Comparison of the metabolic profile of pecan nuts cultivars [Carya illinoinensis (Wangenh.) K. Koch] by NMR spectroscopy.

Pecan nuts (Carya illinoinensis (Wangenh.) K. Koch) contain the highest number of phytochemicals of all nuts, are a natural source of unsaturated fatty acids and other nutrients and can be considered an important addition to the Mediterranean diet al.though several studies have been carried out on pecans, employing several analytical techniques, no systematic study of the metabolic profile is available in literature. In this study, the metabolic profile of pecan nuts of three different cultivars was analysed by Nuclear Magnetic Resonance Spectroscopy. The cultivars compared were Wichita, Stuart, and Sioux, all grown in Italy in the same pedoclimatic conditions. 31 metabolites were identified and 28 were quantified and the three species were differentiated based on multivariate PCA analysis. The differences among them, and the levels of scutellarein and GABA, in particular, were attributed to the adaptation of the plants to the climate in their original areas.

Time Domain NMR Approach in the Chemical and Physical Characterization of Hazelnuts (Corylus avellana L.).

'Tonda Gentile Romana' and 'Tonda di Giffoni' (Corylus avellana L.) are two Italian hazelnut cultivars, recognized under the quality labels "Protected Designation of Origin" (PDO) and "Protected Geographical Indication" (PGI), respectively. Hazelnut seeds are characterized by a complex microstructure and the presence of different physical compartments. This peculiarity has been studied and evidenced by Time Domain (TD) Nuclear Magnetic Resonance (NMR) experiments. This technique allowed the assessment of the presence of different diffusion compartments, or domains, by evaluating the distribution of the spin-spin relaxation time (T2).The aim of this research was to develop a method based on 1H NMR relaxometry to study the mobility in fresh hazelnut seeds ('Tonda di Giffoni' and 'Tonda Gentile Romana'), in order to determine differences in seed structure and matrix mobility between the two cultivars. TD-NMR measurements were performed from 8 to 55 °C in order to mimic post-harvest processing as well the microscopic textural properties of hazelnut. The Carr-Purcell-Meiboom-Gill (CPMG) experiments showed five components for 'Tonda Gentile Romana' and four components for 'Tonda di Giffoni' relaxation times. The two slowest components of relaxation (T2,a about 30-40% of the NMR signal, and T2,b about 50% of the NMR signal) were attributed to the protons of the lipid molecules organized in the organelles (oleosomes), both for the 'Tonda Gentile Romana' and for the 'Tonda di Giffoni' samples. The component of relaxation T2,c was assigned to cytoplasmic water molecules, and showed a T2 value dominated by diffusive exchange with a reduced value compared to that of pure water at the same temperature. This can be attributed to the water molecules affected by the relaxation effect of the cell walls. The experiments carried out as a function of temperature showed, for 'Tonda Gentile Romana', an unexpected trend between 30 and 45 °C, indicating a phase transition in its oil component. This study provides information that could be used to strengthen the specifications underlying the definitions of "Protected Designation of Origin" (PDO) and "Protected Geographical Indication" (PGI).

Efficacy of sodium hypochlorite in the degradation antineoplastic drugs by NMR spectroscopy.

SUMMARY: Antineoplastic drugs are used to treat cancer, having their therapeutic effect by inhibiting the cell division process. Although cancer cells, due to their rapid growth, are more sensitive to the toxic effects of chemotherapeutic agents, healthy cells and tissues may also be damaged. Many studies show acute and chronic toxicity both in patients treated with chemotherapy and in exposed workers. In fact, exposure to these substances can also be linked to the formation of different types of secondary tumors. The International Agency on Research on Cancer (IARC) included some antineplastic drugs in Group 1 (carcinogenic to humans), in Group 2A (probable carcinogens for In recent years, many studies have evidenced the presence of antineoplastic drug contamination on work surfaces, materials and floors and based on these observations, international and national guidelines have been published to limit occupational exposure, with particular attention to procedures post-preparation of chemotherapy to limit as much as possible the accumulation of contaminated residues. The aim of the following study is to determine the effectiveness of the degradation of four antineoplastic drugs: 5-fluorouracil, azacitidine, cytarabine and irinotecan using a low concentration of sodium hypochlorite solution (0.115%). The analytical platform used to monitor the degradation course of the substances under examination was hydrogen nuclear magnetic spectroscopy (1H NMR). In the same experimental conditions the effectiveness of the degradation of the same antineoplastic drugs with a 99.9% ethanol solution was also evaluated. The study showed that the best degradation efficiency (> 90% ) is obtained with the hypochlorite solution after 15 minutes.

