Yachaj (Hive Bee)
01-11-03 14:08
No 397222
      Psilocybine crystallization for beginners  Bookmark   

The crystallization of psilocybine hydrochloride is much easier than previously thought. The most important existing techniques are at the end of this post. Problem of those is that they use chromatography as final purification step.

But that can be omitted!

The magic step probably was in the addition of the few drops of hydrochloric acid to the alcoholic extract.  This produces the psilocybin HCl salt, which is insoluble in acetone and which forms nice big crystals.

It is also important to give each step in the protocol enough time. This is most obvious in the last step, the double acetone wash. The dark residue crystallizes by itself in the acetone after 12 hours!

I looked into this method to see if the green-blue-dark part of the extract can be separated from the psychoactive part. In this method that is not possible. The growing crystals absorb the pigment and then become transparent. The identity of the blueing reaction of psilocybian mushrooms still is a mystery to me.

Summary of the (tested&proven) method:

- harvest&dry the mushrooms/mycelial tissue
- turn it into powder (blender works fine)
- extract overnight with a mixture of ethanol and water (denatured 140 proof is fine)
- filter (vacuum filtration or 0.2um filter mounted on syringe)
- add a few drops of hydrochloric acid (aim at pH3)
- evaporate down to 1/10th of the volume (can be done in a large tupperware container) and put extract in a (tall) vial
- remove fats&resins with a solvent which is not miscible with water (cigarette lighter gas, paint thinner, naphta etc.). Just add the solvent to the extract, mix (slowly, not vigorous) and let it stand for a couple of hours. The solvent floats on top of the extract and can be removed by syringe or by freezing the water and pouring off the solvent.
- remove the rest of the gunk by adding acetone to the extract. Mix slowly and let stand. Again two layers will form. On the bottom of the vial is a dark layer. On top of that floats the acetone which is yellowish to greenish. Remove the toplayer by syringe or pipette.
- add new acetone. Mix slowly. Now the dark layer becomes real sticky. Let stand for a few hours and remove the acetone.

Final step: collect all the dark sticky residue and dry it slowly. Large transparent crystals will form. The more slowly the evaporation the bigger the crystals. The potency of the mushrooms can now be determined by weighing the crystals.

What a relief - Albert Hofmann's approach to use chomatography for the crystallization can be omitted. It is not a difficult technique but it needs a large volume of solvents and very specialized ingredients and reagens. But now I am absolutely sure that large crystals will form after a double acetone wash and simply allow the residue to stand overnight.   

Further reading:

American patents 3183172 and 3192111.
Can be viewed at http://patft.uspto.gov/netahtml/srchnum.htm

Popularized version:

URL: http://nepenthes.lycaeum.org/Plants/shrooms/shroom1.html

Albert Hofmann's
article 'History Of The Basic Chemical Investigations On The Sacred
Mushrooms Of Mexico', which appeared in the now hopelessly difficult to
obtain book TEONANACATL of 1976 (cut, paste&see

"In order to preserve the possibly very sensitive, active principles, we
used only neutral solvents and the extractions were carried out at room
temperature. After the extraction of the finely-ground mushrooms with
chloroform, with benzene, and with acetone, the whole activity was still in
the mushroom material. The active principles were easily and completely
extracted with methanol. From the residue of this extract, inactive
constituents could be eliminated by treatment with chloroform. The remaining
easily water-soluble preparation was purified by precipitation of a
concentrated solution in water with ethanol. The activity remained in the
filtrate. The residue of the evaporated filtrate contained the active
principles enriched a hundred fold compared to the dried mushrooms. A
further concentration of the active constituents was possible by paper
chromatography. Using Whatman-I-paper with water-saturated butanol as
solvent, four zones were obtained, the nature of which was determined by
cutting the chromatograms into small strips, extracting the single strips
with methanol, and weighing the residues. In one of the four bands, the
whole activity was found in the form of an easily water-soluble,
halogen-containing powder. After treatment with silvercarbonate, elimination
of silver ions with H2S, and concentration of the aqueous solution in vacuo,
the substance crystallized in fine white needles. With the few milligrams
obtained in this way we made several tests. The new psychotropic principle,
which was named 'psilocybin,' elicited a violet color with Van Urk-reagent,
characteristic for indole derivatives.
For the subsequent isolation experiments, we could rely on this color test.
When paper chromatograms prepared as described above, were sprayed, after
having been dried, with a solution of p-dimethylaminobenzaldehyde in benzol
and put in an atmosphere of dry HCl gas, psilocybin produced a violet spot
with Rf 0.1. A weaker spot with a blue color and Rf 0.5 was observed,
corresponding to a second active principle, which we named 'psilocin.'"[end

01-11-03 23:19
No 397311
      Maybe you are crystallizing sugars...  Bookmark   

Maybe you are crystallizing sugars... What makes you sure it's psilocybin? Sounds strange...
(Hive Bee)
01-12-03 09:43
No 397458
      Maybe Sugars., But....  Bookmark   

Maybe sugars, but SWIM tried a very simular process (unresearched, just giving it a try).  The residu after a MeOH pull was very sticky and gooie apon evaping the MeOH.  A few ml of Vodka were added to the goo along with a drop of HCl.  A Acetone flash was then done 2x followed by the addition of a few more ml of MeOH.  Neadle like crystals formed after the MeOH evaped off the pie pan.  A tine sample (when scraped with a razor blade, 1/3 the lenth of the blade was used as the sample) was bio-assayed.  A very nice experance was noted for a slightly longer time frame compared to injestion of just caps/stems.

