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A Simple and suitable analytical method for validation of Didanosine (DDI) by reverse phase high performance liquid chromatographic (RP-HPLC) method. The method was performed on a RP-HPLC (Agilent 1120 LC Germany) model, C-18 Zorabax column, 4.6 mm X 250 mm, 5µm particle size. The mobile phase was mixture of methanol and 0.01M sodium acetate buffer adjusted to the pH 6.5 (75:25) at flow rate of 1.2 ml/min. UV detection was performed at 250 nm and the linearity was found to be 10 to 60 µg/ml with a correlation coefficient of 0.9998. The retention time for DDI was 5.17 min. The method showed good recovery and the relative standard deviation of intra and inter day assay results were 99.96% to 100.05 %. This method used for the analysis of different marketed DDI tablets.
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