCAB saponins). The assignments of H-1- and C-13-NMR spectra of the four most abundant GOTCAB saponins confirmed the MS results. ConclusionFour GOTCAB saponins were structurally identified by NMR analysis in the 50% aqueous ethanol extract. Furthermore, LC-MS analyses showed the presence of at least 19 additional polar saponins in these kernel cakes. Copyright (c) 2013 John Wiley & Sons, Ltd. Argania spinosa kernel cakes, byproducts of argan oil extraction process, are known to contain large amounts of saponins. Only a few of them were characterized previously. This was due to the choice of pure ethanol as extracting solvent. The aqueous 50% ethanol improved drastically the extraction of more polar saponins. Four GOTCAB saponins (Glucuronide oleanane-type triterpene carboxylic acid 3,28-bidesmosides) were structurally identified by NMR analyses in this extract. LC-MS analyses showed the presence of at least 19 additional polar saponins.

Introduction\ud\udMyrtus 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. \ud\ud\udObjective\ud\udTo 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. \ud\ud\udMethodology\ud\udUHPLC‐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. \ud\ud\udResults\ud\udThe 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. \ud\ud\udConclusion\ud\udThe 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.

Glucosinolates are an important class of secondary plant metabolites, possessing health-promoting properties. Young broccoli plants are a very good source of glucosinolates with concentrations several times greater than in mature plants. The aim of our study was to develop a liquid chromatography–mass spectrometry and liquid chromatography/tandem mass spectrometry qualitative and quantitative method for the measure of glucosinolates in broccoli sprouts. The described method provides high sensitivity and specificity, allowing a rapid and simultaneous determination of 14 glucosinolates. The proposed method has been validated for eight glucosinolates: glucobrassicin, glucoraphanin, glucoiberin, glucoerucin, progoitrin, gluconapin, sinigrin and glucocheirolin. The linear range was 1–150 µg ml−1, the intra-day and inter-day precision values are within 6% and 8% at the lower limit of quantification, while the overall recovery of the eight glucosinolates was 99 ± 9%. This validated method was used successfully for analysis of glucosinolates content of broccoli sprouts grown in different conditions. Copyright © 2012 John Wiley & Sons, Ltd.

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- β-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.

Ethnopharmacological relevance An investigation of topical anti-inflammatory activity was undertaken on plants used in Central America traditional medicine. Aim of study Four herbal drugs used in the folk medicine of Central America to treat inflammatory skin affections (Acacia cornigera bark, Byrsonima crassifolia bark, Sphagneticola trilobata leaves and Sweetia panamensis bark) were evaluated for their topical anti-inflammatory activity. Materials and methods Petroleum ether, chloroform and methanol extracts were obtained for herbal medicines and then extracts were tested on Croton oil-induced ear dermatitis model in mice. Results Almost all the extracts reduced the Croton oil-induced ear dermatitis in mice and the chloroform ones showed the highest activity, with ID50 (dose giving 50% oedema inhibition) values ranging from 112 μg/cm2 (Byrsonima crassifolia) to 183 μg/cm2 (Sphagneticola trilobata). As reference, ID50 of the non-steroidal anti-inflammatory drug indomethacin was 93 μg/cm2. Conclusions Lipophilic extracts from these species can be regarded as potential sources of anti-inflammatory principles.

