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High-sensitive liquid chromatographic method for determination of neuronal release of serotonin, noradrenaline and dopamine monitored by microdialysis in the rat prefrontal cortex.
J Neurosci Methods. 2004 Dec 30; 140(1-2):163-8.JN

Abstract

A high-sensitive liquid chromatographic method based on precolumn derivatization and fluorescence detection allowing simultaneous determination of serotonin (5-HT), noradrenaline (NA) and dopamine (DA) in brain microdialysis samples is described. 5-HT, NA and DA were derivatized with benzylamine and 1,2-diphenylethylenediamine in the presence of potassium hexacyanoferrate(III) and glycine, which yielded to highly fluorescent and stable benzoxazoles. The derivatized samples were separated on a microbore column (150 mm x 1.0mm i.d., packed with C18 silica, 5 microm) within 60 min. The mobile phase consisted of acetonitrile-Briton-Robinson buffer (pH 7.2) (32:68, v/v) containing 5 mM Na2EDTA and 5 mM octanesulfonic acid sodium salt. The detection limits (signal-to-noise ratio of 3) for 5-HT, NA and DA were 76, 42 and 95 amol/10 microl injected on-column, respectively. Microdialysis samples were collected at 10-min intervals from the probes implanted in the prefrontal cortex of awake rats. The basal levels of 5-HT, NA and DA were 7.3 +/- 0.7, 5.3 +/- 0.31 and 8.1 +/- 0.47 fmol/5 microl (mean +/- S.E.M., n = 5). Following 90-min perfusion with tetrodotoxin (1 microM) or calcium-free Ringer solution, the DA and NA levels were reduced to about 15 and 20%, respectively and the 5-HT levels to 45 and 60% of the basal levels, respectively. Reserpine, 12h after a dose of 5mg/kg i.p., reduced the extracellular 5-HT, NA and DA concentrations to about 34, 39 and 32% of the basal levels, respectively. In conclusion, the preset microdialysis/analytical method enables simultaneous monitoring of basal and pharmacologically reduced neuronal release of 5-HT, NA and DA in the rat brain.

Authors+Show Affiliations

Department of Neuroscience, Karolinska Institutet, Retzius väg 8, S-171 77 Stockholm, Sweden.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

15589346

Citation

Yoshitake, Takashi, et al. "High-sensitive Liquid Chromatographic Method for Determination of Neuronal Release of Serotonin, Noradrenaline and Dopamine Monitored By Microdialysis in the Rat Prefrontal Cortex." Journal of Neuroscience Methods, vol. 140, no. 1-2, 2004, pp. 163-8.
Yoshitake T, Yoshitake S, Fujino K, et al. High-sensitive liquid chromatographic method for determination of neuronal release of serotonin, noradrenaline and dopamine monitored by microdialysis in the rat prefrontal cortex. J Neurosci Methods. 2004;140(1-2):163-8.
Yoshitake, T., Yoshitake, S., Fujino, K., Nohta, H., Yamaguchi, M., & Kehr, J. (2004). High-sensitive liquid chromatographic method for determination of neuronal release of serotonin, noradrenaline and dopamine monitored by microdialysis in the rat prefrontal cortex. Journal of Neuroscience Methods, 140(1-2), 163-8.
Yoshitake T, et al. High-sensitive Liquid Chromatographic Method for Determination of Neuronal Release of Serotonin, Noradrenaline and Dopamine Monitored By Microdialysis in the Rat Prefrontal Cortex. J Neurosci Methods. 2004 Dec 30;140(1-2):163-8. PubMed PMID: 15589346.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - High-sensitive liquid chromatographic method for determination of neuronal release of serotonin, noradrenaline and dopamine monitored by microdialysis in the rat prefrontal cortex. AU - Yoshitake,Takashi, AU - Yoshitake,Shimako, AU - Fujino,Kaoru, AU - Nohta,Hitoshi, AU - Yamaguchi,Masatoshi, AU - Kehr,Jan, PY - 2004/01/15/received PY - 2004/04/02/accepted PY - 2004/12/14/pubmed PY - 2005/4/13/medline PY - 2004/12/14/entrez SP - 163 EP - 8 JF - Journal of neuroscience methods JO - J Neurosci Methods VL - 140 IS - 1-2 N2 - A high-sensitive liquid chromatographic method based on precolumn derivatization and fluorescence detection allowing simultaneous determination of serotonin (5-HT), noradrenaline (NA) and dopamine (DA) in brain microdialysis samples is described. 5-HT, NA and DA were derivatized with benzylamine and 1,2-diphenylethylenediamine in the presence of potassium hexacyanoferrate(III) and glycine, which yielded to highly fluorescent and stable benzoxazoles. The derivatized samples were separated on a microbore column (150 mm x 1.0mm i.d., packed with C18 silica, 5 microm) within 60 min. The mobile phase consisted of acetonitrile-Briton-Robinson buffer (pH 7.2) (32:68, v/v) containing 5 mM Na2EDTA and 5 mM octanesulfonic acid sodium salt. The detection limits (signal-to-noise ratio of 3) for 5-HT, NA and DA were 76, 42 and 95 amol/10 microl injected on-column, respectively. Microdialysis samples were collected at 10-min intervals from the probes implanted in the prefrontal cortex of awake rats. The basal levels of 5-HT, NA and DA were 7.3 +/- 0.7, 5.3 +/- 0.31 and 8.1 +/- 0.47 fmol/5 microl (mean +/- S.E.M., n = 5). Following 90-min perfusion with tetrodotoxin (1 microM) or calcium-free Ringer solution, the DA and NA levels were reduced to about 15 and 20%, respectively and the 5-HT levels to 45 and 60% of the basal levels, respectively. Reserpine, 12h after a dose of 5mg/kg i.p., reduced the extracellular 5-HT, NA and DA concentrations to about 34, 39 and 32% of the basal levels, respectively. In conclusion, the preset microdialysis/analytical method enables simultaneous monitoring of basal and pharmacologically reduced neuronal release of 5-HT, NA and DA in the rat brain. SN - 0165-0270 UR - https://www.unboundmedicine.com/medline/citation/15589346/High_sensitive_liquid_chromatographic_method_for_determination_of_neuronal_release_of_serotonin_noradrenaline_and_dopamine_monitored_by_microdialysis_in_the_rat_prefrontal_cortex_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0165-0270(04)00293-6 DB - PRIME DP - Unbound Medicine ER -