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 00Zen
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Registered: 01/08/02
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 Enhanced Psilocybin Production (7.5 G Psilocybin)   2
#514231 - 01/08/02 10:08 AM (21 years, 10 months ago) |
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A method for the production of 7.5 grams of psilocybin in less than a week and a half.
Typical Ps.C. cultures grown by various organic substrates achieve .4 - .6 alkaloid yields. Typical nutrient enhanced substrates achieve slightly higher yields of alkaloids. Liquid cultures can effectively achieve 1 - 1.1 alkaloid yields by making essential nutrients and carbohydrates easily available.
This is the general potency of this strain; attempts to further improve nutrient uptake will most likely only result in a higher biomass yield. In the instance of the introduction of tryptamine to yield 3% psilocin alkaloid, I view it as a novel biosynthesis mechanism, operating externally of the natural model of the organism.
Total biomass, although affected accumulatively, is proportional to carbohydrate uptake. I have heard unofficial reports of rapid growth with dextrose as well as organic honey, which is composed of multiple complex sugars. Perhaps a more efficient mixture of carbohydrates then just glucose could be developed.
Many funguses grow rapidly in an acidic environment (3 - 5 pH). Ours displays higher yields of alkaloids and biomass between 4 - 4.6 pH but loses durability and suffers from tissue damage and possible cellular ruption due to solution agitation during the process of aeration. I assume that the growth acceleration that occurs at a lower pH is because nutrients are able to permeate the cellular equilibrium easier. Taking advantage of this requires a device capable of aerating the solution without disturbing the mycelium.
This method of mycelium harvest is far superior to fruited bodies in relation to alkaloid yields, processing, extraction, and overall time.
Lets Pretend:
Glucose (C6H12O6) [1000 g]
Ammonium Succinate (NH4OOC-CH2-CH2-COO) [200 g]
Yeast Extract (Organic Compound) [100 g]
Magnesium Sulfate (MgSO4-7H2O) [100 g]
Potassium Phosphate (KH2PO4) [20 g]
Thiamine Hydrochloride (C12H17ClN4OS HCl) [600 mg]
Ferrous Sulfate (FeSO4-7H2O) [500 mg]
Cupric Sulfate (CuSO4-5H2O) [100 mg]
Ammonium Heptamolybdate ((NH4)6Mo7O24-4H2O) [10 mg]
Manganese Chloride (MnCl2-4H2O) [7 mg]
Zinc Sulfate (ZnSO4-7H2O) [6 mg]
Water (H2O) [200 L]
Prepared in 55 Gallon (211L) drum and adjust to pH 5.5 with hydrochloric acid.
a. Lid is sealed and capped with a filtered pressure release valve.
b. Drum and solution is heat sterilized.
c. Cap is swabbed with sterile gauze and H2O2 and removed.
d. Solution inoculated with 1L of precultured Mycelium.
e. Insertion aerator and cap with filtered relief check valve.
f. Solution is kept at 30C with an electric blanket.
g. Solution is aerated for 7 days with filtered air.
h. 235 Ounces (14.5 lbs) of mycelium are strained from solution with cloth
i. Mycelium dried over calcium chloride to yield 750g dry weight.
j. Biomass is ground to a fine powder and re-added to drum with 75 L of methanol.
k. Sealed with cap fitted with large egg whisk style blender attached to the bottom.
l. Heated to 40C with electric blanket for 1-4 hours depending on level of agitation.
m. Solution is filtered with cloth.
n. Solution is re-filtered in a funnel with inert filtration medium.
o. Cap is fitted with a condenser (.5 - 1" coiled copper tubing inside 4-5 ft 6" PVC pipe allowing for coolant flow around coil) and digital thermometer to measure solution temp.
p. Sealed heat element is placed under drum to maintain 65C.
q. Solution is distilled to 1-5 liters.
r. Solution transferred to distillation apparatus and distilled in 1 L batches.
s. Yielding 20-40 grams of residue.
t. Residue is developed in a 100 ml H2O and 500 ml Naptha matrix.
