Detection of four different amino acid neurotransmitters in cultured rat neurons and the culture medium by precolumn derivatization high-performance liquid chromatography.
Neuroreport
; 27(7): 495-500, 2016 May 04.
Article
em En
| MEDLINE
| ID: mdl-26999360
We validated and used a high-performance liquid chromatography procedure for the determination of four different amino acid neurotransmitters in cultured rat neurons and used culture medium. Samples were derivatized using 2,4-dinitrofluorobenzene and the amino acids were separated on a C18 column. The method yielded good reproducibility and sensitivity for the quantification of the four free amino acid neurotransmitters, with average recovery factors of 80.25-118.43%, an intraday precision of 0.09-0.17%, and an interday precision of 0.62-0.74%. The assay method can be readily utilized as a precise, sensitive, and highly accurate method for the determination of concentrations of the four amino acid neurotransmitters in cultured rat neurons and used culture medium. Using the described methods, we found that the aspartate, glutamic acid, glycine, and γ-aminobutyric acid concentrations (µmol/g protein) in cultured rat neurons were 25.23±0.81, 35.16±0.32, 77.56±4.51, and 62.87±3.12, respectively, whereas their concentrations (µM) in the used culture medium were 18.18±0.82, 24.27±1.01, 107.18±9.56, and 35.78±2.98, respectively.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Cromatografia Líquida de Alta Pressão
/
Meios de Cultivo Condicionados
/
Aminoácidos
/
Neurônios
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Neuroreport
Assunto da revista:
NEUROLOGIA
Ano de publicação:
2016
Tipo de documento:
Article