Metabolomic analysis of human urine by UHPLC-HRMS/MS : development of a general method and optimisation of the workflow from the sample preparation to the statistical analysis - Normandie Université Accéder directement au contenu
Communication Dans Un Congrès Année : 2022

Metabolomic analysis of human urine by UHPLC-HRMS/MS : development of a general method and optimisation of the workflow from the sample preparation to the statistical analysis

Résumé

Metabolomic is the study of metabolic profiles obtain from biological matrix. Comparing the profiles of healthy people with those of people exposed to a specific contaminant using metabolomic tools allow the identification of effects biomarkers linked to the contaminant. Several methods are used to obtains metabolic profiles, using different analytical systems or different separation methods. No general methods exist yet for such analysis.The aim of this project is to develop a general method of metabolomic analysis of Human urine for the analysis of biobank samples. Each steps of the workflow must be optimized to collect as much data as possible while being reliable and relevant. The optimizable steps are sample preparation, chromatographic separation, high-resolution mass spectrometry analysis, data pre-treatment, statistical analysis and biological interpretation of the identified biomarkers. This development was made on a group of healthy volunteers of 11 men and 10 women.5 mobile phases usually used for UHPLC in the literature were tested with a QTOF detector. Samples were injected one or several times to estimate replication importance. Three normalisation methods were tested, based on creatinine, osmolarity and total signal. Two ionisation modes were used, combined with reverse phase and HILIC separations. The association of these conditions were evaluated using Galaxy Workflow4Metabolomics. The results were evaluated according to the total numbers of features, and results repeatability. Several treatment methods were then tested with specifics parameters. At the end of these testing, a separation between men and women have been obtained.Finally, a reflexion about generic statistical tools used in metabolomic is made. Biomarkers are usually selected looking at the most variation in the profiles. According to molecular epidemiology these markers may not be the most interesting ones. New statistical tools could suit more for the detection of relevant markers.

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Chimie
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Dates et versions

hal-03846745 , version 1 (10-11-2022)

Identifiants

  • HAL Id : hal-03846745 , version 1

Citer

Quentin Vandoolaeghe, Claire Lopez-Piffet, Valérie Bouchart, Raphael Delepee. Metabolomic analysis of human urine by UHPLC-HRMS/MS : development of a general method and optimisation of the workflow from the sample preparation to the statistical analysis. Journée Normande de Recherche Biomédicale 2022, Jun 2022, Rouen, France. ⟨hal-03846745⟩
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