Development and Validation of a stability indicating method for the simultaneous determination of Atenolol and Hydrochlorothiazide by HPLC

Abstract

A simple, selective, rapid, precise and stability indicating high performance liquid chromatographic method was developed and validated for the simultaneous estimation of Atenolol and Hydrochlorothiazide in pharmaceutical Tablet dosage form.

Materials and Methods: The method involves the use of easily available inexpensive laboratory reagents.  The mobile phase consisted of 50:50 % (v/v) of Methanol & 0.1% v/v orthophosphoric acid operated on isocratic mode. The flow rate is 0.6 ml/min. Chromatographic separation of  Atenolol and Hydhrochlorothiazide was performed on Phenomenox Prodigy C₁₈ column (150 X 4.6 mm id, ODS 2, 5µm). The wavelength of detection is 271 nm. The injection volume is 20µL. The retention time of Atenolol and Hydhrochlorothiazide are 2.68 ± 0.10 minutes and 3.49 ± 0.10 respectively. The run time of analysis is 5 minutes.

Results: A linear response was observed over the concentration range 10.00-100.00 ?g/mL of Atenolol and the concentration range 1.00-10.00 ?g/mL of Hydrochlorothiazide. Limit of detection and limit of quantitation for Atenolol were 0.1?g/mL and 1 ?g/mL, and for Hydrochlorothiazide were 0.05 ?g/mL and 0.5 ?g/mL, respectively. The method was successfully validated in accordance to ICH guidelines acceptance criteria for linearity, accuracy, precision, specificity, robustness. The influence of acid, alkaline, oxidative Stress and photolytic stress conditions on both the drugs was studied. Results indicated complete degradation in alkaline medium for Atenolol and Hydrochlorothiazide

Conclusion: The analysis concluded that the method was selective for simultaneous estimation of Atenolol and Hydrochlorothiazide can be potentially used for the estimation of these drugs in combined dosage form.