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A simple and rapid HPLC‐UV method for the determination of retigabine in human plasma
Author(s) -
Franco Valentina,
Baruffi Katia,
Gatti Giuliana,
Marchiselli Roberto,
Fattore Cinzia,
Canevini Maria Paola,
Crema Francesca,
Perucca Emilio
Publication year - 2018
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.4168
Subject(s) - chromatography , chemistry , phosphoric acid , calibration curve , high performance liquid chromatography , extraction (chemistry) , detection limit , acetonitrile , methanol , elution , chromatographic separation , quantitative analysis (chemistry) , analytical chemistry (journal) , organic chemistry
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 C 18 Chromolith 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 −1 concentration 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.