u. Solution is agitated for 1 hour at room temperature.
v. Non-polar proteins and oils are removed via sep funnel separation of naptha layer.
x. Water is evaporated from extract with a blower using calcium chloride at 30C in 2 hours.
y. Yielding 10-20 grams of extract containing 7.5 g of psilocybin.
z. 375 doses with a street value of over $7000.00
I digress:
If boiling 75 L of methanol isn't your idea of time saving, cost efficient or fun, you can opt to process your ground-dried biomass in 75 (10 g / 1 L) batches. The major benefit being that you can reclaim most your solvent, eliminating the need to buy bulk methanol. The downside is the time involved, even under vacuum this is going to take a little while. Under optimum conditions we could still complete the entire process in less than two weeks.
Perhaps a reflux percolator could be implemented to percolate the bulk material with 1 L methanol while collecting extracts in the distillation chamber. (i.e. the methanol condenses from the distillation chamber into a reservoir layered with: ground biomass and perculate, inert filtrate medium, and filter material; then drains back into the distillation chamber.) Percolation should be complete within 4-24 hours. Fastest results would be seen if condenser displaced about 20C. Alternatively the reservoir could act as a condenser with the addition of a cooling coil to displace vapor temperature and instigate methanol condensation while passing thru the biomass perculate. Biomass should be free of alkaloids upon completion and can be tested for their presence using Keller's reagent. Remaining methanol is distilled off.
Under optimum conditions the entire process could take less than a week and a half.
-Zen
Ref: Hoffmann, Albert "Obtaining Psilocybin and Psilocin from Fungal Material" Sandoz Ltd., US patents 3183172, 1959
Ref: Hofmann, Albert "Method of Inducing Therapeutic Tranquilization with Psilocybin and Psilocin" Sandoz Ltd., US patents 3192111, 1959
Ref: Catalfomo, P. and V.E. Tyler, Jr. "The Production of Psilocybin in Submerged Culture of Psilocybe Cubensis" LLoydia 27:53-63, 1964
Ref: Gartz, Jochen "Synthetic Nutrient Medium for Fungal Manufacture of Indole Alkaloids" Ger. (East) DD 255,749 (cl. C12P15/00), 13 Apr 1988
Edited by 00Zen (01/08/02 10:10 AM)
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 Humidity
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Last seen: 19 years, 10 months
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 Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#514292 - 01/08/02 11:32 AM (21 years, 10 months ago) |
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I read through your entire message, but I find it hard to follow. Are these select quotes from the references at the bottom?
In reply to:
Typical Ps.C. cultures grown by various organic substrates achieve .4 - .6 alkaloid yields. Typical nutrient enhanced substrates achieve slightly higher yields of alkaloids. Liquid cultures can effectively achieve 1 - 1.1 alkaloid yields by making essential nutrients and carbohydrates easily available.
So, Liquid Cuture Fermentation is more productive than solid state substrate fermentation, according to one of the articles.
This goes against what everyone here at the shroomery reports about liquid mycellium (the consensus is that liquid mycellium is not very potent). This might be because you need a lot more liquid culture to equal the same amount of nutrients as a solid culture (For example: mabey you need 5 gallons of liquid culture to equal one quart jar of bird seed).
Does the artical give any numbers on the amount of each substrate used in each potency comparison?
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_____________________________________________________________________________________
"The greatest enemy of knowledge is not ignorance, it is the illusion of knowledge." -Stephen Hawking
Edited by Humidity (01/08/02 11:34 AM)
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Humidity
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: Humidity]
#514302 - 01/08/02 11:40 AM (21 years, 10 months ago) |
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Also could you elaborate on the liquid cultures more and leave out the extraction stuff. Airation devices are cheap and easiy to find at home brew stores. Does anyone have any good info on large scale liquid fermentation, like how you sterililze everything, ect?
