RESUMO
A simple, rapid and selective high-performance thin-layer chromatographic (HPTLC) method has been developed and validated for simultaneous determination of three withanolides (withaferin A, withanone and withanolide A) and three phenolic acids (caffeic acid, ferulic acid and benzoic acid) from different parts (root, stem and leaf) of Withania somnifera and its two commercially available polyherbal formulations. The extraction efficiency of withanolides and phenolic acids were tested using two solvents, chloroform and methanol, respectively. HPTLC separation was performed on silica coated aluminium plates Si 60F254; using toluene, ethyl acetate and acetic acid (60:40:4). The samples were quantitated at 231â¯nm. The purity and identity of peaks of all the six analytes were confirmed by matching Rf values and UV-spectrum with authentic standards. The identity of three withanolides was further confirmed by positive ion electrospray ionization mass spectrometry (ESI-MS/MS) analyses. The developed method was validated for sensitivity, linearity, reproducibility, accuracy, the limit of detection (LOD) and limit of quantification (LOQ) following the guidelines of the International Conference on Harmonization (ICH). The method was found to be linear (râ¯>â¯0.99) in the range of 50-2000â¯ng/band for benzoic acid and 50-1000â¯ng/band for the other five studied metabolites. This simple and accurate HPTLC method provided enhanced resolution of studied analytes as compared to other phytoconstituents present in W. somnifera extracts. It has also been successfully applied in the analysis and quantification of two polyherbal formulations containing W. somnifera plant parts.
Assuntos
Cromatografia em Camada Fina/métodos , Hidroxibenzoatos/química , Extratos Vegetais/química , Withania/química , Vitanolídeos/química , Cromatografia em Camada Fina/economia , Espectrometria de Massas por Ionização por ElectrosprayRESUMO
The present study explores the preparation, characterization and the role of phenolic acid tethered fibrous protein in the management of induced oxidative stress studied under in vitro conditions. In brief, the biomaterial is prepared by engineering the fibrous protein with dihydroxy and trihydroxy phenolic acid moieties and subjected to characterization to ensure the tethering. The resultant biomaterial studied for its efficacy as a free radical scavenger using polymorphonuclear (PMN) cells with induced oxidative stress and also as an agent for cell migration using fibroblasts cells. Results revealed that induced oxidative stress in PMN cells after exposure to UVB radiation managed well with the prepared biomaterial by reducing the levels of superoxide anion, oxygen and hydroxyl radicals. Further, the protein and the phenolic acid interaction supports the cell migration as evidenced from the scratch assay. In conclusion, though phenolic acids are well known for their antimicrobial and antioxidant potential, indenting these acids directly to the wounds is not sensible, but tethering to protein explored the scavenging activity as expected. The present study infers that phenolic acid engineered protein has a significant role in managing the imbalance in the redox state prevailing in wounds and supports the healing at appreciable level.
Assuntos
Hidroxibenzoatos/química , Hidroxibenzoatos/farmacologia , Neutrófilos/efeitos dos fármacos , Neutrófilos/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Escleroproteínas/química , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Compostos de Bifenilo/metabolismo , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Espaço Extracelular/efeitos dos fármacos , Espaço Extracelular/metabolismo , Espaço Extracelular/efeitos da radiação , Radical Hidroxila/metabolismo , Metaloproteinases da Matriz/metabolismo , Neutrófilos/citologia , Neutrófilos/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Picratos/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxidos/metabolismo , Suínos , Raios UltravioletaRESUMO
The blood-brain barrier (BBB) is a major obstacle for drug delivery to the brain. Predicted, focal opening of the BBB through intra-arterial infusion of hyperosmolar mannitol is feasible, but there is a need to facilitate imaging techniques (e.g. MRI) to guide interventional procedures and assess the outcomes. Here, we show that salicylic acid analogues (SAA) can depict the brain territory supplied by the catheter and detect the BBB opening, through chemical exchange saturation transfer (CEST) MRI. Hyperosmolar SAA solutions themselves are also capable of opening the BBB, and, when multiple SAA agents were co-injected, their locoregional perfusion could be differentiated.
