RESUMO
A new pulse sequence for obtaining 19 F detected DOSY (diffusion ordered spectroscopy) spectra of fluorinated molecules is presented and used to study fluoropolymers based on vinylidene fluoride and chlorotrifluoroethylene. The performance of 19 F DOSY NMR experiments (and in general any type of NMR experiment) on fluoropolymers creates some unique complications that very often prevent detection of important signals. Factors that create these complications include: (1) the presence of many scalar couplings among 1 H, 19 F and 13 C; (2) the large magnitudes of many 19 F homonuclear couplings (especially 2 JFF ); (3) the large 19 F chemical shift range; and (4) the low solubility of these materials (which requires that experiments be performed at high temperatures). A systematic study of the various methods for collecting DOSY NMR data, and the adaptation of these methods to obtain 19 F detected DOSY data, has been performed using a mixture of low molecular weight, fluorinated model compounds. The best pulse sequences and optimal experimental conditions have been determined for obtaining 19 F DOSY spectra. The optimum pulse sequences for acquiring 19 F DOSY NMR data have been determined for various circumstances taking into account the spectral dispersion, number and magnitude of couplings present, and experimental temperature. Pulse sequences and experimental parameters for optimizing these experiments for the study of fluoropolymers have been studied. Copyright © 2016 John Wiley & Sons, Ltd.
RESUMO
Anthocyanin constituents in black raspberries (Rubus occidentalis L.) were investigated by HPLC-DAD, and their involvement as potent, significant antioxidants in black raspberries was demonstrated by three common antioxidant assays (FRAP, DPPH, ABTS) in this study. Five anthocyanins were present in black raspberries: cyanidin 3-sambubioside, cyanidin 3-glucoside, cyanidin 3-xylosylrutinoside, cyanidin 3-rutinoside, and pelargonidin 3-rutinoside. Their identities and structures, with particular emphasis on cyanidin 3-xylosylrutinoside, were confirmed by NMR spectroscopy. Two of these anthocyanins, cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside, predominated, comprising 24-40 and 49-58%, respectively, of the total anthocyanins in black raspberries. On the basis of both potency and concentration, cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside were found to be the significant contributors to the antioxidant systems of black raspberries. These findings indicate that these two anthocyanin compounds may function as the primary phenolic antioxidants in black raspberries. These two compounds exhibit potential biological activities that may be exploited in conjunction with other naturally occurring bioactive compounds in black raspberry fruit-based products used in clinical trials for the treatment of various types of cancer.
Assuntos
Antocianinas/análise , Antioxidantes/análise , Frutas/química , Rosaceae/química , Antocianinas/química , Cromatografia Líquida de Alta Pressão , Dieta , Espectroscopia de Ressonância Magnética , Estrutura MolecularRESUMO
Black raspberry (Rubus occidentalis L.) (BR) fruit extracts with differing compound profiles have shown variable antiproliferative activities against HT-29 colon cancer cell lines. This study used partial least-squares (PLS) regression analysis to develop a high-resolution (1)H NMR-based multivariate statistical model for discerning the biological activity of BR constituents. This model identified specific bioactive compounds and ascertained their relative contribution against cancer cell proliferation. Cyanidin 3-rutinoside and cyanidin 3-xylosylrutinoside were the predominant contributors to the extract bioactivity, but salicylic acid derivatives (e.g., salicylic acid glucosyl ester), quercetin 3-glucoside, quercetin 3-rutinoside, p-coumaric acid, epicatechin, methyl ellagic acid derivatives (e.g., methyl ellagic acetyl pentose), and citric acid derivatives also contributed significantly to the antiproliferative activity of the berry extracts. This approach enabled the identification of new bioactive components in BR fruits and demonstrates the utility of the method for assessing chemopreventive compounds in foods and food products.
Assuntos
Frutas/química , Espectroscopia de Ressonância Magnética/métodos , Metabolômica/métodos , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Rosaceae/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , HumanosRESUMO
Tacticity has an enormous influence on the physical and chemical properties of polymers. There is considerable work using 1D NMR and empirical rules to study the stereosequences in polymers. This work shows that 1H/13C/19F 3D NMR experiments can provide superior resolution and atomic connectivity information, so that unambiguous resonance assignments can be made for poly(vinyl fluoride) (PVF). Compared to prior work on 3D NMR studies of stereosequence effects in fluoropolymers, the 3D NMR pulse sequence used in this work is based on single quantum coherence transfer, which eliminates the complicated splitting patterns resulting from evolution of multiple-quantum coherence. In addition, selective excitation of the 19F nuclei of interest significantly reduces the folding of peaks from other spectral regions. This greatly simplifies the spectra and makes the assignment of resonances much easier. Based on these results, it is possible to assign the 19F resonances to the pentad level. For example, consider the resonances of mm-centered sequences, which are not well resolved in 19F-19F COSY 2D NMR spectrum. 1H/13C/19F 3D NMR data provide clear evidence for all of the three pentad structures: mmmm, mmmr, and rmmr. Examples showing the resonance assignments of head-to-tail sequences are presented.