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_____________________________________________________________________________________
"The greatest enemy of knowledge is not ignorance, it is the illusion of knowledge." -Stephen Hawking
Edited by Humidity (01/08/02 11:52 AM)
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00Zen
Stranger
Registered: 01/08/02
Posts: 22
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: Humidity]
#514377 - 01/08/02 01:03 PM (21 years, 10 months ago) |
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The statements are my summation of the references and potential aspects of efficiency increase in this method. The references are a bit more technically oriented yet clearly defines a solid foundation for this method.
While the only real necessity for mycelium production is glucose or similar carbohydrate, alkaloid production requires the presence of essential nutrients not only as precursor components, but to effectively utilize carbohydrates, promote cellular development, and to regulate vital enzyme pathways. I believe from the comparison of key nutrient needs in this fungus and typical plant nutrients that it is well adapted, although not limited to, obtaining all of its necessary nutrients from the break down of organic plant matter. This is why some people would not detect the production of psilocybin until the mycelium has been transplanted from a nutrient deficient solution to an organic substrate.
The guidelines for nutrient solutions are that they contain nutrients in amounts that are proportional to the tissue composition and in a total solution concentration that does not damage the tissue. In general all aspects of this solution contribute to the production of psilocybin as well as promote rapid mycelium growth. The third reference gives a more or less complete nutrient-subtraction medium study that yields further insight into the effects of each nutrient on alkaloid production.
It should be notated that the above methods yields are actually obtained with 2000 g of glucose to yield a 1.02 percent alkaloid content by day 7, unfortunately this carries a side effect whereby the alkaloid content diminishes to 0.15 by day 11. A solution with 1000 g of glucose would yield only 0.52 percent alkaloid content but continues to full growth at day nine with no diminishing of potency.
-Zen
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Anonymous
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 Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#514470 - 01/08/02 02:23 PM (21 years, 10 months ago) |
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Methanol and the Naptha are both highly combustible. I hope you're not doing this in a residential area where a mistake could mean burning down your neighbor's home as well as your own which could entail the loss of life. I agree with humidity, "leave out the extraction stuff."
The purchase of 75 L of methanol would seem to be a red flag for some nefarius governmental agencies and "375 doses with a street value of over $7000.00" - prompts me to suggest that you might also add a disclaimer: "following these instructions may result in jack booted thugs breaking down your door in the middle of the night with a no-knock raid and shooting you because you were startled and jumped out of bed in a 'threatening manner.'"
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00Zen
Stranger
Registered: 01/08/02
Posts: 22
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#514510 - 01/08/02 03:03 PM (21 years, 10 months ago) |
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In a perfect world and sterile environment each component could be heat or chemically sterilized previous to solution preparation. In a less than perfect world the elaboration of steps (a.) and (b.) reveals that the container and solution is heated under 15 psi pressure to achieve 250 F capable of destroying organic contaminants.
As implied by step (c.) all apparatus and surface areas are chemically sterilized with sterile gauze and H2O2, or possibly bleach solution, at each step. This still requires a fairly sterile environment or perhaps a rudimentary glove box apparatus.
Introduction of H2O2 to the solution could help to further prevent contamination, possibly to the point of making this process a breeze. I do not have any empirical data regarding H2O2 to support this statement; any references or significant data in regards would be greatly appreciated.
For aeration I am inclined to use a high output aquarium air pump with inline HEPA filters (charcoal might suffice) to drive multiple air stones at the bottom of the drum.
-Zen
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00Zen
Stranger
Registered: 01/08/02
Posts: 22
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#514697 - 01/08/02 06:25 PM (21 years, 10 months ago) |
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*Checks the sign on the classroom door to make sure it still says, "ADVANCED Mushroom Cultivation"*
I'm assuming that since this is the advanced forum you would have the common sense not to expose flammable materials to an open heat source, drink poisonous solvents, or decide Naptha makes an excellent eye wash. I'm assuming this in the same manner that I'd assume you are capable of dealing with the potent biological toxins that could be present in a contaminated fungal culture.