Assuntos
Barreira Hematoencefálica/diagnóstico por imagem , Meios de Contraste/química , Imagem por Ressonância Magnética Intervencionista/métodos , Imagem de Perfusão/métodos , Ácido Salicílico/química , Animais , Barreira Hematoencefálica/metabolismo , Encéfalo/irrigação sanguínea , Encéfalo/diagnóstico por imagem , Meios de Contraste/administração & dosagem , Hidroxibenzoatos/administração & dosagem , Hidroxibenzoatos/química , Infusões Intra-Arteriais , Concentração Osmolar , Perfusão , Ácidos Ftálicos/administração & dosagem , Ácidos Ftálicos/química , Ratos Sprague-Dawley , Ácido Salicílico/administração & dosagem , Salicilato de Sódio/administração & dosagem , Salicilato de Sódio/química , SoluçõesRESUMO
The antiproliferative effects of a purified white grape pomace extract (PWGPE), as well as of some phenolic standards on colon cancer cells were examined. The phenolic composition of the PWGPE was determined by rapid resolution liquid chromatography/mass spectrometry (RRLC/MS). The PWGPE had 92.6, 43.3 and 6.01mg/g of flavanols, flavonols and phenolic acids, respectively and, along with pure catechin, epicatechin, quercetin and gallic acid, they were all found capable of inhibiting cellular proliferation. PWGPE (100µg/ml) inhibited the proliferation of cells by 52.1% at 48h, whilst catechin, epicatechin, quercetin and gallic acid (60µg/ml) inhibited growth by 65.2%, 62.2%, 81.0% and 71.0%, respectively, at 72h. The PWGPE is an interesting source of phenolic compounds with antiproliferative properties, that could be of interest in the food and pharmaceutical industries.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Neoplasias Colorretais/prevenção & controle , Flavonoides/análise , Hidroxibenzoatos/química , Espectrometria de Massas/métodos , Extratos Vegetais/química , Vitis/química , Catequina/análise , Flavonóis/análise , Humanos , Quercetina/análiseRESUMO
Spent coffee is the main byproduct of the brewing process and a potential source of bioactive compounds, mainly phenolic acids easily extracted with water. Free and bound caffeoylquinic (3-CQA, 4-CQA, 5-CQA), dicaffeoylquinic (3,4-diCQA, 3,5-diCQA, 4,5-diCQA), caffeic, ferulic, p-coumaric, sinapic, and 4-hydroxybenzoic acids were measured by HPLC, after the application of three treatments (alkaline, acid, saline) to spent coffee extracts. Around 2-fold higher content of total phenolics has been estimated in comparison to free compounds. Phenolic compounds with one or more caffeic acid molecules were approximately 54% linked to macromolecules such as melanoidins, mainly by noncovalent interactions (up to 81% of bound phenolic compounds). The rest of the quantitated phenolic acids were mainly attached to other structures by covalent bonds (62-97% of total bound compounds). Alkaline hydrolysis and saline treatment were suitable to estimate total bound and ionically bound phenolic acids, respectively, whereas acid hydrolysis is an inadequate method to quantitate coffee phenolic acids.
