Two-Dimensional Gas Chromatographic and Mass Spectrometric Characterization of Lipid-Rich Biological Matrices-Application to Human Cerumen (Earwax).

Earwax is a readily accessible biological matrix that has the potential to be used in disease diagnostics. However, its semisolid nature and high chemical complexity have hampered efforts to investigate its potential to reveal disease markers. This is because more conventional methods of analysis such as gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry yield unsatisfactory results due to the presence of many nonvolatile and/or coeluting compounds, which in some cases have very similar mass spectrometric profiles. In addition, these routine methods often require the sample to be saponified, which dramatically increases the complexity of the analysis and makes it difficult to determine which compounds are actually present versus those that are produced by saponification. In this study, two-dimensional GC mass spectrometry (GC × GC-MS) was successfully applied for the characterization of the chemical components of earwax from healthy donors using nonpolar (primary) and midpolar (secondary) columns without saponification. Over 35 of the compounds that were identified are reported for the first time to be detected in unsaponified earwax. The resulting GC × GC-MS contour plots revealed visually recognizable compound class clusters of previously reported groups including alkanes, alkenes, fatty acids, esters, triglycerides, and cholesterol esters, as well as cholesterol and squalene. The application of GC × GC-MS revealed results that provide a foundation upon which future studies aimed at comparing healthy donor earwax to that from individuals exhibiting various disease states can be accomplished.

Opportunities in phytochemistry, ecophysiology and wood research via laser ablation direct analysis in real time imaging-mass spectrometry.

Analysis of wood transects in a manner that preserves the spatial distribution of the metabolites present is highly desirable to among other things: (1) facilitate ecophysiology studies that reveal the association between chemical make-up and environmental factors or climatic events over time; and (2) investigate the mechanisms of the synthesis and trafficking of small molecules within specialised tissues. While a variety of techniques could be applied to achieve these goals, most remain challenging and impractical. Laser ablation direct analysis in real time imaging-mass spectrometry (LADI-MS) was successfully used to survey the chemical profile of wood, while also preserving the small-molecule spatial distributions. The tree species Entandrophragma candollei Harms, Millettia laurentii DeWild., Pericopsis elata (Harms) Meeuwen, Dalbergia nigra (Vell.) Benth. and Dalbergia normandii Bosser & R.Rabev were analysed. Several compounds were associated with anatomical features. A greater diversity was detected in the vessels and parenchyma compared with the fibres. Analysis of single vessels revealed that the chemical fingerprint used for timber identification is mainly determined by vessel content. Laser ablation direct analysis in real time imaging-mass spectrometry offers unprecedented opportunities to investigate the distribution of metabolites within wood samples, while circumventing the issues associated with previous methods. This technique opens up new vistas for the discovery of small-molecule biomarkers that are linked to environmental events.

A chemometric strategy for forensic analysis of condom residues: Identification and marker profiling of condom brands from direct analysis in real time-high resolution mass spectrometric chemical signatures.

The rapid and accurate identification of condom-derived lubricant traces takes on heightened importance in sexual assault cases where the assailant has used a condom in order to avoid leaving behind incriminating DNA evidence. Previous reports have demonstrated that a variety of techniques can be used to confirm that a given residue is condom-derived, based on the detection of spermicides, slip agents and/or other common additives. However, limited success has been achieved in differentiating brands from among a broad range of condom types. In this study, the utility of direct analysis in real time-high resolution mass spectrometry (DART-HRMS) combined with chemometrics, for the rapid and accurate attribution of brands to condom residues of various types, was explored and developed. A database of condom residue spectra comprised of 110 different condom types representing 16 brands was generated, with the spectra serving as representative fingerprints for each brand. The spectral fingerprints were subjected to pre-processing prior to the application of Partial Least Squares-Discriminant Analysis (PLS-DA) which was used to generate a classifier that permitted identification of condom brands with an accuracy of 97.4%. An additional criterion was imposed on the PLS-DA to provide the confidence level and credibility of each prediction. The effect of time since deposition, the presence of contaminants and the influence of residue transfer on the prediction accuracy of the model were also assessed. The results from Sparse Discriminant Analysis (SDA) and PLS-DA were followed by application of the Student's t-test to determine m/z values representative of small-molecule markers that were most important for defining brand classes. The m/z values revealed by the two methods were found to be consistent in indicating which masses were representative of markers. The SDA method also provided low-dimensional views of the discriminative directions for classification of condom residues, thereby enabling easy visualization of the relationship between the indicated m/z values and brand discrimination. The results further revealed a subset of 14 m/z values that were observed in all 110 condoms representing the 16 brands, and some of these may serve as potential universal small-molecule condom markers. Overall, the results show that the DART-HRMS database of condom residue spectra can be used to identify residues based on differences in chemical components peculiar to each brand. The database can be readily expanded to include more condoms.