sodium-dodecyl-sulfate and ammonium-bicarbonate

Paracoccidioidomycosis is an important systemic mycosis caused by thermodimorphic fungi of the Paracoccidioides genus. During the infective process, the cell wall acts at the interface between the fungus and the host. In this way, the cell wall has a key role in growth, environment sensing and interaction, as well as morphogenesis of the fungus. Since the cell wall is absent in mammals, it may present molecules that are described as target sites for new antifungal drugs. Despite its importance, up to now few studies have been conducted employing proteomics in for the identification of cell wall proteins in Paracoccidioides spp. Here, a detailed proteomic approach, including cell wall-fractionation coupled to NanoUPLC-MS

Topics: Bicarbonates; Cell Wall; Chromatography, Liquid; Fungal Proteins; Gene Expression Regulation, Fungal; Gene Ontology; Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); Liquid-Liquid Extraction; Metabolic Networks and Pathways; Molecular Sequence Annotation; Mycelium; Paracoccidioides; Phosphopyruvate Hydratase; Proteomics; Sodium Dodecyl Sulfate; Tandem Mass Spectrometry

A sensitive method for the determination of three cationic alkaloids (berberine, palmatine and jatrorrhizine) from human plasma samples was developed by micelle to solvent stacking (MSS) in capillary zone electrophoresis (CZE). In MSS, the sample preconcentration mainly relies on the reversal in the effective electrophoretic mobility of the analytes at the boundary zone between the sample and CZE background solution (BGS). Under the optimized conditions, the sensitivity enhancement factors achieved in terms of corrected peak area were in the range from 47 to 53 for the alkaloids. The limits of detection (LODs) (S/N=3) for berberine, palmatine and jatrorrhizine were 0.01, 0.01 and 0.02μg/mL, respectively. The intraday (n=6) and interday repeatabilities (n=12) expressed as the relative standard deviations (RSDs) were less than 6.9% in terms of peak height and less than 7.3% in terms of corrected peak area, respectively. The recoveries of the method for the three alkaloids were in the range of 95.9-101.5% with peak height as the quantitative signal, and 92.6-103.6% with corrected peak area as the quantitative signal, respectively. The MSS-CZE method proved to be suitable for the analysis of the alkaloids in human plasma samples.

Topics: Berberine Alkaloids; Bicarbonates; Cations; Electrophoresis, Capillary; Humans; Limit of Detection; Methanol; Micelles; Reproducibility of Results; Sodium Dodecyl Sulfate