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Clonazepam quantification in human plasma by high-performance liquid chromatography coupled with electrospray tandem mass spectrometry in a bioequivalence study

✍ Scribed by Luiz E. Cavedal; Fabiana D. Mendes; Claudia C. Domingues; Anil K. Patni; Tausif Monif; Simrit Reyar; Alberto dos S. Pereira; Gustavo D. Mendes; Gilberto De Nucci

Publisher
John Wiley and Sons
Year
2007
Tongue
English
Weight
244 KB
Volume
42
Category
Article
ISSN
1076-5174

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✦ Synopsis

Abstract

A rapid, sensitive and specific method for quantifying clonazepam in human plasma using diazepam as the internal standard (IS) is described. The analyte and the IS were extracted from plasma by liquid‐liquid extraction using a hexane/diethylether (20 : 80, v/v) solution. The extracts were analysed by high‐performance liquid chromatography coupled with electrospray tandem mass spectrometry (HPLC‐MS‐MS). Chromatography was performed on a Jones Genesis C8 4 µm analytical column (100 × 2.1 mm i.d.). The method had a chromatographic run time of 3.0 min and a linear calibration curve over the range 0.5–50 ng/ml (r2 > 0.9965). The limit of quantification was 0.5 ng/ml. This HPLC/MS/MS procedure was used to assess the bioequivalence of two clonazepam 2 mg tablet formulations (clonazepam test formulation from Ranbaxy Laboratories Ltd and Rivotril from Roche Laboratórios Ltda as standard reference formulation). Copyright © 2006 John Wiley & Sons, Ltd.

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