If there is no h2o pressent in any of the solvents during the extraction, then there should be no sugars present in the final product as MeOH and sugars dont like each other.
(Hive Bee)
01-13-03 12:18
No 397747
      Psilly sugars?  Bookmark   

Thanks for the comment Lilienthal! The crystals taste salty and are psychoactive. But that doesn't mean that they are not partly/largely sugar. OTOH - I have just tried to dissolve some glucose in the 140 proof denatured alcohol which was used for the first extraction. It refused to dissolve. Then I mixed one part of this alcohol with 4 parts of acetone - still nothing precipitated.

I wish it would be possible to perform an A/B. But since psilocybine freebase is polar this would not separate the alkaloid from sugars (hey - but this would separate the psilocybine from the psilocine right? Gotta try that! I want to know if the pigment is associated with the psilocybine or with the psilocine - and I think it is the latter).

The interesting thing about these crystals is that even if they are not psilocybin there is no way (besides chromatography) to be sure of that. I repeat - they are bittersalt, they are psychoactive and the yield is in the 10mg (definately below 50mg) range per gram of PF TEK grown cubensis. They are about everything we want them to be!

But Lilienthal, please continue your commenyts. They are valuable!



(Hive Bee)
01-13-03 13:43
No 397764
      Not sugars - but how about urea?  Bookmark   

After reading this:


I think that the salty crystals might be urea. I can't imagine that no one came up with the described crystallization method for psilocybine HCl if it were that simple. And there is twice as much urea as alkaloids in the mushrooms.

So What is a good method to separate urea from psilocybine? Can urea be a/b'd?


(Chief Bee)
01-13-03 21:55
No 397859
      A/B psilocine, leucopsindigo and psindigo  Bookmark   

This was posted by Yachaj elsewhere, and moved here by me /Rhodium

Indeed it seems that the dark pigment in the mushrooms is associated with psilocine, which can be isolated by A/B.

PDF file, see:


At page 77 of Trout's Notes On Some Simple Tryptamines <www.troutsnotes.com>, someone with the alias theobromus proposed two molecular structures of the dark pigment. They were called Psindigo and Leucopsindigo.

Leucopsindigo is drawn as a combination of two psilocine molecules which are attached to eachothers 5th position.

Psindigo looks the same, but the two molecules are attached with a double bond and their 4-OH group is replaced by a double bond & oxygen or 4=O

(Hive Bee)
02-26-03 05:54
No 411796
      ethanol extract  Bookmark   

I was reading somewhere that psilocybin dissolved quite well in alcohols (ethanol, methanol) and psilocin preferred water, and took way longer to come out of the tissue than psilocybin. Bearing this in mind, SWIM has devised an experiment where fresh p. cyanescens are soaked whole in ethanol (96%) for three days. He is going to send me info about his results soon.

I know naaaathing.
(Hive Bee)
02-27-03 03:13
No 412123
      aqueous solutions of alcohols aren't the best...  Bookmark   

"The occurence and extraction of indole derivatives in six species from four genera of higher fungi were investigated. By using pure methanol for extraction of the mushrooms analysis revealed the highest concentrations of psilocybin and baeocystin. The psilocin content of the species was higher by using aqueous solutions of alcohols than with methanol alone but was an artificial phenomenon caused by enzymatic destruction of psilocybin. The extraction with dilute acetic acid yielded better results than with the water containing alcohols. The simlpe one-step procedure with methanol for the quantitative extraction is still the safest method to obtain the genuine alkaloids from funghal biomass."

aqueous solutions of alcohols aren't the best route.


(Hive Bee)
02-27-03 03:24
No 412129
      damn, if need be I'll find that shulgin ref.  Bookmark   

On DMT world, of all places, somebody posted a letter from shulgin responding to the question of extractions from psilocybes, and the conclusion I got was that the psilocybin came out easy with 70%+ alcohol (the rest aqueous), but psilocin prefers to be extracted with water.

SWIM's busy doing an extraction with 96% ethanol, leaving it soak for three days - and at the close to three day mark the mushrooms are sinking to the bottom of the solution, a golden tint has occurred to the solution btw, he's leaving them one more, well to be precise, about another half a day, by then all the shrooms will have sunk.

Psilocybin is easy to extract, but psilocin is much more resistant. SWIM is not sure how to dry the alcohol out now, but he suspects that a goodly proportion of the psilocin will be blue by the time it's dry crystals. Still, if 50% of the psilocin is extracted effectively, that is better than just settling for dry ethanol extract of only psilocybin. SWIM's idea is to take the dry evaporation product and add to an equal amount of ascorbic acid to help prevent oxidation, and store in caps in the freezer.

I know naaaathing.
02-27-03 21:49
No 412362
      extract info  Bookmark   

the following message was written to me via pm on another site and includes some info on psilocin as well as psilocybin.  here it is :

Sure I can help! I'm glad you asked 

Ethanol is NOT the best solvent in the world to use for 2 reasons:
a. it contains water (making it a bitch to evap and work without heat)
b. ethanol does not extract psilocybin very well... (water does, better at least, though it still isn't so great. acidic water works well, but it's still water)

The best advice I can give you is this: Find some pure methanol. Read my journal threads on kitchen chemistry and sources.

You will need the vacuum filter for a different purpose (one which very few people know )...