Capparis spinosa L. (Capparidaceae), also known as caper, is widely known for its very aromatic flower buds (capers),that are largely employed as a flavouring in cooking. Capparis species are regarded as a potential source of important bioactive compounds, in fact, due to their botanical relationship with Brassica species; they contain glucosinolates, secondary plant metabolites, that have been studied for their potential anticarcinogenic properties. In addition, the presence of other numerous beneficial compounds such as polyphenols, alkaloids, lipids, vitamins and minerals have been reported. The aim of this study was to individuate and determinate the principal bioactive compounds occurring in different part (leaves, buds and flowers) of wild and cultivated C. spinosa collected from different area of Sardinia (Italy). Ultra-high performance liquid chromatography-triple quadrupole/linear ion trap tandem mass spectrometry methods were used for identification and simultaneous determination of 27 bioactive molecules. Analysis of different samples revealed qualitative and quantitative differences in the content of flavonoids, glucosinolates, anthocyanins and phenolic acids. In particular, glucocapparin resulted the most abundant with values ranging from 112 to 364 mg/100 g Fresh Weight (FW); followed by rutin with highest value of 126 mg/100 g FW, 4-hydroxyglucobrassicin with highest value of 42 mg/100 g FW and isorhamnetin 3-O-rutinoside with highest value of 24 mg/100 g FW. Based on this metabolomic targeted approach, quantitative results were treated by principal component analysis to explore and visualise correlation and discrimination among collections of C. spinosa samples. Copyright © 2016 John WileySons, Ltd.

Benefits to health from a high consumption of fruits and vegetables are well established and have been attributed to bioactive secondary metabolites present in edible plants. However, the effects of specific health-related phytochemicals within a complex food matrix are difficult to assess. In an attempt to address this problem, we have used elicitation to improve the nutraceutical content of seedlings of Brassica oleracea grown under controlled conditions. Analysis, by LC-MS, of the glucosinolate, isothiocyanate and phenolic compound content of juices obtained from sprouts indicated that elicitation induces an enrichment of several phenolics, particularly of the anthocyanin fraction. To test the biological activity of basal and enriched juices we took advantage of a recently developed in vitro model of inflamed human intestinal epithelium. Both sprouts’ juices protected intestinal barrier integrity in Caco-2 cells exposed to tumor necrosis factor α under marginal zinc deprivation, with the enriched juice showing higher protection. Multivariate regression analysis indicated that the extent of rescue from stress-induced epithelial dysfunction correlated with the composition in bioactive molecules of the juices and, in particular, with a group of phenolic compounds, including several anthocyanins, quercetin-3-Glc, cryptochlorogenic, neochlorogenic and cinnamic acids.

A new combined “untargeted” and “targeted” metabolomics approach to investigate Rosmarinus officinalis L. plants was proposed by using the merging of high-performance thin-layer chromatography (HPTLC) assay and liquid chromatography mass spectrometry (LC–MS) with principal component analysis (PCA). Rapid fingerprints of Rosmarinus extracts were obtained and compared by using HPTLC technique; targeted compounds were identified and quantified by LC–MS/MS (MRM), while the antioxidant capacity of extracts was evaluated by methods based on the ability to scavenge radicals such as ABTS+* and DPPH*, since the metabolite expression is generally correlated with the biological activity. By HPTLC fingerprints, a rapid visual comparison of secondary metabolites in Rosmarinus samples from different places of collection was obtained and PCA was performed providing an overview of the capacity of the HPTLC variables to discriminate samples. HPTLC fingerprint assisted by PCA results a reliable untargeted approach for the discrimination and classification of different samples on the basis of their geographical area, from west to east of Sardinia. The ‘targeted’ approach was performed by developing LC–MS/MS (MRM) method allowing the identification and quantification of 12 compounds belonging to the family of flavonoids and phenolic acids. The combination of quantitative data with PCA allowed to discriminate La Maddalena Rosmarinus sample from others, in a correlated way with antioxidant activities results, showing values significantly higher. A positive Pearson correlation between antioxidant activity measured with DPPH and ABTS and total phenolic contents, finally confirmed that antioxidant activity depends on phenolic amounts.

