Title:Simple and Rapid LC-MS/MS Method for Determination of Perampanel
in Human Plasma and Application to Bioequivalence Study
Volume: 18
Issue: 10
Author(s): Pan Gao and Qiao-gen Zou*
Affiliation:
- Department of Biotechnology and Pharmaceutical Sciences, Nanjing Technology University, Nanjing 210009, PR China
Keywords:
Perampanel, LC-MS/MS, human plasma, bioequivalence study, protein precipitation, pharmacokinetic.
Abstract:
Background: Perampanel (PER) is a third-generation antiepileptic drug (AED). Several
methods have been developed for the quantification of perampanel in plasma. The pharmacokinetic
characteristics of perampanel in healthy Chinese ubjects have not been comprehensively
reported.
Objective: A simple, fast and sensitive LC-MS/MS method was established and validated for the
quantification of perampanel in human plasma and its application to a bioequivalence study.
Methods: Chromatographic separation was accomplished on a ZORBAX Eclipse XDB-Phenyl
column (4.6 mm × 75 mm, 3.5 μm) using a binary gradient with mobile phase (A) (water containing
5 mmol/L ammonium acetate and 0.1% formic acid and (B) acetonitrile-water (95:5, v/v) at a
flow rate of 0.9 mL/min and sample preparation was by one-step protein precipitation via acetonitrile.
Results: The total run time in this study was 4.5 min and the retention time of perampanel and
perampanel-d5 (internal standard) were 2.30 min and 2.32 min, respectively. The method was developed
and validated over the concentration range of 2.00-500 ng/mL for perampanel, with a
correlation coefficient greater than 0.9992. The inter-day precision was 3.1%-3.8% and accuracy
was 98.9%-103.5%. The intra-day precision was 2.4%-6.8% and the accuracy was 97.6%-
104.9%. The extraction recovery ranged from 99.23%-103.84% and the matrix effect was not
significant. Perampanel was proved to be stable in solution and human plasma under different
tested conditions. The validated method was successfully applied to a randomized, open-label, 2-
period, crossover bioequivalence study in healthy Chinese subjects, and the results indicated that
bioequivalence was achieved for 2 formulations of the 4-mg perampanel tablet under both fasting
and fed conditions, and both treatments were safe and well-tolerated by all study subjects.
Conclusion: The validated method was successfully applied to a bioequivalence study of perampanel
in human plasma and has achieved satisfactory results.
