methanol extraction

[benzyme]

the issue is oxidation of the 4-position OH.
psilocin is less stable than bufotenine due to intramolecular forces, sterics
(when you look at the molecules in three dimensions, this becomes apparent)

 

[Ambivalent]

this also looks like a good method to obtain fairly pure psilocin. i am interested if the d.ether could be substituted for another solvent. and maybe the evaporation step on the end could be substituted with some other step. would bringing it back to acidic solution hurt ?
Experimental

A representative sample of 2 to 10g of dried mushrooms is ground to a fine powder by mortar and pestle. The powder is mixed with 100 mL of dilute acetic acid in a 250-mL beaker. The pH is readjusted to pH 4 with glacial acetic acid. After standing 1 h, the beaker is placed in a boiling water bath for 8 to 10 min or until the internal temperature of the acid mixture reaches 70�C. The beaker is removed and cooled to room temperature under running water. The acid mixture is separated from the mushroom powder by suction filtration using glass wool. The filtrate is brought to pH 8 with concentrated ammonium hydroxide and quickly extracted with two 50-mL portions of diethyl ether. Gentle mixing instead of shaking should be used to prevent an emulsion. The ether is dried over sodium sulfate, filtered, and evaporated under nitrogen with no applied heat.
Crude psilocin will appear as a greenish residue. Recrystallization from chloroform/heptane (1:3) yields white crystals. The resulting powder can then be submitted to infrared and mass spectral analyses.
Results and Discussion

This method permits rapid isolation of psilocin from hallucinogenic mushrooms by co-extraction of both psilocin and psilocybin. Dilute acetic acid is an excellent solvent for this purpose, because both compounds are very soluble in acetic acid11 and very little of other interfering substances are extracted, It is most likely some other compounds are co-extracted but are removed from psilocin in the ether extraction from the aqueous base. Psilocybin is completely dephosphorytated to psilocin by heating the acid extract. After addition of the base, extraction into ether should be performed promptly, because of decomposition of psilocin at a greater pH than 712. The extraction and dephosporylation steps produce reasonably pure psilocin from a small amount of mushroom material. Two grams of mushrooms will often be sufficient to obtain an infrared spectrum of psilocin (Fig. 1). Smaller mushrooms exhibits provide ample psilocin for mass spectral analysis (Fig. 2).
This method has been used in our laboratory for six months and has given excellent results in separating psilocin from methanol-soluble compounds. Other identification techniques such as gas chromatography and microcrystalline tests are possible on psilocin extracted by this method.

 

[benzyme]

what you cited is the aforementioned A/B method, from the 1981 paper by Casale et. al.

A/B with psilocybin/psilocin is tricky, it has two groups with different pKa's. it's a suitable method for analysis, but may or may not yield an active product.

I actually neglected the pyrrole nitrogen, which also has its own pKa.

chemicalize: psilocin
chemicalize: psilocybin

(click on manage calculations, pKa)

Gentle mixing instead of shaking should be used to prevent an emulsion. The ether is dried over sodium sulfate, filtered, and evaporated under nitrogen with no applied heat.
Crude psilocin will appear as a greenish residue. Recrystallization from chloroform/heptane (1:3) yields white crystals. The resulting powder can then be submitted to infrared and mass spectral analyses.

yea, I've tried the method listed in that paper before.. three times.

 

[Ambivalent]

(click on manage calculations, pKa)

yea, I've tried the method listed in that paper before.. three times.

everything looks doable, even for a kitchen lab chemist/enthusiast. i was wondering if the "evaporation under nitrogen atmosphere" step can be substituted with some other method to get the psilocin out of the organic solvent.
will it be stable enough to bring it back from the np layer to aqueous acidic solution ? i have no experience how fast oxidation of psilocin occurs under normal conditions.

 

[benzyme]

that's the problem, and the reason nitrogen is used to purge oxygen from the solvent (to prevent autooxidation).
when the amine is deprotonated, the hydroxy is also deprotonated to some extent, especially past a pH of 8.
overshoot it, and everything may as well be tossed out.

that hydroxy-group has to keep its proton (hydrogen) to remain active.

 

[Ambivalent]

than, is it a good idea to react a concentrated solution of psilocin with acetic anhydride to achieve a more stable molecule ? i hope i am not crossing some strict line with this question.

 

[benzyme]

acetic anhydride is a rather watched chemical, as you already know, but you've got the right idea. that's all I can say about that.

 

[Ambivalent]

i understand, and thanks for answering me anyway. it involves a watched chemical yes, otherwise i find this conversion/synthesis example to be almost analogous of the harmine/thh conversion, so i dared to ask for an opinion.

 

[benzyme]

the harmine/thh conversion is a reduction reaction, hydrogenation. what you're inquiring about is an [acetyl] addition reaction (acetylation). two different animals.

 

[futura]

and proteins, which is probably what the green goo is that people get.
got some of that too, it was just separated out by the centrifugal force

i'd conservatively estimate 40% - 45% psilocybin. experiments with re-x are in order

nevertheless, it seems alcoholic extractions are the way to go, to preserve the activity. A/B seems to render product inactive by altering the charge state of the OH, which is ever-susceptible to oxidation.

Would a defat with Anhydrous Acetone remove most of the sugars and fats??