Moringa oleifera is a medicinal plant and an excellent dietary source of micronutrients (vitamins and minerals) and health-promoting phytochemicals (phenolic compounds, glucosinolates and isothiocyanates). Glucosinolates and isothiocyanates are known to possess anti-carcinogenic and antioxidant effects and have attracted great interest from both toxicological and pharmacological points of view, as they are able to induce phase 2 detoxification enzymes and to inhibit phase 1 activation enzymes. Phenolic compounds possess antioxidant properties and may exert a preventative effect in regards to the development of chronic degenerative diseases. The aim of this work was to assess the profile and the level of bioactive compounds in all parts of M. oleifera seedlings, by using different MS approaches. First, flow injection electrospray ionization mass spectrometry (FI-ESI-MS) fingerprinting techniques and chemometrics (PCA) were used to achieve the characterization of the different plant's organs in terms of profile of phenolic compounds and glucosinolates. Second, LC-MS and LC-MS/MS qualitative and quantitative methods were used for the identification and/or determination of phenolics and glucosinolates in M. oleifera.

Glucosinolates are a class of secondary plant metabolites particularly occurring in Cruciferae with potential health-promoting properties, as their hydrolysis products, isothiocyanates, possess chemopreventive and antioxidant activities. In the present study, we systematically studied the in vitro redox behaviour of 15 glucosinolates, by using a range of analytical methods measuring different activities: (i) radical scavenging activity toward peroxyl and toward ABTS radical (chain-breaking activity); (ii) capacity in modulating the in vitro resistance of human low-density lipoprotein (LDL) catalysed by copper (chelating and chain-breaking activity). Data obtained from different assays were compared and analysed by principal component analysis (PCA). PCA allowed us to identify a big cluster of glucosinolates (10 out 15 tested) that do not possess any antioxidant capacity; while, the other five glucosinolates showed moderate and specific antioxidant capacity. Notably, sinalbin and gluconasturtiin were highly active in scavenging ABTS radical and in protecting LDL from copper-catalysed oxidation, respectively. The overall results of this study indicate that just few glucosinolates can act as antioxidants.

Argania spinosa kernel cakes, obtained from argan oil extraction process, are known to contain large amounts of saponins. Only a few have been characterised previously, due to the use of pure ethanol as extracting solvent. The use of aqueous 50% ethanol improved the extraction of more polar saponins.Identification of polar saponins in kernel cakes of Argania spinosa by liquid chromatography-mass spectrometry and NMR techniques.Defatted kernel cakes were first extracted with ethanol and then twice with 50% aqueous ethanol. Individual crude extracts were analysed with an ion-trap mass spectrometer in negative mode electrospray MS and MS/MS modes. NMR experiments were run under standard conditions at 300 K on a Bruker DRX-600 spectrometer.The LC-MS base peak chromatogram of saponins from pure ethanol extract was dominated by 11 large and several small peaks but the UV chromatogram showed only two peaks, corresponding to the main neutral saponins found previously in Argania: arganine A and B. In 50% aqueous ethanol extracts, numerous other saponins were detected. Many of them were glucuronide oleanane-type triterpene carboxylic acid 3,28-O-bidesmosides (GOTCAB saponins). The assignments of (1) H- and (13) C-NMR spectra of the four most abundant GOTCAB saponins confirmed the MS results.Four GOTCAB saponins were structurally identified by NMR analysis in the 50% aqueous ethanol extract. Furthermore, LC-MS analyses showed the presence of at least 19 additional polar saponins in these kernel cakes.

Objective Polyphenols are chemicals derived from plants known to possess antioxidant and anti-inflammatory properties. High intake of fruit and vegetables is believed to be beneficial to human health. Various studies have suggested that dietary polyphenols may protect against cancer and cardiometabolic and neurodegenerative diseases. Nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) are neurotrophins that play key roles in brain cell development, growth, and survival. The aim of this study was to investigate whether or not administration of olive ( Olea europaea L.) polyphenols could have an effect on NGF and BDNF content and the expression of their receptors, TrkA and TrkB, respectively, in the mouse brain. Methods NGF and BDNF were measured by enzyme-linked immunosorbent assay. TrkA and TrkB were measured by Western blotting. Results We found NGF and BDNF elevation in the hippocampus and olfactory bulbs and a decrease in the frontal cortex and striatum. These data were associated with potentiated expression of TrkA and TrkB in the hippocampus and olfactory bulbs but no differences between groups in the striatum and frontal cortex. Polyphenols did not affect some behavioral mouse parameters associated with stressing situations. Conclusions Altogether, this study shows that olive polyphenols in the mouse may increase the levels of NGF and BDNF in crucial areas of the limbic system and olfactory bulbs, which play a key role in learning and memory processes and in the proliferation and migration of endogenous progenitor cells present in the rodent brain.