Secondly I am presenting a method of enhanced psilocybin production that is a culmination of advanced techniques to yield a large quantity of pure psilocybin. Isolation of active alkaloids is imperative to the testing of subtle alkaloid yield differentials when trying to develop a more efficient nutrient solution. Not to mention in this instance Psilocybin is the only desired product. I can understand your fascination with the nutrient mediums, but I am not going to neglect other aspects of the procedure. I am more then happy to elaborate on all aspects of this procedure as they are brought to my attention.
Thank you for pointing out the volatility of these solvents. Though we are heating Methanol to its boiling point in step (p.) we do not approach its flash point. All vapors are condensed back to a liquid state and solution temperature is constantly monitored via digital equipment via step (o.). For additional security all heat sources in steps (f.) (l.) and (p.) are sealed in high temp. plastic to prevent flash of vapors that could develop from faulty seals or other potential human error.
In steps (t.) (u.) and (v.) Naptha is used to "wash" the extra proteins and oils that make up over 50% of our crude extract, yielding a relatively pure extract of psilocybin with only trace amounts of water soluble mycelium by product. While more volatile then Methanol, Naptha is not heated during this process and is commonly found in a rather high purity in lighter fluid. Naptha lighter fluid that evaporates cleanly off the face of a mirror without any trace of residue is suitable for our needs.
Pure methanol is obtainable OTC from several sources, including sports car service centers; model hobbyist shops, camping stores, and often times paint supply of hardware stores. There are also many sources of not so pure methanol that would need to first be distilled such as Heet fuel dryer and some brands of windshield wiper fluid, but it is generally easier to just use a pure source of methanol. Depending on extraction technique used, as described, you could effectively use as little as 1 L of methanol.
(z.) is merely a determination of units and definition of product value, such as an equivalence to "3-5 lbs primo dried caps d00d" not an open invitation for "jack booted thugs" to violate your residence, unless you make it into one.
Since you mentioned it, I live in a large house centered on a 25-acre residential lot. It would take a small nuclear explosion to burn down my closest neighbors home.
-Zen
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slither
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Registered: 02/02/01
Posts: 365
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 Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#514719 - 01/08/02 06:56 PM (21 years, 10 months ago) |
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It seems are more potent species like cyan's or azures would be better suited for something like this. It would be an interesting experiment. I'll read through it later when I have the time. I have 3 2' x 4' flow hoods here, eberbach containers, digital hotplate stirrer, lots of cultures, and just got my floating stir bar in the mail today 
I had read something similar to this previously, but this seems far more detailed, glad to see this topic brought up by someone that seems to know what they're talking about. I'll be back.
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Anonymous
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen] 1
#514883 - 01/08/02 09:46 PM (21 years, 10 months ago) |
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Good EDIT!!!!!!!!!
Great thread. Though I am found of actually producing mushroom fruit bodies, but the mad scientists are gonna love you.
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00Zen
Stranger
Registered: 01/08/02
Posts: 22
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: ]
#514918 - 01/08/02 10:39 PM (21 years, 10 months ago) |
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First post, first sentence right after the title summary.
"Typical Ps.C. cultures.." referring to the cultivation of Psilocybe Cubensis Mushrooms. All following information is directly related to the cultivation and processing of this species of fungus, or related conversation.
Yes psilocybin is a drug with significant street and clinical value; it is the major active ingredient of most Psilocybe and psychedelic mushrooms. Currently the clinical price of psilocybin is higher than the street value.
A patent yielding a method to rapidly produce high quantities of pure psilocybin could be worth a fortune in a year or two due to worldwide drug law reforms. Even the FDA have been changing their view points on these mushrooms and there active ingredients, they are currently doing voluntary human testing with pure psilocybin but the price from the FDA approved labs has been as high as 5 times the street value.
My hobbies include music, psychology, mycology and chemistry.
I did invent an improved novel method to produce MA via electro catalytic reduction about 2 years ago, it implemented environmentally safe chemicals and ultrasonic agitation to allow for a supplement of costly Pd/C catalyst, I believe it was quite popular and is still archived at the Hive.