Grind dry mushies to a powder. Seal the powder in a jar (make sure the level of the powder is AT MOST only half the volume of the jar) and freeze it until you are ready to use it. Refrigerate some methanol. Take the jar of powder out of the freezer... QUICKLY open the methanol jar, then the powder jar, and swiftly add the methanol to the powder about an inch (small jar) or half inch (large jar) above the level of the powder, and QUICKLY seal the mixture jar.
*I say "quickly" because cold jars will gather water condensate from the surrounding air... this will mix with alcohol... this will really suck later. It is best to do all of these procedures in a dessicated glove box (just a bowl of Ca chloride set in the glove box for an hour prior to work).*

Swirl this slurry (methanol+powder) around for about half an hour, relieving pressure every couple minutes (warming alcohol will expand gaseously, so pressure relief is necessary for safety, though I doubt the jar will actually pop or anything...). Let the slurry settle and pour the alcohol through a funnel, filtered by either:
a. vacuum filtration through filter paper (this may clog, though)
b. a layer of dense fabric (high knit sheet or shirt) with a coffee filter on top. This will be squeezed to remove last liquids.

The leftover crud (filter cake) can be extracted by repeating the methanol extraction outlined, or it can be thrown out. You will probably want to re-extract the first few times, until you get the hang of doing this perfectly.

Evaporate the methanol by blowing over the jar with a fan... this, too should be done with dessicated, de-oxygenated air, but the apparatus is too complex to build for description here... it may be in my kitchen chemistry chat posts ??

As you evaporate, you MAY see crystals... maybe not. Either way, continue evaporating to dryness.

This residue may be eaten in doses of about 3 times that you would take of pure psilocin (30mg residue = 10mg psilocin)... this depends on the strength of the shroom (that's why crystal would be better, because you would KNOW how much goodie you are eating). If you have crystals, just measure them directly as weight of psilocin.

BTW, this is a mixture of psilocin and pilocybin... I only say psilocin for the sake of brevity, in that psilocin is the more potent molecule by weight. Most of the mix will be psilocybin, but it is better to err to the lower end of dose.

Here's the big trick to decent purification of these compounds:
Dissolve the residue and/or impure crystals in room temp. petroleum ether (in this case, you can use zippo fluid). You may notice that crystals don't dissolve well, though they might (because of impurities). Only add enough pet. ether to get the stuff suspended/dissolved as you stir... if crystals won't dissolve, DON'T add more pet. ether. Crystals not dissolving is GOOD!

Close the container you are doing this in and put it in the fridge. When it gets to fridge temp, put it in the freezer. (Slow cooling forms bigger crystals ). Leave in freezer until you see no more crystal growth (overnight, usually).

Stir up the mix (still freezing cold) with a cold stirring rod, suspending the crystals in the liquid. Swiftly pour this into the vacuum filter apparatus and flip it on. Rinse the container with a bit of more freezing cold pet ether, and dump this into the filter. You want to get every last crystal, right? 

Leave the vacuum on until the crystals are dried by the air (again, preferably dessicated and de-oxygenated). This is a relatively pure crystalline mix of psilocin and psilocybin.

Depending on experience and perfectionism, the yeild from a single methanol extraction (no re-extraction of the mushy filter cake) of this caliber is approximately 95%. Most losses occur from oxidation, over use of solvents, bad temperature control, and transfers (leftover drops in containers).

To store this pure product, get some pure ascorbic acid (vitamin C) crystals. Dissolve some of the vit. C in twice it's volume of 190pf ethanol. Now, dissolve your cin/cybin crystals in this acidic ethanol.

Put this mixture in a dark container (something opaque) in the freezer.

That's it! 

P.S. Do you mind if I post this answer? I get this question all the time and I'm tired of typing it... 
(Hive Bee)
02-27-03 23:46
No 412386
      fantastic  Bookmark   

That method sounds excellent. Sadly SWIM, living in the land of OZ, has no idea how to find pure methanol other than chemsupplies. Would dried (MgSO4 baked techique) ethanol work well too?

So what I gather from reading your post, that naptha, at least only a little naptha, won't dissolve the goodies, but will strip contaminants and result in crystals. Is it absolutely essential to vacuum filter to clear this off? would gravity filtration in coffee filter work okay too? The naptha my SWIM gets is pretty clean.

I know naaaathing.
(Hive Addict)
02-28-03 06:14
No 412474
      MgSO4 cannot dry ethanol well.  Bookmark   

MgSO4 cannot dry ethanol well.

MgSO4 can, however, dry isopropyl alcohol well (99%), and I dont see why it cant be used for this extraction purpose. Well maybe it doesnt evaporate as quickly.

What about cold acetone? This should work as well but may extract more resin and less psilocybin.

Chemistry is hard to learn, but its worth it.
(Hive Bee)
02-28-03 06:31
No 412479
      MgSO4 doesn't do etOH?  Bookmark   

Oh. Do you know why that is, seems odd.

SWIM's ethanol extract is now drying out, and a white crystalline precipitate is floating around in it. SWIM had this thought that maybe if this precipitate were caught in a filter that it might turn out to be a good way to isolate whatever it is, since it is obviously less soluble than whatever else is in there. SWIM was told many years ago that blue crystals could be made from mushrooms using 'metho' which over here is 96% bittering agent denatured ethanol. Oh, SWIM's method was to soak whole mushrooms in enough ethanol to cover them for three days.

