Reversed-phase high-performance liquid chromatography method for impurity profiling of generic drug Calcium Orotate

Abstract

New reversed-phase high-performance liquid chromatography method has been developed and validated for the identification of impurity in calcium orotatedihydrate (CaOD) drug. The developed method has been achieved superior resolution between the impurity and CaOD using Alltima C-18 column (250 X 4.6 mm, 5 μm). The method has been established by isocratic mobile phase comprising of 0.01 mol/L of potassium dihydrogen orthophosphate, pH adjusted to 2.7 with orthophosphoric acid and acetonitrile in a ratio of 98:2 v/v. The eluted impurity at the retention time of 6.02 min has so far not been reported in earlier methods of analysis. According to International council for harmonization guidelines, the method has been validated with respect to accuracy, precision, specificity, linearity, range, robustness, system suitability, limit of detection and limit of quantification. The percentage recovery of CaOD and uracil is found in the range between 98% and 102%. The linearity data has shown that the method is found to be linear for CaOD and uracil from 25% to 150% level and the correlation coefficient is found to be more than 0.999. The reported method is not affected by small but deliberate changes in the parameters of the method.