Assuntos
Coffea/química , Fenóis/química , Extratos Vegetais/química , Sementes/química , Resíduos/análise , Antioxidantes/química , Antioxidantes/isolamento & purificação , Ácidos Cafeicos/química , Ácidos Cafeicos/isolamento & purificação , Ácido Clorogênico/química , Ácido Clorogênico/isolamento & purificação , Hidroxibenzoatos/química , Hidroxibenzoatos/isolamento & purificação , Fenóis/isolamento & purificação , Extratos Vegetais/isolamento & purificaçãoRESUMO
Persimmon juice is emerging in the global juice market as a new wholesome commercial juice that could effectively complement a healthy diet, given the epidemiological evidence linking a diet rich in fruits and vegetables with reduced incidences of chronic diseases. However, little data are available on the persimmon-juice composition or on the effect of the technological treatment employed for its production. The present work performs a complete qualitative analytical characterization through high-performance liquid chromatography coupled to electrospray time-of-flight mass spectrometry (HPLC-DAD-ESI-TOF/MS) of the diverse persimmon juices produced under different technologies in a pilot plant (clarification, astringency removal, flash vacuum expansion, centrifugation and pasteurization) in order to evaluate the effect of the different production procedures on the polar chemical profile of persimmon juice. Persimmon-juice extracts have been found to be a source of sugars, protein derivatives, organic acids, vitamins, and polyphenols, including simple polyphenols (phenolic acids and flavonoids) and polymerized flavan-3-ols. A marked influence of processing on the composition of the juices has been noticed. Extracts 3 and 7 (undergoing the combinations of clarification and centrifugation, and astringency removal, centrifugation and pasteurization, respectively) contained more polyphenols, which may help reduce risk of chronic diseases.
Assuntos
Bebidas/análise , Carboidratos/química , Cromatografia Líquida de Alta Pressão/métodos , Diospyros/química , Flavonoides/análise , Frutas/química , Hidroxibenzoatos/química , Polifenóis/análise , Carboidratos/análiseRESUMO
The biological properties of polyphenol (PP) depend on its bioaccessibility and bioavailability. Therefore, part of PP released from the food matrix in the gastrointestinal tract through enzymatic hydrolysis is at least partially absorbed. The aim of this study is to determine the bioaccessibility of PP associated with dietary fiber (DF) and the kinetics release of PP in mango (Mangifera indica L.) 'Ataulfo' by-products by an in vitro model. Soluble and insoluble DF values were 7.99 and 18.56% in the mango paste and 6.98 and 22.78% in the mango peel, respectively. PP associated with soluble and insoluble DF was 6.0 and 3.73 g GAE per 100 g in the paste and 4.72 and 4.50 g GAE per 100 g in the peel. The bioaccessibility of PP was 38.67% in the pulp paste and 40.53% in the peel. A kinetics study shows a release rate of 2.66 and 3.27 g PP min(-1) in the paste and peel, respectively. The antioxidant capacity of the paste increased as digestion reached a value of 2.87 mmol TE min(-1) at 180 min. The antioxidant capacity of the peel had its maximum (28.94 mmol TE min(-1)) between 90 and 120 min of digestion; it started with a value of 2.58 mmol TE min(-1), and thereafter increased to 4.20 mmol TE min(-1) at 180 min. The major PPs released during the digestion of paste were gallic and hydroxybenzoic acids, while in the peel, they were hydroxycinnamic and vanillic acids. It was concluded that these phenolic compounds are readily available for absorption in the small intestine and exert different potential health benefits.