RESUMO
Buffaloberry (Shepherdia argentea [Pursh] Nutt.) has historically been used as an important food source by North American indigenous peoples, but its commercial production has been limited. These plants produce fruits rich in carotenoid and phenolic antioxidants, which may have health benefits that may make buffaloberry commercially valuable. Here, we examined these constituents in the fruit of 7 Dakota-grown buffaloberry selections. Primary carotenoids were determined by liquid chromatography-mass spectral analysis and by nuclear magnetic resonance spectroscopy to be lycopene (0.27 ± 0.02 g/kg FW) and methyl apo-6'-lycopenoate (MA6L; 0.32 ± 0.03/kg FW). MA6L comprised the greatest proportion (55%) of carotenoid antioxidants, but its role in human nutrition is still to be evaluated. The fruit contained high total phenolics concentrations (9.06 ± 0.71 g gallic acid equivalents/kg FW). Hydrophilic antioxidant capacity among the 7 selections averaged 49.0 ± 6.6 mmol trolox equivalents/kg FW, respectively, as measured by ferric reducing ability of plasma assay. The soluble solids and titratable acids concentrations were 21% and 2.2%, respectively. This species is adapted to poor soils and can tolerate drier climates. In the Dakotas, buffaloberry flourishes on the American Indian Tribal Reservations, yielding copious amounts of health-beneficial fruit for fresh and processing markets, making it a potentially valuable new crop for marginal lands.
Assuntos
Antioxidantes/análise , Carotenoides/análise , Elaeagnaceae/química , Frutas/química , Cromatografia Líquida de Alta Pressão , Licopeno , Espectroscopia de Ressonância Magnética , North Dakota , South Dakota , Espectrometria de Massas em TandemRESUMO
Nonanthocyanin secondary metabolites potentially contributing to the antiproliferative bioactivity of black raspberry ( Rubus occidentalis L.) fruits were extracted in ethyl acetate and isolated by semipreparative and analytical HPLC and analyzed by NMR, HPLC-ESI-MS, and ESI-MS/MS techniques. Here we present complete and partial structures of a variety of the chemical entities such as quercetin 3-glucoside, quercetin 3-rutinoside, myricetin glucoside, dihydrokaempferol glucoside, benzoic acid ß-d-glucopyranosyl ester, 3,4-dihydroxybenzoic acid, epicatechin, caffeic acid, p-coumaric acid, p-coumaryl glucoside, p-coumaryl sugar ester, ellagic acid, methyl ellagic acid acetylpentose, methyl ellagic acid valerylpentose, trans-piceid, phloretin glucoside (phloridzin), dihydrosinapic acid, salicylic acid ß-d-glucopyranosyl ester, a salicylic acid derivative without attached sugar, p-alkylphenyl glucoside, and a citric acid derivative. To our knowledge, 15 of these compounds were not previously reported in black raspberry fruits.
Assuntos
Frutas/química , Extratos Vegetais/química , Rosaceae/química , Cromatografia Líquida de Alta Pressão , Frutas/metabolismo , Espectroscopia de Ressonância Magnética , Extratos Vegetais/metabolismo , Rosaceae/metabolismo , Metabolismo Secundário , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em TandemRESUMO
A process was developed to ascertain the bioactive components of black raspberry (Rubus occidentalis L.) fruit extracts by relating chemical constituents determined by high-field nuclear magnetic resonance (NMR) spectroscopy to biological responses using partial least-squares regression analysis. To validate our approach, we outlined relationships between phenolic signals in NMR spectra and chemical data for total monomeric anthocyanin (TMA) content and antioxidant capacity by the ferric-reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assays. Anthocyanins, cyanidin 3-O-rutinoside (Cy 3-rut), cyanidin 3-O-(2(G))-xylosylrutinoside (Cy 3-xylrut), and cyanidin 3-O-glucoside (Cy 3-glc), were significant contributors to the variability in assay results, with the two most important NMR bins corresponding to the methyl peaks in Cy 3-rut (6''') and/or Cy 3-xylrut (6(IV)). Many statistically important bins were common among assay models, but differences in structure-activity relationships resulted in changes in bin ranking. The specificity of these results supported the application of the process to investigate relationships among health-beneficial natural products and potential biological activity.