Spatially Correlated Nuclear Magnetic Resonance Profiles as a Tool for Precision Agriculture.

Nuclear magnetic resonance (NMR) profiling, sample georeferentiaton, and geostatistics are applied to evaluate the spatial variability of metabolic expression of durum wheat in fields managed by precision agriculture. Durum wheat at three different vegetation stages, grown in two different places of the Basilicata region, in Italy, is analyzed by NMR. The spatial variability, within each field, of metabolites, quantified by NMR, is evidenced by appropriate geostatistic tools through the definition of a suitable metabolic index. Metabolic maps are compared to highlight the effects of soil and farming strategies.

Farming for Pharming: Novel Hydroponic Process in Contained Environment for Efficient Pharma-Grade Production of Saffron.

Soilless cultivation of saffron (Crocus sativus) in a controlled environment represents an interesting alternative to field cultivation, in order to obtain a standardized high-quality product and to optimize yields. In particular, pharma-grade saffron is fundamental for therapeutic applications of this spice, whose efficacy has been demonstrated in the treatment of macular diseases, such as Age-related Macular Degeneration (AMD). In this work, a hydroponic cultivation system was developed, specifically designed to meet the needs of C. sativus plant. Various cultivation recipes, different in spectrum and intensity of lighting, temperature, photoperiod and irrigation, have been adopted to study their effect on saffron production. The experimentation involved the cultivation of corms from two subsequent farm years, to identify and validate the optimal conditions, both in terms of quantitative yield and as accumulation of bioactive metabolites, with particular reference to crocins and picrocrocin, which define the 'pharma-grade' quality of saffron. Through HPLC analysis and chromatography it was possible to identify the cultivation parameters suitable for the production of saffron with neuroprotective properties, evaluated by comparison with an ISO standard and the REPRON® procedure. Furthermore, the biochemical characterization was completed through NMR and high-resolution mass spectrometry analyses of saffron extracts. The whole experimental framework allowed to establish an optimized protocol to produce pharma-grade saffron, allowing up to 3.2 g/m2 harvest (i.e., more than three times higher than field production in optimal conditions), which meets the standards of composition for the therapy of AMD.

Territorial origin of olive oil: representing georeferenced maps of olive oils by NMR profiling.

Proton NMR profiling is nowadays a consolidated technique for the identification of geographical origin of food samples. The common approach consists in correlating NMR spectra of food samples to their territorial origin by multivariate classification statistical algorithms. In the present work, we illustrate an alternative perspective to exploit territorial information, contained in the NMR spectra, which is based on the implementation of a geographic information system (GIS). Nuclear magnetic resonance spectra are used to build a GIS map permitting the identification of territorial regions having strong similarities in the chemical content of the produced food (terroir units). These terroir units can, in turn, be used as input for labeling samples to be analyzed by traditional classification methods. In this work, we describe the methods and the algorithms that permit to produce GIS maps from NMR profiles and apply the described method to the analysis of the geographical distribution of olive oils in an Italian region. In particular, we analyzed by 1 H NMR up to 98 georeferenced olive oil samples produced in the Abruzzo Italian region. By using the first principal component of the NMR variables selected according to the Moran test, we produced a GIS map, in which we identified two regions incidentally corresponding to the provinces of Teramo and Pescara. We then labeled the samples according to the province of provenience and built an LDA model that provides a classification ability up to 99% . A comparison between the variables selected in the geostatistics and classification steps is finally performed. Copyright © 2016 John Wiley & Sons, Ltd.

Performance Assessment in Fingerprinting and Multi Component Quantitative NMR Analyses.

An interlaboratory comparison (ILC) was organized with the aim to set up quality control indicators suitable for multicomponent quantitative analysis by nuclear magnetic resonance (NMR) spectroscopy. A total of 36 NMR data sets (corresponding to 1260 NMR spectra) were produced by 30 participants using 34 NMR spectrometers. The calibration line method was chosen for the quantification of a five-component model mixture. Results show that quantitative NMR is a robust quantification tool and that 26 out of 36 data sets resulted in statistically equivalent calibration lines for all considered NMR signals. The performance of each laboratory was assessed by means of a new performance index (named Qp-score) which is related to the difference between the experimental and the consensus values of the slope of the calibration lines. Laboratories endowed with a Qp-score falling within the suitable acceptability range are qualified to produce NMR spectra that can be considered statistically equivalent in terms of relative intensities of the signals. In addition, the specific response of nuclei to the experimental excitation/relaxation conditions was addressed by means of the parameter named NR. NR is related to the difference between the theoretical and the consensus slopes of the calibration lines and is specific for each signal produced by a well-defined set of acquisition parameters.

