Amanita compounds chemistry & isolation

[Antoncho]

Hi bees!

Recently, during one of my visits to "Ethnobotanics and Ethnomycology" forum (

http://www.entheogen.ru/

) I was asked a curious question.

One guy named Omar was interested in how to extract muscimol from Amanita mushrooms - and specifically, to get rid of the accompanying muscarine which is thought to bee responsible for fly agaric's toxic effects.

And, frankly speaking, i didn't have a slightest clue    

So - i thought i'd ask you, oh my sophisticated friends

Here are the main Q's that need to bee answered:

1. As i said, how to sep muscimole and muscarine. The former is a primary amine and the 2nd is a quternary amine salt.

Muscimol:



Muscarine:




2. How stable is the molecule itself? Is it stable, you think, to strong acids/bases etc.? Light? Heat?
Generally, what healthy precautions should bee taken? Any ideas?




Hope some of you bees will have something to share




Antoncho

[roger2003]

Maybe, in the references you can find some informations

Muscarine  is found in the fly mushroom, Amanita muscaria, and is a hallucinogenic choline analogue. Its structure was confirmed by X-ray analysis [93]. Muscarine is a parasympathomimetic drug. L-Arabinose was converted into muscarine by EUGSTER [94]. Total synthesis of (+)-muscarine was accomplished by ADAMS [95].
(+)-Muscarine  [300-54-9], C9H20ClNO2 Mr 210: mp 180 – 181 °C (from ethanol).


[93]  F. Jellinek, Acta Crystallogr. 10 (1957) 277.
[94]  C. H. Eugster, Adv. Org. Chem. II (1960), 427.
[95]  J. Adams, M.-A. Poupart, L. Grenier, Tetrahedron Lett. 30 (1989) 1753, and references cited therein.

[Antoncho]

... but .... WHAT are the active species in Amanitas?

Very confused. Some refs say it's muscimol and ibotenic acid.

Others say muscimol is a sedative comp'd, highly potentiating action of narcotics...

OTOH, other sources state that muscarine is a toxin, which is responsible for the unwanted side-effects (nausea etc).



Anyone has a good clue as to the recent state of affairs regarding this issue?


Antoncho


P.S. Literature on the subject is VERY scarce. Search in patent databases hardly gave anything, just this:

Patent FR1427775

- synthesis of muscimol

Patent GB1114116

- synth of ibotenic acid

... nothing was found on xtraction from Amanitas - xcept the 2nd patent where isolation of ibotenic acid is performed, using quite complicated methods and equipment.

But one fact was unquestionnably established - muscimol is absolutely stable to both strong alkalis and acids; in the 1st patent, they boil it for several hours 1st in methanolic KOH (creating muscimol's Me-ether), then in aq. HCl to demethylate

[GC_MS]

Here are some possibly helpful references for you and/or your friend:

Michelot D, Melendez-Howell LM. Amanita muscaria: chemistry, biology, toxicology, and ethnomycology. Mycol Res. 2003 Feb;107(Pt 2):131-46 -

Medline (PMID=12747324)

-

Rhodium's page

(https://www.thevespiary.org/rhodium/Rhodium/pdf/amanita.muscaria.review.pdf)

Professor Buzz. The synthesis of muscimol -

Rhodium's page

(https://www.thevespiary.org/rhodium/Rhodium/chemistry/muscimole.html)

Davis DP, Williams SR. Amanita muscaria. J Emerg Med. 1999 Jul-Aug;17(4):739 -

Medline (PMID=10431970)

Becker H. Components of Amanita muscaria. Pharm Unserer Zeit. 1983 Jul;12(4):111-8 -

Medline (PMID=6684299)

Chilton WS, Ott J. Toxic metabolites of Amanita pantherina, A. cothurnata, A. muscaria and other Amanita species.
 Lloydia. 1976 Mar-Jun;39(2-3):150-7 -

Medline (PMID=985999)

Scotti de Carolis A, Lipparini F, Longo VG. Neuropharmacological investigations on muscimol, a psychotropic drug extracted from Amanita muscaria. Psychopharmacologia. 1969;15(3):186-95 -

Medline (PMID=5389124)

Eugster CH. Chemistry of active materials from the fly agaric (Amanita muscaria). Fortschr Chem Org Naturst. 1969;27:261-321 -

Medline (PMID=4907242)

Eugster CH. Isolation, structure, and syntheses of central-active compounds from Amanita muscaria (L. ex Fr.) hooker. Psychopharmacol Bull. 1967 Dec;4(3):18-9 -

Medline (PMID=5629522)

Bowden K, Drysdale AC, Mogey GA. Constituents of Amanita muscaria. Nature. 1965 Jun 26;206(991):1359-60 -

Medline (PMID=5891274)

Eugster CH, Muller GF, Good R. The active ingredients from Amanita muscaria: ibotenic acid and muscazone. Tetrahedron Lett. 1965 Jun;23:1813-5 -

Medline (PMID=5891631)

Bowden K, Drysdale AC. A novel constituent of Amanita muscaria. Tetrahedron Lett. 1965 Mar;12:727-8 -

Medline (PMID=14291871)

Takemoto T, Nakajima T, Sakuma R. Isolation of a flycidal constituent "IBOTENIC ACID" from Amanita muscaria and A. pantherina. Yakugaku Zasshi. 1964 Dec;84:1233-4 -

Medline (PMID=14266561)

Gonmori K, Yoshioka N. The examination of mushroom poisonings at Akita University. Leg Med (Tokyo). 2003 Mar;5 Suppl 1:S83-6 -

Medline (PMID=12935559)

- DOI:

