uneasyone (Stranger)
07-17-03 10:35
No 448065
      tryptamine cubensis---->psilocin hcl extraction     

I have been dreaming of using this tech to grow my maza or tazmanian strain http://www.shroomery.org/index/par/7953. Due to the high psilocin content of this method and being that i want a reasonably pure water soluble product that dosen't have to pass through the digestive system to be active i think this may be a good start. I am thinking of first washing the dried and powderd mushies with acetone maybe even cloroform or both. Then adding a proper amount of 140 proof grain alcohol back to the dried crushed material. Then adding a few drops of hcl to drop the ph to 3. I think i should probally let this sit in the fridge over night filter and repeat extraction with more alcohol and hcl always keeping liquids. After this i think i should probally combine both extracts defat with an equal amount pet ether separate layers and keep bottom layer. Ok this is where i really need advice i'm thinking that i could add baking soda or lye in small amonts until i bring the ph back to 7 or 8. Evaporate alcohol under vacume and warm water bath until a relativly dry product is formed. Then add more pet ether until disolved or mostly disolved. This is where i'm counting on doctor shulgin http://www.cognitiveliberty.org/shulgin/adsarchive/extraction.htm according to him psilocin is almost insoluble in pure ethanol and most insoluble in pure methanol. Although i'm pretty sure urea will disolve so my idea is to take an equal amount of probally methanol drop the ph to 3 with hcl and add it to my pet ether. I'm hoping the urea will be dissolved in the bottom methanol/ethanol layer of the solution. I'm not sure if the psilocin will stay in the pet either or fall out to the bottom of the jar which would be great. If it stays in the ether i think i will be able to pull it out by gassing the pet ether with hcl gas to precipitate the psilocin salt I hope this will yeild a descent product. All this is just an idea i've never pulled an extraction like this before and welcome all ideas and opinions of those who know more than I so feel free to critcize my idea or offer advice. If you can help me i thank you if not have nice day anyway.

uneasy1
 
 
 
 
    neohippy
(Stranger)
07-17-03 22:00
No 448175
      phosphorous addition     

Couldn't you add a phosphorous compound like phosphoric acid
to allow the mycelium to convert the psilocin into psilocybin before extraction from the mushrooms?
I think I read that the reason why it makes so much psilocin is that it runs out of the phosphorous compound.
 
 
 
 
    uneasyone
(Stranger)
07-17-03 22:47
No 448178
      i'm not really wanting to extract psilocybin     

i don't really want to convert the psilocin to psilocybin i'm dreaming of a fast acting water soluble product that i can ensufulate (snort) laugh and that my main lining crank headed buddies can inject i think psilocybin has to be broken down into psilocin inside the stomach to become active so it wouldn't really be ideal for either method of ingestion

uneasy1
 
 
 
 
    Lilienthal
(Moderator)
07-18-03 02:01
No 448223
      Nobody knows where psilocybin is ...     

Nobody knows where psilocybin is dephosphorylated, but it probably takes place everywhere in the body.
neohippy: Psilocin can be extracted without problems. It would also be nice to support 'what you have read' with links or references crazy.
 
 
 
 
    neohippy
(Newbee)
07-18-03 02:22
No 448227
      shulgin     

>>neohippy: Psilocin can be extracted without problems. It
>>would also be nice to support 'what you have read' with
>>links or references

i was going on what the other person posted where shulgin said ethanol or methanol wont extract psilocin.

http://www.cognitiveliberty.org/shulgin/adsarchive/extraction.htm

if he wanted to extract psilocin, please provide the link or reference on that as i believe that is what he wanted.

The following was found at http://www.jug-or-not.com/shroom/tips.html

"As many may have already figured out, there just isn't enough phosphorus in the substrate to support psilocybin production...quite simply, it is used up. My answer to this was to add a small amount of phosphoric acid (H3PO4)to the water at the same time you add the tryptamine HCl, and adjust to neutral to wide-base Ph paper using NaOH."
 
 
 
 
    Lilienthal
(Moderator)
07-18-03 05:14
No 448257
      It has been shown in Planta Med.     

It has been shown in Planta Med. 56, 327 (1989) that the best solvent for extraction of psilocin is 80% ethanol while 90 - 100% methanol is best for psilocybin extraction under their special conditions. It is no problem to extract the biomass twice with different solvents. Or to use contineuos extraction etc.

