In vivo on-line HPLC-microdialysis: simultaneous detection of monoamines and their metabolites in awake freely-moving rats.
An on-line microbore high performance liquid chromatographic--electrochemical (HPLC--EC) detection method has been developed for the simultaneous measurement of dopamine (3,4-dihydroxyphenethylamine, DA), its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC), and homovanillic acid (3-methoxy-4-hydroxyphenylacetic acid, HVA), as well as serotonin (5-hydroxytryptamine, 5-HT) and its metabolise 5-hydroxyindole-3-acetic acid (5-HIAA) in the striatum of awake and freely moving rats. Our method was capable of detecting DOPAC, 5-HIAA, HVA, DA and 5-HT at retention time of 3, 5, 6.5, 9, and 24 min, respectively, in repeated on-line microdialysis sampling. Analysis was performed using a 150 x 1 mm 5 microm C18 microbore column attached directly to a thin-layer radial flow electrochemical cell (UniJet) comprising a 6 mm glassy carbon working electrode. In order to accommodate signal outputs (due to a varying level in the neurochemicals under investigation), an amperometric detector was equipped with a sensitivity programmable controlling software for automatically switching sensitivity inattentively and repeatedly for each collection cycle.
Chaurasia
C S
CS
Department of Pharmacy and Administrative Sciences, College of Pharmacy and Allied Health Professions, St. John's University, Jamaica, New York 11439, USA.
Chen
C E
CE
Ashby
C R
CR
D015102:3,4-Dihydroxyphenylacetic Acid / Q000032:analysis / Q000378:metabolism; D000097:Acetonitriles / Q000737:chemistry; D000818:Animals; D015306:Biogenic Monoamines / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D003342:Corpus Striatum / Q000378:metabolism; D004298:Dopamine / Q000032:analysis / Q000378:metabolism; D004563:Electrochemistry; D005260:Female; D006719:Homovanillic Acid / Q000032:analysis / Q000378:metabolism; D006897:Hydroxyindoleacetic Acid / Q000032:analysis / Q000378:metabolism; D017551:Microdialysis / Q000295:instrumentation / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012701:Serotonin / Q000032:analysis / Q000378:metabolism; D014851:Wakefulness
D000097:Acetonitriles; D015306:Biogenic Monoamines; D015102:3,4-Dihydroxyphenylacetic Acid; D012701:Serotonin; D006897:Hydroxyindoleacetic Acid; D004298:Dopamine; D006719:Homovanillic Acid; C032159:acetonitrile