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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

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Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

Establishing UPLC–MS/MS method for quantifying amanitines in rat plasma successfully enabled toxicokinetic study of Lepiota brunneoincarnata extracts.

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Acknowledgements

This work was supported by the Natural Science Research of Jiangsu Higher Education Institutions of China (Grant No. 20KJB340001) and the National Natural Science Foundation of China (Grant No. 82002028, 81772020 and 81922041).

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Author notes

  1. Zhengsheng Mao, Youjia Yu, and Hao Sun have contributed equally to this work.

Authors and Affiliations

  1. Department of Forensic Medicine, Nanjing Medical University, Nanjing, 211166, Jiangsu, People’s Republic of China

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

  2. Department of Emergency, Jiangsu Province Hospital, The First Affiliated Hospital of Nanjing Medical University, Nanjing, People’s Republic of China

    Hao Sun & Jinsong Zhang

  3. Key Laboratory of Targeted Intervention of Cardiovascular Disease, Collaborative Innovation Center for Cardiovascular Disease Translational Medicine, Nanjing Medical University, Nanjing, 211166, Jiangsu, People’s Republic of China

    Feng Chen

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  1. Zhengsheng Mao
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  2. Youjia Yu
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Correspondence to Zhengsheng Mao, Jinsong Zhang or Feng Chen.

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All animal care and handling procedures used in this study were approved by the Animal Care and Ethical Committee of Nanjing Medical University.

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Mao, Z., Yu, Y., Sun, H. et al. 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 Toxicol 40, 111–118 (2022). https://doi.org/10.1007/s11419-021-00607-5

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  • DOI: https://doi.org/10.1007/s11419-021-00607-5

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