Department of Forensic Medicine, Division of Molecular Techniques, Faculty of Medicine, Wroclaw Medical University, Sklodowskiej-Curie 52, 50369, Wroclaw, Poland.
Department of Forensic Medicine, Faculty of Medicine, Wroclaw Medical University, 4 J. Mikulicza-Radeckiego Street, 50345, Wroclaw, Poland.
Arch Toxicol. 2024 Dec;98(12):3987-4012. doi: 10.1007/s00204-024-03867-3. Epub 2024 Sep 28.
Both postmortem toxicological and medical-forensic examinations are very important in the case of analyzing various types of chemical substances. Hydroxyzine (HZ) is a first-generation antihistamine drug with a sedative effect that disrupts cognitive function and affects the ability to drive motor vehicles. Enzymatic oxidation of the hydroxy-methyl group to the carboxyl group leads to the formation of its main metabolite-cetirizine (CZ). CZ is the active substance of antiallergic drugs. Because it does not cross the BBB (blood-brain barrier) easily, it is less likely to cause drowsiness or affect memory and impair cognitive function. Therefore, in criminal studies, it is often important what medication had been taken by a person involved, e.g., in a car accident, HZ or CZ. The analysis of both antihistamine drugs is challenging, as usually very low concentrations of the compound of interest need to be determined. Thus, an ultra-sensitive UHPLC-QqQ-MS/MS method was developed for simultaneous determination of HZ and CZ in biological fluid samples. The lower limit of quantification (LOQ) for HZ and CZ was calculated as 0.345 and 0.3696 ng/mL, respectively. Together with a reduced sample volume to 200 μL, it makes the developed method suitable for a sensitive multidrug forensic toxicological analysis. Samples were extracted with simple and fast liquid-liquid extraction (ethyl acetate, pH 9). The present method for the determination of HZ and CZ in human blood proved to be simple, fast, selective, and sensitive. The quantification by LC-MS/MS was successfully applied to the samples coming from 28 authentic biological fluids (blood, urine, vitreous humor, bile and stomach content), both antemortem and postmortem. The performed studies confirm that the developed method is characterized by a high extraction efficiency. Its accuracy, reproducibility, simplicity, and selectivity suggest its application in clinical, toxicological, and forensic laboratories.
在分析各种化学物质的案例中，法医毒理学和医学法医检查都非常重要。羟嗪（HZ）是一种具有镇静作用的第一代抗组胺药，会破坏认知功能并影响驾驶机动车辆的能力。羟甲基基团的酶促氧化作用导致其主要代谢物-西替利嗪（CZ）的形成。CZ 是抗过敏药物的活性物质。由于它不易穿过 BBB（血脑屏障），因此不太可能引起嗜睡或影响记忆和损害认知功能。因此，在刑事研究中，通常很重要的是要了解涉案人员服用了哪种药物，例如在车祸中，是 HZ 还是 CZ。分析这两种抗组胺药具有挑战性，因为通常需要测定非常低浓度的目标化合物。因此，开发了一种超灵敏的 UHPLC-QqQ-MS/MS 方法，用于同时测定生物体液样品中的 HZ 和 CZ。HZ 和 CZ 的定量下限（LOQ）分别计算为 0.345 和 0.3696 ng/mL。与将样品体积减少到 200 μL 相结合，使得所开发的方法适用于灵敏的多药物法医毒理学分析。样品用简单快速的液液萃取（乙酸乙酯，pH 9）提取。用于测定人血液中 HZ 和 CZ 的本方法简单、快速、选择性好且灵敏。LC-MS/MS 定量成功应用于来自 28 份真实生物体液（血液、尿液、玻璃体、胆汁和胃内容物）的样品，包括生前和死后。进行的研究证实，所开发的方法具有高提取效率。其准确性、重现性、简单性和选择性表明它可应用于临床、毒理学和法医实验室。
