Development of a method for qualitative detection of muscimole and muscarine in capsules containing fly agaric and panthera muscular by HPLC-MS/MS method

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Introduction. The growing incidence of acute poisoning with fly agarics is becoming a serious medical and social problem. The capsules containing crushed dried raw materials of red fly agaric (Amanita muscaria (L.) Lam.) and/or panther fly agaric (Amanita pantherina (DC.) Krombh.) are most often found on trading platforms. The raw materials contain psychoactive compounds such as muscimol, ibotenic acid and 1-Methyl-2,3,4,9-tetrahydro-1H-beta-carboline-3-carboxylic acid, which are the reason for their recreational use. The development of methods for the qualitative detection of target substances in capsule extracts is of interest to modern toxicology, since chemical toxicological analysis (CTA) is an essential element of clinical laboratory diagnostics. Obtaining chromatograms and mass spectra of native molecules during capsule analysis will simplify the development of a method for analyzing biological fluids in the future for CTA.

The aim of the study is to develop a method for the qualitative analysis of capsules containing dried raw materials of red and panther mushrooms using liquid tandem chromatography-mass spectrometry.

Material and methods. Red and panther mushroom capsules from the Lesnoy Lekar online store (Russia) were taken as objects of study. Detection and identification of target substances in capsule extracts were carried out on a Nexera XR liquid chromatograph with an LCMS-8050 tandem mass spectrometer (Shimadzu, Japan) using Shim-pack FC-ODS, Shim-pack VP-ODS C8-Phenyl, Shim-pack GIST C18-Aqua HP columns, and a Shimadzu electronic library of mass spectra.

Results. It was found that the optimal method for isolating substances from capsules is methanol extraction and filtration using filter nozzles. Studies have shown that the use of the Shim-pack GIST C18-Aqua HP column and gradient elution mode allows achieving the best parameters for chromatographic separation of muscimol and muscarine.

Conclusions. A method has been developed for isolating muscarine and muscimol from capsules containing crushed dried raw materials of red and panther fly agaric and their qualitative detection using a Nexera XR modular liquid chromatograph with an LCMS-8050 tandem mass spectrometer (Shimadzu, Japan). The Shim-pack GIST C18-Aqua HP column (3 mm × 150 mm, 3 μm) (Shimadzu, Japan) is optimal for analyzing these highly polar hydrophilic toxicants. The following were determined in the methanol extract: muscarine – retention time 3.48 min, muscimol – 2.35 min (the spectrum of ion products had a convergence with the certified library of the device of more than 80%). The obtained data can be of value for clinical and analytical toxicology, and will be further used to develop methods for detecting toxicants in biological objects.

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作者简介

E. Evdokimova

St. Petersburg State Chemical and Pharmaceutical University of the Ministry of Health of the Russian Federation; Scientific and Clinical Center of Toxicology named after Academician S.N. Golikov of the Federal Medical and Biological Agency

编辑信件的主要联系方式.
Email: innova.2000@mail.ru
ORCID iD: 0009-0001-6759-526X
SPIN 代码: 4063-1760

Junior Research Scientist

俄罗斯联邦, Professora Popova str., 14, lit. A, municipal district Aptekarsky Island, St. Petersburg, 197022; Bekhtereva str., 1, St. Petersburg, 192019

O. Balabanova

St. Petersburg State Chemical and Pharmaceutical University of the Ministry of Health of the Russian Federation; St. Petersburg Research Institute of Emergency Medicine named after. I.I. Dzhanelidze

Email: innova.2000@mail.ru
ORCID iD: 0000-0002-8636-9858
SPIN 代码: 6286-3797

Ph.D. (Med.), Lecturer at the Department of Pharmaceutical Chemistry 

俄罗斯联邦, Professora Popova str., 14, lit. A, municipal district Aptekarsky Island, St. Petersburg, 197022; Budapestskaya str., 3, lit. A, St. Petersburg, 192242

D. Pshennikova

St. Petersburg Research Institute of Emergency Medicine named after. I.I. Dzhanelidze

Email: innova.2000@mail.ru
ORCID iD: 0009-0004-2531-410X
SPIN 代码: 3810-1510

Clinical Laboratory Diagnostics Doctor 

俄罗斯联邦, Budapestskaya str., 3, lit. A, St. Petersburg, 192242

E. Kuznetsova

Scientific and Clinical Center of Toxicology named after Academician S.N. Golikov of the Federal Medical and Biological Agency

Email: innova.2000@mail.ru
ORCID iD: 0009-0008-7670-553X

Junior Research Scientist 

俄罗斯联邦, Bekhtereva str., 1, St. Petersburg, 192019

O. Strelova

St. Petersburg State Chemical and Pharmaceutical University of the Ministry of Health of the Russian Federation

Email: olga.strelova@pharminnotech.com
ORCID iD: 0000-0001-6737-1023
SPIN 代码: 1519-7618

Dr.Sc. (Pharm.), Professor, Head of the Department of Pharmaceutical Chemistry

俄罗斯联邦, Professora Popova str., 14, lit. A, municipal district Aptekarsky Island, St. Petersburg, 197022