NMR-Based Metabolomic Comparison of Brassica oleracea (Var. italica): Organic and Conventional Farming.

Brassicaceae family provides several crops which are worldwide known for their interesting phytochemical profiles, especially in terms of content of glucosinolates. These secondary metabolites show several beneficial effects toward consumers' health, and several studies have been conducted to identify cultivation factors affecting their content in crops. One of the agronomic practices which is attracting growing interest is the organic one, which consists in avoiding the use of mineral fertilizers as well as pesticides. The aim of this study is to define the metabolic profile of Brassica oleracea (var. italica) and to compare the samples grown using organic and conventional fertilization methods. The hydroalcoholic and organic extracts of the samples have been analyzed by NMR spectroscopy. Forty-seven metabolites belonging to the categories of organic acids, amino acids, carbohydrates, fatty acids, sterols, and other molecules have been identified. Thirty-seven metabolites have been quantified. Univariate and multivariate PCA analyses allowed to observe that the organic practice influenced the nitrogen transport, the carbohydrate metabolism, the glucosinolate content and the phenylpropanoid pathway in B. oleracea (var. italica).

Stability of the Meat Protein Type I Collagen: Influence of pH, Ionic Strength, and Phenolic Antioxidant.

The water-holding capacity (WHC) is among the key factors in determining the quality of meat and its value, which is strongly influenced by the content and quality of the connective tissue proteins like collagen. Therefore, the factors that influence the proteins' stability, e.g., pH, ionic strength, and the antioxidants which are used to increase the meat shelf-life, also affect the WHC. The interaction of collagen, whose structure is strongly influenced by the interaction with water molecules, can be studied following the behavior of water diffusion by low-resolution 1H NMR experiments. The present study is addressed to study the collagen stability as a function of pH, ionic strength, and phenolic antioxidants like catechin. The experimental study demonstrated how the 1H NMR time domain (TD) experiments are able to evaluate the hydration properties of collagen, not only as a function of ionic strength and pH, but also in determining the ability of catechin to interact both on the surface of the collagen fibrils and inside the fibrillar domain.

NMR-based metabolic study of leaves of three species of Actinidia with different degrees of susceptibility to Pseudomonas syringae pv. actinidiae.

Bacterial canker of Actinidia, caused by the bacterium Pseudomonas syringae pv. actinidiae (Psa), is the most serious disease of these plants worldwide. Leaves of three species of Actinidia, namely A. chinensis var. chinensis, A. chinensis var. deliciosa and A. arguta, having different degrees of tolerance to Psa, were analyzed by Nuclear Magnetic Resonance spectroscopy. Aqueous extracts of leaves were studied and several metabolites, classified as organic acids, amino acids, carbohydrates, phenols and other metabolites, were identified by 1D and 2D NMR experiments and quantified. The metabolic profiles of these species were compared through univariate statistical analysis ANOVA and multivariate PCA. Levels of metabolites with known antibacterial activity, such as caffeic and chlorogenic acids, were observed to be higher in the A. arguta samples. Moreover, these metabolites have different Pearson correlation patterns among the three Actinidia species, suggesting a difference at the phenylpropanoid biosynthetic pathway.

Role of catechin on collagen type I stability upon oxidation: a NMR approach.

The study focuses on the understanding, at molecular level, the mechanism of interaction between protein and flavonoids. Collagen and catechin interactions were investigated by NMR in solution and solid state. The effect of catechin on the stability of collagen to oxidation was also explored. Collagen was treated with two concentrations of catechin solutions. Oxidation was carried out by incubation of collagen solution with three oxidation systems: Fe(II)/H2O2, Cu(II)/H2O2, and NaOCl/H2O2. The effects of oxidation systems were evaluated by high resolution 1 D and 2 D proton spectroscopy and solid state NMR (13C CP MAS) experiments. Interactions between collagen and catechin preferentially occur between catechin B ring and the amino acids Pro and Hyp of collagen. Results showed that both iron and copper oxidation systems were able to interact with collagen by site specific attack. Moreover, catechin protects collagen proline from oxidation by metal/H2O2 systems, preventing copper and iron approach to collagene molecule;this behaviour was more evident for the copper/H2O2 system.

A syn-ent-labdadiene derivative with a rare spiro-ß-lactone function from the male cones of Wollemia nobilis.

