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

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 deficithyperkinetic 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.

Urinary Neurotransmitters Are Selectively Altered in Children With Obstructive Sleep Apnea and Predict Cognitive Morbidity

“…21 It also is unclear how factors other than OSA (eg, medications and supplements or even other concomitant disorders such as attention-defi cit/hyperactivity disorder) that alter neurotransmitter levels in the CNS will affect neurotransmitters in the urine and vice versa. [22][23][24] Further limitations of the present study include a relatively small number of subjects, particularly among those who underwent neurocognitive testing; the fact that none of the control subjects were tested for cognitive function; the absence of a post hoc validation cohort for confi rmatory purposes; and, fi nally, the need for determination of the effects of treatment on the changes reported in this study.…”

Development and validation of a specific and sensitive LC-MS/MS method for quantification of urinary catecholamines and application in biological variation studies

“…Additionally, analytical specificity has been generally neglected in method development and very limited interfering compounds have been tested so far [27][28][29][30][31][32]. A variety of substances, described in Table 3, did not affect the catecholamine analysis (<6 % percent difference).…”

“…Liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) is rapidly emerging as a new technology for catecholamine quantification after employing different sample purification methods [26][27][28][29][30][31][32]. Selective quantification of catecholamines in the presence of drugs that commonly interfere with HPLC analysis could be achieved by diphenylborate (PBA)-based liquid-liquid extraction or solid phase extraction (SPE) with individual Bond Elut Plexa cartridge followed by subsequent LC-MS/MS analysis [27,28].…”

Glass/PDMS hybrid microfluidic device integrating vertically aligned SWCNTs to ultrasensitive electrochemical determinations

“…Serotonin has been determined by several techniques, which integrate separation methods-such as liquid chromatography [36][37][38] and capillary electrophoresis 39 -or direct assays using potentiometry 27,29,40 and sensor-based applications. 41,42 In general, as verified in Table 1, our set up shows a LOD lower than those reported in the literature even when a preconcentration step was employed.…”