Targeted Lipidomics of Drosophila melanogaster During Development.

Lipids play critical roles in developmental processes, and alterations in lipid metabolism are linked to a wide range of human diseases, including neurodegeneration, cancer, metabolic diseases, and microbial infections. Drosophila melanogaster, more commonly known as the fruit fly, is a powerful organism for developmental biology and human disease research. We have previously developed a comprehensive biochemical tool, based on liquid chromatography-mass spectrometry (LC-MS), to probe the dynamics of lipid remodeling during D. melanogaster development. This chapter introduces a step-by-step protocol for extracting and analyzing lipids across all developmental stages (embryo, larvae, pupa, and adult) of D. melanogaster. The targeted semi-quantitative approach offers a comprehensive coverage of more than 400 lipid species spanning the lipid classes, glycerophospholipids, sphingolipids, triacylglycerols, and sterols.

Enhanced microextraction of endocrine disrupting chemicals adsorbed on airborne fine particulate matter with gas chromatography-tandem mass spectrometric analysis.

A novel double-microextraction approach, combining dispersive liquid-liquid microextraction (DLLME) and vortex-assisted micro-solid-phase extraction (VA-µ-SPE) was developed. The procedure was applied to extract endocrine disrupting chemicals (EDCs) consisting of three phthalate esters (PEs) and bisphenol A (BPA) associated with PM2.5 (airborne particulate matter with aerodynamic diameter ≤ 2.5 µm). Gas chromatography-tandem mass spectrometry (GC-MS/MS) was used for determination of the analytes. These analytes were first ultrasonically desorbed from PM2.5 in a 10% acetone aqueous solution. DLLME was used to first preconcentrate the analytes; the sample solution, still in the same vial, was then subjected to VA-µ-SPE. The synergistic effects provided by the combination of the microextraction techniques provided advantages such as high enrichment factors and good cleanup performance. Various extraction parameters such as type and volume of extractant solvent (for DLLME), and type of sorbent, extraction time, desorption solvent, volume of desorption solvent and desorption time (for µ-SPE) were evaluated. Multi-walled carbon nanotubes were found to be the most suitable sorbent. This procedure achieved good precision with intra- and inter-day relative standard deviations of between 1.93 and 9.95%. Good linearity ranges (0.3-100 ng/mL and 0.5-100 ng/mL, depending on analytes), and limits of detection (LODs) of between 0.07 and 0.15 ng/mL were obtained. The method was used to determine the levels of PEs and BPA in ambient air, with concentrations ranging between below the limits of quantification and 0.48 ng/m3. DLLME-VA-µ-SPE-GC-MS/MS was demonstrated to be suitable for the determination of these EDCs present in PM2.5.

Sodium dodecyl sulfate-multi-walled carbon nanotubes-coated-membrane solid phase extraction of glucocorticoids in aqueous matrices.

A membrane-based solid phase extraction (SPE)-ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed for the determination of nine glucocorticoids in water. This new hybrid SPE approach involved the deposition of sodium dodecyl sulfate (SDS)-multi-walled carbon nanotubes (MWCNTs) on a piece of polypropylene membrane that served as the extraction device. Hitherto, such a sample preparation procedure has not been applied to the analysis of water contaminants before. The use of the surfactant helped to disperse the MWCNTs effectively so that they were coated uniformly onto the polypropylene membrane. This increased the overall extraction efficiency of the procedure. Characterisation of the SDS-MWCNTs material was performed using transmission electron microscopy and scanning electron microscopy. The membrane device did not require a pre-conditioning step. The most favourable extraction parameters such as type of surfactant, percentage of surfactant, type of desorption solvent, stirring rate, desorption time, extraction time, temperature, salting-out effect, pH and diameter of MWCNTs were obtained. The method showed linearity ranges from 0.2 to 100 ng mL-1 for hydrocortisone, dexamethasone, cortisone acetate and beclomethasone dipropionate, and 0.5-100 ng mL-1 for the rest of the analytes. Limits of detection ranging from 0.019 to 0.098 ng mL-1, and limits of quantification ranging from 0.065 to 0.326 ng mL-1, were obtained for the analytes. The intra-day repeatability was between 1.77 and 3.56% while the inter-day reproducibility was between 2.69 and 9.53%, respectively. The method was used to analyse glucocorticoids as contaminants in the canal water samples.