An undescribed labdane-like diterpene with a rare spiro-ß-lactone function was identified from the ethanol extract of the male cones of the coniferous tree Wollemia nobilis. This spirolabdadienolide (IUPAC name: syn-ent-8(17),13-labdadien-19,18-olid-15-oic acid methyl ester; trivial name: wollemolide), was isolated by means of traditional and high performance chromatography techniques and structurally elucidated through NMR and MS. In addition, six further known metabolites were evidenced in the extract. Wollemolide, which may be considered an additional chemotaxonomic marker, and 4'-O-methyl-scutellarein, a simple flavonoid, had not been isolated in our previous phytochemical study on the same plant organ. This demonstrates how the molecular pattern of a plant species is in continuous movement and changes with the passing of time according to the climate of the year.

Phytochemical profile of Euphorbia peplus L. collected in Central Italy and NMR semi-quantitative analysis of the diterpenoid fraction.

In this work, the phytochemical profile of the ethanolic extract of Euphorbia peplus L. collected in Central Italy, was reported. This specimen had never been studied before and the analysis was accomplished by means of Column Chromatography for the separation procedure and by means of NMR Spectroscopy and Mass Spectrometry for the identification step. In particular, fourteen compounds were evidenced belonging to five different classes of natural compounds i.e. triterpenoids (pentacyclic and saponin), peculiar diterpenoids (jatrophanes and pepluanes), flavonoids (flavonols), caffeoyl-quinic acids and rare disaccharides. In addition to this, a semi-quantitative analysis on the diterpenoid fraction, by means of NMR Spectroscopy, was also performed in order to provide the real quantities of these compounds in the same fraction and in the total extract. Due to the pronounced chemo variability observed in Euphorbia spp., the availability of a reliable and quick analytical technique, such as that reported in the present study, could be a useful tool in the standardization of plant materials to be used in pharmacological studies or for ethnomedicinal purposes. The technical details for both the general phytochemical analysis and the specific quantitative one, were inserted in this paper. Moreover, the chemotaxonomic and ethnopharmacological relevance of these compounds was also discussed.

Monitoring of pistachio (Pistacia Vera) ripening by high field nuclear magnetic resonance spectroscopy.

The metabolic profiling of pistachio (Pistacia vera) aqueous extracts from two different cultivars, namely 'Bianca' and 'Gloria', was monitored over the months from May to September employing high field NMR spectroscopy. A large number of water-soluble metabolites were assigned by means of 1D and 2D NMR experiments. The change in the metabolic profiles monitored over time allowed the pistachio development to be investigated. Specific temporal trends of amino acids, sugars, organic acids and other metabolites were observed and analysed by multivariate Partial Least Squares (PLS) analysis. Statistical analysis showed that while in the period from May to September there were few differences between the two cultivars, the ripening rate was different.

(1)H NMR-Based Metabolomics Reveals a Pedoclimatic Metabolic Imprinting in Ready-to-Drink Carrot Juices.

Carrots are usually consumed in their native form or processed into many different products. Carrot juice is a popular beverage consumed throughout the world and is attracting increasing attention due to its nutritional value, being a natural source of bioactive compounds. Ready-to-drink carrot juices produced in the same factory were analyzed by (1)H nuclear magnetic resonance (NMR) spectroscopy. The juices were made from carrot roots of the same cultivar grown in three different geographical areas in Italy. More than 30 compounds have been identified and quantified, and the data was subjected to univariate ANOVA and multivariate analyses. Clear geographical-dependent clustering was observed, and the metabolic profiles were related to the different pedoclimatic conditions. The proposed phytoprofiling approach could be employed on an industrial scale to evaluate finished products involving different sites of supply of the raw material, thus improving both the quality and uniformity of the juices.

Nuclear Magnetic Resonance-Based Metabolic Comparative Analysis of Two Apple Varieties with Different Resistances to Apple Scab Attacks.

Apple scab, caused by the fungus Venturia inaequalis, is the most serious disease of the apple worldwide. Two cultivars (Malus domestica), having different degrees of resistance against fungi attacks, were analyzed by (1)H and (13)C nuclear magnetic resonance (NMR) spectroscopy. Aqueous and organic extracts of both apple flesh and skin were studied, and over 30 metabolites, classified as organic acids, amino acids, carbohydrates, phenolic compounds, lipids, sterols, and other metabolites, were quantified by means of one-dimensional (1D) and two-dimensional (2D) NMR experiments. The metabolic profiles of the two apple cultivars were compared, and the differences were correlated with the different degrees of resistance to apple scab by means of univariate analysis. Levels of metabolites with known antifungal activity were observed not only to be higher in the Almagold cultivar but also to show different correlation patterns in comparison to Golden Delicious, implying a difference in the metabolic network involved in their biosynthesis.

