Identification and Analytical Characterization of a Novel Synthetic Cannabinoid-Type Substance in Herbal Material in Europe.

The rapid diffusion of new psychoactive substances (NPS) presents unprecedented challenges to both customs authorities and analytical laboratories involved in their detection and characterization. In this study an analytical approach to the identification and structural elucidation of a novel synthetic cannabimimetic, quinolin-8-yl-3-[(4,4-difluoropiperidin-1-yl) sulfonyl]-4-methylbenzoate (2F-QMPSB), detected in seized herbal material, is detailed. An acid precursor 4-methyl-3-(4,4-difluoro-1-piperidinylsulfonyl) benzoic acid (2F-MPSBA), has also been identified in the same seized material. After extraction from the herbal material the synthetic cannabimimetic, also referred to as synthetic cannabinoid receptor agonists or "synthetic cannabinoids", was characterized using gas chromatography-mass spectrometry (GC-MS), 1H, 13C, 19F and 15N nuclear magnetic resonance (NMR) and high-resolution tandem mass spectrometry (HR-MS/MS) combined with chromatographic separation. A cheminformatics platform was used to manage and interpret the analytical data from these techniques.

Can we trust untargeted metabolomics? Results of the metabo-ring initiative, a large-scale, multi-instrument inter-laboratory study.

The metabo-ring initiative brought together five nuclear magnetic resonance instruments (NMR) and 11 different mass spectrometers with the objective of assessing the reliability of untargeted metabolomics approaches in obtaining comparable metabolomics profiles. This was estimated by measuring the proportion of common spectral information extracted from the different LCMS and NMR platforms. Biological samples obtained from 2 different conditions were analysed by the partners using their own in-house protocols. Test #1 examined urine samples from adult volunteers either spiked or not spiked with 32 metabolite standards. Test #2 involved a low biological contrast situation comparing the plasma of rats fed a diet either supplemented or not with vitamin D. The spectral information from each instrument was assembled into separate statistical blocks. Correlations between blocks (e.g., instruments) were examined (RV coefficients) along with the structure of the common spectral information (common components and specific weights analysis). In addition, in Test #1, an outlier individual was blindly introduced, and its identification by the various platforms was evaluated. Despite large differences in the number of spectral features produced after post-processing and the heterogeneity of the analytical conditions and the data treatment, the spectral information both within (NMR and LCMS) and across methods (NMR vs. LCMS) was highly convergent (from 64 to 91 % on average). No effect of the LCMS instrumentation (TOF, QTOF, LTQ-Orbitrap) was noted. The outlier individual was best detected and characterised by LCMS instruments. In conclusion, untargeted metabolomics analyses report consistent information within and across instruments of various technologies, even without prior standardisation.

Mass spectrometry fingerprinting coupled to National Institute of Standards and Technology Mass Spectral search algorithm for pattern recognition.

A new analytical strategy based on mass spectrometry fingerprinting combined with the NIST-MS search program for pattern recognition is evaluated and validated. A case study dealing with the tracing of the geographical origin of virgin olive oils (VOOs) proves the capabilities of mass spectrometry fingerprinting coupled with NIST-MS search program for classification. The volatile profiles of 220 VOOs from Liguria and other Mediterranean regions were analysed by secondary electrospray ionization-mass spectrometry (SESI-MS). MS spectra of VOOs were classified according to their origin by the freeware NIST-MS search v 2.0. The NIST classification results were compared to well-known pattern recognition techniques, such as linear discriminant analysis (LDA), partial least-squares discriminant analysis (PLS-DA), k-nearest neighbours (kNN), and counter-propagation artificial neural networks (CP-ANN). The NIST-MS search program predicted correctly 96% of the Ligurian VOOs and 92% of the non-Ligurian ones of an external independent data set; outperforming the traditional chemometric techniques (prediction abilities in the external validation achieved by kNN were 88% and 84% for the Ligurian and non-Ligurian categories respectively). This proves that the NIST-MS search software is a useful classification tool.

A nuclear magnetic resonance (1H and 13C) and isotope ratio mass spectrometry (delta13C, delta2H and delta18O) study of Andalusian olive oils.

We have determined delta(13)C, delta(2)H and delta(18)O isotopic abundances in Andalusian olive oils. In addition, the fatty acid composition and the distribution of isomers at positions 1,3 and 2 of glycerol were determined by (1)H and (13)C nuclear magnetic resonance (NMR) spectroscopy, respectively. Isotopic results obtained for four series of oil samples extracted from olives harvested in the 2004/05 and 2005/06 seasons are discussed in terms of olive variety, ripeness, geographical origin, fatty acid composition and growing altitude. A distinction was also established between olives grown in irrigated and in dry land by studying selected samples of the previous series and others from the 2005/06, 2006/07, 2007/08 and 2008/09 seasons. The results showed that olive ripeness does not influence the abundance of any of the three isotopes studied. On the other hand, the olive variety influences the abundance of the oxygen and hydrogen isotopes, and also, less markedly, that of carbon. No clear-cut effect of height or latitude on isotope values is observed, probably because the olive variety also changes with height and latitude, thus masking such influences. The oil samples from dryland-grown olives had increased delta(13)C values relative to irrigation-grown olives. In addition, no definite relationship appears to exist between isotope distribution and fatty acid composition. Finally, oil samples from olives harvested in the 2005/06 season in Italy could be distinguished from those from Spain in terms of their isotopic values (delta(2)H mainly).

