A simple and rapid HPLC-UV method for the determination of retigabine in human plasma

Valentina Franco, Katia Baruffi, Giuliana Gatti, Roberto Marchiselli, Cinzia Fattore, Maria Paola Canevini, Francesca Crema, Emilio Perucca

Research output: Contribution to journalArticlepeer-review


A simple and rapid high-performance liquid chromatographic method with ultraviolet detection was developed for the quantitative determination of retigabine, known also as ezogabine, in human plasma. The assay uses a simple solid-phase extraction for sample preparation and direct injection of the extract into the chromatograph. Flupirtine is used as an internal standard. Chromatographic separation is achieved on a C18Chromolith column (Chromolith Performance, 100 × 4.6 mm i.d.), using as mobile phase water/acetonitrile/methanol (72:18:10 v/v/v) mixed with 0.1% of 85% phosphoric acid. Isocratic elution is conducted at a flow rate of 1.5 mL min-1. The total duration of a chromatographic run is 7 min. Calibration curves are linear over the 25-2000 ng mL-1concentration range, with a limit of quantitation of 25 ng mL-1. Other performance characteristics include high precision (intra- and inter-day coefficients of variation ≤12.6%) and high accuracy (99.7%-108.7%). The method is suitable for the investigation of concentration-response relationships in patients receiving therapeutic doses of retigabine.

Original languageEnglish
JournalBiomedical Chromatography
Publication statusE-pub ahead of print - Dec 12 2017


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