Simple and rapid validated HPLC-fluorescence determination of perampanel in the plasma of patients with epilepsy

Susan Mohamed; Carmina Candela; Roberto Riva; Manuela Contin

doi:10.1016/j.plabm.2017.11.003

Simple and rapid validated HPLC-fluorescence determination of perampanel in the plasma of patients with epilepsy

Pract Lab Med. 2017 Nov 26:10:15-20. doi: 10.1016/j.plabm.2017.11.003. eCollection 2018 Mar.

Authors

Susan Mohamed¹, Carmina Candela^{1

2}, Roberto Riva^{1

2}, Manuela Contin^{1

2}

Affiliations

¹ IRCCS-ISNB Institute of Neurological Sciences of Bologna, Italy.
² Department of Biomedical and Neuromotor Sciences University of Bologna, Bologna, Italy.

Abstract

We present a simple and fast high-performance liquid chromatography method with fluorescence detection for the determination of the antiepileptic drug perampanel in human plasma. The chromatographic separation was performed on a Kinetex PFP (100 × 2.6 mm, 4.6 µm) column, using a mobile phase of sodium acetate 0.03 M pH 3.7 and acetonitrile (40/60, v/v), at a flow rate of 0.8 mL/min. Total chromatography time for each run was 5 min. Sample preparation (250 µL) involved only one simple precipitation step by acetonitrile spiked with mirtazapine as internal standard. The method was validated over a concentration range of 20-1000 ng/mL and successfully applied to measure perampanel concentrations in plasma samples obtained from patients with epilepsy. This assay combines the high specificity of fluorescence detection with a very simple and fast sample pretreatment and can offer real advantages over existing methods in terms of simplicity and transferability to a therapeutic drug monitoring setting.

Keywords: AED, antiepileptic drug; AMPA, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid; Antiepileptic drugs; CBZ, carbamazepine; Clinical pharmacokinetics; Epilepsy; HPLC-F; HPLC-F, high performance liquid chromatography-fluorescence detector; IS, internal standard; LLE, liquid-liquid extraction; LLOD, lower limit of detection; LLOQ, lower limit of quantification; MIR, mirtazapine; MS/MS, tandem mass spectrometer; OXC, oxcarbazepine; PER, perampanel; PHT, phenytoin; Perampanel; QC, quality control; TDM, therapeutic drug monitoring; VPA, valproic acid.