An easy-to-use liquid chromatography method with fluorescence detection for the simultaneous determination of five neuroactive amino acids in different regions of rat brain.

摘要: Abstract Introduction: To comprehend the normal function and pathological characteristics of certain neurological disorders it is important to evaluate neuroactive amino acids levels in animal models. Methods: This work describes a simple liquid chromatography-fluorescence detection (LC-FLD) method for simultaneous determination aspartic acid (Asp), glutamic (Glu), glutamine (Gln), taurine (Tau) γ-aminobutyric (GABA), using methyl-L-arginine as internal standard, samples rat brain tissue. The five target analytes (Asp, Glu, Gln, Tau GABA) were determined single chromatographic run less 11 min after derivatization step with o-phthalaldehyde. derivatives separated on reversed-phase C18 column detected by fluorescence at excitation emission wavelengths 340 448 nm, respectively. Results: was validated accordance international guidelines bioanalytical methods validation presented limits quantification range 25–50 ng mL−1 calibration curves coefficients (r2) equal or higher than 0.9920. In addition, precision (coefficient variation, %) accuracy (bias, meet established criteria, stability sample handling storage conditions demonstrated. Discussion: Unlike other similar assays, current diluted biological matrix, which advantageous order ensure process integrity. Moreover, this LC-FLD successfully applied compounds interest different tissue regions (frontal cortex, amygdala, hippocampus, cerebellum striatum). Thus, assay represents useful tool support multiple nonclinical studies field neurosciences, requiring quantitative profiling pattern analysis acids.