RESUMEN
The metabolite profile of fresh Goji berries from two cultivars, namely Big Lifeberry (BL) and Sweet Lifeberry (SL), grown in the Lazio region (Central Italy) and harvested at two different periods, August and October, corresponding at the beginning and the end of the maturation, was characterized by means of nuclear magnetic resonance (NMR) and electrospray ionization Fourier transform ion cyclotron resonance (ESI FT-ICR MS) methodologies. Several classes of compounds such as sugars, amino acids, organic acids, fatty acids, polyphenols, and terpenes were identified and quantified in hydroalcoholic and organic Bligh-Dyer extracts. Sweet Lifeberry extracts were characterized by a higher content of sucrose with respect to the Big Lifeberry ones and high levels of amino acids (glycine, betaine, proline) were observed in SL berries harvested in October. Spectrophotometric analysis of chlorophylls and total carotenoids was also carried out, showing a decrease of carotenoids during the time. These results can be useful not only to valorize local products but also to suggest the best harvesting period to obtain a product with a chemical composition suitable for specific industrial use. Finally, preliminary studies regarding both the chemical characterization of Goji leaves generally considered a waste product, and the biological activity of Big Lifeberry berries extracts was also investigated. Goji leaves showed a chemical profile rich in healthy compounds (polyphenols, flavonoids, etc.) confirming their promising use in the supplements/nutraceutical/cosmetic field. MG63 cells treated with Big Lifeberry berries extracts showed a decrease of iNOS, COX-2, IL-6, and IL-8 expression indicating their significant biological activity.
Asunto(s)
Antioxidantes/química , Lycium/química , Extractos Vegetales/química , Carotenoides/química , Ácidos Grasos/química , Frutas , Humanos , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Metabolómica , Polifenoles/químicaRESUMEN
Many plants of the genus Allium are widely cultivated and consumed for their nutraceutical and health-enhancing bioactive components effective in many metabolic and infectious diseases. In particular, Allium sativum L. (garlic), the most economically important Allium species, is known to present volatile, comparatively polar sulfur-containing compounds responsible for both the typical garlic aroma and antimicrobial property. More recently, the (moderately) polar portion of garlic metabolome, rich of polyphenols and amino acids, is gaining increasing interest as a source of antioxidants and primary nutrients. In this study, we have explored the chemical diversity of eight different hydroalcoholic extracts obtained by microwave-assisted extraction of white and red crop A. sativum and wild Allium triquetrum, Allium roseum, and Allium ampeloprasum, all originating from the Mediterranean Basin. The aim is to appraise their potential dietetic and healing value through an in-depth chemical characterization and contribute to preserve and exploit natural resources. The multimethodological method applied here is based on an untargeted metabolic profiling by means of high-resolution electrospray ionization Fourier-transform ion cyclotron resonance (ESI FT-ICR) mass spectrometry. More than 850 by ESI(+) and 450 by ESI(-) putative metabolites have been annotated covering all main classes of primary and secondary metabolites, including amino acids, alkaloids, organic and fatty acids, nucleotides, vitamins, organosulfur compounds, and flavonoids. The pigment and polyphenol components have been separated and quantified by a targeted chromatographic high-performance liquid chromatography-photodiode array detector (HPLC-PDA) and CIEL*a*b* colorimetric assay, showing characteristic yellow and red components in each extract, related to a different milieu of anthocyanins and flavonoids as assigned by high-resolution mass spectrometry (MS).
Asunto(s)
Ajo/química , Metaboloma , Alcaloides/análisis , Aminoácidos/análisis , Cromatografía Líquida de Alta Presión/métodos , Ácidos Grasos/análisis , Flavonoides/análisis , Metabolómica/métodos , Polifenoles/análisis , Espectrometría de Masa por Ionización de Electrospray/métodos , Vitaminas/análisisRESUMEN
Celery is a widely used vegetable known for its peculiar sensorial and nutritional properties. Here, the white celery (Apium graveolens L.) "sedano bianco di Sperlonga" PGI ecotype was investigated to obtain the metabolic profile of its edible parts (blade leaves and petioles) also related to quality, freshness and biological properties. A multi-methodological approach, including NMR, MS, HPLC-PDA, GC-MS and spectrophotometric analyses, was proposed to analyse celery extracts. Sugars, polyalcohols, amino acids, organic acids, phenols, sterols, fatty acids, phthalides, chlorophylls, tannins and flavonoids were detected in different concentrations in blade leaf and petiole extracts, indicating celery parts as nutraceutical sources. The presence of some phenols in celery extracts was here reported for the first time. Low contents of biogenic amines and mycotoxins confirmed celery quality and freshness. Regarding the biological properties, ethanolic celery extracts inhibited the oxidative-mediated DNA damage induced by tert-butylhydroperoxide and scavenged DPPH and ABTS radicals.