Nuclear magnetic resonance profiling of wine blends.

Nuclear magnetic resonance (NMR) profiling is used for characterization of monocultivar binary wine mixtures. Classification and quantification of the relative amount of wine in the mixture are made in two steps. First, each sample is classified as a mixture of a determined type by solving the appropriate classification problem using NMR profiles. The relative amount of the two corresponding monovarietal wines is then evaluated by multilinear regression of a selected set of NMR variables. Linear discriminant analysis (LDA), used in the classification step, gives a very good separation among the different mixture classes. On the other hand, a single layer artificial neural network, used to solve the multilinear problem, gives the relative amount of wine type in the mixture with a precision of about 10%.

Geographical origin of durum wheat studied by 1H-NMR profiling.

(1)H-Nuclear magnetic resonance (NMR) spectroscopy was used to detect metabolic profiles of wheat flour samples of different geographical and botanical origin. The NMR profiles were analyzed by multivariate statistical techniques in order to establish the origin of the samples. A linear model, able to discriminate among three different locations, was built achieving a prediction level of about 80% of correctly assigned samples. The principal classes of compounds responsible for the geographic origin discrimination were individuated in aromatic compounds and amino acids. The statistical modeling also indicated that botanical origin information is very poor in the NMR profiles of the analyzed wheat samples.

Identification of milk mixtures by 1H NMR profiling.

Nuclear magnetic resonance profiling, combined with a single-layer artificial neural network, is used for the evaluation of the content of mixtures of different kinds of milk. In particular, aqueous fractions of cow and sheep milk mixtures are analyzed by (1) H NMR. The spectral differences are highlighted by an analysis of the variance and a principal component analysis. The species classification problem is solved by a linear discriminant analysis. The quantification of the relative amount of the milk of two different species is then achieved by solving the appropriate multilinear problem.

Novel thermosensitive calcium alginate microspheres: physico-chemical characterization and delivery properties.

The system described in this paper was obtained by soaking calcium alginate (CaAlg) microspheres in a water solution of poly-[(3-acrylamidopropyl)-trimethylammonium chloride-b-N-isopropylacrylamide] [poly(AMPTMA-b-NIPAAM)], a new block co-polymer recently synthesized by atom transfer radical polymerization (ATRP). The block co-polymer is characterized by a lower critical solution temperature (LCST) of 41 degrees C in aqueous 0.1 M NaCl solution, and can be anchored on the CaAlg microspheres by means of polyion interactions. Polycations (permanently positively charged blocks) and polyanions (free alginate carboxylic groups) interact, leading to microspheres with thermosensitive properties. As an effect of interaction with the microspheres the LCST of the co-polymer is lowered to 36-38 degrees C. In this temperature range a colloidal water suspension of the microspheres collapses, forming macroscopic aggregates. The new system shows, at human body temperature, an improved ability to carry and deliver both hydrophobic and hydrophilic molecules in comparison with unmodified CaAlg microspheres. The release properties of the microspheres loaded with different model drugs can be appropriately modulated by the amount of the poly(AMPTMA-b-NIPAAM). Furthermore, the microspheres show the interesting capability of retaining the activity of a loaded enzyme (horseradish peroxidase), used as a model protein. The results obtained indicate that the proposed drug delivery system may be suitable for drug depot applications.

Influence of dialkyne structure on the properties of new click-gels based on hyaluronic acid.

