Fast liquid chromatography separation and multiple‐reaction monitoring mass spectrometric detection of neurotransmitters

Hammad, Loubna A.; Neely, Matthew; Bridge, Bob; Mechref, Yehia

doi:10.1002/jssc.200900158

Cited by 14 publications

(7 citation statements)

References 38 publications

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“…Furthermore, intra- and inter-assay variations are reduced, which increases the reproducibility of results. LC-MS/MS methods without automated sample preparation [27, 28] have sensitivities and specificities that can be compared with the XLC-MS/MS method described here and are increased when compared to conventional HPLC techniques. The supplementary advantage compared to other LC-MS/MS methods is that our method is completely automated, which means a great reduction in total analysis time including sample clean-up.…”

Section: Discussionmentioning

confidence: 99%

Automated mass spectrometric analysis of urinary and plasma serotonin

et al. 2010

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Serotonin emerges as crucial neurotransmitter and hormone in a growing number of different physiologic processes. Besides extensive serotonin production previously noted in patients with metastatic carcinoid tumors, serotonin now is implicated in liver cell regeneration and bone formation. The aim was to develop a rapid, sensitive, and highly selective automated on-line solid-phase extraction method coupled to high-performance liquid chromatography–tandem mass spectrometry (XLC-MS/MS) to quantify low serotonin concentrations in matrices such as platelet-poor plasma and urine. Fifty microliters plasma or 2.5 μL urine equivalent were pre-purified by automated on-line solid-phase extraction, using weak cation exchange. Chromatography of serotonin and its deuterated internal standard was performed with hydrophilic interaction chromatography. Mass spectrometric detection was operated in multiple reaction monitoring mode using a quadrupole tandem mass spectrometer with positive electrospray ionization. Serotonin concentrations were determined in platelet-poor plasma of metastatic carcinoid patients (n = 23) and healthy controls (n = 22). Urinary reference intervals were set by analyzing 24-h urine collections of 120 healthy subjects. Total run-time was 6 min. Intra- and inter-assay analytical variation were <10%. Linearity in the 0–7300 μmol/L calibration range was excellent (R2 > 0.99). Quantification limits were 30 and 0.9 nmol/L in urine and plasma, respectively. Platelet-poor serotonin concentrations in metastatic carcinoid patients were significantly higher than in controls. The urinary reference interval was 10–78 μmol/mol creatinine. Serotonin analysis with sensitive and specific XLC-MS/MS overcomes limitations of conventional HPLC. This enables accurate quantification of serotonin for both routine diagnostic procedures and research in serotonin-related disorders.

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Section: Discussionmentioning

confidence: 99%

Automated mass spectrometric analysis of urinary and plasma serotonin

et al. 2010

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“…Simultaneous bioanalysis of monoamines in blood plasma were done with UPLC/MSMS according to the previously described method (Hammad et al ., 2009) with minor modifi-cations. To 50 μl aliquots of plasma, 10 μl of 100 ng/ml dopamine- d4 (as internal standard) was added.…”

Section: Methodsmentioning

confidence: 99%

Impact of High Fat Diet-induced Obesity on the Plasma Levels of Monoamine Neurotransmitters in C57BL/6 Mice

Kim

Bae

Lim

2013

Biomolecules and Therapeutics

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Obesity is one of the most serious health problems in developed countries. It negatively affects diverse aspects of human wellbeing. Of these, a relationship between obesity and depression is widely recognized but biomarkers for assessment of obesityassociated mood changes in animal obesity models are rarely known. Here we explored the link between obesity and the plasma levels of monoamine neurotransmitters involved in mood control using a sensitive UPLC/MSMS technique in high fat diet (HFD)- induced obesity model in male C57BL/6 mice to explore the potential utility of plasma tests for obesity-associated mood change. HFD (60% of total calories, 8 weeks) induced significantly higher weight gains in body (+37.8%) and fat tissue (+306%) in male C57BL/6 mice. Bioanalysis of serotonin, dopamine and norepinephrine in plasma at 8 weeks of HFD revealed that serotonin decreased significantly in the obese mice when compared to normal diet-fed mice (2.7 ± 0.6 vs 4.3 ± 2.0 ng/ml, N=8). Notably, a negative correlation was found between the levels of serotonin and body weight gains. Furthermore, principal component analysis (PCA) with the individual levels of neurotransmitters revealed that plasma levels of dopamine and serotonin could apparently differentiate the obese mice from lean ones. Our study demonstrated that blood plasma levels of neurotransmitters can be employed to evaluate the mood changes associated with obesity and more importantly, provided an important clue for understanding of the relationship between obesity and mood disorders.

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“…Therefore, other detection modes, such as mass spectrometry (MS), have been used (Perry et al, 2009;Uutela et al, 2009). MS has recently made a big impact on the determination of this type of compounds (Bourcier et al, 2006;Tareke et al, 2007;Eckstein et al, 2008;Hammad et al, 2009;Cai et al, 2010). MS provides several advantages over conventional methods, because structural information can be given, and moreover, better sensitivity and selectivity can be achieved (Keski-Rahkonen et al, 2007;Buck et al, 2009;de Jong et al, 2010).…”

Section: Introductionmentioning

confidence: 99%