From the bark of Guazuma ulmifolia, a plant used as anti-inflammatory remedy, the flavanocoumarin epiphyllocoumarin (1) along with epiphyllocoumarin-[4β→8]-(-)-epicatechin (2) and epiphyllocoumarin-[4β→8]-(-)-epicatechin-[4β→8]-(-)-epicatechin (3), never reported before, have been isolated. The structures of the proantocyanidins 2-3 have been elucidated by extensive NMR and ESI-MS analysis. On the basis of the flavane nature of the isolated compounds and considering the strong anti-STAT1 activity reported for epigallocatechin 3-O-gallate (EGCG), the affinity of compounds 1-3 for STAT1 has been evaluated by Surface Plasmon Resonance. Compounds 1-2 showed affinity for STAT1 comparable to that exerted by EGCG. In order to confirm this result, molecular docking studies to evaluate the interactions of compounds 1-3, phyllocoumarin (4), the reference compound EGCG (5), and its related compound (+)-gallocatechin 3-O-gallate (6) with STAT1 have been carried out. Furthermore the ability of compounds 1-3 to inhibit STAT1 activation has been evaluated using a nuclear extract obtained from the human monocytic leukemia cell line TPH-1. Flavanocoumarins 1-2 inhibited STAT1-DNA binding.

Phytochemical investigation of the methanolic extract of Byrsonima crassifolia's bark led to the isolation of 8 known phenolic compounds 5-O-galloylquinic acid, 3-O-galloylquinic acid, 3,4-di-O-galloylquinic acid, 3,5-di-O-galloylquinic acid, 3,4,5-tri-O-galloylquinic acid, (+)-epicatechin-3-gallate along with (+)-catechin and (+)-epicatechin. Due to their biological value, in the present study, a high-performance liquid chromatography–tandem mass spectrometry (LC–MS/MS) method, working in multiple reaction monitoring (MRM) mode, has been developed to quantify these compounds. B. crassifolia bark resulted in a rich source of phenolic compounds and particularly of galloyl derivates. The proposed analytical method is promising to be applied to other galloyl derivatives to quantify these bioactive compounds in raw material and final products.

Liquid chromatography electrospray mass spectrometry (LC-ESI/MS) has been applied to the full characterization of saponins and phenolics in hydroalcoholic extracts of roots of liquorice (Glycyrrhiza glabra). Relative quantitative analyses of the samples with respect to the phenolic constituents and to a group of saponins related to glycyrrhizic acid were performed using LC-ESI/MS. For the saponin constituents, full scan LC-MS/MS fragmentation of the protonated (positive ion mode) or deprotonated (negative ion mode) molecular species generated diagnostic fragment ions that provided information concerning the triterpene skeleton and the number and nature of the substituents. On the basis of the specific fragmentation of glycyrrhizic acid, an LC-MS/MS method was developed in order to quantify the analyte in the liquorice root samples. Chinese G. glabra roots contained the highest levels of glycyrrhizic acid, followed by those from Italy (Calabria).

