Development and validation of TLC-densitometric method for the quantification of a steroidal drug, danazol in its pharmaceutical formulations

A reliable and sensitive thin-layer chromatographic (TLC)-densitometric method is developed for the analysis of danazol in its pharmaceutical formulations and validated as per the International Conference on Harmonization (ICH) guideline. Moreover, extraction efficiency of danazol from its pharmaceutical formulations was also optimized by using various extraction methodologies. The method involves a simple extraction procedure under thermomixing at 40°C for 20 min. TLC aluminum sheets precoated with silica gel 60F-254 were employed as stationary phase. The solvent system consisted of hexane-acetone (6:4, v/v) as mobile phase, which showed compact spots of danazol (RF = 0.55 ± 0.005). Densitometric analysis of danazol was carried out in the absorbance mode at 291 nm. The linear regression analysis data for the calibration plots showed good linear relationship with r = 0.996 ± 0.003 in the concentration range of 200–1200 ng spot−1. The mean value of slope and intercept was 7.949 ± 0.78 and 2460 ± 19.72, respectively. The limits of detection (LOD) and quantitation (LOQ) were 1.19 and 3.61 ng spot−1, respectively. The method has been successfully applied for the qualitative and quantitative analysis of danazol in pharmaceutical formulations and can be employed for routine analysis of danazol in bulk drugs and its pharmaceutical dosage forms.