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Simultaneous Analysis of Irbesartan and Hydrochlorothiazide: An Improved HPLC Method with the Aid of a Chemometric Protocol
Institute for Pharmaceutical Chemistry, University of Belgrade-Faculty of Pharmacy, Vojvode Stepe 450, 11 000 Belgrade, Serbia
Institute for Pharmaceutical Chemistry, University of Tuzla-Faculty of Pharmacy, Univerzitetska 8, 18 000 Tuzla, Bosnia and Herzegovina
Institute of Otorhinolaryngology, Clinical Centre of Serbia, Pasterova 2, 11 000 Belgrade, Serbia
* Author to whom correspondence should be addressed.
Received: 23 February 2012; in revised form: 12 March 2012 / Accepted: 13 March 2012 / Published: 16 March 2012
Abstract: Experimental design method was used for HPLC determination of irbesartan and hydrochlorothiazide in combined dosage forms. The traditional approach for optimization of experiments is time-consuming, involves a large number of runs and does not allow establishing the multiple interacting parameters. The main advantages of the experimental design method include the simultaneous screening of a larger number of factors affecting response and the estimation of possible interactions. On the basis of preliminary experiments, three factors-independent variables were selected as inputs (methanol content, pH of the mobile phase and temperature) and as dependent variables, five responses (resolution, symmetry of irbesartan peak, symmetry of hydrochlorothiazide peak, retention factor of irbesartan and retention factor of hydrochlorothiazide) were chosen. A full 23 factorial design, where factors were examined at two different levels (“low” and “high”) was used to determine which factors had an effect on the studied response. Afterwards, experimental design was used to optimize these influent parameters in the previously selected experimental domain. The novelty of our method lies in the optimization step accomplished by Derringer¢s desirability function. After optimizing the experimental conditions a separation was conducted on a Supelcosil C18 (150 mm × 4.6 mm, 5 mm particle size) column with a mobile phase consisting of methanol-tetrahydrofuran-acetate buffer 47:10:43 v/v/v, pH 6.5 and a column temperature of 25 °C. The developed method was successfully applied to the simultaneous separation of these drug-active compounds in their commercial pharmaceutical dosage forms.
Keywords: HPLC; experimental design; irbesartan; hydrochlorothiazide
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Vujić, Z.; Mulavdić, N.; Smajić, M.; Brborić, J.; Stankovic, P. Simultaneous Analysis of Irbesartan and Hydrochlorothiazide: An Improved HPLC Method with the Aid of a Chemometric Protocol. Molecules 2012, 17, 3461-3474.
Vujić Z, Mulavdić N, Smajić M, Brborić J, Stankovic P. Simultaneous Analysis of Irbesartan and Hydrochlorothiazide: An Improved HPLC Method with the Aid of a Chemometric Protocol. Molecules. 2012; 17(3):3461-3474.
Vujić, Zorica; Mulavdić, Nedžad; Smajić, Miralem; Brborić, Jasmina; Stankovic, Predrag. 2012. "Simultaneous Analysis of Irbesartan and Hydrochlorothiazide: An Improved HPLC Method with the Aid of a Chemometric Protocol." Molecules 17, no. 3: 3461-3474.