An UHPLC-TOF-MS method for quantifying novel brominated anticancer compound bromophenol-thiosemicarbazone hybrid and its application to in vivo pharmacokinetic study

Abstract

Background: Bromophenol-thiosemicarbazone hybrid was a novel synthetic brominated anticancer compound with two bromine atoms. Bromophenol-thiosemicarbazone hybrid showed considerable selective inhibitory activity against PARP1 (IC50 = 29.5 nmol/L). UHPLC-TOF-MS was used to establish a new method to quantify bromophenol-thiosemicarbazone hybrid bio-samples for indispensable quantitation analysis in further extensive pre-clinical studies. Methods: Chromatographic and mass spectrometry parameters were optimized for quantitative method establishment. Improved protein-precipitated method was applied to the extraction of bromophenol-thiosemicarbazone hybrid in rat plasma samples. Furthermore, this proposed method was applied to an intravenous bolus dose to male rats. Results: Mobile phase was consisted of water for A and acetonitrile for B with 25 mmol/L formic acid in both A and B. The flow rate was 0.30 mL/min, and the run time of bromophenol-thiosemicarbazone hybrid was 4.0 min. A Thermo Fisher Accucore 2.6 μm C18 column (50 × 2.1 mm i.d.; San Jose, USA) was used for chromatographic separation. High resolution mass spectrometry was used to quantify samples by exact mass number of compound which was operated on negative ionization mode. Linear dynamic range of the established method was widely with 13.7–10000 nmol/L. Pharmacokinetics properties of bromophenol-thiosemicarbazone hybrid were shown in the results. Conclusion: This method was reliable and reproducible from sample preparation to analysis and storage stability under the investigated conditions. It may be useful for analysis of halogenated compounds and brominated compounds in ultra-performance liquid chromatography-mass spectrometry.