Five impurities in posaconazole: detection, genotoxicity prediction and action-mechanism analysis

Abstract

Genotoxic impurities are a class of highly potent mutagenic substances that typically needs to be controlled down to low ppm level in pharmaceuticals. A novel LC-MS/MS limit test method (single impurity ≤16.7 ppm) for detecting five impurities in Posaconazole has been established and partially validated. The chromatographic separation of analytes was conducted on an YMC-Triart C18 column and gradient elution was performed with 0.1% formic acid aqueous solution and acetonitrile as mobile phase A and B, respectively. Five impurities were carried out in ESI+ and MRM with the following mass transition pairs: m/z 390.2→296.2 for BSKZ029, m/z 300.4→136.2 for BSKZ117, m/z 207.0→162.2 for BSKZ135, m/z 209.0→91.0 for BSKZ155 and m/z 499.1→258.1 for BSKZ160. The established method exhibited good specificity, well robustness and high sensitivity with the LOD and LOQ was 1.34-1.56 ng/mL and 4.01-4.69 ng/mL, respectively. The genotoxicity of these impurities were evaluated by two (Q)SAR prediction methodologies, and results showed that BSKZ135 and BSKZ029 exhibited no genotoxicity, while BSKZ155, BSKZ117 and BSKZ160 possessed potent mutagenicity risk. Furthermore, the possible action-mechanisms of which were analyzed and proposed. This study has instructive significance to the analysis and genotoxicity research of genotoxic impurities, and also provides technical support for the supervision of the drug administration.