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Derivatization chemistries for determination of serotonin, norepinephrine and dopamine in brain microdialysis samples by liquid chromatography with fluorescence detection
Author(s) -
Yoshitake T.,
Kehr J.,
Todoroki K.,
Nohta H.,
Yamaguchi M.
Publication year - 2006
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.560
Subject(s) - derivatization , chemistry , microdialysis , monoamine neurotransmitter , chromatography , dopamine , serotonin , biogenic amine , norepinephrine , fluorescence , 3,4 dihydroxyphenylacetic acid , high performance liquid chromatography , homovanillic acid , medicine , biochemistry , receptor , physics , extracellular , quantum mechanics
The present paper provides an overview on currently developed derivatization chemistries and techniques for determination of monoamine neurotransmitters serotonin (5‐HT), norepinephrine (NE) and dopamine (DA) in microdialysis samples by microbore liquid chromatography with fluorescence detection. In mild alkaline conditions, 5‐hydroxyindoles and catecholamines react with benzylamine (BA), forming highly fluorescent 2‐phenyl‐4,5‐pyrrolobenzoxazoles and 2‐phenyl(4,5‐dihydropyrrolo) [2,3‐ f ]benzoxazoles, respectively. However, for derivatization of DA a higher fluorescence intensity was achieved for reaction with 1,2‐diphenylethylenediamine (DPE) rather than with BA, therefore for simultaneous determination of 5‐HT, NE and DA in brain microdialysates, a two‐step derivatization with BA followed by DPE was developed. The detection limits for 5‐HT, NE and DA were 0.2, 0.08 and 0.13 fmol, respectively, in an injection volume of 20 µL, which corresponds to concentrations of 30, 12 and 19.5 p m , respectively in standard solution prior to derivatization. The experimental data presented demonstrate the ability of the technique to simultaneously monitor neuronally releasable pools of monoamine neurotransmitters in the rat and mouse brains at basal conditions and following pharmacological treatments or physiological stimuli. These techniques play an important role in drug discovery and clinical investigation of psychiatric and neurological diseases such as depression, schizophrenia and Parkinson's disease. Copyright © 2005 John Wiley & Sons, Ltd.