{"title":"比较三种广泛应用于布氏锥虫代谢组分析的提取方法","authors":"Fanta Fall, Lieven Desmet, Lúcia Mamede, Laura Schioppa, Pascal de Tullio, Michel Frédérich, Bernadette Govaerts, Joëlle Quetin-Leclercq","doi":"10.1002/cpz1.1043","DOIUrl":null,"url":null,"abstract":"<p><i>Trypanosoma brucei (Tb)</i> is the causative agent of human African trypanosomiasis (HAT), also known as sleeping sickness, which can be fatal if left untreated. An understanding of the parasite's cellular metabolism is vital for the discovery of new antitrypanosomal drugs and for disease eradication. Metabolomics can be used to analyze numerous metabolic pathways described as essential to <i>Tb. brucei</i> but has some limitations linked to the metabolites’ physicochemical properties and the extraction process. To develop an optimized method for extracting and analyzing <i>Tb. brucei</i> metabolites, we tested the three most commonly used extraction methods, analyzed the extracts by hydrophilic interaction liquid chromatography high-resolution mass spectrometry (HILIC LC-HRMS), and further evaluated the results using quantitative criteria including the number, intensity, reproducibility, and variability of features, as well as qualitative criteria such as the specific coverage of relevant metabolites. Here, we present the resulting protocols for untargeted metabolomic analysis of <i>Tb. brucei</i> using (HILIC LC-HRMS). © 2024 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Culture of <i>Trypanosoma brucei brucei</i> parasites</p><p><b>Basic Protocol 2</b>: Preparation of samples for metabolomic analysis of <i>Trypanosoma brucei brucei</i></p><p><b>Basic Protocol 3</b>: LC-HRMS-based metabolomic data analysis of <i>Trypanosoma brucei brucei</i></p>","PeriodicalId":93970,"journal":{"name":"Current protocols","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2024-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Comparison of Three Widely Employed Extraction Methods for Metabolomic Analysis of Trypanosoma brucei\",\"authors\":\"Fanta Fall, Lieven Desmet, Lúcia Mamede, Laura Schioppa, Pascal de Tullio, Michel Frédérich, Bernadette Govaerts, Joëlle Quetin-Leclercq\",\"doi\":\"10.1002/cpz1.1043\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><i>Trypanosoma brucei (Tb)</i> is the causative agent of human African trypanosomiasis (HAT), also known as sleeping sickness, which can be fatal if left untreated. An understanding of the parasite's cellular metabolism is vital for the discovery of new antitrypanosomal drugs and for disease eradication. Metabolomics can be used to analyze numerous metabolic pathways described as essential to <i>Tb. brucei</i> but has some limitations linked to the metabolites’ physicochemical properties and the extraction process. To develop an optimized method for extracting and analyzing <i>Tb. brucei</i> metabolites, we tested the three most commonly used extraction methods, analyzed the extracts by hydrophilic interaction liquid chromatography high-resolution mass spectrometry (HILIC LC-HRMS), and further evaluated the results using quantitative criteria including the number, intensity, reproducibility, and variability of features, as well as qualitative criteria such as the specific coverage of relevant metabolites. Here, we present the resulting protocols for untargeted metabolomic analysis of <i>Tb. brucei</i> using (HILIC LC-HRMS). © 2024 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: Culture of <i>Trypanosoma brucei brucei</i> parasites</p><p><b>Basic Protocol 2</b>: Preparation of samples for metabolomic analysis of <i>Trypanosoma brucei brucei</i></p><p><b>Basic Protocol 3</b>: LC-HRMS-based metabolomic data analysis of <i>Trypanosoma brucei brucei</i></p>\",\"PeriodicalId\":93970,\"journal\":{\"name\":\"Current protocols\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2024-05-06\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.1043\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpz1.1043","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0