Project description:In a previous work using (1)H-NMR we reported encouraging steps towards the construction of a robust expert system for the discrimination of coffees from Colombia versus nearby countries (Brazil and Peru), to assist the recent protected geographical indication granted to Colombian coffee in 2007. This system relies on fingerprints acquired on a 400 MHz magnet and is thus well suited for small scale random screening of samples obtained at resellers or coffee shops. However, this approach cannot easily be implemented at harbour's installations, due to the elevated operational costs of cryogenic magnets. This limitation implies shipping the samples to the NMR laboratory, making the overall approach slower and thereby more expensive and less attractive for large scale screening at harbours. In this work, we report on our attempt to obtain comparable classification results using alternative techniques that have been reported promising as an alternative to NMR: GC-MS and GC-C-IRMS. Although statistically significant information could be obtained by all three methods, the results show that the quality of the classifiers depends mainly on the number of variables included in the analysis; hence NMR provides an advantage since more molecules are detected to obtain a model with better predictions.
Project description:Lipids composed of condensed isoprenyl units connected to glycerol backbones by ether linkages are a distinguishing feature of Archaea. Data suggesting that fatty acids with linear hydrocarbon chains are present in some Archaea have been available for decades. However, lack of genomic and biochemical evidence for the metabolic machinery required to synthesize and degrade fatty acids has left the field unclear on this potentially significant biochemical aspect. Because lipids are energy currency and cell signaling molecules, their presence in Archaea is significant for understanding archaeal biology. A recent large-scale bioinformatics analysis reignited the debate as to the importance of fatty acids in Archaea by presenting genetic evidence for the presence of enzymes required for anabolic and catabolic fatty acid metabolism across the archaeal domain. Here, we present direct biochemical evidence from gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) spectroscopy for the presence of fatty acids in two members of the Crenarchaeota, Sulfolobus solfataricus and Ignicoccus hospitalis. This is the first report providing biochemical data for the existence of fatty acids in these Crenarchaeota, opening new discussions on energy balance and the potential for the discovery of new thermostable enzymes for industry.
Project description:The volatiles in coffee play an important part in the overall flavor profile. In this study, GC-TOF/MS and GC×GC-TOF/MS were used to detect the volatile organic compounds (VOCs) in coffee samples of three different brands at three states (bean, powder, and brew). The differences between the two methods in characterizing VOCs were analyzed using the Venn diagram and PCA (principal component analysis). The important aroma-contributing compounds were further compared and analyzed. The results of the venn diagrams of different coffee samples showed that most VOCs existed in 2-3 kinds of coffee. The PCA of VOCs in different coffee samples showed that the VOCs detected by GC-TOF/MS could distinguish the coffee samples in the different states. GC×GC-TOF/MS was suitable for the further identification and differentiation of the different brands of coffee samples. In addition, pyridine, pyrrole, alcohols, and phenols greatly contributed to distinguishing coffee in three states, and alcohols greatly contributed to distinguishing the three brands of coffee.
Project description:Isotope labeling measurements using mass spectrometry can provide informative insights on the metabolic systems of various organisms. The detailed identification of carbon positions included in the fragment ions of dicarboxylic and tricarboxylic acids in central carbon metabolism is needed for precise interpretation of the metabolic states. In this study, fragment ions containing the carbon backbone cleavage of dicarboxylic and tricarboxylic in the Krebs cycle were investigated by using gas chromatography (GC)-electron ionization (EI)-MS and GC-EI-MS/MS. The positions of decarboxylation in the dicarboxylic and tricarboxylic acids were successfully identified by analyses using position-specific 13C-labeled standards prepared by in vitro enzymatic reactions. For example, carboxyl groups of C1 and C6 of trimethylsilyl (TMS)- and tert-butyldimethylsilyl (TBDMS)-derivatized malic and citric acids were primarily cleaved by EI. MS/MS analyses were also performed, and fragment ions of TBDMS-citric and α-ketoglutaric acids (αKG) with the loss of two carboxyl groups in collision-induced dissociation (CID) were observed.
Project description:Chlorinated paraffins (CPs) are high-volume chemicals used worldwide in various industries as plasticizers, lubricants, and flame retardants. CPs are produced by chlorination of alkane mixtures which leads to complex products of thousands of homologs and congeners. Classic mass spectrometric analyses of CPs allow determining carbon chain lengths and degrees of chlorination while information on the substitution patterns cannot be derived. Therefore, we performed different one- and two-dimensional nuclear magnetic resonance spectroscopy (NMR) experiments, elemental analysis (EA), and gas chromatography coupled with electron capture negative ion mass spectrometry (GC/ECNI-MS) for the analysis of ten technical CP products with 42%, 52%, and 70% chlorine content from four producers. Slight differences in chlorine content but varying chain length compositions were observed for similarly labeled products from different manufacturers. Two-dimensional heteronuclear spectral quantum coherence (HSQC) measurements helped to evaluate ten structural elements in the products and confirmed the presence of geminal chlorine atoms in primary and secondary carbons in products with 70% chlorine. The variation of signal groups increased with increasing chlorine content of the products. Two-dimensional heteronuclear multiple bond coherence (HMBC) analysis of one sample and GC/ECNI-MS measurements indicated the presence of impurities (e.g., C9-CPs, iso-alkanes) in different technical CP products. These methods could in future allow for better distinction of CP mixtures, and an improved trace-back of environmental CPs to the source, based on specific structural features. Additionally, further structural characterization could help in the development of more accurate analysis processes. Graphical Abstract.
