Stability Indicating Method for Clenbuterol Hydrochloride Using UV-Spectrophotometry and TLC

Abstract

A simple, accurate and precise stability indicating UV Spectrophotometric method was developed and validated for Clenbuterol HCl. Stability indicating TLC method has been also developed. Furthermore, isolation of degradation product was done by TLC and characterization of degradation product was done by MS/MS. Stability indicating capability is established by forced degradation of Clenbuterol HCl to acid, base, thermal, oxidation and photo degradation as per ICH guidelines Q1A (R2). Detection was carried out at 247 nm wavelength. Degradation was observed in acidic, basic, thermal, oxidation and photolytic conditions. The method was validated as per ICH guidelines Q2 (R1). The linearity of the proposed method was found to be in the range of 2-12 μg/ml concentrations with correlation coefficient was (r2)=0.9916. The % recovery was found to be 100.59%.Stability indicating TLC method was developed in acid, base, thermal, oxidation and photolytic conditions. Separation was achieved on a Precoated Silica Gel 60F254 Plates using Toluene: Ethanol: Ammonia (18:6:0.15 v/v/v) as mobile phase. Degradation was observed in acidic condition at 1 hr using 5 N HCl as a stressor at room temperature. Spot of Clenbuterol HCl was observed at retension factor (Rf) = 0.55 and acidic degradation product at Rf = 0.82. Then characterization of degradation product was done by MS/MS and in that methylation was observed. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating method.