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 (IC 50 = 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.

中文翻译：

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一种用于定量新型溴化抗癌化合物溴酚-氨基硫脲杂化物的 UHPLC-TOF-MS 方法及其在体内药代动力学研究中的应用

背景溴苯酚-氨基硫脲杂化物是一种新型合成溴化抗癌化合物，具有两个溴原子。溴酚-氨基硫脲杂化物对 PARP1 显示出相当大的选择性抑制活性 (IC 50 = 29.5 nmol/L)。UHPLC-TOF-MS 用于建立一种新方法来量化溴苯酚-缩氨基硫脲混合生物样品，以便在进一步广泛的临床前研究中进行必不可少的定量分析。方法 优化色谱和质谱参数以建立定量方法。将改进的蛋白质沉淀法应用于大鼠血浆样品中溴酚-氨基硫脲杂化物的提取。此外，该提议的方法被应用于雄性大鼠的静脉推注剂量。结果 流动相 A 为水，B 为乙腈，A 和 B 中的甲酸均为 25 mmol/L。流速为 0.30 mL/min，溴苯酚-氨基硫脲混合物的运行时间为 4.0 min。Thermo Fisher Accucore 2.6 μm C18 色谱柱（50 × 2.1 mm 内径；美国圣何塞）用于色谱分离。高分辨率质谱用于通过在负电离模式下操作的化合物的精确质量数来量化样品。所建立方法的线性动态范围很广，为 13.7–10000 nmol/L。结果显示了溴酚-氨基硫脲杂化物的药代动力学特性。结论 该方法在研究条件下从样品制备到分析和储存稳定性均可靠且可重现。