A sensitive and rapid HPLC-ECD method for the simultaneous analysis of norepinephrine, dopamine, serotonin and their primary metabolites in brain tissue.
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.
Alburges
M E
ME
Department of Pharmaceutical Sciences, North Dakota State University, College of Pharmacy, Fargo 58105.
Narang
N
N
Wamsley
J K
JK
D000818:Animals; D015306:Biogenic Monoamines / Q000032:analysis*; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000032:analysis; D004563:Electrochemistry; D007202:Indicators and Reagents; D008297:Male; D009638:Norepinephrine / Q000032:analysis; D051381:Rats; D012701:Serotonin / Q000032:analysis
D015306:Biogenic Monoamines; D007202:Indicators and Reagents; D012701:Serotonin; D004298:Dopamine; D009638:Norepinephrine