A Validated Stability Indicating RP-HPLC Method for Determination of Phenobarbital and Acefylline Piperazine in Bulk Drug and Combined Dosage Form

Abstract

A simple, sensitive, precise and stability indicating RP-HPLC method was developed for
determination of Phenorbarbital and Acefylline Piperazine in bulk drug and in combined
syrup. Isocratic, HPLC method , using Thermo Hypersil C18 gold column with mobile
phase of Acetonitrile - 0.1% orthophosphoric acid (30:70,v/v) was investigated to
separate the drugs from their stressed degredation products. The flow rate was 1.5
ml/min. Detection was carried out at a wavelength of 240 nm. The method was
validated for precision, accuracy, robustness and recovery. Phenobarbital and Acefylline
Piperazine were exposed to acidic, basic, neutral, oxidative and photolytic stress
conditions and the stressed samples were analyzed by the proposed method. Good
linear relationship in the concentration range of (1-100) µg/ml and (4-250) µg/ml for
Phenobarbital and Acefylline Piperazine respectively with correlation coefficient of
0.9999 for both drugs. Intra- and inter-day precision were less than 1% for both
analytes. the method was tested for specifity for determination of target analytes in
presence of excipients like Methylparaben and Propylparaben also The stressed sample
chromatograms demonstrate the specificity of the proposed method for the
determination of target analytes in presence of degradants.