HPTLC method for determination of nevirapine in pharmaceutical dosage form

Abstract

A sensitive, selective, precise and stability-indicating high-performance thin-layer chromatographic method of analysis of nevirapine both as a bulk drug and in formulations was developed and validated. The solvent system consisted of toluene–carbon tetrachloride–methanol–acetone–ammonia (3.5:3.5:2.0:1.0:0.05, v/v/v/v/v). Densitometric analysis of nevirapine was carried out in the absorbance mode at 289 nm. This system was found to give compact spots for nevirapine ( R f value of 0.44±0.02). Nevirapine was subjected to acid and alkali hydrolysis, oxidation, dry heat and wet heat treatment and photodegradation. The drug undergoes degradation under acidic, basic conditions and oxidation. Also the degraded products were well resolved from the pure drug with significantly different R f values. Linearity was found to be in the range of 30–1000 ng/spot with significantly high value of correlation coefficient. The linear regression analysis data for the calibration plots showed good linear relationship with r 2=0.998±0.002 in the working concentration range of 300 ng/spot to 1000 ng/spot. The mean value of slope and intercept were 0.073±0.005 and 36.78±1.50, respectively. The method was validated for precision, robustness and recovery. The limit of detection and quantitation were 5 and 10 ng/spot, respectively. As the method could effectively separate the drug from its degradation products, it can be employed as a stability indicating one. Moreover, the proposed HPTLC method was utilized to investigate the kinetics of acid degradation process. Arrhenius plot was constructed and activation energy was calculated.