Military Institute of Forensic Medicine, Central Military Hospital Prague, Prague, Czech Republic.
Int J Legal Med. 2012 Jul;126(4):519-24. doi: 10.1007/s00414-011-0599-9. Epub 2011 Jul 13.
Ibotenic acid and muscimol are substances which mostly participate in psychotropic properties of Amanita pantherina and Amanita muscaria. They are rapidly absorbed from the gastrointestinal tract and readily excreted in urine. The poisoning with A. pantherina is in the majority of cases accidental because it can be easily mistaken for the edible species (Amanita rubescens, Amanita spissa and Macrolepiota procera). Intoxication with A. muscaria is mostly intentional for recreational purposes. Prognosis of the poisoning is generally good; lethal cases are rare. Mushroom poisoning is often proved by microscopic examination of spores in the stomach or intestinal content. Authors of this article introduce an instrumental method of proving A. pantherina or A. muscaria poisoning. The article describes the isolation of ibotenic acid and muscimol from urine, the derivatization step and the determination of these compounds by gas chromatography/mass spectrometry. Isolation of these alkaloids from urine was performed on a strong cation exchanger (Dowex® 50W X8), and the elution and derivatization of the alkaloids were made in one step with ethyl chloroformate in aqueous solution of sodium hydroxide with the addition of ethanol and pyridine. Cycloserine was used as internal standard. By this method, concentrations of ibotenic acid and muscimol in the urine of four persons intoxicated with A. pantherina were determined. In this study, mass spectra of derivatized ibotenic acid and muscimol are shown, and validation of the method is described.
异羟肟酸和麦司卡林是鹅膏蕈氨酸和鹅膏蕈碱主要参与的精神活性物质。它们从胃肠道迅速吸收，并易于在尿液中排泄。食用 A. pantherina 中毒的情况多数为意外，因为它很容易被误认为是可食用的物种（Amanita rubescens、Amanita spissa 和 Macrolepiota procera）。食用 A. muscaria 中毒多数是为了娱乐目的。中毒的预后通常较好；致死病例很少见。蘑菇中毒通常通过在胃或肠内容物中检查孢子的显微镜检查来证明。本文作者介绍了一种证明 A. pantherina 或 A. muscaria 中毒的仪器方法。文章描述了从尿液中分离异羟肟酸和麦司卡林、衍生化步骤以及通过气相色谱/质谱法测定这些化合物。这些生物碱从尿液中的分离是在强阳离子交换剂（Dowex® 50W X8）上进行的，生物碱的洗脱和衍生化在一步中用乙基氯甲酸酯在氢氧化钠的水溶液中进行，加入乙醇和吡啶。环丝氨酸用作内标。通过这种方法，测定了四名食用 A. pantherina 中毒者尿液中的异羟肟酸和麦司卡林的浓度。在本研究中，展示了衍生化异羟肟酸和麦司卡林的质谱，并描述了方法的验证。
