ANALYTICAL METHOD VALIDATION FOR ASSAY OF ALPROSTADIL BY HPLC USING INTERNAL STANDARD

Abstract

An isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the assay of Alprostadil using an Internal Standard. The successful separation of from its synthetic impurities and degradation products formed under stress conditions was achieved using 4.6mm x 25cm, Packing L1 or equivalent maintained at 25°C with a mobile phase of Methanol, acetonitrile and 0.1 M mono basic potassium phosphate (2:1:2) adjust with phosphoric acid to pH of 3.0. The mobile phase flow rate was 1.0 mL/min, and the detection wavelength was 200 nm. The developed HPLC method was validated with respect to linearity, accuracy, precision, specificity, and robustness. The developed HPLC method to determine the assay of alprostadil can be used to evaluate the quality of regular production samples. It can be also used to test the stability samples of Alprostadil.