I know naaaathing.
(Hive Addict)
02-28-03 23:01
No 412628
      MgSO4 cannot produce absolute ethanol because...  Bookmark   

MgSO4 cannot produce absolute ethanol because absoulte ethanol is so hygroscopic. Methanol is found everywhere. You just not looking hard enough!

P.S. The bottles you describe, biojammer, arent exactly pure alcohols. Try using it directly in the al/hg and you will find out why!

Chemistry is hard to learn, but its worth it.
(Hive Bee)
03-29-03 14:35
No 422260
      The urea is still in here  Bookmark   

dlagwagon is only partly right. Right is that the endproduct is psychoactive. But this method doesn't separate the urea from the psilly.

Furthermore the vacuum filtration and the anhydrous methanol are here more complicated than needed.

Lemme explain.

You can extract in 140 proof ethanol (=70 percent alcohol, 30 percent water) because most of the water can be removed later on with acetone and what remains is not a problem. And psilocybin dissolves faster in 140 proof than in 200.

Just add the liquid to the mushroom powder in a jar, stir, close the jar and put it in the fridge for the night. Then pour the liquid through an old white t-shirt or so in a 2nd jar. Fill a syringe with it, attach a filter and empty the syringe in jar no. 3

The filtration can be done with a disposable 0.2 micrometer liquid filter for syringes (handpowdered, so no electricity or adspirator is needed) because you do not need to save the insoluble particles. The filter can be discarded.

Evaporate the filtered liquid until it has just not begun to crystallize out. A lowtek approach is to do that in a large oven dish (lot of surface), and blow it with a fan.

The remaining liquid is put in a small tall vial. It may be handy and/or optional to add a drop of HCl at this point. Then you are sure that whatever alkaloid is in there becomes a more-or-less stable, polar salt.

Add naphta or pet ether, shake, allow layers to separate and use a pipette to remove the topleyer.

Now add acetone and repeat this procedure. Twice.

The dark granular stuff which remains is a mixture of alkaloids. Among those are the alkaloids you want.

Add 190 proof ethanol, close the vial and put it in boiling water. Wait until the vial is of the same temperature, take it out of the water and allow it to cool down to slightly below the boiling point of the ethanol (so you can safely open the lid) Suck up the liquid in a syringe, filter it and allow it to cool down slowly. White psilly crystals precipitate, urea stays in solution (as long as you don't evaporate the liquid too far of course).

Remove the liquid with a pipette or syringe. Save the crystals.

Normally the water in this mixture would cause problems for obtaining dry crystals, but not here. Psilocybin does not dissolve very well in cold water (at 20 centigrade you need 120 parts of water to dissolve only 1 part of psilocybin).

(Hive Bee)
03-29-03 15:23
No 422268
      yep  Bookmark   


ah, it's magically appeared higher up now...

well anyway, yes, this is correct, it seems that the low solubility of psilocybin in ethanol is actually advantageous because that is how you can separate the urea out of it (man that urea sux eeww..! *holds nose* laugh)

Frankly I don't see the point of extracting psilocin anyway, it breaks down so damn fast. someone said that aqueous alcohols makes enzymes damage psilocybin further up the thread, this is wrong, the enzymes damage the psilocin, according to shulgin the psilocybin is best extracted with water... But for a method to separate the urea off it, absolute alcohol seems to be the thing. And extracting with water you'll end up with urea in the end product and it will be a lot more involved than just using ethanol's insolvency with psilocybin to precipitate it out.

I want to find out whether freezing the solution without boiling down is enough to precipitate the psilocybin crystals. It certainly didn't need to be boiled down all that much when I did it, in fact I'm almost inclined to say that it just needed to be cooled (I used an electric fan). Hopefully swim will have an opportunity to test out this theory some day soon. eating things full of urea can't be good for the old body odor, and also, wouldn't using ethanol also separate the baeocystin out of it as well? Being able to get the psilocybin fully by itself would be a good thing, everything else in mushrooms is not as good or benign. baeocystin, I believe, is responsible for the runny nose/immune depletion/watery eyes thing that happens with really high dose cubensis.

Acetone might be rather good as a substitute to using the freezer, it would probably force the psilocybin into precipitating faster... though the freezer is probably the cheaper option. I can get 96% alcohol easy around here, and I found it pulled all of the alkaloids out of whole cyanescens in three days soaking.

I don't think the process even needs to be as complex as the one you describe here yachaj. Psilocybin is very water soluble, it does not need to be acidified, it doesn't really need to be washed when pulled with absolute ethanol, except perhaps with a salty water wash through the filter. I think that freezing the ethanol will reduce solubility enough. I will have to test this.

It means buckle your seatbelts dorothy, because kansas ... is going bye bye
(Hive Bee)
03-31-03 14:37
No 422730
      A bit of topic.  Bookmark   

When I consume mushrooms I prefer cooking tea of them, and removing the mushroomflesh. Lately I have been experimenting with higher dosages, but I can't seem to reach a higher level.

Could this have to do with psilocybin/cin solubility in water? Maybe the solution gets saturated and no more alkaloids dissolve in the water? I use maybe 6-8grams of mushrooms for about 2dl of water.
03-31-03 15:35
No 422746
      No, probably not.  Bookmark   

No, probably not.
(Hive Bee)
04-01-03 12:37
No 422986
      No boiling down 2  Bookmark   

Urushibara wrote:
> Frankly I don't see the point of extracting psilocin anyway, it breaks down so damn fast.

