Development of a monolithic phase with amino groups and nanoparticles for the determination of carbidopa, levodopa, benserazide, dopamine and 3-O-methyldopa in plasma samples from patients with Parkinson\'s disease by HILIC-MS / MS in column switching mode

Author(s)

Caroline Fernandes Grecco

• Document type: Master's Dissertation
• Press: Ribeirão Preto.
• Institution: Universidade de São Paulo (USP). Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto (PCARP/BC)
• Defense date: 2019-06-14

Examining board members

Maria Eugenia Queiroz Nassur; Yassuko Iamamoto; Dosil Pereira de Jesus; Eduardo Carasek da Rocha

• Advisor: Maria Eugenia Queiroz Nassur

Abstract

Parkinson\'s disease (PD) is an idiopathic neurodegenerative disease that affects about 3% of the Brazilian population aged over 65 years. It can also be seen in individuals less than 40 years old (Parkinsonism). PD is caused by deficiency of the neurotransmitter dopamine in the nigrostriatal and cortical pathway, compromising mainly in the motor system. Levodopa is an efficient prodrug (decarboxylated by the aromatic amino acid decarboxylase in dopamine) to control PD symptoms. Levodopa (LD) has been associated with peripheral dopa decarboxylase inhibitors, such as carbidopa or benserazide, to increase the availability of LD in the central nervous system and to reduce the adverse effects of dopamine formed in the peripheral system. Prolonged PD treatment with LD can lead to fluctuations in motor performance and dyskinesia. We have developed and characterized a hybrid organic-inorganic monolithic column containing amino groups and mesoporous nanoparticles and used it in the first dimension in the column-switching mode to determine carbidopa, levodopa, benserazide, dopamine, and 3-O-methyldopa in plasma samples from PD patients by employing hydrophilic interaction liquid chromatography coupled to tandem mass spectrometry (HILIC-MS/MS). We chemically and morphologically characterized the hybrid organic-inorganic monolithic column containing amino groups and mesoporous nanoparticles by Fourier-transform Infrared Spectroscopy (FTIR), Scanning Electron Microscopy (SEM), and Transmission Electron Microscopy (TEM). FTIR analysis indicated that the functional groups were incorporated into the monolithic phase structure, whereas the SEM and TEM analyses respectively confirmed the in-situ polymerization (the monolithic phase is chemically bound to the capillary inner surface e) and the hexagonal morphology of the synthesized nanoparticles. The HILIC-MS/MS method in the column-switching mode based on the hybrid monolithic column containing amino groups and mesoporous nanoparticles in the first dimension showed adequate linearity with concentrations ranging from LOQ to 2.00 ug mL-1 and coefficients of determination (R2) greater than 0.9910. The intra and interassay accuracy values (EPR) ranged from -10.97% to 12.29% and -9.33% to 10.41%, respectively. The intra and interassay precision values (CV) ranged from 3,45% to 13.35% and 7.65% to 17.61%, respectively. The method presented LOQ values of 33 ng mL-1 for carbidopa, 22 ng mL-1 for levodopa, 170 ng mL-1 for benserazide, 1.2 ng mL-1 for dopamine, and 10 ng mL-1 for 3-O-methyldopa. According to the evaluated analytical validation parameters, the HILIC-MS/MS method in the column-switching mode can adequately determine carbidopa, levodopa, benserazide, dopamine, and 3-O-methyldopa in plasma samples from PD patients (AU)

• FAPESP's process: 17/03726-3 - Development of a monolithic phase with amino groups and addition of nanoparticles for the determination of carbidopa, levodopa, Benserazide, dopamine and 3-O-methyldopa in plasma samples of patients with Parkinson's Disease by MEPS/HILIC-MS/MS
• Grantee: Caroline Fernandes Grecco
• Support Opportunities: Scholarships in Brazil - Master