Catecholamine analysis with microcolumn LC-peroxyoxalate chemiluminescence reaction detection.
BACKGROUND
Plasma catecholamines (CAs) are widely used as an index of sympathetic nervous system activity. In addition, CAs are known to be metabolized by catechol-O-methyltransferase (COMT) to produce their 3-O-methyl metabolites. We previously established a sensitive determination method of CAs and their 3-O-methyl metabolites using HPLC-peroxyoxalate chemiluminescence (POCL) reaction detection system. In this study, a microcolumn (100 x 1.0 mm I.D.) was used for separation to obtain higher sensitivity and shorter analysis time.


METHODS
The system included automated precolumn ion-exchange extraction of amines, followed by separation on an ODS column, coulometric oxidation, fluorescence derivatization with ethylenediamine, and finally POCL reaction detection.


RESULTS
The detection limits for CAs and their 3-O-methyl metabolites were 0.3-2.0 fmol. The analysis time was about 35 min, about half that of previously reported results. The method developed was used in monitoring changes in CAs and 3-O-methyl metabolite concentrations in human plasma during exercise.


CONCLUSIONS
The simultaneous determination method for concentrations of CAs and their 3-O-methyl metabolites in human plasma was developed using micro LC-peroxyoxalate chemiluminescence detection. We were successful in quantitating the changes in plasma CAs and their 3-O-methyl metabolites during exercise.
Tsunoda
Makoto
M
Laboratory of Bio-Analytical Chemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan. makotot@mol.f.u-tokyo.ac.jp
Nagayama
Masatoshi
M
Funatsu
Takashi
T
Hosoda
Saichi
S
Imai
Kazuhiro
K
D001794:Blood Pressure / Q000502:physiology; D002395:Catecholamines / Q000097:blood* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D015444:Exercise / Q000502:physiology; D006801:Humans; D008163:Luminescent Measurements / Q000295:instrumentation / Q000379:methods*; D010070:Oxalates / Q000737:chemistry; D015203:Reproducibility of Results; D013997:Time Factors
D002395:Catecholamines; D010070:Oxalates; C044622:peroxyoxalate