Abstract
Author(s): Ravisankar P, Anusha Rani Kand Pavan G
The present work describes a simple, sensitive, accurate and rapid reverse phase high
Performance liquid chromatographic method for simultaneous estimation and validation of
Atenolol (ATN) and Hydrochlorothiazide (HCTZ) in tablet dosage form. The chromatographic
separation was accomplished on Welchrom RP-C18 Column (250 mm X 4.6 mm; 5μm), Shimadzu
LC-20AT Prominence Liquid Chromatograph and with a mixture of 10 mM Phosphate buffer (pH
3.0): acetonitrile (50:50, v/v). The flow rate was fixed at 1.0 mL/minute and the entire separation
was carried out at an ambient temperature for the HPLC system, the analysis was performed
using Shimadzu SPD-20A Prominence UV-Visible detector at 235 nm. The anti-hypertensive
agents, ATN and HCTZ were separated within 4 minutes. ATN and HCTZ showed retention times
of 2.257 min and 3.367 min respectively. The calibration plots were linear over the concentration
range of 5-25 μg/mL for Atenolol (r2 = 0.9999) and 2.5-12.5 μg/mL for Hydrochlorothiazide (r2 =
0.9999). The method was very sensitive with regard to LOD and LOQ for Atenolol and
Hydrochlorthiazide were found to be 0.0296 μg/mL and 0.6028 μg/mL; and 0.0899 μg/mL and
1.8269 μg/mL respectively. The method was statistically validated for its linearity, precision,
accuracy, specificity, Robustness and ruggedness. The optimized method proved to be specific,
robust and accurate for the quality control of ATN and HCTZ in bulk drug and pharmaceutical
formulations.