The biggest plus of extraction&purification is that the potency of the mushroom (or mycelial tissue) is less important. Without extraction you will always need to keep a close eye on the cultivation technique (substrate, temperature) as well as the mushroom species. If you don't you never know how many are needed. Psilocybin is easy - one gram is always the same.

>I want to find out whether freezing the solution without boiling down is enough to precipitate the psilocybin crystals.

Re-read the method I described: 'boiling down' was not a part of the process. Initial extraction was perfomed at 4 centigrade.

> baeocystin, I believe, is responsible for the runny nose/immune depletion/watery eyes thing that happens with really high dose cubensis.

Where did you get that info? All I know about baeocystin is that it is a very minor component in (sub)tropical psilocybians like cubensis. The only difference between baeocystin and psilocybin is a lone little methyl group at the tail. Perhaps that it causes a very subtle difference in psychoactive terms. But what you are saying is that baeocystine causes lots of undesired effects and psilocybin does not.

I also don't think that the solubility of baeocystin and psilocybin is very different (I guess it is not possible to do that with only some ethanol)

>Psilocybin is very water soluble

Not very. Psilocybin is soluble in 20 parts of boiling water and in 120 parts of water at 20 centigrade. dH2O can be used for recrystallization but I don't know if that is advantageous.

A gram of mushrooms may contain 10mg of psilocybin. It is easy to extract that in an ounce of water. But to get it out of the water you will need to reduce the volume. Hence my choice for a mixture of water and ethanol: it evaporates faster in a shallow dish. An additional plus is that mushroom enzymes, which break down psilocybin are more active in water than in 140 proof ethanol.

04-05-03 07:35
No 424021
      What about
(Rated as: full of typos!)

What about hydrolyzing the psilocybin to psilocin then doning A/B on the psilocin (if i reamember corectly you can a/b psilocin but not psilociybin). you might be able to do the hydrolyzing whith the shrooms in solotion like happens in your tummy.then work on from there. i know psilocin dreaks down quickly, but then what is N2,Vacume and Vitemine C for anyways.
(Hive Bee)
04-05-03 21:09
No 424135
      okay... physprops of psilocin - it is very...  Bookmark   

okay... physprops of psilocin - it is very unstable, oxidises very easily.. hence if extracting it is too much effort (as shulgin certifies that it is - it can take three days) why bother.

regarding the extraction temperature, if we are dealing with absolute ethanol (ie ~96% ethanol, the rest water) what about doing the extraction at -18 to -20°C? aka the temperature of a freezer?

okay, re baeocystin and unwanted side effects... maybe indeed it is not baeyocystin that causes the problem effects I mentioned. But I base my assertion on the experience of 4-20 medium sized mushrooms... generally the effects I mentioned occured at around 5g of dried cubensis mushrooms (from qld australia). I have never heard of anyone talking about such effects from psilocybin, hence I suspect baeocystin.


I was comparing ethanol solubility to water solubility. I can't quote figures, but ethanol is *way* less able to dissolve psilocybin than H2O. I was basing this information on what I read at 'ask dr shulgin' regarding methods discovered by czech chemists.

oh, and fyi on dried shrooms, I don't know what kind of psilocybin mushrooms you are referring to saying 10mg/g dry, afaik, dry cubensis (afaik amazonian strain) only contain about 5mg/g.

In my early days of psilo consumption, my guide used to rate 1.5g of dried cubensis as a baseline dose, 3g as a full dose and 5g as a guaranteed psychedelic dose. In line with mckenna's figures.

the figures I have heard quoted for mg doses of psilocybin are between 12 and 15mg for a full psychedelic dose.

I would like to see a 2g dried cubensis blow someone's mind. I dare you.

It means buckle your seatbelts dorothy, because kansas ... is going bye bye
(Hive Bee)
04-07-03 22:10
No 424575
      RFT=even more important than UTFSE  Bookmark   

MENH2, I mentioned the A/B of PSOH in this thread already! Read before you post!

 See http://www.tacethno.com/info/psilocybe/casale_1985_jfs_30_247.pdf

As you can see, this method uses a lot more solvents than the PSOP crystallization method by M/P (mixed phase) extraction which I am proposing in this thread.

(Hive Bee)
04-07-03 22:36
No 424579
      Cubensis of 1 percent plus exists  Bookmark   

Urushibara wrote:

>I don't know what kind of psilocybin mushrooms you are >referring to saying 10mg/g dry, afaik, dry cubensis (afaik >amazonian strain) only contain about 5mg/g

That is the potency on bulk substrates. But see this:

Cubensis on rye:


Similar results have been reached on a brown rice medium. Two grams of dried young cubensis will most certainly knock many people out of their socks.

> if we are dealing with absolute ethanol (ie ~96% ethanol, > the rest water)

I think 140 proof (70 percent) extracts the PSOP faster

> what about doing the extraction at -18 to -20°C?

No idea. But I don't see a concrete problem which is solved by doing so. I think it is more useful to compare results of the extraction at 4 centigrade to an extraction at roomtemperature. If the latter leads to the same results faster I think I prefer a rt extraction above one which needs to be done in a freezer.

The less equipment you need the better!

(Hive Addict)
06-15-03 02:38
No 440056
      After a long time lost, a bird flies up ...  Bookmark   

After a long time lost, a bird flies up through the long awaited crowd of onlookers...
A document was read by this chemist who went to school for a long time that SWIM ran into. 

