In this study, a simple and reliable LC–MS/MS method was first proposed for the simultaneous determination of TUG‐891 and its metabolites TUG‐891‐alcohol, TUG‐891‐aldehyde, and TUG‐891‐acid in rat plasma. The analytes and fasiglifam (internal standard) were extracted from plasma samples with acetonitrile and separated using an Acquity BEH C₁₈ column (1.7 μm, 2.1 × 50 mm) with water containing 0.05% ammonium hydroxide and acetonitrile containing 0.05% ammonium hydroxide as the mobile phase. A Q‐Exactive Orbitrap mass spectrometer in full‐scan mode was used for mass detection, and the data analysis was obtained using a mass extraction window of 5 ppm. The calibration curves exhibited excellent linearity (correlation coefficient > 0.9981) in the concentration range of 0.5–1000 ng/mL. The lower limit of quantification was 0.5 ng/mL for all analytes. The intra‐ and inter‐day precision was less than 11.31%, and the accuracy ranged from −11.50 to 9.50%. The extraction recovery of the analytes from rat plasma was greater than 82.31%, and no obvious matrix effect was found. The established method was further applied to the pharmacokinetic study of TUG‐891, TUG‐891‐alcohol, TUG‐891‐aldehyde, and TUG‐891‐acid in rat after a single dose of 5‐mg/kg treatment of TUG‐891. The results demonstrated that TUG‐891 was rapidly metabolized into its metabolites and the systemic exposures of the metabolites were much higher than those of TUG‐891.

中文翻译：

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使用LC-HRMS同时测定大鼠血浆中的TUG-891及其代谢物，并将其应用于临床前药代动力学研究。

在这项研究中，首次提出了一种简单可靠的LC-MS / MS方法，用于同时测定大鼠血浆中的TUG-891及其代谢物TUG-891-酒精，TUG-891-醛和TUG-891-酸。用乙腈从血浆样品中提取分析物和法西格坦（内标），并使用Acquity BEH C _{18进行分离}色谱柱（1.7μm，2.1×50 mm）以含有0.05％氢氧化铵的水和含有0.05％氢氧化铵的乙腈为流动相。使用全扫描模式的AQ-Exactive Orbitrap质谱仪进行质量检测，并使用5 ppm的质量提取窗口进行数据分析。校准曲线在0.5–1000 ng / mL的浓度范围内表现出极好的线性（相关系数> 0.9981）。所有分析物的定量下限为0.5 ng / mL。日内和日间精度均低于11.31％，精度范围为-11.50至9.50％。从大鼠血浆中提取分析物的回收率大于82.31％，未发现明显的基质效应。既定方法进一步应用于TUG‐891，TUG‐891‐醇，TUG‐891‐醛的药代动力学研究 单剂量5 mg / kg TUG-891治疗后大鼠中的TUG-891-酸和TUG-891-酸。结果表明，TUG-891被迅速代谢成其代谢产物，且代谢产物的全身暴露量远高于TUG-891。