My plan is to:

Grind up dried Cubensis into a fine powder.

Soak in Anhydrous Acetone for 24 hours.

Vacum Filter and dry.

Soak in Anhydrous Methanol for 24 hours.

Vacum filter and discard powder.

Fan evaporate Methanol in 9x13 glass baking pan leaving Psilocybin Crystals.

Wash crystals in 150 degree Farenheit Water to boil off remaining methanol(Methanol boils at 149 Deg. F)

Evaporate water leaving Psilocybin Crystals.

Dissolve crystals in known quantity of Everclear for easy dosage and storage.

Crystals plus methanol should total about 1 ml per gram of starting material.

So if you started with 20 grams of cubes, put all crystals into measuring vial and add Everclear to reach 20ml mark and there you have it...1 ml = 1 Gram cubes!!

I keep in 1 ml vials in the freezer...some will crystalize out on bottom of vial...

When ready to dose just take as many vials out of freezer as dose you want, take cap off and empty and then drop drop vials into glass of very warm water till all crystals melt...

Swallow water and away you go!!

No stomach upset as your not trying to digest nearly undigestable Cubensis cells!!

 

[futura]

the basic approach is grinding dry mushrooms and soaking in alcohol. this is allowed to sit in the freezer for a few day, then transferred to room temp. this may be cycled (optional). then, heat the slurry in a 40 C water bath for half an hour. filter, and evap
(if possible, use reduced pressure).

Why the heat(40 C water bath for half an hour??) I thought heat was bad??

 

[benzyme]

40 C is no problem with psilocybin. I was probably thinking about the acidic conversion/extraction of psilocin when I posted that. acid-catalyzed dephosphorylation requires low heat. a hot water bath, in general, helps increase solubility of alkaloids in solution

 

[InMotion]

@futura - "Defat" with acetone will pull the actives.

The main key here is air exposure, the bane to almost all amines especially aryl amines and tertiary amines(commonly used in radical reactions). UV Light is also not the friend of activated phenol groups in which psilocin is.

In my opinion if one doesn't have the proper equipment or means to remove solvent with very little atmosphere it may be best to find satisfaction in an ethanol tincture or a water based tea. Tincture keeps best cold and in the dark, ascorbic acid can be added to stop the initiation phase of some undesirable radical reactions.

 

[futura]

@futura - "Defat" with acetone will pull the actives.
quote]

What can I use to "Defat" the Mushroom Powder before doing a methanol extraction?

I plan on defatting,then Vacum drying, then doing an Anhydrous Methanol extraction, Vacum Purging Methanol, scraping up results from filter paper and dissolving in Everclear(1ml per 1 gram of the Mushroom Powder I started with) and storing in 1 ml vials stored in freezer. When ready to dose will just take cap off vial and pour contents into 8oz warm water with a little citric acid, swirl vial with water to clean out any crystals and down the hatch!!

Any thoughts or suggestions on this??

 

[Orion]

Defatting with acetone won't pull the actives at all, go right ahead! Just make sure it is anhydrous acetone.

 

[Trickster]

About a year ago 30 g of cubes were extracted with hot methanol. The extract has been dried under vacuum. It looked like rather homogeneous thick brown goo. For some reason the extract has been forgotten for almost a year. It has been kept in a fridge ~5C). To my amazement when I opened the glass there was a huge crystal closely resembling that of salt. There is also dark brown sludge in the glass. I separated them and saved both. A photo of the crystal together with 1 euro cent is attached. I did not try to remove the goo completely.
Some notes on its physical properties. It weighs 1.75 g. It does not melt under rt. It is rather translucent and looks like a single crystal. A few words about the liquid. It does not mix with ethanol or methanol. It is easily dissolved in hot water and quite active.
I would appreciate your comments, comrades.

 

[endlessness]

very interesting... got a thiele tube to check for mp?

When you say the liquid is active, do you mean the brown goo? Or the crystal is active? Did you check for solubility of crystal on any solvent.

 

[Trickster]

very interesting... got a thiele tube to check for mp?

Nope. I could order if from a glassblower but I have no idea what liquid I can use in it with the psilocybin mp around 220C.

When you say the liquid is active, do you mean the brown goo?

Yes. I moved the goo to a smaller beaker trying to scrape off every little bit. Then I washed the old beaker with warm water and me and my gf had a wonderful mild trip (~1.8g of dry cubes by my subjective experience). Coming down right now.

Or the crystal is active?

Didn't test it yet.

Did you check for solubility of crystal on any solvent.

I thought of that. What would be a definitive solubility test? DCM or Benzene compared to Methanol of water?

 

[Orion]

If it does not dissolve in methanol surely it cannot be psilocybin ?

 

[Trickster]

I think, this crystal could not be pure psilocybin. The math is simple. There were 30 g of dry cubes. Even if I got lucky and there was 1% of psilocybin in there, there could be only 300 mg of psilocybin, not 1750 mg. On top of that, the remaining goo is still very active. Still it is very interesting what is in there. Maybe I should do a bioassay, trying 20 mg of the crystal substance first.

If it does not dissolve in methanol surely it cannot be psilocybin ?

Right. It should dissolve in methanol and should not in benzene. Will test in a few days.