Isolation and Characterization of Novel Degradation Products of Eliglustat Tartrate Using 2D-NMR and HRMS: Development and Validation of Stability Indicating RP-UPLC Method for Quantification of Assay and Characterized Impurities

Abstract

Eliglustat tartrate is an oral inhibitor of glucosylceramide synthase used in the therapy of Gaucher’s disease. The drug was subjected to forced degradation study using various stress parameters like acidic, basic, oxidation, photolysis and thermal conditions. The drug showed significant degradation under acidic and peroxide stress conditions, however the drug remained stable under basic, neutral, thermal and photolytic stress conditions. The present research reports the characterization of degradation products of Eliglustat by liquid chromatography-tandem mass spectrometry(HRMS) and 2D-NMR techniques along with the development and validation of stability indicating ultra performance liquid chromatographic method for determination and quantification of Eliglustat drug in presence of characterized impurities. Two novel degradation products (DP1 & DP-2) were found under stress conditions. These degradation products were isolated using preparative high performance liquid chromatography. The structure of the DP-1 was identified as the cis- diastereoisomer of Eliglustat and DP-2 as the N-Oxide impurity of Eliglustat which were not reported in any of the literature. The chromatographic method was developed on UPLC to achieve separation of degradation products and Eliglustat using Acquity; BEH; C-18; 100 x 2.1 mm; 1.7 μm column within a short run time of 8.0 minutes. The method was validated for specificity, precision, linearity and accuracy. The quantitation limits obtained were in the range of 0.3-3.0 μg ml-1 for the characterized impurities. This method is compatible to LCMS analysis which enables to identify the unknown impurities formed in the process.