Abstract
Purpose: This paper presents an improved kinetic-spectrophotometric procedure for determining clonazepam (CZP) in pharmaceutical formulations and human serum.Methods: The method is based on ligand-exchange reaction. The reaction was followed spectrophotometrically by measuring the rate of change of absorbance at 425 nm in ethanolic sodium hydroxide solution.Results: The optimum operating conditions for reagent concentrations and temperature were established. Linear calibration curve was obtained in the range of 0.32 - 4.10 μg mL-1. The optimized conditions yielded a theoretical detection limit of 0.24 μg mL-1 based on the 3.3So criterion, where S0 is standard deviation of the calibration line. The interference of certain drugs, foreign ions and amino acids on the reaction rate were studied in order to assess the selectivity of the method.Conclusion: The developed method is sensitive, accurate and reproducible and could be used for routine anlysis of clonazepam in pharmaceutical preparations and serum samples.Keywords: Clonazepam, Ligand-exchange reaction, Kinetic spectrometry, Validation, Pharmaceutical preparation, Human serum
Highlights
Clonazepam is an anticonvulsant agent widely used in the treatment of epilepsy in adults and children [1]
Several methods for the assay of this compound have been reported by many authors, e.g., spectrometric methods [4,5,6], potentiometric methods [7,8], voltammetry [9], polarography [10], high-pressure liquid chromatography (HPLC) [10,11,12], gas chromatography coupled with mass spectrometry (GC-MS) [11,13]
The mixture was centrifuged at 3500 rpm for 5 min, filtered through a 0.45 μm membrane filter (Millipore) directly into a 50 mL volumetric flask and made up to volume with ethanol to obtain a solution of theoretical CZP concentration of 40.0 μg mL-1
Summary
Clonazepam is an anticonvulsant agent widely used in the treatment of epilepsy in adults and children [1]. As a result of the therapeutic importance and widespread use of this compound, the literature contains many reports dealing with its determination. The British Pharmacopoeia [2] recommends non-aqueous titration with perchloric acid for the determination of clonazepam. For dosage forms, high-pressure liquid chromatography (HPLC) is recommended by US Pharmacopoeia [3]. Several methods for the assay of this compound have been reported by many authors, e.g., spectrometric methods [4,5,6], potentiometric methods [7,8], voltammetry [9], polarography [10], HPLC [10,11,12], gas chromatography coupled with mass spectrometry (GC-MS) [11,13]
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