Asunto(s)
Apium/química , Fitoquímicos/análisis , Apium/metabolismo , Aminas Biogénicas/análisis , Cromatografía Líquida de Alta Presión , Ecotipo , Flavonoides/análisis , Micotoxinas/análisis , Fenoles/análisis , Extractos Vegetales/química , Hojas de la Planta/química , Hojas de la Planta/metabolismoRESUMEN
Many essential oils (EOs) are screened as potential sources of antimicrobial compounds. EOs from the genus Satureja have recognized biological properties, including analgesic, anti-inflammatory, immunomodulatory, anticancer, and antimicrobial activity. This study aimed to obtain a metabolite profile of commercial essential oil of S. montana L. (SEO) and to evaluate its antimicrobial properties, both alone and combined with gentamicin towards Gram-negative and Gram-positive bacterial strains. Untargeted analyses based on direct infusion Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and on GC-MS have provided a high metabolome coverage, allowing to identify carvacrol, cymene and thymol as the major components of commercial SEO. SEO exerted an antimicrobial activity and induced a synergistic interaction with gentamicin against both reference and clinical bacterial strains. A significant reduction of Escherichia coli, Staphylococcus aureus and Listeria monocytogenes biofilm formation was induced by SEO. As a result of SEO treatment, clear morphological bacterial alterations were visualized by scanning electron microscopy: L. monocytogenes and S. aureus showed malformed cell surface or broken cells with pores formation, whereas E. coli displayed collapsed cell surface. These results encourage further studies about bactericidal and antibiotic synergistic effect of SEO for combined therapy in clinical setting as well as in agricultural systems.
Asunto(s)
Antiinfecciosos/farmacología , Gentamicinas/farmacología , Aceites Volátiles/farmacología , Extractos Vegetales/farmacología , Aceites de Plantas/farmacología , Satureja/química , Biopelículas/efectos de los fármacos , Línea Celular , Supervivencia Celular/efectos de los fármacos , Cimenos , Combinación de Medicamentos , Sinergismo Farmacológico , Cromatografía de Gases y Espectrometría de Masas , Bacterias Gramnegativas/citología , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/citología , Bacterias Grampositivas/efectos de los fármacos , Humanos , Pruebas de Sensibilidad Microbiana , Monoterpenos/aislamiento & purificación , Monoterpenos/farmacología , Aceites Volátiles/química , Aceites de Plantas/química , Timol/aislamiento & purificación , Timol/farmacologíaRESUMEN
A multi-methodological approach was applied to study red sweet peppers (Capsicum annuum L.) ecotype "Cornetto di Pontecorvo" grown in a greenhouse or in open field. This approach includes morphological analysis, chemical composition determination, and biological activity evaluation of different extracts from pepper fruits. Untargeted analyses, namely NMR spectroscopy and mass spectrometry, allowed the comprehensive pepper metabolite profile of pepper pulp, peel and seeds hydroalcoholic and organic extracts to be determined, showing the presence of sugars, organic acids, amino acids and other secondary metabolites. Targeted analyses, such as HPLC-PDA, HPLC-TLC and spectrophotometric analyses allowed polyphenols, tannins, flavonoids and pigments content to be determined. Samples quality and freshness were verified by the low content of biogenic amines and mycotoxins, as determined using HPLC-FLD and HPLC-MS, respectively. Preliminary biological results demonstrated the ability of the organic extracts to inhibit α-amylase, a key enzyme in the control of glucose metabolism.
Asunto(s)
Capsicum/química , Análisis de los Alimentos/métodos , Extractos Vegetales/farmacología , Antifúngicos/farmacología , Aminas Biogénicas/análisis , Capsicum/metabolismo , Cromatografía Líquida de Alta Presión , Flavonoides/análisis , Calidad de los Alimentos , Frutas/química , Espectroscopía de Resonancia Magnética , Espectrometría de Masas , Micotoxinas/análisis , Polifenoles/análisis , Semillas/química , Taninos/análisis , alfa-Amilasas/antagonistas & inhibidoresRESUMEN
Coriander (Coriandruim sativum L., Apiaceae) is known for its antimicrobial activity and the aim of this study was to investigate the effect of its essential oil (CDO) against multidrug resistant uropathogenic Escherichia coli (UPEC). CDO was able to inhibit the growth of UPEC strains and propidium iodide uptake, - and electron microscopy examination suggested that bacterial structural modifications occurred. The presence of CDO reduced the MIC of gentamicin. E.coli adhesion efficiency on cell monolayers and abiotic surfaces was not affected by subMIC oil concentrations; furthermore, CDO showed cytotoxic activity towards the HEp-2 tumor cell line. These findings contribute to the knowledge about essential oils as sources of potential antimicrobial agents against uropathogenic E. coli and encourage further investigations.
