Identification and characterization of new degradation products of belinostat using UHPLC-Q-TOF-MS/MS and <i>in silico</i> toxicity prediction
Lovekesh Mehta, Tanveer Naved, Parul Grover, Monika Bhardwaj, Debaraj Mukherjee, Dushyanth R. Vennapu
Abstract
Belinostat is a histone deacetylase inhibitor used for the management of hematological malignancies and solid tumors. In the current study, stability-indicating ultra-performance liquid chromatography (RP-UPLC) and developed highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) for identification, quantification and characterization of belinostat and its degradation products. The separation was carried out with Acquity BEH C-18 (100 mm × 2.1 mm, 1.7 µm) column using mobile phase comprising of 0.05% ortho-phosphoric acid in water and acetonitrile with gradient elution with 5 µL injection volume at 220 nm detection wavelength. The analysis was carried out at a 0.25 mL/min flow rate along with 45 °C column temperature throughout the analysis. The optimized method was completely validated as per ICH guideline Q2 (R1). Drug substance was treated with different stress conditions for a different time period as indicated by ICH recommended guidelines. Belinostat was sensitive to acidic, basic and oxidative degradation conditions. High-resolution mass spectrometry coupled with QTOF was used to identify and characterize degradation products of belinostat. The most plausible mechanism for all degradation products was established for the first time. TOPKAT (TOxicity Prediction by Komputer Assisted Technology) software was used to evaluate the in silico toxicity of belinostat and its degradation products.