参考

  1. Voynova M., Shkondrov A., Kondeva Burdina M. et al. Toxicological and pharmacological profile of Amanita muscaria (L.) Lam. – a new rising opportunity for biomedicine. Pharmacia. 2020; 67(4): 317–323. doi: 10.3897/pharmacia.67.e56112.
  2. Turkia Mika. Psycholytic Dosing or «microdosing» of Amanita Muscaria (red Fly Agaric) Mushrooms – a Retrospective Case Study. PsyArXiv. 2023. Nov. 7. doi: 10.31234/osf.io/dve46 4.
  3. Meisel E. M., Morgan B., Schwartz M. et al. Two Cases of Severe Amanita Muscaria Poisoning Including a Fatality. Wilderness Environ Med. 2022; 33(4): 412–416. doi: 10.1016/j.wem.2022.06.002.
  4. Vendramin A., Brvar M. Amanita muscaria and Amanita pantherina poisoning: two syndromes. Toxicon. 2014; 90: 269–272. doi: 10.1016/j.toxicon.2014.08.067.
  5. Stebelska K. Fungal hallucinogens psilocin, ibotenic acid, and muscimol: analytical methods and biologic activities. Ther Drug Monit. 2013; 35(4): 420–442. doi: 10.1097/FTD.0b013e31828741a5.
  6. Flament E., Gaulier J., Paret N. et al. Application to several suspected poisoning cases of a validated analytical method for the determination of muscarine in human biological matrices using liquid chromatography with high-resolution mass spectrometry detection. Drug Test Anal. 2024; 16(4): 331–338. doi: 10.1002/dta.3542.
  7. Ginterová P., Sokolová B., Ondra P. et al. Determination of mushroom toxins ibotenic acid, muscimol and muscarine by capillary electrophoresis coupled with electrospray tandem mass spectrometry. Talanta. 2014; 125: 242–247. doi: 10.1016/j.talanta.2014.03.019.
  8. Kim E.J., Kwon O.S., Hur C.G. et al. Muscimol Directly Activates the TREK-2 Channel Expressed in GABAergic Neurons through Its N-Terminus. Int. J. Mol. Sci. 2021; 22(17): 9320. doi: 10.3390/ijms22179320.
  9. Zhu S., Noviello C.M., Teng J. et al. Structure of a human synaptic GABAA receptor. Nature. 2018; 559(7712): 67–72. doi: 10.1038/s41586-018-0255-3.
  10. Trueb L., Carron P.N., Saviuc P. Intoxication par les champignons [Mushroom poisoning]. Rev Med Suisse. 2013; 9(394): 1465–1470, 1472. French. doi: 10.53738/rev-med.2013.9.394.1465.
  11. Xu X.M., Zhang J.S., Huang B.F. et al. Determination of ibotenic acid and muscimol in plasma by liquid chromatography-triple quadrupole mass spectrometry with bimolecular dansylation. J. Chromatogr. B. Analyt. Technol. Biomed. Life Sci. 2020; 1146: 122128. doi: 10.1016/j.jchromb.2020.122128.
  12. Tsujikawa K., Kuwayama K., Miyaguchi H. et al. Determination of muscimol and ibotenic acid in Amanita mushrooms by high-performance liquid chromatography and liquid chromatography-tandem mass spectrometry. J. Chromatogr. B. Analyt. Technol Biomed. Life Sci. 2007; 852(1-2): 430–435. doi: 10.1016/j.jchromb.2007.01.046.
  13. Flament E., Guitton J., Gaulier J. M. et al. Human Poisoning from Poisonous Higher Fungi: Focus on Analytical Toxicology and Case Reports in Forensic Toxicology. Pharmaceuticals (Basel). 2020; 13(12): 454. doi: 10.3390/ph13120454.
  14. Soyka M. Paranoid-halluzinatorische Psychose bei Intoxikation mit rotem Fliegenpilz [Paranoid hallucinatory psychosis following intoxication with fly agaric]. Nervenarzt. 2023; 94(9): 849–851. doi: 10.1007/s00115-023-01511-6.
  15. Lee M., Dukan E., Milne I. Amanita muscaria (fly agaric): from a shamanistic hallucinogen to the search for acetylcholine. J. R. Coll. Physicians Edinb. 2018; 48(1): 85–91. doi: 10.4997/JRCPE.2018.119.
  16. Государственная фармакопея Российской Федерации XV издания. [State Pharmacopoeia of the Russian Federation XV edition. (In Russ.)].
  17. Su Y. T., Liu J., Yang D. N. et al. Determination of ibotenic acid and muscimol in species of the genus Amanita section Amanita from China. Toxicon. 2023; 233: 107–257. doi: 10.1016/j.toxicon.2023.107257.
  18. Ginterová P., Sokolová B., Ondra P. et al. Determination of mushroom toxins ibotenic acid, muscimol and muscarine by capillary electrophoresis coupled with electrospray tandem mass spectrometry. Talanta. 2014; 125: 242–247. doi: 10.1016/j.talanta.2014.03.019
  19. Merová B., Ondra P., Staňková M. et al. Determination of muscarine in human urine by electrospray liquid chromatographic-mass spectrometric. J Chromatogr B Analyt Technol Biomed Life Sci. 2011; 879(25): 2549–2553. doi: 10.1016/j.jchromb.2011.07.009

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2. Fig. 1. Scheme of the conversion of ibotenic acid to muscimol

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3. Fig. 2. Ion chromatogram containing muscarine, Shim-pack FC-ODS (LC-QqQ)

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4. Fig. 4. Ion chromatogram containing muscimol, Shim-pack FC-ODS (LC-QqQ)

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5. Fig. 3. Spectrum of product ions of muscarine. Collision energy (CE) – 15

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6. Fig. 5. Spectrum of product ions of muscimol. Collision energy (CE) – 15

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7. Fig. 6. Chromatograms on the Shim-pack VP-ODS C8-Phenyl column: A and Б – of red fly agaric extract containing muscimol from capsules and muscarine from capsules respectively; B and Г – of panther fly agaric extract containing muscimol from capsules and muscarine from capsules respectively

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8. Fig. 7. Chromatograms on the Shim-pack VP-ODS C8-Phenyl column: A and Б – of red fly agaric extract containing muscimol from capsules and muscarine from capsules respectively; B and Г – of panther fly agaric extract containing muscimol from capsules and muscarine from capsules respectively

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