1H NMR-based urinary metabolic profiling reveals changes in nicotinamide pathway intermediates due to postnatal stress model in rat.

The maternal separation protocol in rodents is a widely recognized model of early life stress allowing acute and chronic physiological consequences to be studied. An (1)H NMR-based metabolomic approach was applied to urines to evaluate the systemic metabolic consequences of maternal separation stress in female rats after the beginning of weaning and 4 weeks later when the rats were reaching adulthood. Furthermore, because maternal separation is considered as a model mimicking the inflammatory bowel syndrome, the lactulose/mannitol test was used to evaluate the influence of postnatal maternal separation on gut permeability and mucosal barrier function by (1)H NMR spectroscopy analysis of urine. The results showed no statistical differences in gut permeability due to maternal separation. The application of ANOVA simultaneous component analysis allowed the contributions of physiological adaptations to the animal's development to be separated from the metabolic consequences due to postnatal stress. Systemic metabolic differences in the maternally separated pups were mainly due to the tryptophan/NAD pathway intermediate levels and to the methyladenosine level. Urinary NMR-based metabolic profiling allowed us to disentangle the metabolic adaptive response of the rats to postnatal stress during the animal's growth, highlighting the metabolic changes induced by weaning, gut closure, and maturity.

Metabolic profile of different Italian cultivars of hazelnut (Corylus avellana) by nuclear magnetic resonance spectroscopy.

High-resolution proton NMR spectroscopy was performed on three Italian hazelnut cultivars, Tonda di Giffoni, Mortarella and Tonda Gentile Romana, and it allowed to define their metabolic profile. The hazelnuts were grown in the same pedoclimatic conditions in the Monti Cimini (Latium) area. The samples were obtained by using a modified Bligh-Dyer extraction protocol which did not give rise to artefacts arising from the demolition of macromolecular structures such as proteins and polysaccharides. Metabolites belonging to different chemical classes (amino acids, organic acids, carbohydrates, lipids and miscellaneous compounds) were identified and quantified. The three cultivars were discriminated by means of univariate (ANOVA) and multivariate (PCA) statistical analysis.

NMR methodologies in the analysis of blueberries.

An NMR analytical protocol based on complementary high and low field measurements is proposed for blueberry characterization. Untargeted NMR metabolite profiling of blueberries aqueous and organic extracts as well as targeted NMR analysis focused on anthocyanins and other phenols are reported. Bligh-Dyer and microwave-assisted extractions were carried out and compared showing a better recovery of lipidic fraction in the case of microwave procedure. Water-soluble metabolites belonging to different classes such as sugars, amino acids, organic acids, and phenolic compounds, as well as metabolites soluble in organic solvent such as triglycerides, sterols, and fatty acids, were identified. Five anthocyanins (malvidin-3-glucoside, malvidin-3-galactoside, delphinidin-3-glucoside, delphinidin-3-galactoside, and petunidin-3-glucoside) and 3-O-α-l-rhamnopyranosyl quercetin were identified in solid phase extract. The water status of fresh and withered blueberries was monitored by portable NMR and fast-field cycling NMR. (1) H depth profiles, T2 transverse relaxation times and dispersion profiles were found to be sensitive to the withering.

Direct interaction of hydrophilic gold nanoparticles with dexamethasone drug: loading and release study.

Water-soluble gold nanoparticles functionalized by sodium 3-mercapto-1-propansulfonate (Au-3MPS) were synthesized with different Au/thiol molar ratios for their ability to interact with biomolecules. In particular, a synthetic glucocorticoid steroid, i.e. dexamethasone (DXM) was selected. Herein, the formation of the Au-3MPS/DXM bioconjugate is reported. Au-3MPS nanoparticles show a plasmon resonance at 520 nm, have a spherical morphology and average size of 7-10 nm. The total number of gold atoms was estimated to be about 10600, with a surface component of 8800 atoms and a number of thiol ligands of about 720, roughly one anchored thiol every 10 surface gold atoms. The drug-nanoparticle interaction occurs through the fluorine atom of DXM and Au(I) atoms on the gold nanoparticle surface. The 3MPS ligands closely pack apart each other to leave room for the DXM, that lies at the gold surface in an unusual, almost parallel feature. The loading efficiency of DXM on Au-3MPS was assessed in the range 70-80%, depending on the thiol content. Moreover, our studies confirmed the drug release of about 70% in 5 days. Thanks to their unique properties, i.e. high water solubility, small size and almost monodispersity, Au-3MPS display high potential in biotechnological and biomedical applications, mainly for the loading and release of water insoluble drugs.

