Rapid detection of α-amanitin and β-amanitin in rat plasma by ultra-performance liquid chromatography-tandem mass spectrometry and its application to the toxicokinetic study of Lepiota brunneoincarnata.

Zhengsheng Mao, Youjia Yu, Hao Sun, Yue Cao, Qiaoyan Jiang, Chunyan Chu, Yang Sun, Shuainan Huang, Jinsong Zhang, Feng Chen

Forensic toxicology 2022 Jan

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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. © 2021. Japanese Association of Forensic Toxicology.

Citation

Zhengsheng Mao, Youjia Yu, Hao Sun, Yue Cao, Qiaoyan Jiang, Chunyan Chu, Yang Sun, Shuainan Huang, Jinsong Zhang, Feng Chen. Rapid detection of α-amanitin and β-amanitin in rat plasma by ultra-performance liquid chromatography-tandem mass spectrometry and its application to the toxicokinetic study of Lepiota brunneoincarnata. Forensic toxicology. 2022 Jan;40(1):111-118