Development of a liquid chromatography-tandem mass spectrometry method for assaying cenobamate in rat plasma

Abstract

Cenobamate is a candidate drug that is being evaluated in a phase 3 clinical trial as an epilepsy treatment. In the present study, we developed and validated a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for assaying cenobamate in rat plasma. Acetonitrile was used for plasma protein precipitation, whereas carisbamate was used as the internal standard. A Spursil C₁₈ column and 10 mM ammonium formate and acetonitrile (60:40, v/v) were used for chromatographic separation. Detection was done using a triple quadrupole mass spectrometer by multiple reaction monitoring at transitions of m/z 268.06 ➜ 198.00 for cenobamate and m/z 216.09 ➜ 198.10 for carisbamate. The standard curve (r = 0.9946) was linear over a concentration range of 10 – 5000 ng/mL. Intra- and inter-day precision values were <14.97 and 14.86%, respectively, whereas intra- and inter-day accuracy values were less than 3.37 and 7.13%, respectively. Matrix effect, extraction recovery, and process efficiency were 98.59, 93.97, and 92.58%, respectively. Furthermore, cenobamate remained stable in rat plasma samples following three freeze-thaw cycles, storage at room temperature for 6 h, long-term storage at −20 °C for 4 weeks. The LC-MS/MS method was successfully used for studying the pharmacokinetics of cenobamate in rats.

Graphical Abstract

Keywords:

• Cenobamate
• LC-MS/MS
• validation
• pharmacokinetics
• protein precipitation

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was financially supported by the research fund of Chungnam National University under Grant.