Determination of dopamine norepinephrine, serotonin and their major metabolic products in rat brain by reverse-phase ion-pair high performance liquod chromatography with electrochemical detection

https://doi.org/10.1016/0091-3057(85)90490-3Get rights and content

Abstract

A method is described for the separation and quantitation of catecholamines, serotonin, and their major metabolites with use of reverse-phase, ion-pair liquid chromatography with electrochemical detection. This method employs columns packed with a microparticulate C-18 resin, octyl sodium sulfate as the ion-pairing agent, and isocratic elution with a citrate-phosphate buffer containing methanol. Conditions are described for the separation of norepinephrine, normetanephrine, 3-methoxy-4-hydroxyphenylglycol, epinephrine, metanephrine, dopamine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, serotonin, and 5-hydroxyindole-3-acetic acid and for their quantitation in extracts of rat brain tissue.

Cited by (63)

  • Novel and sensitive reversed-phase high-pressure liquid chromatography method with electrochemical detection for the simultaneous and fast determination of eight biogenic amines and metabolites in human brain tissue

    2014, Journal of Chromatography A
    Citation Excerpt :

    HPLC combined with ECD (HPLC-ECD) is still considered the method of choice given its potential as a specific and sensitive technique capable of automation, its low cost of analysis, and often simpler sample pretreatment requirements in comparison to for example HPLC with FD [5]. Many applications using ion-exchange reversed-phase HPLC with ECD (RP-HPLC-ECD) have been developed for the determination of these endogenous molecules in biological samples [6–11], but often only a limited number of compounds could be analysed in a single chromatographic run. We previously reported the development, optimization and validation of an ion-exchange reversed-phase HPLC with ECD method to determine eight biogenic amines and metabolites in mouse brain tissue [12].

  • Polymelamine modified edge plane pyrolytic graphite sensor for the electrochemical assay of serotonin

    2014, Talanta
    Citation Excerpt :

    The determination of 5-HT is informative in the diagnosis of various diseases and hence makes it the subject of biologically and pharmacologically oriented research [23]. Several techniques have been employed for the determination of serotonin such as high performance liquid chromatography, capillary electrophoresis, mass and fluorescent spectrometry, and capillary liquid chromatography coupled with mass spectrometry [24–28]. However, most of these techniques require heavy and expensive instruments with sophisticated process, tedious time consuming pretreatment and derivatization.

  • Innovative green supercritical fluid chromatography development for the determination of polar compounds

    2012, Journal of Chromatography A
    Citation Excerpt :

    Polar compounds have been widely studied by many chromatographic techniques. The most common method is reverse-phase liquid chromatography in conjunction with ion-pairing reagents to improve the retention of polar compounds [12–14]. These mobile phases contain at least one organic solvent, such as acetonitrile or methanol, and one ion-pairing agent such as trifluoroacetic acid (TFA), sodium octanesulfonate or hexanesulfonic acid.

  • Biogenic amines and their metabolites in mouse brain tissue: Development, optimization and validation of an analytical HPLC method

    2010, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
    Citation Excerpt :

    Also, a miniaturized LC system (column size: 150 mm × 1 mm, C18, 3 μm, flow rate: 0.05 ml/min) has replaced the conventional system (column size: 250 mm × 4.6 mm; C18, 5 or 10 μm, flow rate: 1 ml/min), yielding higher mass sensibility, requiring a small injection volume, significantly reducing mobile phase consumption rendering HPLC–ECD one of the most suitable methods for analysis of biogenic amines and metabolites [26]. Simultaneous determination of biogenic amines and their metabolites in brain tissue using HPLC–ECD, thought straightforward and feasible, seemed poorly achievable and limited to 2–5 compounds in previous studies [27–35]. This is most likely due to the difficulty in association of very early eluting of MHPG peak and very late eluting peak of 5-HT in the same chromatogram.

View all citing articles on Scopus
View full text