Development, validation and application of a GC–MS method for the simultaneous detection and quantification of neutral lipid species in Trypanosoma cruzi

Abstract

The development and validation of an analytical method for the simultaneous analysis of five neutral lipids in Trypanosoma cruzi epimastigotes by GC-MS is presented in this study. The validated method meets all validation parameters for all components and the chromatographic conditions have been optimized during its development. This analytical method has demonstrated good selectivity, accuracy, within-day precision, recovery and linearity in each of the established ranges. In addition, detection and quantification limits for squalene, cholesterol, ergosterol and lanosterol have been improved and it is worth highlighting the fact that this is the first time that squalene-2,3-epoxide validation data have been reported. The new validated method has been applied to epimastigotes treated with compounds with in vitro anti-T.cruzi activity. This new methodology is straightforward and constitutes a tool for screening possible sterol biosynthesis pathway inhibitors in Trypanosoma cruzi, one of the most studied targets in Chagas disease treatment. Therefore, it is an interesting and useful contribution to medicinal chemistry research.

Exceptional Solvent Tolerance inYarrowia lipolyticaIs Enhanced by Sterols

“…Three biological replicates of wildtype and YlCW001 cells were grown in 0% and 8% (v/v) [EMIM][OAc] until mid-late exponential phase and cell pellets were stored at −20°C. Sterol quantification was performed with the equivalent cell mass of 5 OD 600nm as previously described (84) using a HP-5MS 30 m × 0.25 mm i.d. × 0.25 μm column (Agilent Technologies, USA) for separation of sterols.…”

Multifunctional organometallic compounds for the treatment of Chagas disease: Re(i) tricarbonyl compounds with two different bioactive ligands

“…Ergosterol biosynthesis in Trypanosoma cruzi has been validated as a chemotherapeutic target in several steps of its route, including inhibition of the enzymes squalene-2,3-epoxidase (catalyzes the conversion of squalene to squalene-2,3-epoxide), lanosterol synthase (catalyzes the conversion of squalene-2,3-epoxide to lanosterol), lanosterol 14-α-demethylase (catalyzes the conversion of lanosterol to 4,4-dimethyl-5α-cholesta-8,14,24,trien-3-β-ol), among others. 64,65…”

“…Terbinafine inhibits squalene-2,3-epoxidase but also lanosterol 14-α-demethylase, leading to accumulation of squalene and lanosterol. 65…”

The Quantitation of Squalene and Squalane in Bronchoalveolar Lavage Fluid Using Gas Chromatography Mass Spectrometry

“…SQE has been quantified in various matrices including olive oil, human hair, human serum, and incense smoke by means of gas chromatography flame ionization detection and gas chromatography-mass spectrometry (GC-MS) ( Wu et al, 2016 ; Santivanez-Veliz et al, 2017 ; Budge and Barry, 2019 ; Pacetti et al, 2019 ; Aresta et al, 2020 ). Qualitative confirmation of SQE in human BAL fluid ( Kanaji et al, 2008 ) has been reported using GC-MS.…”