Determination of catecholamines and endogenous related compounds in rat brain tissue exploring their native fluorescence and liquid chromatography

摘要: The profiling analysis of catecholamines and their metabolites in brain tissue offers a crucial key to understand functions the body opportunity follow up neural diseases. A rapid simple liquid chromatography-fluorescence detection (LC-FLD) method was developed validated for simultaneously measuring several endogenous related compounds rat samples. target analytes measured this bioanalytical assay were levodopa (L-DOPA), dopamine (DA), norepinephrine (NE), epinephrine (E), 3-O-methyldopa (3-O-MD), homovanillic acid (HVA), being 3,4-dihydroxybenzylamine (DHBA) used as internal standard (IS). six (L-DOPA, DA, NE, E, 3-O-MD HVA) can be determined single chromatographic run less than 12min, all (analytes IS) detected using native fluorescence monitored at excitation/emission wavelengths 279nm/320nm, respectively. conditions experimentally optimized then validation parameters (linearity, limits quantification detection, precision accuracy, recovery, stability selectivity) examined. In accordance with international guidelines Food Drug Administration European Medicines Agency described herein exhibited range 2-25ngmL-1, linearity wide concentration ranges (r2≥0.994), acceptable (coefficient variation ≤8.76%) accuracy (bias ±14.65%) levels. Since procedure does not involve pre-purification or derivatization sample, absolute recovery found around 100%. Moreover, LC-FLD successfully applied determination interest samples different regions (cerebellum, amygdala, cortex, hippocampus, striatum, mesencephalon, medulla oblongata, substantia nigra ventral tegmental area). Hence, represents valuable tool support pre(non)clinical studies broad field neurosciences, requiring quantitative these bioamines metabolites.