Analytical method development and validation for simultaneous estimation of clonidine hcl and chlorthalidone in bulk and tablet dosage form by rp hplc method

Abstract

A simple, accurate and precise HPLC method for simultaneous determination of Clonidine HCl and Chlorthalidone in pure and tablet dosage form has been developed.HPLC of Waters (Model: Alliance 2695) with Phenomenex Luna C18 (4.6 mm I.D. × 250 mm, 5 µm) column was used for chromatographic separation. It contains waters injector and PDA Detector (Deuterium). Mobile phase consists of Methanol: Water (65:35% v/v) and flow rate adjusted was 1ml/min. Wavelength selected for detection was 220nm and injection volume was 10 µl. By using the developed method, retention time of Clonidine HCl and Chlorthalidone was found to be 3.2min and 5.4min respectively. The method has been validated for linearity, accuracy and precision.  Linearity of Clonidine HCl and Chlorthalidone were in the range of 75–375μg/ml and 15–75μg/ml respectively. The percentage recoveries obtained for Clonidine HCl and Chlorthalidone were found to be in range of 99.3 – 99.6%. LOD and LOQ were found to be 12.5µg/ml and 38.1µg/ml for Clonidine HCl 3.7and 11.4µg/ml for Chlorthalidone. The developed HPLC method offers several advantages such as rapidity, usage of simple mobile phase and easy sample preparation steps. Further, improved sensitivity makes it specific and reliable for its intended use. Hence, this method can be applied for the analysis of pure drug and pharmaceutical dosage forms. From the present study it can be concluded that the proposed method is simple, sensitive, precise, specific, accurate andreproducible.Results ofvalidation parameters demonstrated that the analytical procedure is suitable for its intended purpose and meets the criteria defined in ICH Q2R1.