I thought psilocybin was inactive anyway because it couldn't pass the blood/brain barrier? I'm sure someone mentioned psilocybin gets converted to the active psilocin in the gut.

Psilocybin is a prodrug - it gets converted to psilocin in the body. It is far better to have psilocybin extracted than psilocin because the former is far more stable from oxidation and decomposition whereas the latter is a fairly susceptible molecule that requires some delicate handling


But how about psilocybin salts? psilocybin can theoretically form salts and some psilocybin form in the mushrooms may very well be ethanol soluble. I believe the ethanol quote refers to the freebase psilocybin.

I didnt know psilocybin could form salts, being a phosphate ester (and thus I didnt know you could even call it 'freebase' psilocybin), and specially because I thought acidic solutions (which would be necessary for the salt to form) would actually necessarily dephosphorylate it to psilocin....

and also in lorax's plan, he used plain alcohol soak on fungal mass, so how could it be forming a salt anyways?





yes but this is not really relevant to my point. Even if psilocybin does get metabolized to psilocin before being absorbed, its metabolic product is active, which means ingesting psilocybin is going to be psychoactive, and thus leaving psilocybin behind on a fungal mass is still waste of precious material

Ah, tough issue; psilocybin is a zwiterion, (just like the amino acids that make up proteins) and therefore has a net positive charge in low pH, a net negative charge at high pH and a net zero charge at the isoelectric point, or pI.

This is because it has an amine group that can theoretically be protonated (so psilocybin becomes positively charged) and can theoretically form salts like psilocybin carbonate, but also has the phosphate group that can be deprotonated (so psilocybin becomes negatively charged) and can theoretically form salts like calcium psilocybinate, sodium psilocybinate etc. The amine can have a charge of 0 or +1, where the phosphate group can have a charge of 0, +1 and +2 (ot has two sites of possible deprotonation). Benzyme has calculated theoretically that the net zero pH for psilocybin to be around 4 (he also showed that psilocybin can be found in 5 different states depending on the combination of charges that the phosphate and the amine may have!) If we assume this to be true, then psilocybin is a freebase at pH ~4, positively charged (and potentially salt-forming at pH lower of 4 and negatively charged at pH above 4.
I attached one paper some pages ago where some people showed that psilocybin has a negative charge from pH 4, which totally supports ben's calculations.

SWIM was recently working on am approach for psilocybin isolation form liberty caps (~1% psilocybin) playing around with the pH but he had to leave this project behind because of more important priorities in life! He did an aqueous extraction of fungal biomass using just boiling water. This water had a pH or around 6 at room temperature, indicating that the environment in the mushrooms is slightly acidic, and implying that psilocybin is not at its freebase state, maybe some phosphate salt? And maybe soluble in ethanol? Who knows?

Interesting information Inf, gives me the jumpstart to research more into this all, thanks!

But still my question remains: Even if it theoretically can form a salt, wont it dephosphorylate before that happens? IIRC I had read that plenty of times in publications that acidic conditions do so.... Maybe there are other conditions that can prevent dephosphorylation and yet make it form a salt?

Also, you say that theoretically it can form a salt, but was there ever any analysis that it can in fact do so, and not just theoretically? Is there any publication out there of this happening, and if so, under which conditions?

endlessness quote...
"yes but this is not really relevant to my point. Even if psilocybin does get metabolized to psilocin before being absorbed, its metabolic product is active, which means ingesting psilocybin is going to be psychoactive, and thus leaving psilocybin behind on a fungal mass is still waste of precious material "

I agree that no one wants to waste any of the precious material, especially from the mushroom

I was wondering how freebase psilocybin would be active without getting broken down to psilocin... Or are you just wanting to extract to make oral consumption more palatable?

I would love to see what vaping pure psilocin is like, but due to it being so unstable.. It dosen't seem possible.

Im not sure I understand you, camakazi.. Im not trying to make oral consumption more palatable, im commenting on someone's mushroom extraction. Lets say for the sake of using numbers that 50% of the alkaloids in a mushroom is psilocin, and 50% is psilocybin. If he pulls with ethanol, he might pull those 50% psilocin but the other half of the alkaloids will be left behind. So again, it doesnt matter if psilocybin becomes psilocin in the body, what matters is that it is gonna be psychoactive one way or another (if you take pure psilocybin you will have a psychoactive experience). So im not claiming it is active without getting broken down to psilocin, im just arguing that it is, directly or after being metabolized to psilocin, an active alkaloid and therefore shouldnt be wasted. Did that make more sense now?

