Simple and Highly Sensitive UPLC-ESI-MS/MS Assay for Rapid Determination of Suvorexant in Plasma

Suvorexant is a dual orexin receptor antagonist, recently approved by USFDA for the treatment of insomnia. It is drug-of-abuse and listed in Schedule IV drug of the Controlled Substances Act. In this study, a simple and highly sensitive UPLC-MS/MS assay was developed and fully validated for the determination of suvorexant in rat plasma. Both suvorexant and internal standard (rivaroxaban; IS) were separated on Aquity BEHTM C18 column after the extraction form plasma using diethyl ether as extracting agent. An isocratic mobile phase, consisting of acetonitrile and 10mM ammonium acetate in composition ratio of 85: 15 was eluted at flow rate of 0.3 mL/min. Both suvorexant and IS were eluted within one min with total run time of 1.5 min only. The ionization was performed on electrospray ionization interface in positive mode by multiple reaction monitoring. Precursor to product ion transition of 451.12 > 104.01 for qualifier and 451.12 > 186.04 for quantifier were used for suvorexant whereas 436.10 > 144.93 for IS, respectively. The calibration curves in plasma were linear in the concentration range of 0.33-200 ng/mL (r(2) >= 0.995) having limit of detection and limit of quantification of 0.10 and 0.33 ng/mL, respectively. All the validation parameters results were found to be within the acceptable limits according to the "Scientific Working Group for Forensic Toxicology (SWGTOX) guidelines" and can be implemented for forensic analysis.