Simultaneous estimation of amitryptyline and chlordiazepoxide by RP-HPLC method

Abstract

A simple, accurate, rapid, precise and novel Reverse phase High Pressure liquid chromatographic method (RP-HPLC)
has been developed and validated for simultaneous determination of Amitryptyline & Chlordiazepoxide. Amitriptyline
and Chlordiazepoxide shows λmax at 230nm and 259 nm respectively. The selected and optimized mobile phase was
mixed phosphate buffer (pH 3.0) and Acetonitrile were mixed in the ratio of 55:45 and conditions were flow rate (1.0
ml/minute), wavelength (252 nm), Run time was 12 min. The retention time were found to be 2.857 min and 5.667 min
for Amitryptyline & Chlordiazepoxide respectively. Linearity and range was found to be 25-150 µg/ml for
Amitryptyline and 10-60 µg/ml for Chlordiazepoxide. The proposed chromatographic conditions were found
appropriate for the quantitative determination of the drugs. The method was validated for accuracy, precision,
specificity, linearity, robustness, sensitivity, LOD and LOQ. The proposed method was successfully used for
quantitative analysis of tablets. No interference from any component of pharmaceutical dosage form was observed.
Validation studies revealed that method is specific, rapid, reliable, and reproducible.