Roberto Consonni

Geographical Characterization of Polyfloral and Acacia Honeys by Nuclear Magnetic Resonance and Chemometrics

The importance of geographical origin determination is an increasing and pressing requirement for all foods. Honey is one of the largest studied foods due to its nutritional and medicinal properties in a correct diet. In this paper, a total of 41 honey samples (polyfloral and acacia) from different countries have been analyzed in terms of (1)H NMR spectroscopy coupled with multivariate statistical methods. Unsupervised principal component analysis resulted as an efficient tool in distinguishing (1)H NMR spectra of polyfloral and acacia honey samples and for geographical characterization of the latter ones. Hierarchical projection to latent structures discriminant analysis was successfully applied for the discrimination among polyfloral honey samples of different geographical origins. (13)C NMR spectroscopy was applied to honey samples with the aim to investigate possible sugar isoforms differentiation. Our preliminary data indicated a different isoforms ratio between betaFP and betaFF only for polyfloral Argentinean samples, while Hungarian samples showed resonance shifts for some carbons of alphaFF, betaFP, betaFF, and alphaGP isoforms for both varieties. These data confirmed the potentiality of (13)C spectroscopy in food characterization, especially in sugar-based foods.

Evaluation of saffron (Crocus sativus L.) adulteration with plant adulterants by 1H NMR metabolite fingerprinting

In the present work, a preliminary study for the detection of adulterated saffron and the identification of the adulterant used by means of (1)H NMR and chemometrics is reported. Authentic Greek saffron and four typical plant-derived materials utilised as bulking agents in saffron, i.e., Crocus sativus stamens, safflower, turmeric, and gardenia were investigated. A two-step approach, relied on the application of both OPLS-DA and O2PLS-DA models to the (1)H NMR data, was adopted to perform authentication and prediction of authentic and adulterated saffron. Taking into account the deficiency of established methodologies to detect saffron adulteration with plant adulterants, the method developed resulted reliable in assessing the type of adulteration and could be viable for dealing with extensive saffron frauds at a minimum level of 20% (w/w).

Quantification of Coffea arabica and Coffea canephora var. robusta in roasted and ground coffee blends.

This study reports direct quantification of arabica in roasted and ground coffee blends of Coffea arabica and Coffea canephora var. robusta. (1)H-NMR analysis of water extracts of coffee blends were combined with multivariate statistical analysis to obtain an OPLS model with high predictive capability. This approach allowed to evaluate the composition of coffee blends of unknown arabica and robusta content, on the basis of multiple chemical components. Differences in geographical origin of the analyzed samples did not affected the compositional determination of coffee blends. This approach represents a valid tool in authentication procedures of arabica and robusta blends of roasted and ground coffee.

NMR Characterization of Saccharides in Italian Honeys of Different Floral Sources

The saccharide profiles of 5 different botanical species in 86 Italian honey samples were investigated by ¹H and ¹H-¹³C NMR spectroscopy. Nineteen saccharides were identified in the aqueous extracts, namely, fructose, glucose, gentiobiose, isomaltose, kojibiose, maltose, maltulose, melibiose, nigerose, palatinose, sucrose, turanose, erlose, isomaltotriose, kestose, maltotriose, melezitose, raffinose, and maltotetraose. PCA performed on NMR spectral regions, in particular between 4.400 and 5.700 ppm and the fructose signal at 4.050 ppm, revealed a partial sample grouping. The score contribution plots derived from PCA performed using the mean values for the buckets of the anomeric region for each floral source allowed the identification of saccharides characterizing different honeys. OPLS-DA models were further evaluated to confirm the previous findings. OPLS-DA models were also built to highlight differences between polyfloral and high mountain polyfloral honeys and between high mountain polyfloral and rhododendron honeys, both collected at high altitude; S-plots highlighted the characteristic saccharides.

NMR based geographical characterization of roasted coffee

The increasing attention for food quality and safety led to develop several analytical techniques suitable to address these demands. Coffee has been already demonstrated to represent a worldwide appreciated beverage and its high economical value could induce frauds or adulteration practices involving both compositional and geographical aspects. In the last years, metabolic profiling revealed to be suitable to face the quality determination of food matrices and NMR confirmed its potentiality in metabolites characterization. The present study reports the capability of NMR spectroscopy to investigate the metabolite content of roasted Coffea arabica samples from the three main production areas, America, Africa and Asia. OPLS-DA models performed on (1)H NMR data led to a clear separation of samples according to their origin: fatty acids, chlorogenic acids and lactate and finally acetate and trigonelline resulted the main compounds characterizing the American, African and Asian samples respectively. The analytical approach here presented confirmed the potentiality of the joined NMR analysis and statistical treatments in quality determination of food matrices.