Assuntos
Antioxidantes/metabolismo , Fibras na Dieta/metabolismo , Suplementos Nutricionais , Frutas/química , Mangifera/química , Modelos Biológicos , Polifenóis/metabolismo , Antioxidantes/química , Antioxidantes/economia , Ácidos Cumáricos/análise , Ácidos Cumáricos/química , Ácidos Cumáricos/economia , Ácidos Cumáricos/metabolismo , Fibras na Dieta/análise , Fibras na Dieta/economia , Suplementos Nutricionais/análise , Suplementos Nutricionais/economia , Digestão , Manipulação de Alimentos , Indústria de Processamento de Alimentos/economia , Frutas/economia , Ácido Gálico/análise , Ácido Gálico/química , Ácido Gálico/economia , Ácido Gálico/metabolismo , Humanos , Hidrólise , Hidroxibenzoatos/análise , Hidroxibenzoatos/química , Hidroxibenzoatos/economia , Hidroxibenzoatos/metabolismo , Resíduos Industriais/análise , Resíduos Industriais/economia , Absorção Intestinal , Cinética , México , Polifenóis/análise , Polifenóis/química , Polifenóis/economia , Solubilidade , Propriedades de Superfície , Ácido Vanílico/análise , Ácido Vanílico/química , Ácido Vanílico/economia , Ácido Vanílico/metabolismoRESUMO
BACKGROUND: Grapes are one of the world's staple fruit crops, with about 80% of the yield being utilised for winemaking. Since grape by-products still contain large amounts of secondary metabolites, uses other than as fertilisers might be appropriate. In this study, white grape pomace (WGP) was incorporated in wheat flour at levels of 10, 20 and 30% (w/w) to investigate its influence on rheological, nutraceutical, physical and sensory properties. RESULTS: Farinograph characteristics of dough with different levels of WGP showed a decrease in water absorption from 56.4% (0% WGP) to 45.9% (30% WGP). Addition of WGP reduced hardness and caused a deterioration in brightness and yellowness of all enriched samples. The smallest addition of WGP (10%) caused an approximately 88% increase in total dietary fibre content as compared with the control. The content of phenolic compounds increased from 0.11 mg g⻹ with 0% WGP to 1.07 mg g⻹ with 30% WGP. The most stable phenols were as follows: γ-resorcylic acid < gallic acid < tyrosol < catechin < isovanilic acid. An assay of radical-scavenging activity showed that WGP addition greatly enhanced the antioxidant properties of biscuits. Acceptable biscuits were obtained when incorporating 10% WGP. CONCLUSION: WGP might be utilised for the novel formulation of biscuits as an alternative source of dietary fibre and phenols.
Assuntos
Pão/análise , Fibras na Dieta/análise , Alimentos Fortificados/análise , Frutas/química , Resíduos Industriais/análise , Polifenóis/análise , Vitis/química , Antioxidantes/análise , Antioxidantes/química , Antioxidantes/economia , Pão/economia , Fenômenos Químicos , Fibras na Dieta/economia , Farinha/análise , Farinha/economia , Alimentos Fortificados/economia , Indústria de Processamento de Alimentos/economia , Humanos , Hidroxibenzoatos/análise , Hidroxibenzoatos/química , Hidroxibenzoatos/economia , Resíduos Industriais/economia , Fenômenos Mecânicos , Valor Nutritivo , Pigmentação , Polônia , Polifenóis/química , Polifenóis/economia , Análise de Componente Principal , Sensação , Água/análiseRESUMO
Chlorophyll fluorescence (ChlF) excitation spectra were measured to assess the UV-sunscreen compounds accumulated in fully expanded leaves of three woody species belonging to different chemotaxons, (i.e. Morus nigra L., Prunus mahaleb L. and Lagerstroemia indica L.), grown in different light microclimates. The logarithm of the ratio of ChlF excitation spectra (logFER) between two leaves acclimated to different light microclimates was used to assess the difference in epidermal absorbance (EAbs). EAbs increased with increasing solar irradiance intercepted for the three species. This epidermal localisation of UV-absorbers was confirmed by the removal of the epidermis. It was possible to simulate EAbs as a linear combination of major phenolic compounds (Phen) identified in leaf methanol extracts by HPLC-DAD. Under UV-free radiation conditions, shaded leaves of M. nigra accumulated chlorogenic acid. Hydroxybenzoic acid (HBA) derivatives and hydroxycinnamic acid (HCA) derivatives greatly increased with increasing PAR irradiance under the low UV-B conditions found in the greenhouse. These traits were also observed for the HCA of the two other species. Flavonoid (FLAV) accumulation started under low UV-A irradiance, and became maximal in the adaxial epidermis of sun-exposed leaves outdoors. A decrease in the amount of HCA was observed concomitantly to the intense accumulation of FLAV for both leaf sides of the three species. Judging from the logFER, under low UV-B conditions, larger amounts of HCA are present in the epidermis in comparison to FLAV for the three species. Upon transition from the greenhouse to full sunlight outdoors, there was a decrease in leaf-soluble HCA that paralleled FLAV accumulation in reaction to increasing solar UV-B radiation in the three species. In M. nigra, that contains large amounts of HCA, the logFER analysis showed that this decrease occurred in the adaxial epidermis, whereas the abaxial epidermis, which is protected from direct UV-B radiation, continued to accumulate large amounts of HCA.
