2.3.8. HPLC Method of Triamcinolone Acetonide Determination

A series of triamcinolone acetonide concentrations to obtain a calibration curve were prepared by dissolving the drug in methanol:water mixture 75:25 v/v ranging from 1 to 100 μg/mL. These solutions were analysed using Agilent 1260 Infinity Quaternary System HPLC (Agilent Technologies Ltd., Stockport, UK) connected to UV/Visible spectrophotometer which was set at 240 nm. A reverse phase Thermo Scientific Hypersil Gold C18 column (150 mm × 4.6 mm, 5 μm, Thermo-Scientific, Waltham, MA, USA) held at 30 °C was used for the chromatographic separation. The mobile phase consisted of a mixture of 50:50 v/v water:acetonitrile and was delivered with steady flow rate of 0.8 mL/min. The injection volume was 10 μL. The calibration curve of triamcinolone acetonide showed linearity in the predicted concentration range with regression value of 0.999. The equation (y = 24.688x − 7.4975) was derived from the areas of the triamcinolone acetonide absorbance peak plotted against respective drug concentration. This HPLC method was used to determine the drug contents in samples of prepared matrices as well as drug release from these matrices.