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Updated: Aug 22, 2025

Semi-Targeted Ultra-High-Performance Chromatography Coupled to Mass Spectrometry Analysis of Phenolic Metabolites in Plasma of Elderly Adults
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Development and Validation of a UHPLC-MS/MS-Based Method to Quantify Cenobamate in Human Plasma Samples.

Bruno Charlier1,2, Albino Coglianese1,3, Francesca Felicia Operto1,2

  • 1Department of Medicine, Surgery and Dentistry "Scuola Medica Salernitana", University of Salerno, Baronissi, 84081 Salerno, Italy.

Molecules (Basel, Switzerland)
|November 11, 2022
PubMed
Summary

Cenobamate (CNB), a new antiseizure medication, was quantified in plasma using UHPLC-MS/MS. This method facilitated monitoring CNB levels in non-adult patients, leading to a seizure-free state at 150 mg/day.

Keywords:
UHPLC–MS/MSantiseizure medicationscenobamateliquid chromatographymass spectrometrytherapeutic drug monitoring

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Area of Science:

  • Pharmacology
  • Analytical Chemistry
  • Clinical Pharmacology

Background:

  • Cenobamate (CNB) is a novel antiseizure medication (ASM) approved for uncontrolled partial-onset seizures in adults.
  • Its exact mechanism of action is not fully elucidated, but it involves voltage-gated sodium channels and GABA ion channels.
  • CNB exhibits significant oral bioavailability and interacts with co-administered ASMs and CYP450 substrates, necessitating therapeutic drug monitoring.

Purpose of the Study:

  • To develop and validate a sensitive ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for quantifying cenobamate (CNB) in human plasma.
  • To apply the validated method for evaluating the pharmacokinetic profile of CNB in non-adult patients.
  • To assess the relationship between CNB dosage, plasma concentrations, and seizure control.

Main Methods:

  • Development and validation of a UHPLC-MS/MS analytical method for CNB quantification in human plasma, adhering to current guidelines.
  • Preliminary application of the method to monitor plasma CNB concentrations in two non-adult patients over a two-month period.
  • Pharmacokinetic parameter evaluation based on serial plasma sampling.

Main Results:

  • A validated UHPLC-MS/MS method was successfully established for accurate CNB measurement in plasma.
  • Preliminary pharmacokinetic data from two non-adult patients demonstrated a linear increase in plasma CNB concentrations with escalating dosage.
  • Both patients achieved a seizure-free state when maintained at a daily dose of 150 mg.

Conclusions:

  • The developed UHPLC-MS/MS method provides a reliable tool for therapeutic drug monitoring of cenobamate.
  • This study presents the first pharmacokinetic evaluation of CNB in non-adult patients, showing dose-proportional increases in plasma levels.
  • Achieving a seizure-free state at 150 mg/day highlights the potential efficacy of CNB in this population, warranting further investigation.