Virgin olive oil authentication by multivariate analyses of 1H NMR fingerprints and delta13C and delta2H data.

(1)H NMR fingerprints of virgin olive oils (VOOs) from the Mediterranean basin (three harvests) were analyzed by principal component analysis, linear discriminant analysis (LDA), and partial least-squares discriminant analysis (PLS-DA) to determine their geographical origin at the national, regional, or PDO level. Further delta(13)C and delta(2)H measurements were performed by isotope ratio mass spectrometry (IRMS). LDA and PLS-DA achieved consistent results for the characterization of PDO Riviera Ligure VOOs. PLS-DA afforded the best model: for the Liguria class, 92% of the oils were correctly classified in the modeling step, and 88% of the oils were properly predicted in the external validation; for the non-Liguria class, 90 and 86% of hits were obtained, respectively. A stable and robust PLS-DA model was obtained to authenticate VOOs from Sicily: the recognition abilities were 98% for Sicilian oils and 89% for non-Sicilian ones, and the prediction abilities were 93 and 86%, respectively. More than 85% of the oils of both categories were properly predicted in the external validation. Greek and non-Greek VOOs were properly classified by PLS-DA: >90% of the samples were correctly predicted in the cross-validation and external validation. Stable isotopes provided complementary geographical information to the (1)H NMR fingerprints of the VOOs.

1H NMR-based protocol for the detection of adulterations of refined olive oil with refined hazelnut oil.

A (1)H NMR analytical protocol for the detection of refined hazelnut oils in admixtures with refined olive oils is reported according to ISO format. The main purpose of this research activity is to suggest a novel analytical methodology easily usable by operators with a basic knowledge of NMR spectroscopy. The protocol, developed on 92 oil samples of different origins within the European MEDEO project, is based on (1)H NMR measurements combined with a suitable statistical analysis. It was developed using a 600 MHz instrument and was tested by two independent laboratories on 600 MHz spectrometers, allowing detection down to 10% adulteration of olive oils with refined hazelnut oils. Finally, the potential and limitations of the protocol applied on spectrometers operating at different magnetic fields, that is, at the proton frequencies of 500 and 400 MHz, were investigated.

Determination of origin of Atlantic salmon (Salmo salar): the use of multiprobe and multielement isotopic analyses in combination with fatty acid composition to assess wild or farmed origin.

Variability within the stable isotope ratios in various lipidic fractions and the fatty acid composition of muscle oil has been analyzed for a large sample (171 fish) of wild and farmed Atlantic salmon ( Salmo salar) from 32 origins within Europe, North America, and Tasmania. Sampling was extended over all seasons in 2 consecutive years and included fish raised by different practices, in order to maximize the range of variation present. It is shown that two readily measured parameters, delta 15N measured on choline and delta18 O measured on total oil, can be successfully used to discriminate between fish of authentic wild and farmed origin. However, the certainty of identification of mislabeling in market-derived fish is strengthened by including the percentage of linoleic acid C18:2n-6 in the lipidic fraction. Thus, several apparent misidentifications were found. The combination of these three analytical parameters and the size of the database generated makes the method practical for implementation in official laboratories as a tool of labeling verification.

Control of oenological products: discrimination between different botanical sources of L-tartaric acid by isotope ratio mass spectrometry.

Following previous studies on counterfeit of wines with synthetic ingredients, the possibility of frauds by natural external L-tartaric acid has also been investigated. The aim of this research was to map the stable isotope ratios of L-tartaric acid coming from botanical species containing large amounts of this compound: grape and tamarind. Samples of L-tartaric acid were extracted from the pulp of tamarind fruits originating from several countries and from grape must. delta(13)C and delta(18)O were measured for all samples. Additional delta(2)H measurements were performed as a complementary analysis to help discrimination of the botanical origin. Different isotopic patterns were observed for the different botanical origins. The multivariate statistical analysis of the data shows clear discrimination among the different botanical and synthetic sources. This approach could be a complementary tool for the control of L-tartaric acid used in oenology.

Determination of the geographical origin of green coffee by principal component analysis of carbon, nitrogen and boron stable isotope ratios.

In this study we show that the continental origin of coffee can be inferred on the basis of coupling the isotope ratios of several elements determined in green beans. The combination of the isotopic fingerprints of carbon, nitrogen and boron, used as integrated proxies for environmental conditions and agricultural practices, allows discrimination among the three continental areas producing coffee (Africa, Asia and America). In these continents there are countries producing 'specialty coffees', highly rated on the market that are sometimes mislabeled further on along the export-sale chain or mixed with cheaper coffees produced in other regions. By means of principal component analysis we were successful in identifying the continental origin of 88% of the samples analyzed. An intra-continent discrimination has not been possible at this stage of the study, but is planned in future work. Nonetheless, the approach using stable isotope ratios seems quite promising, and future development of this research is also discussed.