Benzyme: Do you have pics of this yet? Good work man, SWIM is going to be doing a psilocyin extraction soon, and theres nothing like freeze precip for getting nice and pure.

fuck, a shitload of crystal psilocybin would really fuck the party up

So, theoretically, what would good pH levels for the acid and base steps be?

4 and 8, respectively. though don't keep it in base for too long
between 2.5 or 2.6 to 3 is ok too, so vinegar (5% acetic) would work fine, maybe dilute it a bit.

What about chloroform? Harder to come by, but not completely inaccessible. LSD tartrate is precipitated in a tartic acid/chloroform solution.

D-tartaric, specifically (the d-isomer helps preserve the stereochemistry of the intermediate).

chloroform is a harder to come by, as it's used little outside of research; it's been widely replaced by the less toxic methylene chloride, although both are obtainable if one searches thoroughly.

are you sure it was chloroform? I was under the impression that chlorine ions isomerize lsd

You know what, it is because it was not in chloroform. Been a long time since I last looked at Otto Snow's work. He had LSD in methanol, and precipitated it by adding a solution of tartic acid/MeOH. Lol, silly me.

Maybe put the fungus in a sohxlet and extract with MeOH, and then add a solution of tartric acid/MeOH to precipitate the goodies?

I am just trying to take the principals of one method and apply it to another situation. I don't understand this stuff like you do, but I am still hopeful I can figure out something good.

Freebase Psilocin sounds fascinating. Has anyone tried smoking/vaping it yet?

If so, is it effective? How long does it last?

Thanks. I'd love to try this out.

[Edit] I've done my own research on a few other fora and there are those who claim to have vaped freebase Psilocyben/Psilocen.

There are those who claim to have vaporised Psilocin crysrals.



Seems intriguing. I might try this if I ever get around to growing mushrooms.

The Nexus is certainly full of very interesting ideas that's for sure. In this thread a claim is made that the inner salt of psilocin will move into xylene (!).

Well, once in xylene (if the claim is true), Could FASA work? Situation is not quite the same as with DMT where the fumaric acid donates a proton directly to the amine. In the case of psilocin, the amine is already pronated and it is the hydroxy group the one that needs a proton from the acid to become positive overall. However, once that proton is transferred, the psilocin has an ovearall positive charge at the amine group, just like DMT during FASA. Could the fumerate salt form and crash out of the xylene?

I think this is worth a try, right? Maybe try tartaric acid too which is somewhat soluble in acetome (0.02 mol fraction, see attached, which I think is roughly about 40mg for every ml of acetome?, Seems high...).

Update:

10g of dry mushrooms where pulverized and extracted in vitamin C water (pH 4). Water was brought up to 70C to convert psilocybin to psilocin. Extract was filtered and ammonium sulfate (30%) was added. Proteins crashed out and where removed. Resulting clear tawny liquid was brought to a pH of 8.25 with ammonia drops and became darker in color. Xylene was gently mixed in for 5 minutes, and allowed to separate. Between the water and xylene layers a clear oily diffuse layer formed. Oily layer and xylene where separated out and FASA added. Cloudiness was observed and upon shaking a white precipitate formed (picture below).

The precitpitate could be many things and needs to be tested further. One possibility could be that it contains psilocin fumarate. On the other hand maybe it only contains inactive mushroom gunk. Feedback welcome.

Plan is to isolate and cleanup the precipitate as per a standard DMT FASA followed by bioassay. If activity is noticed, send a sample out for analysis.

Update: there seems to be some activity in the dried precipitate (first image). I took 30mg and while not a deep experience I'm pretty sure I felt the effects of a small dose (equivalent to sub gram of dry mushrooms). I don't want to claim that this is certain (could be placebo), so will send out for analysis.

About 200mg were recovered from the first (and only) xylene pull of the 10 dry grams. A bit too much for it to be only psilocin fumarate (would expect about 120mg total for 1% yield if pure and that should take several pulls)

I'm not experienced with FASA, and in fact, this was my first time using it. I followed the wiki info with one dry acetone wash at the end. Seemed to work just like it should.

A sample of the powder dissolved in a little bit of water very easily, so I think there is not a lot of free fumaric acid (pH was above 6 too, seems consistent with a salt). When this sample dried it turned off white and slightly blue-green (second picture), so maybe this kind of water cleanup is not advisable for our hypothetical psilocin fumerate.

Based on this will send out the powder for analysis.

Checked the leftover xylene that had FASA added from the April 17th post. It's turning blue after a few days (image below). I think this is consistent with psilocin moving into the xylene. There must have been some unreacted psilocin.

