Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi

Trypanosoma cruzi, the causative agent of Chagas disease, faces significant metabolic challenges due to fluctuating nutrient availability and oxidative stress within its insect vector. Metabolomic techniques, such as gas chromatography–mass spectrometry (GC–MS), have been widely used to study the ad...

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Main Authors: Michel Silva, Mario Izidoro, Bruno Bonifácio, Sergio Schenkman
Format: Article
Language:English
Published: Bio-protocol LLC 2025-07-01
Series:Bio-Protocol
Online Access:https://bio-protocol.org/en/bpdetail?id=5368&type=0
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author Michel Silva
Mario Izidoro
Bruno Bonifácio
Sergio Schenkman
author_facet Michel Silva
Mario Izidoro
Bruno Bonifácio
Sergio Schenkman
author_sort Michel Silva
collection DOAJ
description Trypanosoma cruzi, the causative agent of Chagas disease, faces significant metabolic challenges due to fluctuating nutrient availability and oxidative stress within its insect vector. Metabolomic techniques, such as gas chromatography–mass spectrometry (GC–MS), have been widely used to study the adaptive mechanisms of the parasite. This article describes a standardized method for the untargeted metabolomics analysis of T. cruzi epimastigote, covering parasite cultivation, sample deproteinization with methanol, metabolite extraction, derivatization with BSTFA, and GC–MS analysis. To ensure robustness and reproducibility, statistical analysis uses univariate tests, as well as multivariate approaches such as principal component analysis (PCA) and partial least squares (PLS) regression. The protocol offers a reliable and sensitive method to study metabolic responses in T. cruzi under environmental stress, with low biological variability and high reproducibility.
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spelling doaj-art-3e551c4bfe3b462089092b4eff6e161b2025-08-20T02:37:20ZengBio-protocol LLCBio-Protocol2331-83252025-07-01151310.21769/BioProtoc.5368Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruziMichel Silva0Mario Izidoro1Bruno Bonifácio2Sergio Schenkman3Department of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, BrazilHospital São Paulo, São Paulo, BrazilDepartment of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, BrazilARIES, Antimicrobial Resistance Institute of São Paulo, São Paulo, BrazilDepartment of Microbiology, Immunology, and Parasitology, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, BrazilARIES, Antimicrobial Resistance Institute of São Paulo, São Paulo, BrazilTrypanosoma cruzi, the causative agent of Chagas disease, faces significant metabolic challenges due to fluctuating nutrient availability and oxidative stress within its insect vector. Metabolomic techniques, such as gas chromatography–mass spectrometry (GC–MS), have been widely used to study the adaptive mechanisms of the parasite. This article describes a standardized method for the untargeted metabolomics analysis of T. cruzi epimastigote, covering parasite cultivation, sample deproteinization with methanol, metabolite extraction, derivatization with BSTFA, and GC–MS analysis. To ensure robustness and reproducibility, statistical analysis uses univariate tests, as well as multivariate approaches such as principal component analysis (PCA) and partial least squares (PLS) regression. The protocol offers a reliable and sensitive method to study metabolic responses in T. cruzi under environmental stress, with low biological variability and high reproducibility.https://bio-protocol.org/en/bpdetail?id=5368&type=0
spellingShingle Michel Silva
Mario Izidoro
Bruno Bonifácio
Sergio Schenkman
Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi
Bio-Protocol
title Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi
title_full Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi
title_fullStr Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi
title_full_unstemmed Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi
title_short Untargeted Metabolomics of Epimastigote Forms of Trypanosoma cruzi
title_sort untargeted metabolomics of epimastigote forms of trypanosoma cruzi
url https://bio-protocol.org/en/bpdetail?id=5368&type=0
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