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Development of the column switching SALLE-UHPLC-MS/MS method to anandamide and 2-arachidonoylglycerol determination in brain samples of rats submitted to an animal model of Parkinsons Disease

Grant number: 18/06068-0
Support Opportunities:Scholarships in Brazil - Master
Effective date (Start): June 01, 2018
Effective date (End): July 31, 2020
Field of knowledge:Physical Sciences and Mathematics - Chemistry - Analytical Chemistry
Acordo de Cooperação: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Maria Eugênia Queiroz Nassur
Grantee:Igor Gustavo de Carvalho Oliveira
Host Institution: Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (FFCLRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil
Associated research grant:17/02147-0 - Single drop chromatography and its coupling to mass spectrometry: instrumental strategies, development of materials, automation and analytical applications, AP.TEM


The endogenous compounds Anandamide (AEA) and 2-araquidonoilglycerol (2-AG), arachidonic acid derivatives, are part of a class of neurotransmitters called endocannabinoids (EC), that together with the cannabinoid cellular receptors (CB1 e CB2) and metabolic enzymes compose the endocannabinoid system (eCB). Some studies indicates that the endogenous levels of these ECs undergo changes in response of physiological, pathological and pharmacological stimuli, making the determination of its concentration in biological samples the subject of study in several pathologies, specially neurodegenerative disorders, such as Parkinson disease (PD). Experimental models in animals have been developed to mimic PD characteristics observed in human's clinic, usually the selective acute injection of neurotoxins, such as the dopamine hydroxylated analogous 6-hydroxydopamine (6OHDA) in rats, is used to create acute neurodegenerative lesions of the nigrostriatal pathways. The liquid chromatography tandem mass spectrometry has been considered the reference technique in endocannabinoids determination in biological samples. The sample preparation step has been required in the development of analytical methods, to eliminate interferes and pre-concentrate the analytes, most of the time present in trace levels in biological samples. The salting-out assisted liquid-liquid extraction (SALLE) is a extraction technique based in the salting-out effect, consisting in the reduction of the mutual miscibility between water and a water miscible organic solvent by the addition of an electrolyte. In consequence of the salt addition, the non-charged organic analytes present in the sample suffers solubility reduction in the water phase and are extracted to the organic phase. The column-switching liquid chromatography has been used to dynamic extraction and online analysis of analytes. In this project, the column-switching SALLE-UHPLC-MS/MS method will be optimized and validated to AEA and 2-AG determination in brain samples of rats induced to Parkinsonism, treated or not with L-dopa and doxycycline. (AU)

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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)
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)
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
OLIVEIRA, Igor Gustavo de Carvalho. Development of the SALLE-UHPLC-MS/MS method to determine anandamide and 2-arachidonoylglycerol in rat brain samples submitted to a Parkinson\'s Disease animal model. 2020. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PCARP/BC) Ribeirão Preto.

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