Asunto(s)
Antibacterianos/farmacología , Coriandrum/química , Aceites Volátiles/farmacología , Aceites de Plantas/farmacología , Escherichia coli Uropatógena/efectos de los fármacos , Antibacterianos/química , Farmacorresistencia Bacteriana Múltiple , Infecciones por Escherichia coli/microbiología , Humanos , Pruebas de Sensibilidad Microbiana , Aceites Volátiles/química , Escherichia coli Uropatógena/crecimiento & desarrolloRESUMEN
The iron(III) protoporphyrin IX complex with imidazole, a biologically relevant ligand, occupying an axial position, has been studied by infrared multiple photon dissociation (IRMPD) spectroscopy. The complex has been delivered in gas-phase by electrospray ionization (ESI), mass selected in an ion trap, and assayed by IRMPD spectroscopy in two complementary frequency regions. The fingerprint range (900-1900 cm(-1)) has been scanned using the Orsay free-electron laser beamline (CLIO), while the X-H (X = C,N,O) stretching region (3000-3600 cm(-1)) has been inspected using a tabletop IR optical parametric oscillator/amplifier (OPO/OPA) laser source. DFT calculations have been performed to obtain a comprehensive pattern of the various potential conformers yielding optimized geometries, relative thermodynamic parameters, and respective IR spectra. The comparison between the IR spectra for representative conformers and the experimental IRMPD features suggests the coexistence of two families of conformers involving different degrees of folding and hydrogen bonding between the two propionic acid functionalities on the periphery of the protoporphyrin IX macrocycle in a ratio depending on environmental conditions such as ESI solvent and temperature. The observed conformational variability of the porphyrin substituents in the naked heme-imidazole complex is consistent with the fine-tuning of the reactivity properties of this important prosthetic group by the specific surroundings in the protein core.
Asunto(s)
Hemo/química , Compuestos Férricos/química , Imidazoles/química , Iones/química , Propionatos/química , Espectrofotometría Infrarroja , Esterol 14-Desmetilasa/química , TermodinámicaRESUMEN
The interest in the radical cations of amino acids is twofold. On the one hand, these species are relevant in enzymatic catalysis and in oxidative damage of proteins. On the other hand, as constituents of peptides and proteins, they aid the mass spectrometric characterization of these biomolecules, yielding diagnostic fragmentation patterns and providing complementary information with respect to the one obtained from even electron ions. The cysteine radical cation has been obtained by S-NO bond cleavage of protonated S-nitrosocysteine and thoroughly characterized by IRMPD spectroscopy, both in the 1000-2000 cm(-1) range (the highly structurally diagnostic, so-called 'fingerprint' range) and in the 2900-3700 cm(-1) spectral range, encompassing O-H and N-H stretching vibrations. In this way the distonic structure in which the charge is on the NH(3) group and the spin is on the sulfur atom is unambiguously demonstrated. This tautomer is a local minimum on the potential energy surface, at 29.7 kJ mol(-1) with respect to the most stable tautomer, a captodative structure allowing extensive delocalization of charge and spin.
Asunto(s)
Cationes/química , Cisteína/química , Gases/química , Cisteína/análogos & derivados , S-Nitrosotioles/química , Espectrofotometría Infrarroja , TermodinámicaRESUMEN
High-resolution Fourier transform ion cyclotron resonance mass spectrometry is employed to gain thorough kinetics and thermodynamics information on the reaction of free and ligated heme-type ions with selected ligands, with the aim of obtaining an insight into the coordination environment of the prosthetic group in a variety of biomolecular ions. Adopting a stepwise approach towards systems of increasing complexity, we examined the reactivity of free gaseous iron(III) protoporphyrin IX ions, Fe(III)-heme(+), of the charged species from microperoxidase-11 (MP11) (covalently peptide bound heme), and of the multiply charged ions from heme proteins, namely, cytochrome c (cyt c) and myoglobin (examples of noncovalently protein bound hemes). Among an array of test compounds allowed to react with Fe(III)-heme(+), OP(OMe)(3) and P(OMe)(3) proved to be similarly efficient ligands in the first addition step, yet displayed markedly distinct reactivity towards heme iron already engaged in axial coordination. The ease with which P(OMe)(3) acts as a second axial ligand is exploited to probe structural and conformational features of biomolecular ions. In this way, circumstantial evidence is gained of a folded conformation of +2 charge state ions from MP11 and an elongated one for the +3 charge state ions. Similarly, both the general reaction pattern and detailed kinetics and thermodynamics data point to a regiospecific addition reaction of P(OMe)(3) directed at the heme iron within multiply charged ions from cyt c. This unprecedented example of ion-molecule reaction which specifically involves a prosthetic group belonging to protein ions stands in contrast to the multiple, nonspecific interactions established by OP(OMe)(3) molecules with the protonated sites of multiply charged cyt c and apomyoglobin ions. This finding may develop and provide sensitive probes of the structure and bonding features of protein ions in the gas phase.