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Development of an LC-MS/MS method for the analysis of serotonin and related compounds in urine and the identification of a potential biomarker for attention deficit hyperactivity/hyperkinetic disorder

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Abstract

Serotonin is a major neurotransmitter and affects various functions both in the brain and in the rest of the body. It has been demonstrated that altered serotinergic function is implicated in various psychiatric disorders including depression and schizophrenia. Serotonin has also been implicated along with dopamine in attention deficit–hyperkinetic disorder (AD-HKD). This study provides a versatile validated method for the analysis of serotonin, hydroxyindole acetic acid and dopamine in urine using LC-MS/MS. This method was then used to quantify these analytes in a test group of 17 children diagnosed with severe AD-HKD. This group was compared to a matched control group to investigate the possibility that one of these compounds may be a potential biomarker for this condition. The developed method provided good linear calibration curves for the multiplex assay of analytes in urine (0.05–3.27 nmol/L; R 2 ≥ 0.9977). Acceptable inter-day repeatability was achieved for all analytes with RSD values (n = 9) ranging from 1.1% to 9.3% over a concentration range of 0.11–3.27 μmol/L in urine. Excellent limits of detection (LOD) and limits of quantitation (LOQ) were achieved with LODs of 8.8–18.2 nmol/L and the LOQs of 29.4–55.7 nmol/L for analytes in urine. Recoveries were in the ranges of 98–104%, 100–106% and 91–107% for serotonin, 5-HIAA and dopamine, respectively. An appropriate sample clean-up procedure for urine was developed to ensure efficient recovery and reproducibility on analysis. Evaluation of matrix effects was also carried out and the influence of ion suppression on analytical results reported. Confirmatory analysis was carried out on a linear trap quadrupole-Orbitrap mass spectrometer to obtain high mass accuracy data of the target analytes in the clinical samples.

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Acknowledgments

We gratefully acknowledge funding from the Irish Research Council for Science, Engineering and Technology (IRCSET) funding M. Moriarty. The Council of Directors, Technological Sector Research-Strand III 2006 Grant Scheme, awarded to Dr A. Furey is also acknowledged for funding the formation of the ‘Team Elucidate’ research group. The Higher Education Authority (Programme for Research in Third-Level Institutions, Cycle 4 (PRTLI IV)) National Collaboration Programme on Environment and Climate Changes: Impacts and Responses is also acknowledged. Thanks also to Dr Paul Corcoran for advice regarding statistical analysis.

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Authors and Affiliations

  1. Team Elucidate, Department of Chemistry, Cork Institute of Technology, Cork, Ireland

    Merisa Moriarty & Ambrose Furey

  2. Department of Biological Sciences, Cork Institute of Technology, Bishopstown, Cork, Ireland

    Aoife Lee & Brendan O’Connell

  3. Child and Adolescent Mental Health Services, Health Service Executive, South, North Cork Area and the National Suicide Research Foundation, Cork, Ireland

    Ann Kelleher & Helen Keeley

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  1. Merisa Moriarty
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  2. Aoife Lee
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Correspondence to Ambrose Furey.

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Moriarty, M., Lee, A., O’Connell, B. et al. Development of an LC-MS/MS method for the analysis of serotonin and related compounds in urine and the identification of a potential biomarker for attention deficit hyperactivity/hyperkinetic disorder. Anal Bioanal Chem 401, 2481–2493 (2011). https://doi.org/10.1007/s00216-011-5322-7

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  • DOI: https://doi.org/10.1007/s00216-011-5322-7

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