If you read the original article (J. Gartz, Planta Med. 55, 249 (1988)) you can see that it is not obvious what caused the low psilocybin content compared to psilocin when he spiked the medium with tryptamine.

Either the same low psilocybin content was seen in the control experiment (but he doesn't give us that data, it's a very poor article!). That would support the view that it was his phosphate deficient medium (but I don't think so, it is cow-dung / rice grain medium). That would also mean that if you use another medium there should be enough phosphate in it.
Or it could be that the added tryptamines are somehow suppressing phosphorylation (my favorite explanation).
 
 
 
 
    neohippy
(Newbee)
07-18-03 22:05
No 448386
      psilocin extraction
(Rated as: good read)
    

An Aqueous-Organic Extraction Method for the Isolation and Identification of Psilocin from Hallucinogenic Mushrooms.
J.F. Casale; Journal of Forensic Sciences 1985 30 (1): 247-250

20 grams of dried psilocybin containing mushrooms are ground into powder. the powder is mixed with 150 ml of dilute acetic acid. pure (glacial) acetic acid is added to pH 4. heat on a boiling water bath until it reaches 70 degrees(approx 10 minutes). the solution is then suction filtered and concentrated. ammonium hydroxide is added to pH 8. extract with 150 ml of ether or any water insoluable solvent that psilocin is soluable in. evaporate under nitrogen or argon to leave psilocin.

Edit: Full article available here: Post 449383 (Rhodium: "Mushroom Aqueous-Organic Psilocin Extraction", Tryptamine Chemistry) /Rhodium
 
 
 
 
    urushibara
(Hive Bee)
07-20-03 21:05
No 448754
      i don't see how a/b will work     

I might be mistaken, but aren't both psilocybin (well, psilocybin especially) and psilocin amphoteric? certainly psilocybin will be, being an ester.

I think that the two pass approach would be the way to go. methanol will remove the psilocin, and though psilocybin is not extracted well with ethanol, I have found that 95% ethanol in sufficient excess will result in precipitated psilocybin after filtering the mixture and cooling it/reducing it.

we can't stop here, this is bat country
 
 
 
 
    uneasyone
(Stranger)
07-20-03 21:20
No 448756
      i'm not sure     

i'm not sure whether it will work or not but i have read other bee's post that said psilocybin can't be extracted with a/b but pilocin can. i would also like to know if urea is soluble in nonpolar solvents like ether if not i could skip a step in my theoretical process and go straight to gassing. but try to remember psilocin extraction is what i'm aiming for unless someone has a viable method to convert psilocybin into psilocin . i want a fast acting product that dosen't have to pass through the digestive system to become active. i want to thank everyone for their advice and ask you all to keep it comin.

uneasy1
 
 
 
 
    urushibara
(Hive Bee)
07-21-03 01:39
No 448779
      dephosphorylation     

I think dephosphorylation occurs in the stomach. I might be wrong. in my experience the way to get consistent fast onset is to take it on an empty stomach. this was with dried cubensis.

what about deliberately providing the fungi with adequate phosphorous to increase the ratio of psilocybin that is formed? psilocybin is easier to extract, just soak/simmer and filter/freeze and there it is. it could be snuffed I suppose, I don't see why that wouldn't work. maybe dissolved in water in a nasal sprayer.

we can't stop here, this is bat country
 
 
 
 
    Lilienthal
(Moderator)
07-21-03 01:58
No 448780
      HELP, the forum is clogged by newbees :-S You...
(Rated as: Take it damn serious!)
    

HELP, the forum is clogged by newbees crazy You all better spend your time reading the textbooks, the literature, and the Hive archives instead of clogging this forum with stupid posts. Or go to the Newbee forum. I think I should use the rating buttons more often... crazy THIS IS NOT A NEWBEE CHAT!!!
 
 
 
 
    Lilienthal
(Moderator)
07-21-03 01:59
No 448781
      Look at the structure of psilocin and ...     

Look at the structures of psilocin and psilocybin to see if they are zwitterionic or not. That has nothing to do with amphoterism or beeing an ester.
 