Acacia nilotica (L.) Del. syn is a species rich in polyphenolic constituents, in which catechins are hypothesized to possess antioxidant properties and to play a role in the anti-inflammatory activity of several plants. Due to the complexity of catechin derivatives, the investigation of this class of natural compounds has been limited by difficulties in their separation. In this paper, rationalization of the phenolics occurring in the 80% EtOH extract of Acacia nilotica pods, on the basis of ESI-MS and ESI-MS/MS profiles, has been proposed. Additionally, an LC-ESI-MS qualitative study has been performed by using a C18 polar endcapped stationary phase. The fragmentation pattern obtained evidenced the presence in A. nilotica pods of galloylated catechin- and gallocatechin derivatives along with galloylated glucose derivatives. The structures were confirmed by NMR, after isolation of the pure compounds. In addition, the radical scavenging activities of extracts and pure compounds were investigated, by using the TEAC assay. Furthermore quantitative analyses were performed by LC-ESI-MS/MS, confirming the interest of this species as a rich source of very strong antioxidant principles.

The major phytochemical constituents, namely, alkaloids, flavonoids and ellagic acid derivatives, of leaves of Camptotheca acuminata were identified using high performance liquid chromatography (HPLC) coupled with electrospray mass spectrometry (ESI-MS) in extracts of plants cultivated in Italy and collected at different growth stages. Alkaloids related to camptothecin were identified and quantified by HPLC coupled with ESI-tandem mass spectrometry (MS/MS) employing, respectively, an ion trap and a triple quadrupole mass analyser. The fragmentation patterns of alkaloids related to camptothecin were analysed and a specific Multiple Reaction Monitoring HPLC-MS/MS method was developed for the quantitative determination of these constituents. The described method provides high sensitivity and specificity for the characterisation and quantitative determination of the alkaloids in C. acuminata.

Six secondary metabolites from the methanolic extract of Sweetia panamensis (Fabaceae) bark were isolated and characterised. Along with the pyrones desmethylangonine β-d-O-glucopyranoside and desmethylangonine β-d-O-glucopyranosyl-(1→6)-O-β-d-glucopyranoside, already reported in this species, 5-O-caffeoylquinic acid (chlorogenic acid), 4-O-caffeoylquinic acid, 3-O-caffeoylquinic acid and the isoflavonoid 5-O-methylgenistein 7-O-β-d-glucopyranoside were isolated for the first time from S. panamensis. Additionally, an LC-ESI-MS qualitative analysis was performed and an ultra performance liquid chromatography (UPLC) method was developed and validated for the determination of these compounds. The UPLC method was applied to the quantitative analysis of plant samples. Pyrones and caffeoylquinic acids resulted to be the main compounds in the extract; in particular desmethylangonine β-d-O-glucopyranosyl-(1→6)-O-β-d-glucopyranoside was the most abundant compound.

Phytochemical investigation of the methanolic extract of Bursera simaruba bark led to the isolation of 11 compounds, including lignans yatein, beta-peltatin-O-beta-D-glucopyranoside, hinokinin and bursehernin, and three natural compounds namely 3,4-dimetoxyphenyl-1-O-beta-D-(6-sulpho)-glucopyranoside, 3,4,5-trimetoxyphenyl 1-O-beta-D-(6-sulpho)-glucopyranoside and 3,4-diidroxyphenylethanol-1-O-beta-D-(6-sulpho)-glucopyranoside. Their structures were established by NMR and ESI/MS experiments. Additionally, an LC-ESI/MS qualitative study on the phenolic compounds and an LC-ESI/MS/MS quantitative study on the lignans found in the methanolic extract of B. simaruba bark were performed to give value to the plant as source of these biological active compounds. Quantitative analyses results confirmed that compounds yatein, beta-peltatin-O-beta-D-glucopyranoside, hinokinin and bursehernin are major compounds in the bark and, in particular, beta-peltatin-O-beta-D-glucopyranoside appears to be the most abundant.

