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1 Acta Chromatographica

1 2011

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Optimization and validation of a new high-performance liquid chromatographic method for analysis of acetazolamide in tablets using Box-Behnken statistical experiment design

Ihssane B, Charrouf M, Abourriche A, Abboud Y, Bouabidi A, Bennamara A, Saffaj T

Acta Chromatographica

2011

Vol. 23, no. 1

41--57

This paper describes the optimization and validation of an analytical method for HPLC analysis of acetazolamide in tablets using Box-Behnken design. This multivariate approach enables substantial improvement of chromatographic performance using fewer experiments, without additional cost of columns or other equipment. By use of quadratic regression analysis, equations were developed describing the behavior of the response as simultaneous functions of the selected independent variables. Accordingly, the optimum conditions were determined. For quality-control samples these were use of a C18 column with acetonitrile-phosphoric acid buffer solution (0.01 M, pH 7.1) 15:85 (ν/ν) as mobile phase at a flow rate of 1.0 mL min-1 Diode-array detection was performed at 266 nm. The method was validated, in accordance with ICH guidelines, for accuracy, precision (intra-day and inter-day coefficient of variation <2.0%), selectivity, and linearity (R2 = 0.9993) over the concentration range 10–300 μg mL. -1 The method is simple, rapid, sensitive, and accurate and the retention time is less than 4 min. The method is therefore suitable for routine quality-control monitoring of acetazolamide in tablets.