Mao Zhengsheng, Yu Youjia, Sun Hao, Cao Yue, Jiang Qiaoyan, Chu Chunyan, Sun Yang, Huang Shuainan, Zhang Jinsong, Chen Feng
Department of Forensic Medicine, Nanjing Medical University, Nanjing, 211166, Jiangsu, People's Republic of China.
Department of Emergency, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing, People's Republic of China.
Forensic Toxicol. 2022 Jan;40(1):111-118. doi: 10.1007/s11419-021-00607-5. Epub 2021 Dec 1.
Lepiota brunneoincarnata is a well-known poisonous mushroom and is responsible for fatal mushroom poisoning cases worldwide. α-Amanitin and β-amanitin are the main amatoxin compounds of Lepiota brunneoincarnata. However, there are no published toxicokinetic studies of Lepiota brunneoincarnata. To study the toxicokinetics of Lepiota brunneoincarnata, we developed an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for determination of α-amanitin and β-amanitin in rat plasma.
UPLC-MS/MS analyses were performed with a triple quadrupole mass spectrometer in positive-ion mode. The sensitivity of α-amanitin and β-amanitin detection was increased by inhibiting the production of [M + Na] adducts. α-Amanitin and β-amanitin were separated and quantified on an UPLC octadecyl silyl column in only 2.5 min.
The linear ranges were 3.0-3000 ng/mL for α-amanitin and 1.8-1800 ng/mL for β-amanitin with a correlation coefficient r > 0.99 for both analytes. The lower limit of quantification of 3.0 ng/mL for α-amanitin and 1.8 ng/mL for β-amanitin was achieved using only 50 μL of rat plasma. The accuracy of α-amanitin and β-amanitin was between - 9.5 and 7.0% with the precision ranged from 2.2 to 12.5%. The developed method was then applied for Lepiota brunneoincarnata toxicokinetic study after intravenous administration of Lepiota brunneoincarnata extracts.
Establishing UPLC-MS/MS method for quantifying amanitines in rat plasma successfully enabled toxicokinetic study of Lepiota brunneoincarnata extracts.
褐肉环柄菇是一种著名的有毒蘑菇，在全球范围内导致致命的蘑菇中毒事件。α-鹅膏毒肽和β-鹅膏毒肽是褐肉环柄菇的主要鹅膏毒素化合物。然而，目前尚无关于褐肉环柄菇的毒代动力学研究报道。为研究褐肉环柄菇的毒代动力学，我们开发了一种超高效液相色谱-串联质谱（UPLC-MS/MS）方法，用于测定大鼠血浆中的α-鹅膏毒肽和β-鹅膏毒肽。
采用三重四极杆质谱仪在正离子模式下进行UPLC-MS/MS分析。通过抑制[M + Na]加合物的产生，提高了α-鹅膏毒肽和β-鹅膏毒肽检测的灵敏度。α-鹅膏毒肽和β-鹅膏毒肽在UPLC十八烷基硅烷柱上仅需2.5分钟即可分离并定量。
α-鹅膏毒肽的线性范围为3.0 - 3000 ng/mL，β-鹅膏毒肽的线性范围为1.8 - 1800 ng/mL，两种分析物的相关系数r均>0.99。仅使用50μL大鼠血浆即可实现α-鹅膏毒肽3.0 ng/mL和β-鹅膏毒肽1.8 ng/mL的定量下限。α-鹅膏毒肽和β-鹅膏毒肽的准确度在-9.5%至7.0%之间，精密度范围为2.2%至12.5%。在静脉注射褐肉环柄菇提取物后，将所建立的方法应用于褐肉环柄菇的毒代动力学研究。
成功建立了用于定量大鼠血浆中鹅膏毒素的UPLC-MS/MS方法，从而能够对褐肉环柄菇提取物进行毒代动力学研究。
