"EMU: Acquisition of Triple Quad (QTRAP) Microflow-LC-MS/MS system for determination of mediators, hormones and metabolites in complex biological samples with minimal analytical quantities"

• Grant number: 23/17113-4
• Support Opportunities: Research Infrastructure Program - Small Medium Sized
• Start date: September 01, 2024
• End date: August 31, 2027
• Field of knowledge: Biological Sciences - Immunology - Applied Immunology
• Principal Investigator: Lúcia Helena Faccioli
• Grantee: Lúcia Helena Faccioli
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
• Associated researchers: Alline Cristina de Campos ; Ana Lúcia da Costa Darini ; Elaine Aparecida Del Bel Belluz Guimarães ; Francisco Silveira Guimaraes ; Francisco Wanderley Garcia de Paula e Silva ; Vera Lúcia Lanchote

Abstract

In recent decades, advances in mass spectrometry technology have resulted in unprecedented developments in biomedical scientific research, increasing knowledge about the functions of different biological systems, as well as their molecular composition. These technological advances have led to the creation of specific research fields, such as lipidomics. In this sense, the "omics" approach to lipids in exploratory (untarget) or quantitative (target) form, intends to correlate data from studies that examine molecular aspects in biological, biotechnological or mimetic systems, applied to diseases or biochemical processes. These methodologies are analytical techniques with great sensitivity and depend on large-scale, state-of-the-art mass spectrometry. Furthermore, depending on the biological matrix and its source or quantity, a micro-flow system in liquid chromatography must be coupled for better separation, identification and quantification of non-polar analytes in minimal quantities. Lipidomics has emerged as a crucial component in the integrative study of the role of genes, proteins and metabolites that fully describe a cellular function. From the specialized literature, two themes emerge as drivers of the current renaissance of the lipid area: 1) new discoveries of biochemical systems that have revealed the specific role of lipid-lipid or lipid-protein interactions in cellular regulation and 2) the prospect of identifying individual lipids in small amounts, obtaining complete scenarios of lipid classes in pathophysiological contexts (biomarkers). In this project, we intend to obtain a mass spectrometer system (triple-quadrupole with QTRAP) for quantitative lipidomics in target mode, coupled to micro-flow liquid chromatography, enhancing analytical sensitivity for rare and low-concentration samples of analytes in complex biological matrices. The main equipment features of this EMU project are: 1) mass range from 5 to 1,250 m/z covering many classes of lipid compounds; 2) 5 ms switching time in MRM mode, performing positive and negative ion analyzes in the same acquisition, without compromising sensitivity; 3) a dynamic range of up to six orders of magnitude spanning analyte concentrations from low-level trace detection to those at high abundance; 4) high-energy dynode detector and a multichannel electron multiplier providing high-quality data for each sample acquisition; 5) MRM acquisition rate of 500 MRM/sec, being able to analyze a wide range of compounds; 6) 12,000 Da/s scanning speed, detecting the crucial analytes in a complex sample. This equipment will be linked to the physical structure and existing equipment in the bi-institutional facility "CEQIL - Center of Excellence in Quantification and Identification of Lipids" of the Faculty of Pharmaceutical Sciences of Ribeirão Preto and the Department of Chemistry of the Faculty of Philosophy, Sciences and Letters of Ribeirão Preto - USP, which will serve the scientific interests of several research groups on the USP- Ribeirão Preto campus (FCFRP; FMRP; FORP and FFCLRP) and other research and teaching units in public and/or private institutions. In this way, this equipment for quantified lipidomics will collaborate with the development of research in the basic areas of chemistry, biochemistry, cell biology and immunology, as well as in the areas of clinical medicine, dentistry and applied pharmacology. Furthermore, it will be an important tool to assist in the training of professionals and specialists in this area of study, providing innovation and international competitiveness for our projects. (AU)