Identification of dabigatran etexilate major degradation pathways by liquid chromatography coupled to multi stage high-resolution mass spectrometry

Abstract

Dabigatran etexilate (DABET) is an oral direct thrombin inhibitor that has been approved for the prevention of blood clot formation. As the active pharmaceutical ingredient (API) may undergo degradation, leading to drug activity loss or to the occurrence of adverse effects associated with degradation products, thorough knowledge of the API's stability profile is required. Since very few studies have been reported on the drug stability profile, a study related to DABET's behaviour under stress conditions was carried out in order to identify its major degradation pathways. DABET was subjected to hydrolytic (acidic and alkaline), oxidative, photolytic and thermal stress, as per ICH-specified conditions. Up to ten degradation products along with dabigatran, the active metabolite of DABET, were formed and detected by reverse phase liquid chromatography in gradient mode (LC) coupled to UV and mass spectrometry (UV-MS). Structures were determined by elemental composition determination and study of the fragmentation patterns, using high-resolution mass spectrometry in multistage mode (HR-MSⁿ). Under hydrolytic stress conditions, O-dealkylation may occur and formation of benzimidic acid derivatives was also observed. DABET was shown to be much less susceptible to photolysis and oxidative stress, even if N-dealkylation was highlighted. In view of the structures identified, various degradation pathways of DABET have been proposed.