Today I attempted to extract psilocin from some mushrooms without success. The process I used was based on this article: Extraction Method for the Isolation and Identification of Psilocin; Casale, J. F., J. Forens. Sci. 30(1), 247-250 (1985). There were to be 2 substitutions made, namely petroleum distillates for ether and hexane. Here are my notes:
- 12.5g dried mushrooms were ground to a fine powder in a blender.
- 12.1g of this powder (~0.4g were lost in grinding) was transferred to a 3/4l flask to which was added a 5% aqueous acetic acid solution. Solution capped and mixed by swirling resulting in a green/grey sludge with pH between 3 and 4, probably being closer to 4. This was left to sit in a dark place.
- Solution mixed each 10 minutes for a total of 7 more times (total time since adding acid, 70mins). NOTE: I added 10 minutes to the extraction time because it should probably be stirred constantly.
- For the last 5 minutes the solution was swirled continuously while I waited for a pot of water to boil.
- The flask was set onto a thick rag in a boiling water bath for 8 minutes with the lid open swirling occasionally and then cooled under running, cold water (~5 mins.) NOTE: During the heating, the muck started to turn a dark blue, from its original green/grey. This was noticed on the surface before shaking.
- The muck was scraped out of the flask and into a conical coffee filter and allowed to drip for 5 minutes after which the remaining liquid was squeezed out. (80ml grey/white cloudy liquid was recovered)
- 20% aqueous NaOH was added to bring the solution up to pH 8 (not sure exactly how much but definitely less than 10ml). A very dark, opaque blue solution resulted. NOTE: This should be done carefully, yet quickly. Carefully because the change is not linear--15 drops might produce no change in one region of the pH scale and 2 drops might bring you from pH7-pH8. Quickly because basic conditions + air = rapid amine destruction.
- NOTE: Here is where everything starts to go wrong...
- 50ml petroleum distillates added. On pouring I noticed that this was a much higher boiling fraction of petroleum distillates than I thought, it was obvious that it probably wasn't extracting anything from the aqueous phase.
- I blitzed down to the gas station and bought some naphtha, came back, separated the two phases with a plastic bag sep. funnel and added 50ml of naphtha to the aqueous phase. This also seemed to be a terrible choice.
- Again separated off the aqueous phase and tried the only other solvent I had on hand, chloroform. This formed a nasty emulsion which I tried, unsuccessfully, to break with salt.
- By this time the aqueous layer was an impenetrable navy blue, almost black and I was out of solvents to try so I gave up.
Possible conclusions:
- High boiling petroleum distillates are probably not very good solvents for psilocin.
- Chloroform is also probably not a very good solvent for psilocin and is probably also not good for extracting aqueous solutions due to its tendency to form shitty, un-bustable emulsions.
Discussion:
I made the judgments of good solvent/bad solvent on the basis of color change alone. This is based on my experience with doing extractions of colored solutions and the fact that Casale obtained crude psilocin as a greenish residue. My experience is that if the product is colored and the solution being extracted is colored, then the extract will also be colored. This isn't necessarily true in all cases though for a variety of reasons. However, I didn't have the motivation to continue with the procedure of evaporating all those nasty solvents if I wasn't pretty sure it was working. I'll be trying this again in the next few days with the "proper" solvents. [Edit: oh yeah, and chloroform eats the hell out of plastic. But maybe only when mixed with a little water and naphtha?]
UPDATE: ROUND 2
This extraction was to go as previously with the following modifications: - 1. Extract the aqueous phase with several small portions of chloroform instead of ether.
- 2. Recrystallize from methanol.
Everything else would be the same as above and as in the article with the exception that I wouldn't be drying the extract under nitrogen, just plain wet air.
- The actual procedure went as planned except the jar in which the mush was soaked cracked in the boiling water bath after only about 2-3 minutes. The working volume of water was now about 3-fold and the temperature of the liquid was now possibly above 70C (as called for in the article) but I didn't have a thermometer so I didn't check. This made it harder to filter and also necessitated the use of much more chloroform than I would have liked.
- The basified aqueous layer was extracted 5x with a total volume of about 200ml chloroform. It remained totally clear and the aqueous phase remained a deep blue. The chloroform was poured into a shallow aluminum baking tray and set outside under a fan for 1 hour.
- At the end of this time, the pan contained a small amount of white residue and a few drops of green oil which was spread out on the pan and allowed to dry under then fan for another 20 minutes or so. The remaining greenish residue was scraped off with a scalpel and put into the bottom of a test tube. It was noticed that a lot of dust was stuck in the drying residue (use filtered air in the future?).
- Boiling methanol was added to the residue with shaking forming a greyish solution but the dust made it difficult to see when/if all of the residue dissolved.
- The test tube was capped and put in a freezer for 2 days.
- As of now, there are no crystals, only dust, in the liquid.
More possible conclusions: - A thicker insulating layer should be placed under a flask in a boiling water bath when using crappy non-Pyrex equipment.
- Always have a thermometer handy.
- Air might be as bad for psilocin as we thought (when someone says psilocin is unstable, it's left unclear what they're comparing it to--compared to graphite, or compared to nitroglycerin? LSD is also somewhat unstable but it manages to hold itself intact for at least a few months in ambient conditions....)
- That blue stuff (whether it is the breakdown products of psilocin or not) isn't soluble in non-polar solvents. This is at least congruent with it being an oxidation product and suggests that there is either significant functionalization, or complete breakdown of at least one of the rings.
- My experience with color carrying through in all parts of the reaction is countered by the case of psilocin extractions.
Questions remaining: - Exactly what are the oxidation products of psilocin. Does anybody have a reference to an article which discusses this? For that matter, what are the metabolites of psilocin in humans?
- Why is psilocin so much more unstable than DMT or any of the 5-substituted DMTs?
- By what mechanism does the oxidation take place? It's fairly certain that it occurs at the 4-position and not at the amine since this behavior is not seen in DMT itself (except at high pH when all amines are easily oxidized. An interesting note, however, is that the blue color isn't seen in this case until the solution is basified, indicating that (if the blue is in fact oxidized psilocin) psilocin is actually stabilized in acid (a fact which is also alluded to by the Casale article).
Edited by ChuangTzu (06/26/05 07:37 AM)
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