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.

Analytical method for authentication of Traditional Balsamic Vinegar of Modena.

(13)C NMR spectroscopy was employed for evaluating glucose and fructose isoforms content in Traditional Balsamic Vinegar of Modena. With the use of reference spectra recorded in water for samples obtained with respect to set rules, a shift determination method for fructose carbon isoforms was introduced to determine the frauds present in unknown Traditional Balsamic Vinegar of Modena samples. No sample preparation yields this approach highly reliable and time saving. Following this approach, for the first time, an objective analytical technique can be used alternatively to the actual procedures for Traditional Balsamic Vinegar certification.

Plant Metabolomics: A Characterisation of Plant Responses to Abiotic Stresses

As with all organisms, plants thrive within a range of environmental conditions that are optimal for their growth and development. They must, however, respond and adapt to conditions that deviate from the optimal, such as low/high temperature, dehydration, high salinity, oxidative stress, heavy metals and nutrient deficiency; these deviations are often responsible for losses in productivity and for spatial (geographical) and temporal (growing season) limitations in the cultivation of crops. Although plants and animals share some responsive mechanisms to unfavourable environmental conditions, plants, as sessile organisms, have developed highly sophisticated and efficient strategies of response. Because of the great interest for both basic and applied research, many scientific endeavours have long addressed the understanding of the mechanisms underlying the stress response and the identification of the specific genes/metabolites that are responsible for tolerance phenotypes. In recent years, the “omics” approaches have allowed high-throughput analyses of the changes that are induced by environmental stresses, confirming data previously obtained with targeted analysis and extending the scope of investigation. It is noteworthy that the metabolomic changes that have been observed in plants subjected to stress conditions depend on different causes; therefore, they have different significance and are expected to differently correlate with tolerance/sensitivity phenotypes. Namely, changes in the metabolome composition due to adverse environmental conditions may depend on i) the stability and catalytic activity of enzymes involved in the production/degradation of specific metabolites, ii) the production of abnormal compounds (or abnormal concentrations of normal compounds) as a result of cell damage, iii) the adjustment of concentration of some metabolites to restore homeostasis and normal metabolic fluxes and iv) the synthesis and/or accumulation of compounds involved in mediating tolerance mechanisms. The main goal of studying metabolic changes during stress responses is to identify metabolites belonging to the (iii) and (iv) groups that are responsible for stress tolerance. Upon exposure to osmotic stress as a result of low temperature, drought and high salinity, plants accumulate a range of osmolytes with the primary function of turgor maintenance.

Relaxometric studies for food characterization: the case of balsamic and traditional balsamic vinegars

NMR spectroscopy is a powerful technique for investigating the structure and composition, as well as the physicochemical properties, of foodstuff. NMR-field cycling modality reports about the relaxation times of solvent molecules as a function of the applied magnetic field strength. In the case of aqueous solutions, this methodology is particularly valuable in assessing the interactions of water molecules with paramagnetic and large-size macromolecular systems. (1)H NMR field cycling relaxometry has been used to characterize traditional balsamic vinegars and balsamic vinegars of Modena. It has been found that the longitudinal relaxation time (T(1)) of the water proton resonance is mainly determined by the water molar fraction and the occurrence of dissolved macromolecules and paramagnetic metal ions. Actually, the observed (1)H nuclear magnetic resonance dispersion (NMRD) profiles appear markedly affected by the formation of paramagnetic macromolecular adducts. It has been shown that counterfeit specimens can be identified on the basis of the comparison of their T(1) and T(2) (transverse relaxation time) values with respect to the corresponding values of genuine samples. For the latter ones, a relationship has been found that relates the observed T(1) to the age of the vinegar.

Metabolite profiles of formula milk compared to breast milk

Breast milk (BM) feeding is the gold standard in neonate nutrition. When BM is not available it can be substituted or integrated with commercial formula milk (FM) usually sold under different brands and formulations. In this work, the low-molecular-weight hydrophilic compounds in milk were studied by gas chromatography electronic impact mass spectrometry (GC-MS), comparing eight different FM brands with BM samples. With the aid of multivariate statistical data analysis, a marked variability among FM brands, especially driven by the presence of prebiotics in their formulation, was highlighted. Quali-quantitative differences were found between FM and BM. Orotic acid and isomaltulose were found exclusively in FM, while phenylalanine and tyrosine levels were high in two FM brands. Moreover, higher levels of malic acid, sugars (glucose, fructose and galactose), and mannitol were detected in FM. On the other hand, BM showed a higher amino acid content. In conclusion, GC-MS proved to be a very sensitive analytical technique for the study of FM, highlighting metabolite differences among FM brands, and between FM and BM, that may have a possible strong impact on neonatal nutrition.