Analytical Lifecycle Management (ALM) and Analytical Quality by Design (AQbD) based Analytical Method Development for Separation of Related Substances in Amodiaquine Hydrochloride along with its Degradation Products and Structural Elucidation by LC-Quadrupole-Time of Flight-Tandem Mass Spectrometry

Abstract

The current study aimed to develop a robust, regulatory-flexible, stability-indicating ultra high performance liquid chromatography (UHPLC) analytical procedure compatible with mass spectrometry for the determination of impurities in amodiaquine hydrochloride using analytical lifecycle management (ALM) and analytical quality by design (AQbD) principles. The analytical target profile (ATP) and critical method attributes (CMAs) for the analytical method were identified. The pH of the mobile phase, flow rate and column oven temperature were identified as critical method parameters (CMPs) through risk assessment studies and they were screened and optimized using the design of experiments (DoE) to generate the design space (DS). Finally, all the impurities were well separated in a 15 min run time by using an Acquity UPLC BEH-C18 column with 20 mM ammonium acetate pH 8.0 as mobile phase A and acetonitrile as mobile phase B with a gradient program of time (min), % of B: 0/15, 1.0/15, 9/55, 12/55, 12.1/15 and 15/15. The flow rate was 0.3 mL min-1 and the column oven temperature were 30 ºC. Evaluated the robustness of the developed analytical method. Degradation products obtained from the forced degradation studies are well separated from process impurities and main compound. Based on the stress studies, impurity-3 was identified as a key degradation product in base degradation, while DP-1, impurity-3, DP-2, DP-3 and DP-4 were in the oxidative degradation. Total 9 impurities out of 4 process related and 5 degradation impurities were well separated in the developed method. Used the same method to LC coupled tandem mass spectrometer for separation and structure elucidation of degradation products. The method validation was carried out in accordance with ICH Q2 (R1) and USP <1225> guidelines.