Developing an UHPLC-MS/MS methodology for lipidomic analysis and monitoring of eicosanoids production in biological samples

Abstract

Lipidomics consists in the qualitative and quantitative study of the lipids composition of a biological system. Among the variety of lipid classes, the eicosanoids such as prostaglandins (PGs), leukotrienes (LTs) and lipoxins (LXs) have been widely studied as they are related to various diseases such as cancer, diabetes, arthritis, asthma, Alzheimer's and infectious diseases such as tuberculosis (TB). However, determining the lipidoma of a biological system represents a big challenge since these compounds are produced at very low concentrations, often in the form of isobaric or isomeric molecules. In this context, UHPLC-MS/MS (ultra efficiency liquid chromatography coupled to tandem mass spectrometry) is a versatile technique that allows a rapid and sensitive analysis with high potential to characterize and quantify eicosanoids in complex biological samples. Hence, our project aims to the development of an analytical methodology employing UHPLC-MS/MS to determine the lipid mediators profile in human serum samples or cellular supernatants. This project is innovator and pioneer in Brazil. Initially, this method will be applied in the study about the production of lipid mediators by patients from the Hospital das Clínicas - Ribeirão Preto, including those with active TB or not. The data obtained will help to understand the relationship between Mycobacterium tuberculosis (Mtb) infection and eicosanoids production as well to find biomarkers to differentiate patients with TB active, latent and non-infected people which is a big challenge in the control of TB. This method will be referenced to different research groups with similar proposals that work with humans or animals. We believe that the lipidomic approach will be crucial for a better understanding of the role of lipid mediators in imune responses in human diseases. (AU)