A sensitive and rapid HPLC‐ECD method for the simultaneous analysis of norepinephrine, dopamine, serotonin and their primary metabolites in brain tissue

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Abstract
This report details a very sensitive, rapid and accurate ion‐pair HPLC‐ECD method for the analysis of biogenic amines and their metabolites in brain tissue. The method described enables detection of picogram amounts of 3‐methoxy‐4‐hydroxy phenethyleneglycol (MHPG) (37 pg), 3,4‐dihydroxy phenylacetic acid (DOPAC) (18 pg), norepinephrine (NE) (12 pg), epinephrine (E) (6 pg), 5‐hydroxyindoleacetic acid (5‐HIAA) (18 pg), 3,4‐dihydroxyphenylethylamine (DA) (6 pg), 3‐methoxy‐4‐hydroxyphenylacetic acid (HVA) (12 pg) and 5‐hydroxytryptamine (5‐HT) (12 pg). The linearity of the method is from 18.75 ng/mL to 300 ng/mL for MHPG; 9.37 to 150 ng/mL for DOPAC and 5‐HIAA; 6.25 to 100 ng/mL for NE, HVA and 5‐HT; and from 3.12 to 100 ng/mL for E and DA. The reproducibility, expressed as coefficient of variance (CV%) within‐run and between‐run groups, was 2.25% and 19.49% for MHPG; 3.84% and 29.84% for DOPAC; 0.89% and 8.97% for NE; 1.26% and 5.61% for E; 1.07% and 28.77% for 5‐HIAA; 2.65% and 10.65% for DA; 5.97% and 24.38% for HVA; and 4.44% and 9.45% for 5‐HT.
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