In Sudan, the roots of Hydnora johannis (Hydnoraceae) are traditionally used for the treatment of dysentery, diarrhea, cholera and swelling tonsillitis. The ethnomedicinal value of H. johannis was investigated through phytochemical study, in vitro antibacterial activity and preliminary cytotoxic tests. Determination of total phenols, flavonoids and proanthocyanidins was carried out using spectrometric methods. The antibacterial activity of the water and ethanolic extracts was determined using the microdilution method. Pure compounds were isolated from the ethyl acetate extract by chromatographic methods and their structures were established by spectroscopic methods. Cytotoxicity assay was performed against selected human mouth epidermoid carcinoma cell line (KB), and non-cancer human fetal lung cell line (MRC-5). Both water and ethanol (70%) extracts were found to contain the same amount of total phenols and proanthocyanidins, whereas the level of flavonoids was higher in the ethanol extract. The water extract was found to possess antibacterial activity against Enterococcus faecalis (MIC value of 16 μg/mL), Bacillus subtilis , B. cereus and Staphylococcus aureus (MIC values of 64 μg/mL) but not against bacteria mainly responsible for diarrhea. This leads to the suggestion that, the mode of action of water extract which is rich in tannins was not connected to their inhibition to the diarrhea bacteria but to their action on the digestive tract. Reduction in potency of the water and ethanol (70%) extracts when fractionated was observed. The ethyl acetate fraction obtained from fractionation of ethanol extract possessed only activity against the two strains of S. aureus with MIC values of 128 μg/mL. In addition, six compounds were isolated from the ethyl acetate extract as cirsiliol (3′,4′,5-trihydroxy-6-7-dimethoxy flavone) ( 1 ), trans 3′5-dihydroxy-4′7-dimethoxydihydroflavonol ( 2 ), oleic acid ( 3 ), vanillin (4-hydroxy-3-methoxybenzaldehyde) ( 4 ), protocatechuic acid (3,4 dihydroxy benzoic acid) ( 5 ) and dl catechin (trans (+) 2-(3,4-dihydroxyphenyl-3,4-dihydro-2H-1-benzopyran-3,5,7-triol)) ( 6 ). Four compounds from the ethyl acetate extract were also identified by GCMS as stigmasterol ( 7 ), oleic acid ( 3 ), myristic acid ( 8 ), and palmitic acid ( 9 ). Little cytotoxicity is reported against the cell lines used. Thus, the safety of this plant in the traditional medicine should be verified by much further testing, including in vivo experiments and clinical studies.

Vinca sardoa (Stern) Pignatti (Apocynaceae) is largely distributed in Sardinia where it is considered a typical endemism. Since the plant is used in traditional folk medicine, the aim of the present work was to identify and quantify the polyphenolic metabolites, due to the well-known importance of polyphenolic compounds, as well as to evaluate antioxidant activity in different parts of the plant. The compounds were identified and determined in the methanol extracts of leaves, flowers, and roots by developing different LC-MS/MS methods. The obtained data show that leaves possess the highest amount of polyphenols, in particular quinic acid (3401 mg/100 g), chlorogenic acid (1082 mg/100 g), caffeoylquinic acid isomer 1 (190 mg/100 g), and robinin (633 mg/100 g). Likewise, antioxidant tests showed that leaves possess the main radical scavenging activities in both ABTS (49.19 ± 3.41 μg/mL, 30.88 ± 3.04 μg/mL at time zero and after 50 min, respectively) and DPPH assays (223.97 ± 30.81 μg/mL, 109.52 ± 12.89 μg/mL at time zero and after 30 min, respectively). Taking into account that leaves differed most from flowers and roots in the content of caffeoylquinic acid and chlorogenic acid, of which antioxidant properties are widely recognized, it is reasonable to assume that these two compounds are involved in the differences described. The relationship between the high polyphenolic content and interesting antioxidant activities, justifies its use in ethnobotany and may be suggest a use of this specie, after removal of the alkaloid fraction, in the pharmaceutical, phytotherapy, and cosmetic industries.