Development and Validation of Hplc Method for Determination of Human Insulin in Pharmaceutical Preparation

A simple and reliable high-performance liquid chromatography (HPLC) method with diode array detection was developed and validated for determination of human insulin in pharmaceutical preparation. The HPLC separation was achieved on a Phenomenex reversed-phase column (250 mm×4.6 mm, 5 m) using a mobile phase of acetonitrile and 0.2 M sodium sulfate buffer (pH 2.4) solution (25:75, v/v) at a flow rate of 1.2 ml min. The diode array detector (DAD) was operated at 206 nm and column temperature was adjusted at 40 C. The method was validated for specificity, linearity, precision, accuracy and limit of quantitation. Also, the method was applied for the quality control of commercial Actrapid HM formulation.