-Zen
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00Zen
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#514981 - 01/08/02 11:41 PM (21 years, 10 months ago) |
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The idea is intriguing slither; those species produce 1% alkaloid content in substrates where Ps. C. would produce only 0.5%. I have never seen a definitive study on the alkaloid yields of various species when cultured in a submerged nutrient. If raising glucose levels directly raises alkaloid yields in these strains as well, the results could be phenomenal, in the 2% range. The typical glucose and phosphate needs of these potent species are probably slightly higher to begin with.
The nutrient solution has been optimized for Ps. C. from a standard fungal growth solution. If these species grow under similar conditions as Ps. C. I don't see where the harm in trying would be. I believe these species have quite a different lifespan this may effect the optimal harvest point. This is typically a few days before maximum growth is achieved.
I've been humoring the idea of putting a digital microscope in a positive pressure glove box with a rack of liquid nutrient slide cultures. Even most low cost digital microscopes are capable of producing vivid, hi res imagery at 400x magnification. A remarkably scalable process, even during small-scale cultivation on a slide the initial life cycle can still be viewed and timed. In the original testing of this solution an array of 30 ml cultures were used so that they could test varying levels of nutrients. Cultures with a higher biomass were generally regarded to have a superior nutrient solution makeup. It would probably be easier to produce determinable amounts of biomass difference or alkaloid yield with larger cultures in the 300ml - 1L range.
The original nutrient solution is as follows:
"Medium no. 1 [ammonium succinate (1 g), Glycine (9 g), Glucose (5 g), yeast extract (.5 g), KH2PO4 (.1 g), thiamine hydrochloride (.003 g), (NH4)6Mo7O24-4H2O (.05 mg), ZnSO4-7H2O (.3 mg), MnCl2-4H2O (.35 mg), FeSO4-7H2O (2.5 mg), CuSO4-5H2O (0.5 mg), MgSO4-7H2O (0.5 g), distilled water, to make (1.0 L), adjust to pH 5.5 with hydrochloric acid] accumulated psilocybin but not psilocin. Maximum production of the former compound occurred on the seventh day (0.52 per cent, dry weight of mycelium), whereas growth attained its maximum (average 112.6 mg, dry weight of mycelium per 30 mL of medium in a 125 mL flask) on the ninth day."
The most important addition is:
"Doubling the normal amount of [glucose] promoted psilocybin accumulation which reached a level of 1.02 percent (dry weight) by the seventh day. After eleven days, this level dropped to 0.15 percent; a similar sudden decline in psilocybin content was also noted in those flasks containing a reduced phosphate concentration."
Nice setup by the way, slither, sounds like you're ready roll. I'll try to put together an acquisitions write up for the necessary nutrients for you. They're all relatively inexpensive common compounds.
-Zen
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slither
addict
Registered: 02/02/01
Posts: 365
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 Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#515016 - 01/09/02 12:45 AM (21 years, 10 months ago) |
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Wow, now I see you've really put alot of thought into this. I've been playing with liquid inoculants for quite some time, but just for inoculation. Haven't had much time to put alot of thought into what you're talking about. Still I think it would be really fun to try on a smaller scale. I believe I could realistically do this contam free with say a 1-2 gallon jar/flask on the hot plate stirrer in front of a hood. Wouldn't take very long at all starting with a clean isolated culture blended in an eberbach. I don't think I'd ever have the need for 55 gals worth, but even for someone that did, it'd probably good to do smaller trials first.
For the more potent variety's I'm sure you'd have to make other changes in the formula, but a good start would probably be sawdust or a wood extract made from boiling hardwoods I'd assume since most of the more potent species are woodlovers. I know Stamets lists a few liquid culture recipe's for woodlovers in either TMC or GGMM, can't seem to find either right now though. It just seems if something like this were to be done it would be much more practical to start with a more potent species. In my experience it's not hard at all to grow out azure mycelium, in fact I've grown it on chicken scratch among other things at room temperature.
