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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


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)

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Scientific publications (12)
(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)
GRECCO, CAROLINE FERNANDES; TUMAS, VITOR; HALLAK, JAIME EDUARDO CECILIO; QUEIROZ, MARIA EUGENIA COSTA. In-tube solid-phase microextraction (sulfopropyl methacrylate-co-ethylene glycol dimethacrylate monolithic capillary) coupled to LC-MS/MS to determine beta amyloid peptides in Alzheimer's cerebrospinal fluid samples. SEPARATION SCIENCE PLUS, v. N/A, p. 12-pg., . (17/02147-0, 20/00126-8, 19/04386-7)
GRECCO, CAROLINE FERNANDES; DE SOUZA, ISRAEL DONIZETI; OLIVEIRA, IGOR GUSTAVO CARVALHO; QUEIROZ, MARIA EUGENIA COSTA. In-Tube Solid-Phase Microextraction Directly Coupled to Mass Spectrometric Systems: A Review. SEPARATIONS, v. 9, n. 12, p. 19-pg., . (17/02147-0, 19/19485-0, 20/00126-8, 20/06526-8, 19/04386-7)
CRUZ, JONAS CARNEIRO; DE SOUZA, ISRAEL DONIZETI; LANCAS, FERNANDO MAURO; COSTA QUEIROZ, MARIA EUGENIA. Current advances and applications of online sample preparation techniques for miniaturized liquid chromatography systems. Journal of Chromatography A, v. 1668, p. 16-pg., . (19/19485-0, 17/02147-0, 20/00126-8)
SOUZA, ISRAEL D.; OLIVEIRA, IGOR G. C.; QUEIROZ, MARIA EUGENIA C.. Innovative extraction materials for fiber-in-tube solid phase microextraction: A review. Analytica Chimica Acta, v. 1165, . (20/00126-8, 18/06068-0, 17/02147-0, 19/19485-0)
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, . (19/19485-0, 18/06068-0, 20/00126-8, 14/50891-1)
CRUZ, JONAS CARNEIRO; DE SOUZA, ISRAEL DONIZETI; GRECCO, CAROLINE FERNANDES; FIGUEIREDO, EDUARDO COSTA; QUEIROZ, MARIA EUGENIA COSTA. Recent advances in column switching high-performance liquid chromatography for bioanalysis. SUSTAINABLE CHEMISTRY AND PHARMACY, v. 21, . (19/19485-0, 19/04386-7, 20/00126-8, 17/02147-0)
COSTA QUEIROZ, MARIA EUGENIA; DE SOUZA, ISRAEL DONIZETI; DE OLIVEIRA, IGOR GUSTAVO; GRECCO, CAROLINE FERNANDES. In vivo solid phase microextraction for bioanalysis. TRAC-TRENDS IN ANALYTICAL CHEMISTRY, v. 153, p. 18-pg., . (20/06526-8, 17/02147-0, 19/04386-7, 19/19485-0, 20/00126-8)
GRECCO, CAROLINE FERNANDES; DE SOUZA, ISRAEL DONIZETI; QUEIROZ, MARIA EUGENIA COSTA. Novel materials as capillary coatings for in-tube solid-phase microextraction for bioanalysis. JOURNAL OF SEPARATION SCIENCE, v. 44, n. 8, p. 1662-1693, . (17/02147-0, 20/00126-8, 19/04386-7, 19/19485-0)
PINTO, MANIA A. L.; DE SOUZA, ISRAEL D.; MIRANDA, LUIS FELIPPE C.; QUEIROZ, MARIA EUGENIA C.. A New Molecularly Imprinted Polymer for In-Tube SPME/UHPLC-MS/MS of Anandamide in Plasma Samples. Journal of the Brazilian Chemical Society, v. N/A, p. 9-pg., . (13/03808-9, 15/07619-1, 17/02147-0, 20/00126-8, 19/19485-0)
DE SOUZA, ISRAEL DONIZETI; QUEIROZ, MARIA EUGENIA COSTA. Advances in sample preparation and HPLC-MS/MS methods for determining amyloid-beta peptide in biological samples: a review. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, v. 415, n. 18, p. 19-pg., . (17/02147-0, 19/19485-0, 20/00126-8)

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