A simple, specific, precise and accurate simultaneous high-performance thin-layer chromatographic method of analysis for hydrocortisone and clotrimazole was developed and validated. The method employed HPTLC glass plates (20x10) mm precoated with silica gel 60 F 254 as the stationary phase. The solvent system consisted of Toluene: Propanol: Ammonia (13:3:0.1v/v) that gave compact spots with R f value of 0.27 ± 0.01 and 0.58 ± 0.02 for hydrocortisone and clotrimazole, respectively. Densitometric analysis of hydrocortisone and clotrimazole was carried out in the absorbance/reflectance mode at 226nm. The calibration curves showed good linear relationship with R 2 = 0.996 + 0.003 and 0.996 + 0.002 in the concentration range of 200-1200 and 200-1000 ng/µl for hydrocortisone and clotrimazole, respectively. The method was validated for precision, recovery and robustness. The LOD & LOQ were found to be 35.31, 107.01 and 34.93, 105.87 ng/µl for hydrocortisone and clotrimazole respectively. Statistical analysis proved that the method is repeatable, specific and accurate for the estimation of the studied drugs. Resolution of hydrocortisone, clotrimazole and their degradation products formed under different stress conditions (acid-base hydrolysis, oxidation and thermal degradation) was successfully achieved with the developed method indicating that it can be employed as a stability indicating method. Hydrocortisone (cortisol);11,17,20-dione, the main physiologic glucocorticoid in humans has an anti-inflammatory and an immunosuppressive effect besides its role on carbohydrate and protein metabolism 1-3 . The anti-inflammatory effect of hydrocortisone is due to reduction of histamine release, reduction of IgG production and decreasing of the activity of neutrophils and macrophages [4][5][6]imidazole is an imidazole derivative with antimycotic activity that is known to inhibit cytochrome P-450, ergosterol biosynthesis, and proliferation of cells and to interfere with cellular Ca 2+ homeostasis 7-9 . BACKGROUND
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