 
 
 
    Rhodium
(Chief Bee)
07-23-03 20:00
No 449383
      Mushroom Aqueous-Organic Psilocin Extraction
(Rated as: excellent)
    

An Aqueous-Organic Extraction Method for the Isolation and Identification of Psilocin from Hallucinogenic Mushrooms
Casale, J. F.
Journal of Forensic Sciences, Vol. 30, No. 1, pp. 247-250 (1985) (../rhodium/chemistry /psilocin.extraction.html)

Abstract

A simple aqueous extraction method for the isolation and identification of psilocin from Psilocybe Cubensis mushrooms is reported. This method employs a dephosphorylation of the phosphate ester to psilocin, which facilitates a greater product yield and simplifies identification. Psilocin extracted by this method is sufficiently concentrated and free of cocontaminants to allow identification by infrared spectroscopy and gas chromatography/mass spectrometry.

The tryptamines are one of four categories of hallucinogenic indoles in more than 20 classes of indole compounds comprising approximately 600 alkaloids1. Considerable research has been conducted with psilocin and psilocybin since their isolation by Hofmann et. al.2 Several extraction techniques1,3-6 have been used to isolate psilocin and psilocybin from more than two dozen species of mushrooms in four genera (Conocybe. Panaeolus, Psilocybe, Stropharia). The techniques that use methanol co-extract other compounds such as urea, ergosterol, ergosteral peroxide, ,-trehalose, baeocystin, and norbaeocystin3,4,7. At present, a useful aqueous extraction procedure has not been reported for psilocin and psilocybin.

The dephosphorylation of psilocybin to psilocin in vivo has been well documented1,8,9 and is thought to account for most or all of its central nervous system activity8. Conversion of psilocybin to psilocin is also necessary for aqueous extraction with organic solvents because of the very low lipid solubility of psilocybin. Extraction of only one compound also permits infrared analysis of the extract.

Concentration and detectability of psilocin and psilocybin are dependent on several variables, including:

1. The absence of glucose, which will prevent the production of psilocybin10.
2. Low levels of ammonium succinate, which will give poor yields of psilocybin10.
3. The growing medium, which requires a pH of less than 710.
4. Timing: maximum production of psilocybin occurs on the seventh day after germination, while maximum production of the mycelium is reached by the ninth day10.
5. Temperature: complete loss of psilocin and psilocybin will occur in harvested mushrooms left at room temperature for an extended period of time3.
6. Oxidation: psilocin will oxidize to a blue product (possibly accounting for the bluing color in the four genera containing psilocin and psilocybin)9.

Because of the increasing popularity of these mushrooms and kits available from drug oriented publications for growing mushrooms containing psilocin and psilocybin in cow manure a simple aqueous extraction procedure has been developed that extracts reasonably pure psilocin from mature mushrooms. This extraction method greatly simplifies the identification of psilocin from those mushrooms by infrared spectroscopy and gas chromatography/mass spectrometry (GS/MS).

Experimental Procedure

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 arc 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). (../rhodium/chemistry /pictures/psilo.xtract.fig1.gif)
Smaller mushrooms exhibits provide ample psilocin for mass spectral analysis (Fig. 2). (../rhodium/chemistry /pictures/psilo.xtract.fig2.gif)

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.

References

[1] Schultes, R. E., "Indole Alkaloids in Plant Hallucinogens" Journal of Psychedelic Drugs, Vol. 8, No. 1, Jan.-March 1976, pp. 7-25.
[2] Hofmann, A., Heim, R., Barck, A., Kobel, H., Frey, A., et al, "Psilicybin [sic] and Psilocin" Helvetica Chimica Acta, Vol. 42, No. 2, 1959, pp. 1557-1572.
[3] Beug, M. W. and Bigwood, J., "Quantitative Analysis of Psilocybin and Psilocin in Psilicybe Baeocystis (Singer and Smith) by High-Performance Liquid Chromatography and by Thin-Layer Chromatography" Journal of Chromatography, Vol 207, No. 3, March 1981, pp. 379-385.
[4] Koike, Y., Wada, K., Kusano, G., Nozoe, S., and Yokoyama, K., "Isolation of Psilocybin from Psilocybe Argentypes and Its Determination in Specimens of Some Mushrooms" Journal of Natural Products, Vol. 44, No. 3, May-June 1981, pp. 362-365.
[5] Ott, J. and Guzmán, G., "Detection of Psilocybin in Species of Psilocybe Panaeolus and Psathyrella" Lloydia, Vol. 39, No. 4, July-Aug. 1976, pp. 258-260.
[6] Guzmán, G. and Ott, J., "Description and Chemical Analysis of a New Species of Hallucinogenic Psilocybe from the Pacific Northwest" Mycologia, Vol. 68, No. 6, Nov. 1976, pp. 1261-1267.
[7] Lenny, A. W. and Paul, A. G., "Raeocystin and Norbaeocystin: New Analogs of Psilocybin form Psilocybe Baeocystis" Journal of Pharmaceutical Sciences, Vol. 57, No. 10, Oct. 1968, pp. 1667-1671.
[8] Horita, A. and Weber, L. J., "Dephosphorylation of Psilocybin in the Intact Mouse" Toxicology and Applied Pharmacology, Vol. 4, No. 6. Nov. pp. 730-737.
[9] Horita, A. and Weber, L. J., "The Enzymatic Dephosphorylation and Oxidation of Psilocybin and Psilocin by Mammalian Tissue Homogenates" Biochemical Pharmacology, Vol. 7, No. 1, 1961, pp. 47-54.
[10] Catalfomo, P. and Tyler, V. E., "The Production of Psilocybin in Submerged Culture by Psilocybe Cubensis" Lloydia, Vol. 27, No. 1, March, 1964, pp. 53-63.
[11] Clarke, E. G. C., Isolation and Identification of Drugs, Pharmaceutical Press, London, 1974, p. 526.
[12] Agurell, S. and Eilsson, L., "Biosynthesis of Psilocybin Part II. Incorporation of Labeled Tryptamine Derivatives" Acta Chemica Scandinavica, Vol. 22, No. 4, 1968, pp. 1210-1218.
 