Hydrogels have been widely used in tissue engineering as a support for tissue formation and/or to deliver drug locally. A novel procedure for the in situ rapid chemical gelation of aqueous solutions of hyaluronan (HA) was employed. HA was functionalised with an arm bearing a terminal azido group (HAAA). When HAAA was mixed with a series of dialkyne reagents of different length, a 1,3-dipolar cycloaddition ("click-chemistry") reaction took place in the presence of catalytic amount of Cu(I) resulting in fast gelation at room temperature. The resulting gels were characterised in terms of degree of cross-linking by (1)H HR-MAS NMR. The kinetic of gelation and the determination of elastic moduli as well as the degree of swelling and the controlled release of a model drug, were studied as a function of chemical nature of the dialkyne group, catalyst concentration, HAAA concentration and temperature. All these variables allowed the swelling ratio and the extent of release of a drug, doxorubicin, entrapped within the gel, to be modulated. In all cases the kinetic of release reached the stationary state within 150 h. The height of the plateau was dependent on the overall (chemical and topological) degree of cross-linking.

A preliminary study of soft cheese degradation in different packaging conditions by 1H-NMR.

This preliminary work deals with the influence of packaging on the degradation of an Italian soft cheese studied by NMR. The NMR profiles of aqueous cheese extracts were acquired as a function of time during storage of cheese inside and outside the original package. From the NMR spectra 15 metabolites are quantified and used in a kinetic degradation model and in a multivariate algorithm in order to evaluate the changes in the metabolic composition of cheese stored under different conditions.

TG, FT-IR and NMR characterization of n-C16H34 contaminated alumina and silica after mechanochemical treatment.

This paper deals with the application of mechanochemistry to model systems composed of alumina or silica artificially contaminated with n-C16H34. The mechanochemical treatment was carried out by means of a ring mill for times ranging from 10 to 40h. Thermogravimetry and infrared and nuclear magnetic resonance spectroscopies were used for the characterization of the mechanochemical products. The results have indicated that, in the case of alumina, almost all the contaminant n-C16H34 undergoes a complex oxidative reaction path whose end products are strongly held on the surface. These end products are most likely made of crosslinked, partially oxidized hydrocarbon chains bond to the solid surface via COO(-) groups. In the case of silica, the hydrocarbon undergoes a different, equally complex reaction path, but to a lower extent. In this case the end products are most probably carbonylic compounds and graphitic carbon. Then, for both solid matrices, the mechanochemical treatment promotes significant modification of the chemical nature of the polluting hydrocarbon with end products much more difficult to remove from the surface. As the systems studied are models of sites contaminated by aliphatic hydrocarbon, the results are worthy of consideration in relation to the mobility of the contaminants in the environment.

Novel hydrogels via click chemistry: synthesis and potential biomedical applications.

A novel procedure for the in situ rapid chemical gelation of aqueous solutions of hyaluronan has been employed. In brief, water-soluble polysaccharide derivatives bearing side chains endowed with either azide or alkyne terminal functionality have been prepared. When the latter two types of derivatives are mixed together in aqueous solution they give rise to a 1,3-dipolar cycloaddition reaction resulting in fast gelation (in the presence of catalytic amounts of Cu(I)) at room temperature. Gel formation has been characterized rheologically and could also be followed qualitatively by means of IR spectroscopy. The resulting gels have been studied in terms of swelling properties and, in particular, NMR spectral features. Carrying out the gelation process in aqueous solutions of benzidamine and doxorubicin, respectively, the polysaccharide networks acted as drug reservoirs. The doxorubicin release resulted in well controllable acting upon the gels degree of cross-linking. Finally, formation of the click-gels using aqueous suspensions of Saccharomices cerevisiae yeast cells allowed the obtainment of scaffolds inside which cells were homogeneously distributed and smoothly adhered to the inner pores surfaces, according to SEM analysis. After 24 h about 60% of the entrapped cells exhibited proliferating activity. Click-gels prepared as detailed herein do have a number of positive features that make them, in perspective, materials of choice for drug release and tissue engineering manipulations.

Wheat flour enzymatic amylolysis monitored by in situ (1)H NMR spectroscopy.

Starch enzymatic degradation caused by endogenous hydrolases is studied by in situ NMR spectroscopy on a set of hard and soft wheat flours. The results obtained by two different techniques (HR-MAS and (1)H NMR in solution) are analyzed in terms of a Michaelis-Menten kinetic phenomenological model taking into account the presence of endogenous enzymes and their eventual inactivation. The parameters resulting from the best fit of all experimental data to the kinetic model equations are submitted to a multivariate statistical analysis to assess the role of the oligosaccharides release in distinguishing between hard and soft wheats.