Application of an LC/HPLC method development and validation for the simultaneous estimation of amitriptyline hydrochloride and chlordiazepoxide in tablet dosage form by using a reverse phase technique

Faraat Alia, G. N. Singha, P.

Abstract

A rapid, simple, sensitive, precise and cost effective RP-HPLC method was developed and validated for the determination of Amitriptyline and Chlordiazepoxide in pure and tablet dosage form. The method involved determination of Amitriptyline and Chlordiazepoxideby resolving on RP-HPLC using Waters C18, 250 mm × 4.6 mm (5μm)reverse‐phase column, utilizing a mobile phase of Methanol:Acetonitrile:0.2M Orthophosphoric acid, pH 4.5, 56:24:20. Mobile phase was delivered with isocratic flow rate of 1.0 ml/minute. Ultra violet detection was carried out at 240 nm simultaneously over the concentration range of 50-1600 ng/ml for Amitriptylineand 10-320 ng/ml for Chlordiazepoxide respectively. Separation was completed within 15 minutes. The mean recoveries were found to be 98.46 ±0.47 % w/w for Amitriptylineand 96.60 ±0.98 % w/w for Chlordiazepoxide respectively. The coefficient (r2) were 0.998 for Amitriptylineand 0.999 for Chlordiazepoxiderespectively. The LOD and LOQ were found to be 25 and 83 ng/ml for AmitriptylineHcland 4.5 and 15 ng/ml for Chlordiazepoxide respectively. The assay percentage of the marketed formulation calculated were 97.47 ± 0.58 % w/w for AmitriptylineHcl and 95.29 ± 0.59 % w/w for Chlordiazepoxide respectively and intra-day and inter-day precision data were found to be < 2 % RSD.Developed method are simple, rapid, accurate, precise, reliable, economical and can be used for routine analysis for quantitative estimation of both components in solid tablet dosage form.