Journal of Liquid Chromatography & Related Technologies ( IF 1.3 ) Pub Date : 2021-05-04 , DOI: 10.1080/10826076.2021.1906271 Lovekesh Mehta 1 , Tanveer Naved 1 , Parul Grover 2 , Monika Bhardwaj 3, 4 , Debaraj Mukherjee 3 , Dushyanth R. Vennapu 5
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.
中文翻译:
使用 UHPLC-Q-TOF-MS/MS 和计算机毒性预测鉴定和表征贝利司他的新降解产物
摘要
Belinostat 是一种组蛋白去乙酰化酶抑制剂,用于治疗血液系统恶性肿瘤和实体瘤。在目前的研究中,稳定性指示超高效液相色谱 (RP-UPLC) 和开发的高灵敏度液相色谱-串联质谱 (LC-MS/MS) 用于对贝立司他及其降解产物进行鉴定、定量和表征。使用 Acquity BEH C-18(100 mm × 2.1 mm,1.7 µm)色谱柱进行分离,流动相包含 0.05%邻苯二甲酸- 磷酸的水和乙腈溶液,在 220 nm 检测波长下以 5 µL 进样体积进行梯度洗脱。在整个分析过程中,分析以 0.25 mL/min 的流速和 45 °C 的柱温进行。根据 ICH 指南 Q2 (R1) 对优化方法进行了完全验证。如 ICH 推荐的指南所示,原料药在不同的时间段内用不同的压力条件处理。Belinostat 对酸性、碱性和氧化降解条件敏感。使用高分辨率质谱联用 QTOF 来鉴定和表征 Belinostat 的降解产物。首次建立了所有降解产物的最合理机制。TOPKAT (TOxicity Prediction by Komputer Assisted Technology) 软件用于评估贝立司他及其降解产物的计算机毒性。