Crystals from Boiling water, mp 220-228°; from boiling methanol, mp 185-195°. uv max (methanol): 220, 267, 290 nm (log E 4.6, 3.8, 3.6). pH 5.2 in 50% aq ethanol. Sol in 20 parts boiling water, 120 parts boiling methanol; difficultly sol. in ethanol. Practically insol in chloroform, benzene. LD50 in mice, rats, rabbits (mg/kg): 285, 280, 12.5 i.v. (Usdin, Efron).

Would that not suggest that a bee would merely use a distillation apparatus with vacuum with some methanol (after separating the retail grade methanol from all the water that's added)?  What's the trouble with that? Whay all the hubub over A/B's and ethanol? According to the notes above, use water...and if a bee is afraid of damaging the active ingredient because of heat, use a vacuum. 

Plates from methanol, mp 173-176°. Amphoteric substance. Unstable in soln, esp. akaline soln. Very slightly sol in water. uv max: 222, 260, 267, 283, 293nm (log E 4.6, 3.7, 3.8, 3.7, 3.6).

According to the document read, and some of the above information posted by several bees, it seems that this active ingredient isn't worth fighting for. 

However, there is concern to SWIM that Baeocystin and Norbaeocystin may be concentrated just like the desired ingredient...is this a factor?  SWIM doesn't really want to die right now.

SWIMs' not too sure about chromatography (in red above), does that have to do with all the hubub? If a bee doesn't have a chromatography column?

Please correct SWIM if there's a problem with the above logic.

It tastes like burning!shockedtongue
(Hive Addict)
06-15-03 10:33
No 440108
      Not chromo...  Bookmark   

Those items in red have nothing to do with chromatography but are the ultraviolet transmission/absorbtion peaks for the compound at specific wavelengths. This will help you to identify the compound.   The device is a uv spectrophotometer, which used ones can be had for $100-200 on auction sites.

Infinite Radiant Light - THKRA
06-15-03 10:42
No 440109
      They are talking about recrystallization of...  Bookmark   

They are talking about recrystallization of already crystalline material and the spectroscopical data of the pure compound - not about chromatographical separation. The quotes have nothing at all to do with purification from plant material.

Baeocystin, norbaeocystin, and psilocin usually are only trace alkaloids. They can be isolated using sophisticated methods, but the yield is extremely low compared to psilocybin. There is not the slightest hint that those trace alkaloids could be more toxic than psilocybin itself, don't worry about your life smile.
(Hive Addict)
06-17-03 04:34
No 440433
      MnkyBoy78  Bookmark   

Looks like this

Maybe sugars, but SWIM tried a very simular process (unresearched, just giving it a try).  The residu after a MeOH pull was very sticky and gooie apon evaping the MeOH.  A few ml of Vodka were added to the goo along with a drop of HCl.  A Acetone flash was then done 2x followed by the addition of a few more ml of MeOH.  Neadle like crystals formed after the MeOH evaped off the pie pan.  A tine sample (when scraped with a razor blade, 1/3 the lenth of the blade was used as the sample) was bio-assayed.  A very nice experance was noted for a slightly longer time frame compared to injestion of just caps/stems.

If there is no h2o pressent in any of the solvents during the extraction, then there should be no sugars present in the final product as MeOH and sugars dont like each other.

seems to have solved the sugar and urea problem as urea is soluble in water (big time...Solubility in Water: 1,193 g/L at 250C), and it's toxic to the gonads...yea yea cool...are you thretening me? I am cornholio!

Is that right?

(Hive Addict)
06-17-03 06:45
No 440449
      In the freezer  Bookmark   

I have about 400mls of etoh and Saturated salt water.. nothing is falling out.. so I'm guessing I should toss it back on the vacuum evaporator and take 50% or so of the solvents off...  maybe the psilocybin will drop then.. I know it's good, as the tlc plate shows the goodies... just can't get it to crystalize yet...

Infinite Radiant Light - THKRA
(Hive Addict)
06-24-03 14:49
No 442156
      Mnkyboy78  Bookmark   

If one ounce of mushrooms were processed, would it bee worth crystallizing and purifying a batch of that size?
06-24-03 16:31
No 442177
      :-S You are (probably) not able to crystallize  Bookmark   

crazy You are (probably) not able to crystallize psilocybin. At least no without sophisticated methods like chromatography.
(Hive Bee)
06-27-03 19:55
No 442952
      And now the standardized extract for dummies!  Bookmark   

Liliental wrote:
"You are (probably) not able to crystallize psilocybin. At least no without sophisticated methods like chromatography."

Fortunately there are less sophisticated methods which work.
One example is this one:

"SWIM tried a very simular process (unresearched, just giving it a try).  The residu after a MeOH pull was very sticky and gooie apon evaping the MeOH.  A few ml of Vodka were added to the goo along with a drop of HCl."

Further simplification:

1) instead of MeOH take 140 proof EtOH (70 percent EtOH, 30 percent water). The advantage of this liquid is that it extracts both PSOH and PSOP at low temperatures, in 12hrs time, but it is not as toxic as MeOH and more commonly available.

2a) evaporate it down until everything is just liquid. Add plenty of 190+ proof EtOH and a few drops of HCl. PSOP precipitates out!