 
 
 
    Rhodium
(Chief Bee)
07-27-03 11:33
No 450198
      Ref #2: Psilocybin und Psilocin aus Rauschpilzen
(Rated as: good read)
    

Here comes Ref #2 from Post 449383 (Rhodium: "Mushroom Aqueous-Organic Psilocin Extraction", Tryptamine Chemistry) (great psilocybin/psilocin crystal images on the last page!)

Psilocybin und Psilocin, zwei psychotrope Wirkstoffe aus mexikanischen Rauschpilzen
Albert Hofmann, et. al.
Helvetica Chimica Acta, Vol 52, Fasciculus 5 1557-1572 (1959) (../rhodium/pdf /psilocybin.und.psilocin.pdf)
 
 
 
 
    Rhodium
(Chief Bee)
08-13-03 19:11
No 453450
      Ref #3: Analysis of Psilocybin and Psilocin
(Rated as: good read)
    

Here comes Ref #3 from Post 449383 (Rhodium: "Mushroom Aqueous-Organic Psilocin Extraction", Tryptamine Chemistry)

Quantitative Analysis of Psilocybin and Psilocin in Psilicybe Baeocystis (Singer and Smith) by HPLC and TLC
Beug, M. W. and Bigwood, J.
Journal of Chromatography, Vol 207, No. 3, March 1981, pp. 379-385. (../rhodium/pdf /psilo.analysis.bigwood.pdf)
 
 
 
 
    Rhodium
(Chief Bee)
08-14-03 14:07
No 453575
      Ref 12: Biosynthesis of Psilocybin
(Rated as: excellent)
    

Here comes Ref #12 from Post 449383 (Rhodium: "Mushroom Aqueous-Organic Psilocin Extraction", Tryptamine Chemistry)

Biosynthesis of Psilocybin Part II. Incorporation of Labeled Tryptamine Derivatives
Agurell, S. and Eilsson, L.
Acta Chemica Scandinavica, Vol. 22, No. 4, pp. 1210-1218 (1968) (../rhodium/pdf /psilocybin.biosynthesis-2.pdf)
 
 
 
 
    Rhodium
(Chief Bee)
02-19-04 10:06
No 489757
      Ref #10:Psilocybin Production in Submerged Culture
(Rated as: excellent)
    

Here comes Ref #10 from Post 449383 (Rhodium: "Mushroom Aqueous-Organic Psilocin Extraction", Tryptamine Chemistry)

The Production of Psilocybin in Submerged Culture by Psilocybe Cubensis
Philip Catalfomo and V. E. Tyler, Jr.
Lloydia 27(1), 53-63 (1964) (../rhodium/pdf /psilocybin.submerged.culture.pdf)


The article has also been referenced in the following posts:
Post 254261 (Zen: "Re: Enhanced Psilocybin Production", Tryptamine Chemistry)
Post 253487 (Zen: "Re: Enhanced Psilocybin Production", Tryptamine Chemistry)
Post 240882 (foxy2: "Re: PC Gulf Coast", Tryptamine Chemistry)


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