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NeonBlack
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#515276 - 01/09/02 09:43 AM (21 years, 10 months ago) |
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Since the object is a pure product, why not run the extract through a chromatography column? That's realy the only way to get a pure crystalline product.I'm sure that after the naptha wash it's pretty clean but why not take the extra step? After putting it through the column I'm sure the yield would be a little less than the 7.5 grams you state, but then you would know exactlty how much psilocybin you had, not just a ballpark figure.
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Food
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 Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#515399 - 01/09/02 12:41 PM (21 years, 10 months ago) |
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Excellent post - but where might one be able to obtain one of these drums you speak of ?
And roughly how much do they cost ?
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--------mushworld.com-----More info than you can throw a stick at-
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slither
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Registered: 02/02/01
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: Food]
#515430 - 01/09/02 01:03 PM (21 years, 10 months ago) |
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I've got a steel 55 gal drum, but I almost doubt it would hold the pressure of 15psi. Plus the prolonged heat that it would take to get the pressure up would carmelize everything in the bottom.
I'd love to see pics of a pressurized lid for one though and the egg beater top, sounds phenominal. I really think large carboys on hot plate stirrers would be alot more practical/realistic since that is how mass mycelial cultures are made in gourmet cultivation. It would be expensive though.
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Humidity
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: slither]
#515554 - 01/09/02 04:17 PM (21 years, 10 months ago) |
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How about a modified hot water heater.
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_____________________________________________________________________________________
"The greatest enemy of knowledge is not ignorance, it is the illusion of knowledge." -Stephen Hawking
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Joshua
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#515629 - 01/09/02 05:41 PM (21 years, 10 months ago) |
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Has your methodology been tested? Where can I volunteer for some of the clinical trials you speak of? Do you have any article references of recent psilocybin/psilocin research? Do you have a method for dosing the resulting crude product?
Joshua
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The Shroomery Bookstore
Great books for inquiring minds!
"Life After Death is Saprophytic!"
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homage_etd
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 Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: 00Zen]
#516222 - 01/10/02 08:39 AM (21 years, 10 months ago) |
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I apologize for sidetracking, but i have a question, lest i inquire here, it would not be answered.
As in the mycelium, do fruiting bodies also fluxuate in psilocybin content so radically between phases of growth?
And if so, at what phase is optimal psilocybin content observed?
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*shrugs*
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00Zen
Stranger
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Re: Enhanced Psilocybin Production (7.5 G Psilocybin) [Re: homage_etd]
#517124 - 01/11/02 05:24 AM (21 years, 10 months ago) |
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The Magic Potion
I found a nice place that likes to mix chemicals for you. I requested price checks for 100g / 500g / 2.5kg / 12kg. Here's a summary of the order I sent off to the lab:
Product: 100 g of concentrated Synthetic Nutrient Solution
Concentration: 100 g will produce 8 L of aqueous nutrient solution.
Acidity: 12.5 g of concentrate should produce 1 L of solution with a pH of 5.5
Quality: Does not need to meet reagent purity, food grade is acceptable.
Make Up: Relevance of definition in the order of Formula, Name, then CAS RN:
(Recommended CAS RN may not be the optimum form of substance)
