Laura Ruth Cagliani

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.

On the Traceability of Commercial Saffron Samples Using 1H-NMR and FT-IR Metabolomics

In previous works on authentic samples of saffron of known history (harvest and processing year, storage conditions, and length of time) some biomarkers were proposed using both FT-IR and NMR metabolomics regarding the shelf life of the product. This work addresses the difficulties to trace back the “age” of commercial saffron samples of unknown history, sets a limit value above which these products can be considered substandard, and offers a useful tool to combat saffron mislabeling and fraud with low-quality saffron material. Investigations of authentic and commercial saffron samples of different origin and harvest year, which had been stored under controlled conditions for different lengths of time, allowed a clear-cut clustering of samples in two groups according to the storage period irrespectively of the provenience. In this respect, the four-year cut off point proposed in our previous work assisted to trace back the “age” of unknown samples and to check for possible mislabeling practices.

Sudan dyes in adulterated saffron ( Crocus sativus L.): Identification and quantification by 1 H NMR

Saffron, the dried red stigmas of Crocus sativus L., is considered as one of the most expensive spices worldwide, and as such, it is prone to adulteration. This study introduces an NMR-based approach to identify and determine the adulteration of saffron with Sudan I-IV dyes. A complete (1)H and (13)C resonance assignment for Sudan I-IV, achieved by two-dimensional homonuclear and heteronuclear NMR experiments, is reported for the first time. Specific different proton signals for the identification of each Sudan dye in adulterated saffron can be utilised for quantitative (1)H NMR (qHNMR), a well-established method for quantitative analysis. The quantification of Sudan III, as a paradigm, was performed in varying levels (0.14-7.1g/kg) by considering the NMR signal occurring at 8.064ppm. The high linearity, accuracy and rapidity of investigation enable high resolution (1)H NMR spectroscopy to be used for evaluation of saffron adulteration with Sudan dyes.

Recent developments in honey characterization

Honey is a very complex matrix to be investigated. Several chemical classes of compounds are present in a very large range of concentrations. A large number of research groups worldwide have focused their attention and studies to improve the knowledge of honey characterization; it appears clear that new developments of technological improvements would help to provide insight on this complex problem. In the meantime, it is also clear that the officially recognized methodologies are no longer sufficient to answer to the more demanding questions concerning honey authenticity. Complicated and time demanding sample preparation, and the requirement for people specialized in pollen analysis have significantly slowed down analysis response, becoming nowadays unaffordable. The emerging new techniques are opening new frontiers in honey characterization and the most promising approach seems to be the multidisciplinary one, focused on the detection of multiple components, with the aid of chemometrics. In the present review recent findings in honey characterization using advanced analytical techniques such as NMR and Raman spectroscopy, mass spectrometry (MS) also coupled with chromatographic techniques, and other methods are presented covering the period between 2010 and 2015.