I think I understand where the confusion arises; I believe that you assume the salt formation to occur at the site of phosphoryl ester formation? Salt formation (if it happens!) happens on different parts of the molecule (either the "positive" end, or the "negative" end, as shown in the diagram below. I hope this clears things up!

EDIT: more on psilocybin from benzyme, where it shows the pH range at which psilocybin exists as zwitterion (blue line). According to this graph at a pH of 4.2 psilocybin is at teh freebase state whereas protonation of amine (positive charge) and deprotonation of the phosphate (negative charge) are at a minimum.


Finallly, re to the psilocybin->psinocin conversion, we are not sure enough of how psilocybin dephosphorylates to psilocin. Incubation with strong acid at hot temperature seems to do it, or that's what anecdotal reports say. At least this is the implication of a mushroom tea with a dash of lemon that comes on faster. Or is it because the lemon tea does not dephosphorylate but instead extracts the actives, thus making it faster to come?

Mushroom extracts with strong acetic acid definitely does it, as some papers describe but again this may be due to a combination of enzymes that get extracted from mushrooms and/or the low pH. Stomach pH is low (around 2) it may dephopshorylate psilocybin; Or it may not, and psilocybin is dephosphorylated by phosphatase enzymes in the body. Or a combination of both? I may be wrong on some of my dephosphorylation assertions as it has been some time since I read articles about this issue. Please chime in if something is wrong or needs amendment!

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No it wouldn't work and it doesn't work. Steps 4 and 5 are the culprit as they are the most problematic ones. But the other steps are fine, even though defat is not a big issue with mushrooms. During the makign of the mushroom tea more things than just alkaloids get into the water. Notably sugars and proteins. These will be present in step 4 when one tries to re-x and will also form crystals. Many people see crystals when doing similar procedures but these crystals/precipitates are often things not psilocybin.

SWIM has tried these things and there's always far more that crash out from a hot methanol and/or ethanol crystallisation. This, or he'll have to assume that he gets yields of ~10% psilocybin crystals, which is horrendously unrealistic!

The main issue is if one can get rid of these things without getting rid of psilocybin....

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No, proteins are often very water-soluble molecules, same goes for sugars. Defat would not be of much help here. One can make an acetone defat on dried tea, but this does nnot really remove much.

Also, what crashes out of ethanol is a mixture of psilocybin, proteins, sugars and whatnot. One step forward, one step backwards on other words.

Ok I understand what your saying, I'll brainstorm and come back with another barrage of questions later lol.

Thanks!

^From here.

The other thread says a/b with acetic/amonia or acetic/sodium carb should work as long as very little heat is used and the ph doesn't get too high in the latter step. It also says that fumaric acid won't be good for psilocin because it doesn't protect the molecule from oxidation and the product would degrade, which is a bummer since FASA is sooo easy. (weird since bufo has an OH group but is relatively stable and the fumarate is just fine, all because of the 4 vs 5 position ). The recommendation in that thread is to use ascorbic acid in solution with psilocin for storage to prevent oxidation.

So assuming a/b gets psilocybin => psilocin, some questions:

1. Could an a/b with FASA yield reasonably isolated psilocin fumarate? Will this salt form at all or long enough to be filtered from the solvent? The logP is damn close to dmt. It could then be freebased and stored with vit-c to avoid oxidation.

2. Is psilocin ascorbate solid? Could it be precipitated from product-rich solvent in a FASA-like fashion? (ascorbic acid dissolved in whatever solvent being used, added drop-wise to psilocin containing solvent until no more clouding appears)


FASA or some salt precipitation to isolate sounds peachy, but this is hinging off the assumption that a/b dephosphorylates.

fumaric acid is an antioxidant. I am such a biochem noob

yeah you are

ascorbic acid is the commonly used antioxidant for these things, n00b.

I hope something comes of this, im getting rather obsessed with obtaining pure psilocybin...

got 300g dry to try this on
is it worth my while to do a comparison - would it be beneficial to the community ?
eg etoh or etoh/water vs methanol
il do it under vaccum - no sonicator tho.

yes

sonication is optional, but evap under reduced pressure helps resolve the product.
I guestimate psilocybin content of an anhydrous alcoholic extraction to be somewhere in the 40% area, the number is probably lower with EtOH containing water. The pictures from Crystals of The Gods is very misleading...once the product is separated from the alcohol, it is not crystalline at all.


I have yet to isolate psilocybin into pure crystals, it's very tricky. the molecule seems to bind water, proteins, and other biomolecules.

Im interested in getting into a potent tonic, not bothered about crystals, but i want it to be pure without other misc alks