10.1016/S1344-6223(02)00127-X

Maurer HH, Schmitt CJ, Weber AA, Kraemer T. Validated electrospray liquid chromatographic-mass spectrometric assay for the determination of the mushroom toxins alpha- and beta-amanitin in urine after immunoaffinity extraction. J Chromatogr B Biomed Sci Appl. 2000 Oct 1;748(1):125-35 -

Montanini S, Sinardi D, Pratico C, Sinardi AU, Trimarchi G. Use of acetylcysteine as the life-saving antidote in Amanita phalloides (death cap) poisoning. Case report on 11 patients. Arzneimittelforschung. 1999 Dec;49(12):1044-7 -

Medline (PMID=10635453)

Jaeger A, Jehl F, Flesch F, Sauder P, Kopferschmitt J. Kinetics of amatoxins in human poisoning: therapeutic implications. J Toxicol Clin Toxicol. 1993;31(1):63-80 - [pubmed]8433416[/pubmed]


Here are some abstracts from PubMed:

Hohn H, Schoenemann J. Cholinergic syndrome with unconsciousness in amanita poisoning. Dtsch Med Wochenschr. 2000 Nov 10;125(45):1366-8 - ABSTRACT: HISTORY AND ADMISSION FINDINGS: A 41-year-old patient was found in his flat in a state of coma. After emergency treatment his vital signs were stable and he was transferred to an acute hospital with possible cannabis intoxication. The patient, a hobby gardener, was previously well and had an adversion to the use of any chemical substances. The main symptom showed a cholinergic syndrome with deep coma. We assumed plant ingestion because of the clinical picture and history. INVESTIGATIONS: The laboratory results were within normal limits apart from a slight rise of the serum creatinine kinase level. The electrocardiogram showed a bradycardia. A drug-screening could not be performed. TREATMENT AND COURSE: The differential diagnosis of plant alkaloids or mushroom toxins were considered due to possible plant ingestion and a cholinergic syndrome. Later the toadstool (Amanita muscaria) was found. After treatment oft the cholinergic syndrome with high doses of atropine primary poison elimination was performed. 24 hours later the patient awoke from his coma. Visual hallucinations persisted for a few days. No organic damage due to the intoxication was found. CONCLUSION: Toxic mushroom ingestion can produce a variety of clinical pictures. Most commonly an anticholinergic syndrome is found, but this was not the case in this patient. The effect of the poison depends on the amount and the preparation, so that no reliable outcome prediction can be made. The drug "poisonous mushroom" is legal and hallucinogenic substances are trendy. As a result clinical signs like those described here will have to be expected in the future.

[Rhodium]

I just wanted to have this citation properly posted for the sake of TFSE - I uploaded the PDF in June, but forgot to post a notice in any of the forums here...

Review:
Amanita muscaria - Chemistry, biology, toxicology, and ethnomycology
Didier Michelot & Leda Maria Melendez-Howell

Mycol. Res. 107(2), 131-146 (2003)

(https://www.thevespiary.org/rhodium/Rhodium/pdf/amanita.muscaria.review.pdf)
DOI:

10.1017/S0953756203007305

Abstract
The fly agaric is a remarkable mushroom in many respects; these are its bearing, history, chemical components and the poisoning that it provokes when consumed. The 'pantherina' poisoning syndrome is characterized by central nervous system dysfunction. The main species responsible are Amanita muscaria and A. pantherina (Amanitaceae); however, some other species of the genus have been suspected for similar actions. Ibotenic acid and muscimol are the active components, and probably, some other substances detected in the latter species participate in the psychotropic effects. The use of the mushroom started in ancient times and is connected with mysticism. Current knowledge on the chemistry, toxicology, and biology relating to this mushroom is reviewed, together with distinctive features concerning this unique species.

[Lilienthal]

Wasn't J. Ott providing Aldrich with muscimol?

[crocodile]

Your post about the "spore bank in Florida" reminds me of someone called Stephen L. Peele (does Peele's Lepiota strike a chord?). I am pretty sure he was never busted (he actually had a DEA licence to work with Psilocybe mushrooms) and he was/is into mushroom extractions and has published several booklets on the subject. He is still active in the field and is still offering spores of many exotic mushrooms.

Hope this helps... PM me if you need more info.

[demorol]

Here is another article for those interested in chemistry of Amanita Muscaria. The paper describes preparation of muscimol and its derivatives, including N-methylmuscimol, N-isopropylmuscimol, N-morpholinomuscimol, and other.
 

Synthesis of N-Substituted Muscimol Derivatives Including N-Glycylmuscimol
Michael FREY, Volker JÄGER

Synthesis 1985, 1100
DOI:

10.1055/s-1985-31439

Abstract

The preparation of muscimol (a potent but toxic GABA neurotransmitter agonist), from dimethyl acetylenedicarboxylate via methyl 3-hydoxyisoxazole-5-carboxylate and the corresponding amide, has been improved and extended to a general synthesis of N-alkyl and N,N-dialkyl derivatives. The efficient route to muscimol itself [muscimol itself costs 60-170 DM/10mg] has enabled a study of its chemistry to be undertaken, a first result of which is its incorporation into a peptide, i.e. the preparation of N-glycylmuscimol.

[Rhodium]

An Improved Synthesis of Muscimol
P. Pevarello & M. Varasi

Synthetic Communications 22(13), 1939-1948 (1992)

(https://www.thevespiary.org/rhodium/Rhodium/djvu/muscimol.djvu)

Abstract
A regiospecific 1,3-dipolar cycloaddition/elimination is the key step of a convenient, gram-scale synthesis of the GABA-agonist Muscimol (1).