Rapid Determination of Norepinephrine, Dopamine, Serotonin, Their Precursor Amino Acids, and Related Metabolites in Discrete Brain Areas of Mice Within Ten Minutes by HPLC with Electrochemical Detection

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Abstract
A rapid and simple chromatographic procedure using HPLC with electrochemical detection is described for simultaneous determination of the substrates from precursor amino acids to metabolites related to synthesis and metabolism of three monoamine neurotransmitters—norepinephrine (NE), dopamine (DA), and 5‐hydroxytryptamine (5‐HT, serotonin)—in discrete brain areas of the mouse. Under the present instrumental and mobile phase conditions, the procedure permits simultaneous determination of three monoamines (NE, DA, and 5‐HT), two precursor amino acids (tyrosine and tryptophan), and four respective metabolites (3‐methoxy‐4‐hydroxyphenylglycol, 3,4‐dihydroxyphenylacetic acid, homovanillic acid, and 5‐hydroxyindoleacetic acid) within 10 min in one chromato graphic run. By varying column temperature, this procedure also permits simultaneous determination of 10–14 monoamine‐related substrates including the nine substrates described above within 15–21 min. The validity of the present procedure is demonstrated by analyzing the effect of an α2‐adrenergic agonist (clonidine) and an α2‐antagonist (yohimbine) in mouse hypothalamus.