Exploring human breast milk composition by NMR-based metabolomics.

Breast milk is a complex fluid evolutionarily adapted to satisfy the nutritional requirements of growing infants. In addition, milk biochemical and immunological components protect newborns against infective agents in the new environment. Human milk oligosaccharides, the third most abundant component of breast milk, are believed to modulate the microbiota composition, thus influencing a wide range of physiological processes of the infant. Human milk also contains a number of other bioactive compounds, the functional role of which has not yet been clearly elucidated. In this scenario, NMR-based metabolic profiling can provide a rapid characterisation of breast milk composition, thus allowing a better understanding of its nutritional properties.

Impact of the Mediterranean fruit fly (medfly) Ceratitis capitata on different peach cultivars: the possible role of peach volatile compounds.

The relationship between susceptibility of different peach cultivars (cvs) to the Mediterranean fruit fly (medfly), Ceratitis capitata, and the volatile composition of ripe fruit of each cv has been investigated, since understanding the fruit-insect interaction mechanism is crucial for developing control strategies for such a pest. Volatile compounds were analyzed by SPME-GC-MS in three cvs highly susceptible to medfly attack (Fair Time, Flaminia, Sicilia Piatta), and in two less susceptible cvs (Percoca Romagnola 7 and Doctor Davis). Among the volatile compounds detected, 88 could be identified. The main differences found in the volatile composition of the cvs, concerned the relative abundance of esters. The least susceptible cvs, above all Percoca Romagnola 7, contained the higher amounts of hexenyl, hexyl, 3-methylbutyl, butyl and 2-methylpropyl esters; among these, some C6 derivatives detected, such as (Z)-3-hexenyl acetate, are known to act as priming agents, enhancing plant defence response to insects. Instead, a lower relative content of methyl esters, such as methyl hexanoate and methyl octanoate, known to act as medfly pheromone and attractant respectively, was found in the least susceptible cvs.

Peach fruit: metabolic comparative analysis of two varieties with different resistances to insect attacks by NMR spectroscopy.

The metabolite profile of aqueous extracts of two peach varieties, Percoca Romagnola 7 and Flaminia, with different susceptibilities to Ceratitis capitata attack was investigated by means of 1D and 2D high-field NMR spectroscopy. Water-soluble metabolites belonging to different classes such as organic acids (citric, fumaric, malic, quinic, shikimic, and succinic acids), sugars (fucose, fructose, fructose-6-phosphate, glucose, glucose-6-phosphate, rhamnose, sucrose, and xylose), amino acids (alanine, asparagine, isoleucine, threonine, and valine) and other metabolites such as myo-inositol, choline, trigonelline, catechin, chlorogenic and neochlorogenic acids, orthophosphate, and α-l-glycerophosphorylcholine were identified. The metabolite profile together with a suitable statistical analysis was used to make a comparison between the two varieties. The levels of glucose, xylose, myo-inositol, choline, isoleucine, and valine were found to be higher in Flaminia than in Percoca Romagnola 7 samples, whereas the levels of fumaric acid, alanine, quinic acid, sucrose, fucose, and chlorogenic and neochlorogenic acid were found to be higher in Percoca Romagnola 7 than in Flaminia samples.

Metabolic profiling and outer pericarp water state in Zespri, CI.GI, and Hayward kiwifruits.

The metabolic profiling of aqueous extracts of Zespri Gold ( Actinidia chinensis ) and CI.GI (a controlled crossbreed from different species of Actinidia deliciosa ) kiwifruits and the water state of the outer pericarp of entire fruits were monitored over the season by means of high-field NMR spectroscopy and T(2) relaxation time measurements, respectively, and compared with the corresponding ones of Hayward kiwifruits previously investigated. A more complete assignment of the (1)H spectrum with respect to that obtained previously was reported: histidine, phenylalanine, quercetin 3-rhamnoside, and epicatechin were identified. Metabolic profiling confirmed Zespri's earlier maturation compared with the two other varieties. The water state of entire kiwifruits was measured nondestructively on fruits attached to the plants or detached from the plants. T(2) relaxation times were found to be sensitive to the kiwifruit developmental stage.