2b) you can also evaporate all the 140 proof. The end result is a piece of brown gunk. This piece can be placed in 190+ proof EtOH (+ a drop of HCl). Don't touch it, just watch. In a few days to a week, lots of white dust separates from the brown gunk and collects on the bottom of the container.

Correct me if I am wrong but my explanation of the phenomenon is as follows. Compare it to a bottle of NaCl saturated water, which is visible because of the thin layer of table salt on the bottom. It looks as if the salt doesn't dissolve because the water has dissolved all the NaCl it can hold. But in reality the salt is constantly dissolving and precipitating at the same speed. So the layer stays the same.

Now compare this to a brown gunk of urea/psop with impurities in a bottle of 190+ proof EtOH with HCl.

Slowly the brown gunk dissolves, releasing both urea and PSOP. Urea stays in solution but the solvent is saturated with PSOP HCl in no time. It precipitates out!

But nevertheless, more PSOP HCl is dissolving from the brown gunk. And precipitates out. The chance for an individual PSOP HCl particle to precipitate back into the brown gunk is much smaller compared to the chance of precipitating on the bottom of the container.

Even more funny: the solvent is saturated with PSOP HCl. That means that, if you know the quantity of PSOP HCl which is dissolved in this solvent, at this pH and temperature, (in mg/ml), then you know exactly how many ml are needed for a dose. And this is totally independent from the potency of the mushrooms. Just prepare the PSOP in EtOH&HCl mixture and you know how many ml are needed.

No chromatography. No scales (except for the 1st time to determine the solubility data). No weird solvents, no rocket science - just vodka (separated into its constituents, EtOH and dihydrogen monoxide). Fat chance that the HCl isn't even needed.


06-27-03 20:18
No 442955
      Oh, no doubt that SWIU got a precipitate.  Bookmark   

Oh, no doubt that SWIU got a precipitate. But I strongly doubt that it is crystallized psilocybin. Psilocybin might co-precipitate with that amorphous mixture of polar stuff, but believe me: it is not that easy. See it as a concentration step, but please don't talk about crystallization in that context (unless you provide strong evidence).

BTW I would not add HCl to, it just makes no sense (psilocybin is already charged because it is zwitterionic) and such strong acidic condition will probably lead to some degradation.

[This is really not meant to discourage you from experimenting and posting here smilesmilesmile]
(Hive Bee)
06-28-03 11:43
No 443095
      but believe me: it is not that easy.  Bookmark   

but believe me: it is not that easy. See it as a concentration step, but please don't talk about crystallization in that context (unless you provide strong evidence)

If it are not crystals, but it is white, reacts as PSOP with the Marquis test and a bioassay, matches the solibility behavior of PSOP (good in water, not in alcohol/acetone/pet ether), begins to desintegrate (becomes brownish) when it is exposed to 50+ centigrade and when it is exposed to air for a few days (just like as is described for PSOP HCl), and it is the main component which remains when the urea is crystallized out then what else than crystalline PSOP can it be?

Of course you have a point if you state that even if it is 99.99999 percent pure it is still not 100, so still not crystalline.

This is really not meant to discourage you from playing with complicated time consuming yield wasting methods, but I see no point in going further. Extraction&purification is needed to prepare a standardized product, so the strain or species or potency of the starting material is no longer a concern.

I think a separation of the PSOH from the PSOP (and the (nor)baeocystin etc.) indeed requires chromatography. But that is not needed for a reliable endproduct of constant quality.

What you want is absolute 100 percent crystallization, with not a single contaminant molecule in a chunk of PSOP. Right? For that I don't think that even chromatography will satisfy your needs.


06-28-03 13:16
No 443098
      My point is: your product is probably a ...  Bookmark   

My point is: your product is a mixture of everything soluble in aqueeous MeOH / EtOH and unsoluble in ethanol. That simply means: Polar compounds, e.g. amino acids, peptides, sugars, nucleotides, or urea, just to name a few. It's probably a mixture of several hundred to thousand compounds, it it most probably NOT pure (= crystallized) psilocybin. It might be one of the main compounds in there, but without checking against pure psilocybin we even don't know that.

The term "crystallization" means "pure preparation consisting of the compound in a defined crystal structure", it doesn't mean "amorphous powder containing a bit (or a bit more) compound" tongue.

It sounds like a great way for concentration, but might not a good procedure for standardization.
(Hive Bee)
07-03-03 11:34
No 444180
      recrystallise it then?  Bookmark   

would it clean it up if it was recrystallised?

you can't keep peace with a gun
07-03-03 12:08
No 444184
      If it is not crystallized in the beginning you  Bookmark   

If it is not crystallized in the beginning you can't re-crystallize. It would be a fractionated precipitation, but I doubt you could purify it much that way.
(Hive Bee)
07-03-03 19:43
No 444266
      recrystallise and wash?  Bookmark   

Maybe some of those things could be washed off?

Of course the only way to get it fully separate is with a liquid chromatograph.

still, considering that even if it's only 10% that means you only need 100-150mg of material, and it isn't likely to vary that much from one patch to the next. easy to cap, quite reliable for dosing (who can measure 10mg anyway?)

you can't keep peace with a gun
(Hive Bee)
07-22-03 15:32
No 449051
      Lilienthal wrote: "your product is a...
(Rated as: good read)

Lilienthal wrote:
"your product is a mixture of everything soluble in aqueous MeOH / EtOH and unsoluble in ethanol"

True. But the undesired components do not interfere enough with the solubility of the major psychoactive component (PSOP) to make the standardized extract unreliable.

