UPLC Transfer and Method Validation of the Determination of Prednisolone Tablets

Abstract

       The method transfer and validation for the determination of Prednisolone in tablets was established using Ultra Performance Liquid Chromatography (UPLC). The separation was achieved on a ZORBAX Eclipse Plus C18 (1.8 μm, 2.1 × 100 mm) at 35 degree Celsius. The mobile phase was the mixture of methanol and water (55:45, v/v) at the flow rate of 0.3 ml per minute with UV detection at 254 nm. Specificity test showed that prednisolone peak was not interfered by its degradation product peaks under stress conditions: acid, base, heat, hydrolysis, oxidation and light. The linearity test showed linear relationship between the area under the peak and the concentration levels in the range of 0.15-0.25 mg per ml with correlation coefficient, r, 0.9999. The percentage mean recovery from three concentration levels was 100.5 (% RSD = 1.17%). The precision test was performed by the assay of six samples within the same day, and different days and analysts gave % RSD of 0.7, 0.6 and 0.7, respectively. The robustness test indicated that the described method was robust when the chromatographic conditions were changed such as column batch, column temperature, composition and flow rate of mobile phase with no effect on the system suitability and the test result. This method takes less analytical time than the method specified in the United States Pharmacopoeia and British Pharmacopoeia. Therefore, the proposed method was suitable for the quantification of prednisolone in tablets and could be used as an alternative method.