Laboratory LAT LUMTOX, La Voulte sur Rhône, France.
CHU Lille, Laboratory of toxicology, Lille, France.
Drug Test Anal. 2024 Apr;16(4):331-338. doi: 10.1002/dta.3542. Epub 2023 Jul 24.
Despite prevention efforts, many cases of mushroom poisoning are reported around the world every year. Among the different toxins implicated in these poisonings, muscarine may induce parasympathetic neurological damage. Muscarine poisonings are poorly reported in the current literature, implying a lack of available data on muscarine concentrations in human matrices. A validated liquid chromatography with high-resolution mass spectrometry detection (Orbitrap technology) method was developed to determine muscarine concentrations in human urine, plasma, and whole blood samples. Muscarine was determined using 100 μL of biological fluids, and precipitation was used for sample preparation. Liquid chromatography-mass spectrometry was performed using an Accucore Phenyl-X analytical column with the electrospray source in positive ion mode. Muscarine was quantitated in parallel reaction monitoring (PRM) mode with D9-muscarine as the internal standard. The method was validated successfully over the concentration range 0.1-100 μg/L for plasma and whole blood and 1-100 μg/L for urine, with acceptable precision and accuracy (<13.5%), including the lower limit of quantification. Ten real cases of suspected muscarine poisoning were successfully confirmed with this validated method. Muscarine concentrations in these cases ranged from 0.12 to 14 μg/L in whole blood, <LOQ to 43 μg/L in plasma, and <LOQ to 1537 μg/L in urine. This work provides a tool of choice for the diagnosis of muscarine poisoning when mushrooms are not available for identification, as well as the possibility of determining toxins in the blood, which is an important step toward understanding the pharmacokinetics of the mycotoxin.
尽管采取了预防措施，但每年仍有许多蘑菇中毒病例在全球范围内报告。在这些中毒事件中涉及的不同毒素中，毒蕈碱可能会引起副交感神经的神经损伤。目前文献中对毒蕈碱中毒的报道较少，这意味着人类基质中毒蕈碱浓度的可用数据不足。开发了一种经过验证的液相色谱-高分辨率质谱检测（Orbitrap 技术）方法，用于测定人尿液、血浆和全血样本中的毒蕈碱浓度。使用 100 μL 生物流体进行毒蕈碱测定，并采用沉淀法进行样品制备。采用 Accucore Phenyl-X 分析柱，电喷雾源在正离子模式下进行液质联用。毒蕈碱以 D9-毒蕈碱为内标，在平行反应监测（PRM）模式下定量。该方法在血浆和全血的浓度范围为 0.1-100μg/L，尿液的浓度范围为 1-100μg/L 时得到了成功验证，具有可接受的精密度和准确度（<13.5%），包括定量下限。该方法成功验证了 10 例疑似毒蕈碱中毒的真实案例。这些病例的全血中毒蕈碱浓度范围为 0.12-14μg/L，血浆中<定量下限至 43μg/L，尿液中<定量下限至 1537μg/L。当无法获取蘑菇进行鉴定时，该方法为毒蕈碱中毒的诊断提供了首选工具，并且有可能确定血液中的毒素，这是理解真菌毒素药代动力学的重要一步。
