High Performance Liquid Chromatography Method Development and Validation for Separation of Liothyronine Sodium Related Substances Using a Quality by Design Approach

Abstract

The application of the Quality by Design (QbD) principles in developing a novel high performance liquid chromatography (HPLC) method for the analysis of liothyronine sodium related substances for finished product using Fusion QbD® software is explored. The effect of various chromatographic parameters including, column stationary phase, initial hold time and gradient time on separations were systematically investigated. Results show that optimal separations of these compounds in a standard solution can be achieved using a X Bridge C18 column (150 mm × 4.6 μm, 5m), maintained at sample temperature 15°C by using pH 2.0 buffer solution as mobile phase-A and methanol as mobile phase-B with a flow rate of 1.0 ml/min at 225 nm of detection and a gradient time of 38 minutes. The injection volume is 200 μl. Pre-validation studies of the method were performed and all the parameters met the acceptance criteria. The results are demonstrated that optimized method is selective, linear, precise, repeatable and accurate.