But hey - look at this. It is about the use of 4-DMCA instead of 4-DMBA as main ingredient in Ehrlich's reagent. 4-DMCA is OTC and most likely useful as indicator when a simple piece of typing paper is used as TLC surface.

From the paper then all different components can be separately recognized, -extracted and purified with only alcohol as solvent.

Of course the results will probably be better (less losses)with more difficult to obtain solvents and real TLC paper, but given the fact that the desired alkaloids can readily be biosynthesized from brown rice or malt agar, a crude typing paper TLC with 4-DMCA as OTC color indicator can be the finishing touch.

(newbees who have no idea about TLC and mushrooms may also read the text http://www.erowid.org/plants/mushrooms/mushrooms_article2.shtml)

Occurence of 5-hydroxylated indole derivatives in Paneolina foenescii (Fries) Kuehner from various origin. T. Stijve, C. Hischenhuber, D. Ashley. Z. Mycol. 50: 361 (1984)

Ehrlich's reagent (Révélateurs pour la chromatographie en couches minces et sur papier, E. Merck, Darmstadt 1975, no 91, p. 32) was initially used because it yielded brightly coloured spots with most of the compounds of interest. However, detection of psilocybin required a few minutes heating at 100 deg. C and under these circumstances co-extracted urea yielded a brightly yellow zone which interfered with the evaluation of serotonin, a major constituent. In addition, sensitivity for tryptophan was poor.

Better results were obtaines using 4-dimethylamino cinnamaldehyde (DMCA) (Fluka no 39421) which was used as a solution of 0,5 g in 10 ml fuming concentrated hydrochloric acid, mixed with 50 ml methanol. This reagent was more sensitive than its benzaldehyde analogue and did not need heating to react with the various indoles. In addition, it produced a different shade of colour with each compound. For example, psilocin turned greenish gray, psilocybin reddish, bufotenin violet, 5-hydroxyindole acetic acid green, serotonin and 5-hydroxytryptophan bright blue, tryptophan purple and tryptamine purple red.

It should be pointed out that these colours may vary with the chemical nature of the TLC adsorbent. For example, psilocybin spots are reddish on Si02 and on SilCel layers, but violet on cellulose.

The reagent proved tobe remarkably sensitive: the detection limitfor serotonin and 5-hydroxytryptophan was 10 ng and for psilocybin and tryptophan 25 ng. At room temperature psiocybin was the last of the indolic ompounds to become visible. Usually, optimal visibility was obtained after 10-15 mm. The reaction could be accelerated by slightly heating with a stream of warm air from a hair-dryer.

Interestingly, the DMCA reagent reacted only very slowly with urea, which yielded a reddish spot only after a few hours, and thus did not interfere with the determination of serotonin.

07-22-03 20:55
No 449093
      Typing paper for TLC makes NO sense!
(Rated as: good read)

Typing paper for TLC makes NO sense! Use silica gel sheets on polyamide with fluorescence indicator instead. Or if you really have to: filter paper. But separation on paper needs other solvent mixtures, takes longer, and gives a way lower resolution. If you don't have problems to buy detection reagents you can also buy TLC sheets. The para-dimethylamino-cinnamic aldehde is more sensitive than the para-dimethylamino-benzaldehyde, but usually you don't need that extra sensitivity and take the cheaper pDMABA.
(Hive Addict)
07-23-03 08:04
No 449173
      read the Van Urk Salkowski paper  Bookmark   

I posted and is on rhodium's site.. that has excellent sensistivity and very discriminating color changes.. the place to start w/ tryptamines...

Infinite Radiant Light - THKRA
(Hive Bee)
07-23-03 14:06
No 449245
      Lilienthal wrote: "My point is: your...  Bookmark   

Lilienthal wrote: "My point is: your product is a mixture of everything soluble in aqueeous MeOH / EtOH and unsoluble in ethanol. That simply means: Polar compounds, e.g. amino acids, peptides, sugars, nucleotides, or urea, just to name a few."

I have just tried to get a precipitate from glucose and of tryptophan in a similar way (shake with 140 proof ethanol, filter off the solids and mix the liquid with 10 times as much 190+ proof ethanol) but didn't get any precipitate.

So Lilienthal is right in general terms. But specifically with cubensis extraction I still think that the white stuff which drops out of the filtered 140 proof ethanol extract at the moment that 190+ proof is added is fairly pure PSOP.

(Hive Bee)
07-24-03 11:56
No 449484
      psilocybin precipitate *would* be quite pure imo  Bookmark   

I'm inclined to agree yachaj - sure, in the sum total of the contents of the solvent before precipitating it there is oodles of junk, there's all sorts of junk (the major crap in there is urea, you can smell it when it's evaporated down), but the use of ethanol is selective for precipitating the psilocybin. most everything that was named is much more soluble in ethanol than psilocybin.

on another note, rhodium just posted on another thread about a method of doing an acid/base extraction on psilocybin that dephosphorylates the psilocybin into psilocin using acetic acid and a little heat. Post 449383 (Rhodium: "Mushroom Aqueous-Organic Psilocin Extraction", Tryptamine Chemistry) it's an acid/base, and the article quoted claims that it is pure enough to fully characterise with it's IR spectra etc.

I put an idea forward in a response there too - would making the psilocin into the ascorbate salt help allieviate stability problems?

we can't stop here, this is bat country