1. Glucose (C6H12O6) [80 g] CAS: [50-99-7]
2. Ammonium Succinate (C4H12N2O4) [8 g] CAS: [2226-88-2]
3. Yeast Extract (Organic Extract) [4 g] CAS: [8013-01-2]
4. Magnesium Sulfate (MgSO4-7H2O) [4 g] CAS: [7487-88-9]
5. Glycine (C2H5NO2) [3 g] CAS: [CAS 56-40-6]
6. Potassium Phosphate (KH2PO4) [800 mg] CAS: [7778-77-0]
7. Thiamine Hydrochloride (C12H17ClN4OS HCl) [24 mg] CAS: [67-03-8]
8. Ferrous Sulfate, Heptahydrate (FeSO4-7H2O) [20 mg] CAS: [7720-78-7]
9. Cupric Sulfate, 5-Hydrate (CuSO4-5H2O) [4 mg] CAS: [7758-98-7]
10. Manganese Chloride, 4-Hydrate (MnCl2-4H2O) [2.8 mg] CAS: [7773-01-5]
11. Zinc Sulfate, Heptahydrate (ZnSO4-7H2O) [2.4 mg] CAS: [7733-02-0]
12. Ammonium Molybdate, 4-Hydrate ((NH4)6Mo7O24-4H2O) [.4 mg] CAS: [12027-67-7]
13. DiHydrogen Oxide (H2O) [146.4 mg - X] CAS: [7732-18-5]
14. Hydrochloric Acid (HCl) [X mg] adjusted to balance solution pH when properly diluted.
In a perfect world, they'd be a perfect supply for new and improved < Mycelium MagicGro(tm) > *j/k*; but it turns out that most labs and chem supplies are about 10 times higher in price then if you just find the right place to look. For instance Magnesium Sulfate sells for $12.50 / 125g at a popular online chem supply. But a quick spin around the bathroom isle in the super market can land you 500g U.S.P. grade MgSO4 for $1.00 in the form of Epsom salts. Another potentially pricey aspects of mass production is all the Glucose, 2 kg of top of the line high octane D-Glucose can run close to $100.00 from a chem supply. The brewers among us have already realized this can be had in good purity from corn sugars at $2.00 per kilo.
Elusive Chemicals - http://chem.com/catalogs/
Essential chemicals such as Ammonium Succinate and Ammonium Molybdate, 4-Hydrate have proven rather elusive to this method of thinking. They are easily and legally available from a chem supply but you're going to pay a bit more for them. Nonetheless, some people don't mind paying for the convinence, in which case the aforementioned link is a good starting point.
Vitamins
The nutrients also contain a large group of minerals that can be found in multi-vitamins. Typically the dosage is the wrong ratio of nutrients, there's myriad amounts of possibly undesirable substances, and in order to get a successful dose of Thiamine you have to find the super thiamine vitamins with 300% DV. I'd imagine a trip to the local GNC or Health Food Store could be very inspiring in the search for other cost effective supplies of these nutrients though. The desired form of these minerals is a Sulfate because it is relatively neutral and doesn't destroy your carefully balance pH. I've been told of on going experiments implementing PDA, high levels of glucose, and multi-vitamins attempting to create a suitable environment for submerged psilocybin production; there has been no definitive data from this source yet.
Containers
After careful discourse on another forum it has become clear that direct distillation of bulk solvents is inefficient and unnecessarily dangerous. A soxhlet extraction apparatus is the ideal embodiment for the task of collecting alkaloids from the bulk-dried mycelium. I wish at this point one of the H2O2 Guru's could step forward and offer a "solution" that would allow us to simply heat sterilize batches of nutrient solution then safely add the solution to a chemically sterilized fermentation vessel. This could allow for nearly unlimited variations of possible vessels. How about those nifty 20-40 gallon clear water jugs they use for drink dispensers? That would give you a chance to pull back the blanket and see the magic at work.
Chromatography
NeonBlack mentioned implementing a chromatography column to separate 7.5 g of pure psilocybin from the 2.5 - 12 g pf undesired crude extract. Unfortunately though this results in an amazingly pure sample of psilocybin it is unable to be crystallized until the ionic state of the material has been changed. (see Hoffman?s original patent for details) The following article outlines the basis of psilocybin chromatography and establishes determinable identifiers for psilocybin, psilocin, baeocystin, as well as three other trace alkaloids, in the preferred solvent system: butanol-acetic acid-water (12:3:5) Anyone wishing to try this procedure should probably look up the related text starting with:
Ref: "Quantitative Analysis of Psilocybin and Psilocin in Psilocybe Baeocytis by
High-Performance Liquid Chromatography and by Thin-Layer Chromatography" Journal of Chromatography, 207 (1981) 379-385
MAPS - http://maps.org/research/index.html#PSILOCYBIN
Multidisciplinary Association for Psychedelic Studies has become a political leader on the fore fronts of drug reevaluation and decriminalization. In their most recent FDA and DEA endorsed studies with psilocybin starting on Nov 27, 2001 MAPS paid $12,250 for the synthesis of one gram of psilocybin. There's also information regarding a few other clinical studies, as well as a whole storehouse of information regarding other various substances.
