Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jan 23;11(2):109.
doi: 10.3390/toxics11020109.

UPLC-MS/MS Based Identification and Quantification of a Novel Dual Orexin Receptor Antagonist in Plasma Samples by Validated SWGTOX Guidelines

Muzaffar Iqbal  1 , Abdullah Alshememry  2 , Faisal Imam  3 , Mohd Abul Kalam  2 , Ali Akhtar  4 , Essam A Ali  1
Affiliations

UPLC-MS/MS Based Identification and Quantification of a Novel Dual Orexin Receptor Antagonist in Plasma Samples by Validated SWGTOX Guidelines

Muzaffar Iqbal et al. Toxics. .

Abstract

Lemborexant (LEM) is a novel dual orexin receptor antagonist (DORA), recently approved for the treatment of insomnia. As with other DORAs, LEM has potential of abuse and therefore placed in Schedule IV class by the United States Drug Enforcement Administration (USDEA). In this study, a sensitive and accurate UPLC-MS/MS assay was developed for the quantification of LEM in human plasma sample using losartan as an internal standard (IS). The chromatographic separation was performed by using gradient elution of mobile phase, comprising of 10 mM ammonium acetate and acetonitrile with a flow rate of 0.3 mL/min. An Acquity UPLC BEH C18 (1.7 μm, 2.1 × 50 mm) column was used for separation of LEM and IS by maintaining the oven temperature of 40 °C. The electrospray ionization in positive mode was used for sample ionization. The precursor to product ion transition of 411.12 > 175.09 (qualifier) and 411.1 > 287.14 (quantifier) was used for detection and quantification of LEM, respectively, in multiple reaction monitoring mode. Being a drug of abuse, the assay was validated according to "Scientific Working Group for Toxicology" (SWGTOX) guidelines, including limit of detection (LOD), limit of quantification (LOQ), precision and bias, calibration model, interferences, carry-over effects, matrix effects, and stability parameters. The LOD and LOQ of the assay were 0.35 and 1.0 ng/mL, respectively. The linear range was between 1-300 ng/mL with correlation coefficient of ≥0.995. The method was also cross validated in rat plasma samples with acceptable ranges of precision and accuracy before its application for pharmacokinetic study in rats.

Keywords: DORA; Lemborexant; SWGTOX; UPLC-MS/MS; abuse; insomnia.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Chemical structure of LEM (a) and losartan (b).
Figure 2
Figure 2
Representative LEM mass spectra of precursor ion (a) and precursor to product ion (b) in ESI positive mode.
Figure 3
Figure 3
Representative MRM chromatogram of LEM and IS in blank plasma (a) and plasma fortified at LOQ level concentration (b). [411.12 > 175.09 → Qualifier ion; 411.12 > 287.14 → Quantifier ion; 423.1 > 207.1 → IS].
Figure 4
Figure 4
Plasma concentration versus time profile of LEM (a) and representative MRM chromatogram of LEM and IS (b) in rat after oral administration of 10 mg/kg LEM. [411.12 > 175.09 → Qualifier ion; 411.12 > 287.14 → Quantifier ion; 423.1 > 207.1 → IS].
See this image and copyright information in PMC

References

    1. Chung K.F., Yeung W.F., Ho F.Y., Yung K.P., Yu Y.M., Kwok C.W. Cross-cultural and comparative epidemiology of insomnia: The Diagnostic and statistical manual (DSM), International classification of diseases (ICD) and International classification of sleep disorders (ICSD) Sleep Med. 2015;16:477–482. doi: 10.1016/j.sleep.2014.10.018. - DOI - PubMed
    1. Asnis G.M., Thomas M., Henderson M.A. Pharmacotherapy Treatment Options for Insomnia: A Primer for Clinicians. Int. J. Mol. Sci. 2015;17:50. doi: 10.3390/ijms17010050. - DOI - PMC - PubMed
    1. Sateia M.J., Buysse D.J., Krystal A.D., Neubauer D.N., Heald J.L. Clinical Practice Guideline for the Pharmacologic Treatment of Chronic Insomnia in Adults: An American Academy of Sleep Medicine Clinical Practice Guideline. J. Clin. Sleep Med. 2017;13:307–349. doi: 10.5664/jcsm.6470. - DOI - PMC - PubMed
    1. Kumar A., Chanana P., Choudhary S. Emerging role of orexin antagonists in insomnia therapeutics: An update on SORAs and DORAs. Pharmacol. Rep. 2016;68:231–242. doi: 10.1016/j.pharep.2015.09.002. - DOI - PubMed
    1. Centre for Drug Evaluation and Research US FDA Approves Eisai’s DAYVIGOtm (Lemborexant) for the Treatment of Insomnia in Adult Patients. NDA 212028 Multi-Disciplinary Review and Evaluation DAYVIGO (Lemborexant) [(accessed on 10 December 2022)];2018 Available online: https://www.accessdata.fda.gov/drugsatfda_docs/nda/2019/212028Orig1s000M....