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Development and validation of a simple, rapid and sensitive LC-MS/MS method for the measurement of urinary neurotransmitters and their metabolites

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Abstract

Neurotransmitters play crucial roles in physiological functions and their imbalances have demonstrated association in the pathology of several diseases. The measurement of neurotransmitters possesses a great potential as a significant clinical tool. This study presents the development and validation of an LC-MS/MS method for simultaneous quantification of multi-class neurotransmitters associated with dopamine, tryptophan and glutamate-γ-aminobutyric acid pathways. A total of ten neurotransmitters and their metabolites (dopamine, epinephrine, metanephrine, tryptophan, serotonin, kynurenic acid, kynurenine, anthranilic acid, GABA, glutamic acid) were determined based on a simple and rapid ‘dilute and shoot’ method using minimal urine volume. The chromatographic separation was achieved using a Poroshell 120 Bonus-RP LC Column in combination with a gradient elution within an 8.5-min time frame. The method exhibited good sensitivity as the limits of quantification ranged between 0.025 and 0.075 μg/mL with acceptable matrix effects (< ± 14.5%), no carryover and good linearity (r 2 > 0.98). The accuracy and precision for all analytes were within tolerances, at < ± 9.9% mean relative error (MRE) and < 8.6% relative standard deviation (RSD), respectively. The method was successfully applied in measuring the neurotransmitter concentrations in urine of healthy donors. Furthermore, the undertaken stability experiments indicated that acidified urine specimens allowed the analytes to be stable for prolonged durations in comparison to those untreated. The study also reveals the performance of the method is unaffected by the absence of expensive deuterated reference standards under the experimental conditions employed which further simplifies the analytical procedures and provides a significant cost saving for running the assay.

The quantification of multi-class neurotransitters associated with the dopamine, tryptophan and GABA-glutamate pathways using a simple ‘dilute and shoot’ LC-MS/MS method.

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Authors and Affiliations

  1. Centre for Forensic Science, University of Technology Sydney, PO Box 123, Broadway, Ultimo, NSW, 2007, Australia

    Jingya Yan & Shanlin Fu

  2. Climate Change Cluster, University of Technology Sydney, PO Box 123, Broadway, Ultimo, NSW, 2007, Australia

    Unnikrishnan Kuzhiumparambil

  3. Department of Clinical Biochemistry, The Children’s Hospital at Westmead, Locked Bag 4001, Westmead, NSW, 2145, Australia

    Sushil Bandodkar

  4. School of Psychology and Illawarra Health and Medical Research Institute, University of Wollongong, Wollongong, NSW, 2522, Australia

    Nadia Solowij

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  1. Jingya Yan
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  4. Nadia Solowij
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  5. Shanlin Fu
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Corresponding author

Correspondence to Shanlin Fu.

Ethics declarations

Ethics approval for collecting human urine samples was obtained from the UTS Human Research Ethics Committee (Ethics Approval No. UTS HREC 2010268A). Informed consent was obtained from all donors who provided urine samples.

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The authors declare that they have no conflict of interest.

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Yan, J., Kuzhiumparambil, U., Bandodkar, S. et al. Development and validation of a simple, rapid and sensitive LC-MS/MS method for the measurement of urinary neurotransmitters and their metabolites. Anal Bioanal Chem 409, 7191–7199 (2017). https://doi.org/10.1007/s00216-017-0681-3

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  • DOI: https://doi.org/10.1007/s00216-017-0681-3

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