An automatic analyzer for catecholamines and their 3-O-methyl metabolites using a micro coulometric flow cell as a postcolumn reactor for fluorogenic reaction.
A coulometric flow cell for a miniaturized LC system was developed. The cell was examined, as 3-O-methyl catecholamines were converted to their relative omicron-quinones for subsequent fluorometric and chemiluminescence detection. Its performance was evaluated in comparison with commercially available amperometric and coulometric detectors in terms of specification of the low dead volume and high conversion efficiency. The fully automated small-bore LC analyzer for simultaneous determination of catecholamines and their 3-O-methyl metabolites included precolumn pretreatment, column switching, column separation, postcolumn oxidative conversion, fluorometric derivatization, and chemiluminescence detection. The detection limits were 0.3-2.0 fmol for catecholamines and their 3-O-methyl metabolites. Because of the high sensitivity, the required volume of rat plasma sample was only 15 microL.
Takezawa
K
K
Graduate School of Pharmaceutical Sciences and Graduate School of Engineering, The University of Tokyo, Japan.
Tsunoda
M
M
Watanabe
N
N
Imai
K
K
D000818:Animals; D002138:Calibration; D002395:Catecholamines / Q000032:analysis* / Q000097:blood / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004563:Electrochemistry; D005470:Fluorometry; D008163:Luminescent Measurements; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley
D002395:Catecholamines