Determination of suvorexant in human plasma using 96-well liquid-liquid extraction and HPLC with tandem mass spectrometric detection

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
S A BreidingerEric J Woolf

Abstract

A method, using liquid chromatography with tandem mass spectrometric detection (LC-MS/MS), was developed for the determination of suvorexant (MK-4305, Belsomra(®)), a selective dual orexin receptor antagonist for the treatment insomnia, in human plasma over the concentration range of 1-1000ng/mL. Stable isotope labeled (13)C(2)H3-suvorexant was used as an internal standard. The sample preparation procedure utilized liquid-liquid extraction, in the 96-well format, of a 100μL plasma sample with methyl t-butyl ether. The compounds were chromatographed under isocratic conditions on a Waters dC18 (50×2.1mm, 3μm) column with a mobile phase consisting of 30/70 (v/v %) 10mM ammonium formate, pH3/acetonitrile at a flow rate of 0.3mL/min. Multiple reaction monitoring of the precursor-to-product ion pairs for suvorexant (m/z 451→186) and (13)C(2)H3-suvorexant (m/z 455→190) on an Applied Biosystems API 4000 tandem mass spectrometer was used for quantitation. Intraday assay precision, assessed in six different lots of control plasma, was within 10% CV at all concentrations, while assay accuracy ranged from 95.6 to 105.0% of nominal. Quality control (QC) samples in plasma were stored at -20°C. Initial within day analysis of QCs after one fre...Continue Reading

References

Apr 9, 2011·Journal of Neurogenetics·Christopher J WinrowJohn J Renger
Oct 2, 2012·ACS Chemical Neuroscience·Corey R Hopkins

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Citations

Nov 9, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Mariah Carson, Sarah Kerrigan
Jan 5, 2018·Journal of Analytical Toxicology·Brian WatersShin-Ichi Kubo
Apr 5, 2017·Journal of Analytical Toxicology·Muzaffar IqbalKhalid A Al-Rashood

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