Simultaneous Estimation Amitriptyline Hydrochloride and Chlordiazepoxide by HPLC Method

Seerla Eswaraiah, Shaik Mohammed Yusuf*, T. Naga Mohan, D. Chinna Raju, E. Vijay Kumar
Srinivasa Institute of Pharmaceutical Sciences, Proddatur, YSR Kadapa, A.P. India

A B S T R A C T
A simple, economic, selective, precise, and accurate Reverse Phase High Performance Liquid Chromatography method for analysis of Amitriptyline HCl & Chlordiazepoxide in tablet dosage form was developed and validated according to ICH guidelines. The quantification of the drug was carried out by using Agilent Zorbin C18 (250 X 4.6 mm, 5μ) column its equivalent in isocratic mode and maintain column at 30°C, using mobile phase comprising of potassium dihydrogen phosphate buffer: Acetonitrile in the ratio of 50:50s v/v with a flow rate of 1.0ml/min and the detection wavelength was carried at 274 nm. The retention time for Amitriptyline HCl & Chlordiazepoxide was found to be 1.975min & 2.773min. The percent assay was found to be 100% & 99%.  The method was validated and the response was found to be linear in the drug concentration range of 50µg/ml to150µg/ml for Amitriptyline HCl and 50µg/ml to 150µg/ml for Chlordiazepoxide. The LOD and LOQ for Amitriptyline HCl were found to be 2.53 µg/ml, 8.44 µg/ml respectively. The LOD and LOQ for Chlordiazepoxide were found to be 1.961 µg/ml, 6.53 µg/ml respectively. This method was found to be good percentage recovery for Amitriptyline HCl and Chlordiazepoxide were found to be 100.00 and 100.00 respectively indicates that the proposed method is highly accurate. The specificity of the method shows good correlation between retention times of standard with the sample so, the method specifically determines the analyte in the sample without interference from excipients of tablet dosage forms. The method was successfully applied to Amitriptyline Hydrochloride and Chlordiazepoxide combination Tablet dosage form.
Keywords: Amitriptyline HCl, Chlordiazepoxide, Zorbin C18, Acetonitrile, Tablet.