Project description:Samples of unifloral sulla (Hedysarum coronarum L.) honey from Sardinia (Italy) were analysed. To investigate the chemical composition of the honey volatiles two solvent systems were used for ultrasonic solvent extraction (USE): 1) a 1:2 (v/v) pentane and diethyl ether mixture and 2) dichloromethane. All the extracts were analysed by GC and GC/MS. These procedures have permitted the identification of 56 compounds that include norisoprenoids, benzene derivatives, aliphatic compounds and Maillard reaction products. Norisoprenoids were the major compounds in both extracts, dominated by vomifoliol (5.3-11.2%; 9.6-14.0%) followed by minor percentages of other norisoprenoids such as α-isophorone, 4-ketoisophorone, 3-oxo-α-ionol or 3-oxo-α-ionone. Other abundant single compounds in the extracts were 3-hydroxy-4-phenylbutan-2-one (0.8-5.4%; 0.6-5.7%) and methyl syringate (3.0-5.7%; 2.2-4.1%). The composition of the volatiles and semi-volatiles in the obtained extracts suggests that sulla honey is quite distinctive relative to the other honeys that have been chemically studied by GC/MS, but no specific markers of the honey botanical origin were found.
Project description:Numerous volatile organic compounds (VOCs) with a large chemical diversity are emitted by plant flowers. They play an important role in the ecology of plants, such as pollination, defense, adaptation to their environment, and communication with other organisms. The Tillandsia genus belongs to the Bromeliaceae family, and most of them are epiphytes. The aromatic profile of the Tillandsia genus is scarcely described. In this study, we use the headspace solid phase microextraction (HS-SPME) coupled with gas chromatography combined with mass spectrometry (GC-MS) method developed in our laboratory to explore the chemical diversity of the VOCs of fragrant flowers of three species of the genus Tillandsia. We were able to identify, for the first time, 66 volatile compounds (monoterpenes, sesquiterpenes, phenylpropanoids, and other compounds). We identified 30 compounds in T. xiphioides, 47 compounds in T. crocata, and 43 compounds in T. caliginosa. Only seven compounds are present in all the species studied. Comparison of the volatile compounds profiles by principal component analysis (PCA) between T. xiphoides, T. crocata, and T. caliginosa species showed a clear difference in the floral emissions of the studied species. Moreover, floral VOCs profiles allowed to differentiate two forms of T. xiphioides and of T. crocata.
Project description:Sixteen organic acids were quantified in peel and pulp of Amber, Laird's Large and Mulligan cultivars of tamarillo using GC-MS. Fourteen of these compounds had not previously been quantified in tamarillo. An untargeted metabolomics approach was used in parallel to identify and quantify 64 more metabolites relative to the internal standard, indicating abundances of glutamic acid, pro-line, aspartic acid and γ-aminobutyric acid as well as lower concentrations of several other essential fatty acids and amino acids. The main findings were that total organic acid concentration was significantly higher (p < 0.05) in pulp than in peel, with the highest concentration seen in Mulligan pulp (219.7 mg/g DW). Remarkably, after citric acid, the potent bactericide itaconic acid was the second most abundant organic acid. At least 95% of organic acids in tamarillo were one of these two acids, as well as cis-aconitic, malic and 4-toluic acids. Differences between cultivar chemotypes were as substantial as differences between tissues. These results suggest that the bitter flavour of the peel does not result from organic acids. The combination of targeted and untargeted metabolomics techniques for simultaneous qualitative and quantitative investigation of nutrients and flavours is efficient and informative.
Project description:Urine is ideal for non-targeted metabolomics, providing valuable insights into normal and pathological cellular processes. Optimal extraction is critical since non-targeted metabolomics aims to analyse various compound classes. Here, we optimised a low-volume urine preparation procedure for non-targeted GC-MS. Five extraction methods (four organic acid [OA] extraction variations and a "direct analysis" [DA] approach) were assessed based on repeatability, metabolome coverage, and metabolite recovery. The DA method exhibited superior repeatability, and achieved the highest metabolome coverage, detecting 91 unique metabolites from multiple compound classes comparatively. Conversely, OA methods may not be suitable for all non-targeted metabolomics applications due to their bias toward a specific compound class. In accordance, the OA methods demonstrated limitations, with lower compound recovery and a higher percentage of undetected compounds. The DA method was further improved by incorporating an additional drying step between two-step derivatization but did not benefit from urease sample pre-treatment. Overall, this study establishes an improved low-volume urine preparation approach for future non-targeted urine metabolomics applications using GC-MS. Our findings contribute to advancing the field of metabolomics and enable efficient, comprehensive analysis of urinary metabolites, which could facilitate more accurate disease diagnosis or biomarker discovery.