Will publish the LC/MS data on the precipitated powder when I have it.

Results from the FASA precipitate are in (grateful thanks to benzyme who ran the anlysis more than once to verify). Note that the masses are shifted in the mass spec below by +2 a.u. For example 258 au in the plot, really means 256, so will be using the notation 258 (256) when reffering to the plot.

The largest peak at 202 (200) is unkown at this time. We are not sure of what it is, psilocin should have been at 206 (204), and a simple oxidized form of psilocin at 204 (202). Would be grateful if anyone had any theories on what this peak is. We don´t know what it is after reviewing the valuable discussions on oxidized psilocin previously done on the Nexus (see this post by Pitubo). Any thoughts on this peak and the peak at 242 (240)?

Among the smaller peaks, there is one at 272 (270) and 258 (256). We are pretty confident these are baeocystin and norbaeocysin. This is great, because it means we have moved actives into the xylene and precipitated them with FASA. This is also consistent with the previous bioassay.



We failed to obtain psiclocin though. This could be either because (1) there was too much time or temperature was to high during basing (original attached paper warns to be quick and cool down the extract before basing), or (2) psilocin did not move into the xylene.

On the other hand, the detection of (nor)baeocystin is very interesting. They moved into xylene, despite not being at the isoelectric point. This implies that these compound moved well into xylene (a surprise, perhaps ionic strength of water affects them drastically via the mechanism suggested by downwardsfromzero). The natural question is if we had not dephosphorilated psiclocybin, would it have moved into xylene as well? The observation hints that (nor)baeocystin did not dephosporilate as easily as psilocybin (perhaps the dephosphorilizing mushroom enzymes are specific to psilocybin).

So for the next round of tests, we are starting with an alkaline extraction in Ca(OH)2. Work by Infundibulum has indicated that the actives stay mostly in the water. The extract is salted with ammonium sulfate to make it slightly acidic (near the pI of psilocybin) and extracted with xylene. It may be important to denaturalize proteins before making acidic so dephosphorilation does not start. This process has been done and a small bioassay gave close eyes visuals, so it will be sent out for analysis.

Another next round of tests involves careful management of temperate and time during the original extraction to avoid psilocin oxidation.

Ideas welcome to help us isolate these elusive zwitterions. Cheers.

Endlessness and nen888 also found a (200) peak years ago when working on acacia extracts. Looks like they determined it was leptocladine. Endlessness pointed out several theoretical secondary peaks with (157) among them.

Could leptocladine be a candidate for what we are getting here at (200) and (158 )?

Also, is leptocladine considered a mutagen (see attached paper)? I wonder what McKenna would say about that and his stoned ape hypothesis

Not sure what it really is, but it would be interesting if some acacias and magic mushrooms shared a ß-carboline.

So this is a brilliant thing to know, and I'm really excited to test if heptane works as well as theorized. However, I remember in other extraction threads that there needs to be a certain pH for this to work. I also remember benzyme talking about a psilo extraction that involved diethyl ether, though I can't for the life remember what that invovled or where I saw it. Maybe in chat?

But back to the issue at hand: does this freeze precip need to be at a certain pH at each stage in order for it to work?

Edit: I also found an instructable for a simple anaerobic chamber, since psilocin need an oxygen free space. This can easily be applied to other oxygen-sensitive compounds, and most of these boxes are made with polypropylene, which resists most chemicals pretty well: http://www.instructables...c-Chamber-aka-glove-box/

I just want to thank you for the contributions in these many threads Loveall. Some of these results seem quite promising. I'm fairly certain of the blue and blue green colours being a good indicator of the presence and degradation of psilocin.

I am lost as to why no psilocin was found in the analysis, even though the xylene clearly shows a pale greyish blue discoloration. Seems to me that psilocin did move into the xylene. Was the psilocin fumarate somehow converted to pure psilocin before analysis or did it remain a salt? Could either one of these degrade into an unknown form before being analysed?

One other thing. You mentioned other actives were present in the extract, is it reasonable to assume these could be the cause of the perceived activity if not placebo ? This could mean that all of the psilocin was still in the xylene but FASA could still produce an active extract free of psilocin (...IF that's the case).

I'm also interested in the idea of not having to dephosphorylate the psilocybin before having migrate into a separate nonpolar phase. This bypasses a lot of problems if possible.