Assuntos
Clorofila/efeitos da radiação , Lagerstroemia/metabolismo , Morus/metabolismo , Fenóis/metabolismo , Epiderme Vegetal/efeitos da radiação , Folhas de Planta/efeitos da radiação , Prunus/metabolismo , Aclimatação , Ácido Clorogênico/química , Ácido Clorogênico/metabolismo , Clorofila/metabolismo , Cromatografia Líquida de Alta Pressão , Ácidos Cumáricos/química , Ácidos Cumáricos/metabolismo , Flavonoides/química , Flavonoides/metabolismo , Efeito Estufa , Hidroxibenzoatos/química , Hidroxibenzoatos/metabolismo , Luz , Fenóis/análise , Epiderme Vegetal/metabolismo , Folhas de Planta/metabolismo , Especificidade da Espécie , Espectrometria de Fluorescência , Fatores de TempoRESUMO
We have developed a new method for the three-dimensional modeling of extended X-ray absorption fine structure (EXAFS) spectra which enables the extraction of the local structure of aqueous metal complexes from spectral mixtures of several components. The new method combines two techniques: Monte Carlo simulation and target transformation factor analysis (TFA). Monte Carlo simulation is used to create random arrangements between the X-ray absorbing metal ion and the ligand atoms, and to calculate the theoretical EXAFS spectrum of each arrangement. The theoretical EXAFS spectrum is then introduced as test spectrum in the TFA procedure, to test whether or not the test spectrum is likely to be a component of the spectral mixtures. This coupled procedure is repeated until the error in the test spectrum is minimized. The new method can thus be used to isolate and refine the structure of complexes from spectral mixtures and to determine their relative concentrations, solely on the basis of an estimate of a ligand structure. The performance of the proposed method is validated using uranium Liii-edge EXAFS spectra of binary mixtures of two uranium(VI) 3,4-dihydroxybenzoic acid complexes.
Assuntos
Absorciometria de Fóton/métodos , Técnicas Biossensoriais , Metais/análise , Compostos Organometálicos/análise , Absorção , Hidroxibenzoatos/análise , Hidroxibenzoatos/química , Ligantes , Metais/química , Modelos Químicos , Método de Monte Carlo , Compostos Organometálicos/química , Compostos de Urânio/análise , Compostos de Urânio/químicaRESUMO
Electron spin resonance (ESR) spectroscopy has been used to assess the antioxidant capacity of eight Scotch whiskeys by measuring the extent by which the original spirits, or pyridine solutions of their residues, reduced Fremy's radical or galvinoxyl radical. All whiskeys displayed antioxidant activity greater than that of a 0.2 mM solution of Trolox in the Fremy's assay and of a 0.1 mM solution of quercetin in the galvinoxyl assay. The relative antioxidant capacities determined according to the two assays were highly correlated and strongly related to the total phenol content as determined by using the Folin-Ciocalteu method. Activity was a consequence of maturation in oak casks with the "newmake" spirit showing no effect. Of 10 aromatic constituents analyzed, activity was most strongly correlated with ellagic acid and gallic acid in both assays. The reductive capacities of four major phenolics were determined, which, in summation, accounted for 31-53% of the total antioxidant activity of the whiskeys. There was no evidence for synergistic interaction between the phenols investigated.