High Perfarmance Liquid Chromatographic Determination of Ascorbic Acid in Pharmaceutical Preparations, Fruit Juices and Human Serum


A high performance liquid chromatographic method for determination of ascorbic acid is described. The analysis is achieved using cosmosil 5C18-MS-II column (250 mm x 4.6 mm i.d., 5 µm particle size) at room temperature. The mobile phase used was acetonitrile (ACN), dichloromethane (DCM), and 0.25% K2HPO4 solution in the ratio (90:5:5)(V:V:V). The flow rate is set to 1.0ml.min-1 with UV-detection at 246 nm. Beer's law is obeyed over the concentration range 0.05-30 µg.ml-1. The method is accurate (relative error % is less than 0.12%), precise (RSD better than ±1.25%), successfully applied to the determination of ascorbic acidin its pharmaceutical preparations, fruit juices, and human serum