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Determination of neurotransmitters and their metabolites using one- and two-dimensional liquid chromatography with acidic potassium permanganate chemiluminescence detection

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Abstract

High-performance liquid chromatography with chemiluminescence detection based on the reaction with acidic potassium permanganate and formaldehyde was explored for the determination of neurotransmitters and their metabolites. The neurotransmitters norepinephrine and dopamine were quantified in the left and right hemispheres of rat hippocampus, nucleus accumbens and prefrontal cortex, and the metabolites vanillylmandelic acid, 3,4-dihydrophenylacetic acid, 5-hydroxyindole-3-acetic acid and homovanillic acid were identified in human urine. Under optimised chemiluminescence reagent conditions, the limits of detection for these analytes ranged from 2.5 × 10−8 to 2.5 × 10−7 M. For the determination of neurotransmitter metabolites in urine, a two-dimensional high-performance liquid chromatography (2D-HPLC) separation operated in heart-cutting mode was developed to overcome the peak capacity limitations of the one-dimensional separation. This approach provided the greater separation power of 2D-HPLC with analysis times comparable to conventional one-dimensional separations.

2D-HPLC separation and permanganate chemiluminescence detection of neurotransmitter metabolites

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Authors and Affiliations

  1. Centre for Chemistry and Biotechnology, School of Life and Environmental Sciences, Deakin University, 75 Pigdons Road, Waurn Ponds, 3216, Victoria, Australia

    Brendan J. Holland, Xavier A. Conlan, Paul G. Stevenson, Neil W. Barnett, Jacqui L. Adcock & Paul S. Francis

  2. School of Psychology, Deakin University, 221 Burwood Highway, Burwood, 3125, Victoria, Australia

    Susannah Tye & Ashlie Reker

Authors
  1. Brendan J. Holland
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  2. Xavier A. Conlan
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  3. Paul G. Stevenson
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  4. Susannah Tye
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  5. Ashlie Reker
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  6. Neil W. Barnett
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  7. Jacqui L. Adcock
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  8. Paul S. Francis
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Corresponding authors

Correspondence to Xavier A. Conlan or Paul S. Francis.

Additional information

Published in the topical collection Analytical Bioluminescence and Chemiluminescence with guest editors Elisa Michelini and Mara Mirasoli.

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Holland, B.J., Conlan, X.A., Stevenson, P.G. et al. Determination of neurotransmitters and their metabolites using one- and two-dimensional liquid chromatography with acidic potassium permanganate chemiluminescence detection. Anal Bioanal Chem 406, 5669–5676 (2014). https://doi.org/10.1007/s00216-013-7514-9

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  • DOI: https://doi.org/10.1007/s00216-013-7514-9

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