Improved high-performance liquid chromatographic method for GABA and glutamate determination in regions of the rodent brain.
A C18 reversed-phase column and isocratic fluorescence HPLC method for the simultaneous detection of glutamate and gamma-aminobutyric acid (GABA) is described. In this article a fast and more efficient method for the extraction of these neurotransmitters in rat brain tissue is also presented. The supernatant was derivatized with o-phthalaldehyde (OPA) and analyzed by HPLC with fluorescence detection. Intraday reproducibility was 97.0% and 96.7% and interday reproducibility was 97.1% and 93.7% for GABA and glutamate, respectively. Recovery assays indicate that the accuracy of the method for GABA is 99.6+/-2.3% and for glutamate is 101.9+/-1.8%. In addition, the time consumed to run a sample is lower than that described by other authors. Mean elution time was 3.10 min and 8.22 min for glutamate and GABA, respectively. Thus, in a total runtime of less than 9 min both neurotransmitters were detected. Moreover, when compared to the current methods, the extraction solution used here allowed a high drawing out of the neurotransmitters, glutamate and GABA, from the hippocampus, thalamus and prefrontal cortex of the rat brain.
de Freitas Silva
Danielle Marra
DM
Departamento de Bioquímica e Imunologia, Laboratório de Neurociência Comportamental e Molecular - LaNeC. Universidade Federal de Minas Gerais, Brazil.
Ferraz
Vany P
VP
Ribeiro
Angela Maria
AM
D000818:Animals; D001921:Brain / Q000033:anatomy & histology / Q000378:metabolism*; D001923:Brain Chemistry / Q000502:physiology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D018698:Glutamic Acid / Q000032:analysis* / Q000378:metabolism; D008297:Male; D009448:Neurochemistry / Q000379:methods*; D017729:Presynaptic Terminals / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D009435:Synaptic Transmission / Q000502:physiology; D013997:Time Factors; D005680:gamma-Aminobutyric Acid / Q000032:analysis* / Q000378:metabolism; D009764:o-Phthalaldehyde / Q000737:chemistry
D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid; D009764:o-Phthalaldehyde