Identification of Suvorexant in Urine Using Liquid Chromatography-Quadrupole/Time-of-Flight Mass Spectrometry (LC-Q/TOF-MS)

J Anal Toxicol. 2017 Apr 1;41(3):224-229. doi: 10.1093/jat/bkw132.


Suvorexant (Belsomra®) is a new hypnotic drug with a novel mechanism of action. In prescribed doses of 10 mg before bedtime, the drug produces rapid onset of sleep by inhibiting the orexin neurons of the arousal system, promoting decreased wakefulness. Suvorexant is a potent and highly selective dual orexin receptor antagonist. Sedative hypnotics are of forensic importance due to their widespread use, potential for additive effects with other central nervous system depressants, impairing effects and potential for misuse. In this report we describe a highly sensitive assay for the identification and quantification of suvorexant in urine. Suvorexant was isolated using liquid/liquid extraction (LLE) and identified using liquid chromatography-quadrupole/time-of-flight mass spectrometry. Suvorexant was quantified using a quadratic calibration model between 5 and 250 ng/mL (R2 = 1.000, n = 6). Processed sample stability was demonstrated for up to 24 h. The limit of detection was 0.5 ng/mL and the limit of quantification (LOQ) was 5 ng/mL. The accuracy, bias and precision of the assay at the LOQ were 99% (81-117%), -1% and 12% (n = 18). Intraassay (n = 5) and interassay (n = 15) precision (% CV) at 10, 50 and 200 ng/mL were ≤8%, and bias ranged from -2% to 4% (98-104% accuracy). No qualitative interferences were detected from matrix, internal standard or 50 common drugs. Matrix effects evaluated at low and high concentrations were -16% and -9%, respectively, and produced CVs of 11% and 5% (n = 20). Suvorexant is a new drug of forensic importance. In this report we describe how a simple acidic/neutral LLE can be used to isolate this lipophilic drug with high recoveries and sound analytical performance.

MeSH terms

  • Azepines / chemistry
  • Azepines / urine*
  • Calibration
  • Chromatography, Liquid / methods*
  • Forensic Toxicology / instrumentation
  • Forensic Toxicology / methods*
  • Humans
  • Limit of Detection
  • Liquid-Liquid Extraction
  • Mass Spectrometry / methods*
  • Molecular Structure
  • Orexin Receptor Antagonists / chemistry
  • Orexin Receptor Antagonists / urine*
  • Reproducibility of Results
  • Sleep Aids, Pharmaceutical / chemistry
  • Sleep Aids, Pharmaceutical / urine*
  • Triazoles / chemistry
  • Triazoles / urine*


  • Azepines
  • Orexin Receptor Antagonists
  • Sleep Aids, Pharmaceutical
  • Triazoles
  • suvorexant