Optimization of nuclear magnetic resonance spectral acquisition methods for metabolomic analysis of bovine serum: process validation

• Grant number: 23/01189-1
• Support Opportunities: Regular Research Grants
• Start date: September 01, 2023
• End date: February 28, 2026
• Field of knowledge: Agronomical Sciences - Animal Husbandry - Animal Production
• Agreement: CNPq
• Principal Investigator: Nara Regina Brandão Cônsolo
• Grantee: Nara Regina Brandão Cônsolo
• Host Institution: Faculdade de Medicina Veterinária e Zootecnia (FMVZ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated research grant: 20/08845-3 - The metabolomic approach to improve Nellore beef tenderness: a concept of metabotype and biomarkers, AP.JP
• Associated scholarship(s): 23/11605-2 - Optimization of nuclear magnetic resonance spectral acquisition methods for metabolomic analysis of bovine serum: process validation, BP.JD

Abstract

Metabolomics analysis is a valuable tool that enables a deep understanding of the phenotype of animals and has been widely used for improving animal production processes. One of the main techniques used in metabolomics studies is Nuclear Magnetic Resonance (NMR) spectroscopy. The main difficulty for analyzing biological samples of animal origin by NMR is the presence of many macromolecules, such as lipids and proteins, which reduce the spectral resolution and hide the metabolites of interest. The most used protocol for removing these macromolecules is ultrafiltration with 3KDa filters. Although effective, it is a laborious and expensive step due to the high unit cost of the filters. Other NMR methods and post-processing steps can remove the broad signals of the macromolecules. However, some of these approaches, such as the pulse sequence called CPMG lead to the distortion of the metabolite signals areas and make accurate quantification impossible. Thus, there is still an inconsistency in the way of biological samples should be prepareed and the spectrum obtained, in order to more faithfully reflect animal metabolism. This project focuses on testing and validating two of the most recently published new techniques (DREAMTIME and FDM) that could replace the sample filtration process without the deficiencies of the CPMG technique. The knowledge produced by this proposal will promote a scientific and technological improvement in the field of metabolomics applied to animal production, and will certainly guide the way in which biological samples should be processed for the project to which it is linked. The results of this project may bring greater efficiency in processing samples and obtaining biofluid spectra, eliminating the need for ultrafilters without prejudice to the quantification of metabolites. (AU)