Trancart Marilène, Penot Mylène, Meesemaecker Gwladys, Boffy Romain, Hanak Anne-Sophie, Calas André-Guilhem, Taudon Nicolas
Toxicology and Chemical Risks Department, French Armed Forces Biomedical Research Institute, Brétigny-sur-Orge, France.
Analytical Development and Bioanalysis Unit, Platforms and Technological Research Department, French Armed Forces Biomedical Research Institute, Brétigny-sur-Orge, France.
J Anal Methods Chem. 2025 Apr 8;2025:9923229. doi: 10.1155/jamc/9923229. eCollection 2025.
Despite the availability of current resources, the development of medical countermeasures remains crucial in combatting the threat posed by chemical warfare agents, such as organophosphorus compounds (OPs), which are toxic nerve agents requiring rapid medical intervention. Within the available therapeutic arsenal, muscarinic antagonists such as atropine are administered to mitigate the effects of excessive cholinergic system stimulation, which leads to respiratory tract obstruction due to hypersecretions and bronchoconstriction. Tiotropium, an FDA-approved bronchodilator, acts as a muscarinic receptor antagonist and could, therefore, serve as a potential alternative. To assess its potential efficacy in attenuating OP-induced respiratory effects in a murine intoxication model, it was necessary to first characterize its pharmacokinetic properties. A liquid chromatography-mass spectrometry method was developed and validated following ICH M10 guidelines for the quantification of atropine and tiotropium in 10 μL of plasma. The sample pretreatment procedure involved solid-phase extraction. Chromatographic separation was achieved using a fully porous sub 2 μm C18 column. The analysis was completed in just 4 min, with analytes identified and quantified using two selected reaction monitoring transitions. The mean extraction recoveries exceeded 90% for both drugs, and no matrix effect was observed. The lower limits of quantification were 0.5 ng/mL for tiotropium and 1.0 ng/mL for atropine, with an upper limit of 1000 ng/mL. The signal-to-concentration ratio demonstrated recoveries of back-calculated concentrations ranging from 94% to 108% (relative standard deviation (RSD) < 9.0%). Within-run and between-run precisions were both below 8% with accuracies ranging from 87% to 110%. This highly specific and sensitive method has proven useful for analyzing samples from pharmacokinetic studies conducted in mice. Following intraperitoneal administration, the AUC for tiotropium was approximately twice that of atropine, while its Tmax was half as long (4.9 vs. 8.2 min). The terminal half-lives were approximately 7.5 min for tiotropium and 9.8 min for atropine.
尽管有现有资源，但开发医学应对措施对于对抗化学战剂（如有机磷化合物（OPs））所构成的威胁仍然至关重要，有机磷化合物是有毒神经毒剂，需要迅速的医学干预。在现有的治疗手段中，会使用毒蕈碱拮抗剂（如阿托品）来减轻胆碱能系统过度刺激的影响，这种过度刺激会因分泌物过多和支气管收缩导致呼吸道阻塞。噻托溴铵是一种经美国食品药品监督管理局（FDA）批准的支气管扩张剂，作为毒蕈碱受体拮抗剂，因此可能是一种潜在的替代药物。为了评估其在小鼠中毒模型中减轻OP诱导的呼吸影响的潜在疗效，首先有必要对其药代动力学特性进行表征。按照国际人用药品注册技术协调会（ICH）M10指南，开发并验证了一种液相色谱 - 质谱法，用于定量测定10μL血浆中的阿托品和噻托溴铵。样品预处理程序包括固相萃取。使用全多孔亚2μm C18柱实现色谱分离。分析仅需4分钟即可完成，使用两个选择反应监测跃迁对分析物进行鉴定和定量。两种药物的平均萃取回收率均超过90%，且未观察到基质效应。噻托溴铵的定量下限为0.5 ng/mL，阿托品的定量下限为1.0 ng/mL，上限为1000 ng/mL。信号与浓度比显示，反算浓度的回收率在94%至108%之间（相对标准偏差（RSD）< 9.0%）。批内和批间精密度均低于8%，准确度在87%至110%之间。这种高度特异且灵敏的方法已被证明可用于分析小鼠药代动力学研究中的样品。腹腔注射后，噻托溴铵的曲线下面积（AUC）约为阿托品的两倍，而其达峰时间（Tmax）仅为阿托品的一半（4.9对8.2分钟）。噻托溴铵的末端半衰期约为7.5分钟，阿托品的末端半衰期约为9.8分钟。
