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

Abstract

New reversed-phase high-performance liquid chromatography (HPLC) method has been developed and validated for the identification of impurity in calcium orotate dihydrate (CaOD) drug. The developed analytical method has been achieved superior resolution between the impurity and CaOD using Alltima C-18 column. The method has been established by isocratic mobile phase condition 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. Further, this method highlights that the eluted impurity in the drug is uracil and confirmed by spiking the standard uracil into CaOD sample. According to International council for harmonization (ICH) guidelines, the developed method has been validated with respect to accuracy, precision, specificity, linearity, range, robustness, system suitability, limit of detection and limit of quantification.