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Development of the methods of the microextraction hyphenated with LC-MS/MS or directly to MS/MS for the determination of biomarkers in biological samples of patient or rats with neurodegenerative diseases

Grant number: 20/00126-8
Support type:Regular Research Grants
Duration: July 01, 2020 - June 30, 2022
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
Principal Investigator:Maria Eugênia Queiroz Nassur
Grantee:Maria Eugênia Queiroz Nassur
Home Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Assoc. researchers:José Alexandre de Souza Crippa ; Vitor Tumas

Abstract

The conclusive diagnosis of neurodegenerative diseases, such as Parkinson's disease (PD) and Alzheimer's disease (AD), has been predominantly clinical and, in these cases, the neurodegeneration is already installed in the patient's brain. Therefore, efforts have been made to identify molecular biomarkers for early diagnosis and monitoring these neurodegenerations progression. High performance liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) has been considered a reference analytical technique for the determination of biomarkers in biological fluids. Due to complexity of biological fluids, LC-MS/MS methods require a sample preparation step to exclude the biological matrix macromolecules and to concentrate the analytes (trace levels). The sample preparation techniques have been simplified, miniaturized (lower sample and organic solvent consumption), and/or hyphenation with chromatographic systems, automation (high-throughput performance). In this context, we can highlight the salting-out assisted liquid-liquid microextraction (microSALLE) and in-tube microextraction (in-tube SPME) hyphenated with LC-MS/MS. The development of capillaries with innovative bifunctional stationary phase for in-tube SPME, such as organic monolithic phases combined with restricted access materials (RAM) and polymerizable zwitterionic ionic liquids (ZILs) has allowed selective pre-concentration of the analytes. In this project, for the evaluation of potential biomarkers for DA, the following methods will be developed for the determination of AD biomarkers in plasma and cerebrospinal fluid samples: (a) in-tube SPME-LC-MS/MS using a organic monolithic capillary with sulfonic groups with RAM covering for the determination of ²-amyloid peptides, (b) in-tube SPME-MS/MS using a polyester(ether)ketone (PEEK) capillary packed with nitinol fibers (SPME support) coated with ZILs for determination ²-amyloid peptides and (c) in-tube SPME-LC-MS/MS using a organic monolithic capillary with cation-exchange groups with RAM covering for determination of homocysteine and homocysteic acid.For the evaluation of PD biomarkers, the microSALLE/LC-MS/MS method will be developed for the determination of anandamide (AEA) and 2-arachidonoylglycerol (2-AG) in parkinsonism-induced rats treated or not with levodopa (L-Dopa). Therefore, in agreement with the state-of-the-art of the miniaturized sample preparation and separation techniques, this project is noteworthy in the field of neuroscience. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
CARVALHO OLIVEIRA, IGOR GUSTAVO; DE SOUZA, ISRAEL DONIZETI; DO NASCIMENTO, GLAUCE CRIVELARO; DEL BEL, ELAINE; COSTA QUEIROZ, MARIA EUGENIA. In-tube solid-phase microextraction directly coupled to tandem mass spectrometry for anandamide and 2-arachidonoylglycerol determination in rat brain samples from an animal model of Parkinson's disease. Journal of Chromatography A, v. 1636, JAN 11 2021. Web of Science Citations: 0.

Please report errors in scientific publications list by writing to: cdi@fapesp.br.