Hello, the diethyl ether extraction paper from Casale is here: https://erowid.org/archive/rhodium/pdf/shroom.psilocin.extraction.pdf

What we are doing here is a fumarate precipitation. Unfortunately, psilocybin is missing from the mass spec result. Good news is that we got confirmation of (nor)baeo and we also got some activity. We are trying to figure out how to get the psilocybin part now.

Thanks Orion. I've been thinking about your post. Maybe the psilocybin precipitated with the proteins and that is why it did not show up at the end. Soooo, I'm doing more tests not removing any percipitates. Will send details out in a minute.

New round of testing:

- Ground 150g fresh mushrooms in Ca(OH)2 with splash of water to make a slurry. pH ~ 12. No clear degradation from a color change. Stable light gray.
- Spread slurry in Pyrex shallow pan and dry at 170C. Chalky flaky powder forms, still light gray. This may denaturalize proteins and trap them in a Ca matrix so they are not in the way (free psilocybin and fewer emulsion chances, but this is all speculation).
- Extracted powder with IPA, add FASA, nothing.
- Extracted powder with denatured alcohol, add FASA, nothing.
- Extracted with xyleme, add FASA, nothing.
- Added Ammonium Sulphate, extract with xyleme, add FASA, nothing.
- Added some water to dissolve Ammonium Sulfate. Strong Ammonia smell from reaction with Ca(OH)2. pH is lowered (pH meter says down to 9.6).
- Xylene extraction. Add fumarate. White precipitate and lots of it . Resulting loose flaky powder is in the mail for analysis. However, could simply be a protein powder with some (nor)baeo.

Next steps: Wait for analysis. If no psilocybin extract with xyline again at pH ~ 4 and send that out if more white precipitate forms during FASA.

Feedback/ideas welcome. What am I missing?

This is similar to how we got (nor)baeo fumarate, however protein precipitates where never discarded this time. There was also a drying step and several solvent washes of the alkaline powder which may or may not be needed (I'm guessing the alcohol and xylene washes are not needed, they we're just tests).

While we wait for results, I tried the following shortcut,


New round of testing:

- Ground 350g fresh mushrooms in Ca(OH)2 with splash of water to make a slurry. pH ~ 12. No clear degradation from a color change. Stable light gray.
- Added Ammonium Sulphate. Strong Ammonia smell from reaction with Ca(OH)2. pH is lowered (pH meter says down to 9.6).
- Xylene extraction atempted. Massive emulsion. Xylene is trapped in a matrix of congealed junk. Was able to pour of a minimal amount of Xylene after waiting for 24 hours. White precipitate forms with FASA.

So this doesn't work. Need to add steps to avoid the horrible emulsion. Not sure which one(s) from the previous post matter, maybe the 170C dry? Something was denaturalized and/or removed to avoid the emulsion. The shortcut presented here is unworkable really.

Chiming in because I love the scientific approach to the psilocin extraction problem.

I did experiments using CHCl3 as a solvent for psilocin base. DCM was difficult to deal with and get a good extraction with minimal emulsions, chloroform was really preferred.

The extraction started with dried fruits, was heated in an acidic state to dephosphorylate as much psilocybin as possible, and was filtered quickly after each extraction to yield what would likely have been delightfully powerful albeit unpleasantly sour mushroom tea.

Basifying with NaOH and extracting with CHCl3 did not yield an appreciable amount of psilocin base.

Basifying with ammonia did result in the extraction of psilocin base, but this was impossible to salt out from the solvent without co-extracting ammonium ions as well. I don't know how ammonia solubility in xylene compares to in chloroform, but I would assume that some is being transferred during salting.

Basifying exclusively with an excess of sodium bicarbonate was also unsuccessful in transferring psilocin to chloroform.

Loading the solution with an excess of sodium bicarbonate, then carefully raising the pH with a tiny amount of NaOH solution to around 9.5-10, then extracting the solution with chloroform, was the most successful method.

The loading of carbonate ions in solution prevents the NaOH from deprotonating the hydroxyl group on psilocin which forms the water-soluble phenolate ion. Well, indolate. I don't know that this is actually what is happening but it is a hypothesis consistent with my observations.

Salting with tartaric acid in water, or citric acid in water, via careful titration, followed by slow/cool evaporation, has been successful in generating salts with high correspondence to dosage numbers published by Shulgin.

I think the preferred method would be to vacuum distill the solvent from the crude psilocin base extract, weigh the flask, and add the corresponding presumed stoichiometric amount of acid-acetone solution, let stir, and vacuum distill the acetone off. Then in a very dry room or glove box, quickly transfer the material into an amber bottle, preferably also with an inert atmosphere and desiccant.