Homage
That's a loaded question, mycelium growth repeatedly displays the same behavior when in the same environment. As soon as the mycelium begin the next leg of their life cycle results begin to vary even in identical environments. The answer I'm going to present is a theory at best that was developed in conversation by my colleagues and I . As presented earlier the mycelium rapidly produces psilocybin but no psilocin until right before growth starts to slow down. In a healthy culture the mycelium hold their potency for awhile even after growth is retarded, in glucose activated cultures suffering from hyper activity the potency drop off is astounding at this point, yet still no presents of psilocin.
It is generally accepted that psilocin is a precursor to psilocybin within the organism, but our data seems to reflect differently. One of my colleagues believes that psilocybin acts as a catalytic agent possibly during the production of spores or other tissues, in a process whereby the phosphate group of the psilocybin is reduced to yield psilocin. It is then feasible that this rather unstable chemical could further breakdown to baeocystin and some of the other trace alkaloids visible under chromatography. This has the implication that there may be only a few enzymatic pathways producing a single active alkaloid, and the other resulting alkaloids are but artifacts of this original alkaloid pathway.
It is my belief that the psilocybin content acts as a timing mechanism, once it has been introduced into an environment that has the essentials for alkaloid production it begins stock piling them like mad. Once the alkaloid content is high enough in a certain area of the mycelium, the mushroom has detected proper nutrients for reproduction, and thus begins to trigger a fruiting response. It is interesting to note that the phosphorous that is essential to psilocybin production is also essential to the production of DNA.
The mushroom continues to produces psilocybin but I think there is an active mechanism during the growth of the fruit that causes the conversion of this alkaloid to psilocin thereby maintaining a relatively level alkaloid content. The fruits seem to have quite a variety of complexity even in a homogenous culture. Due to how well the fruit does or does not function can result in two mushrooms next to each other with radically different potencies. Generally fruits are considered to have the highest psilocybin content right before the veil breaks, after which point there is a noticeable increase in psilocin and a slight drop in psilocybin. The mycelium is amazingly efficient at hoarding psilocybin to start a fruit and some people find the most potent fruits are aborts. The logic behind this is that the mycelium had produced a good portion of the alkaloids the fruit would need but for some reason the fruit faltered yielding a highly potent storehouse of alkaloids that has hardly been touch.
As stated before I think the organism is highly sensitive to it's alkaloid levels as a means of internal timing and basic environmental "sensing". Psilocin build up consecutively raises during each sequential fruiting, this could allow the organism to detect it's age and eventual retire to the end of it's life cycle. Generally all living organisms have a built in ageing mechanism to help promote environmental adaptation.
It's theoretical and speculative but it's my general take on this mystified mushroom.
-Zen
Edited by 00Zen (01/11/02 07:19 AM)
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gray1
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Re: Enhanced Psilocybin Production (7.5 G Psilocyb [Re: 00Zen]
#517236 - 01/11/02 09:09 AM (21 years, 10 months ago) |
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joshua, are you a molecular biologist? nice des pic
it's nice to see the usual gang of mad scientists contributing to this post
a few of my observations
sterilizing 55 gallon drum is no small task and contamination would be costly in both materials and time, scale down to several growth chambers and then combine biomass for extraction if it pleases you. (also, i have no idea how you'd sterilize that bad boy, i work with an industrial size autoclave and it would just barely fit into it, never be able to get to full temperature, and 55 gallons of liquid media weighs a lot., surface hydrogen peroxide isn't really going to cut it, bleach would be more better)
interesting ideas, but i don't really buy into your idea about psilocybin being a growth regulator, differentiator
alkaloids such as these are secondary metabolites and generally end products, while there are always further degradation products of other alkaloids (maybe as negative feedback entities?) secondary metabolites are primarily produced in secondary growth structures ie the mushroom fruits. generally, people consider liquid mycellium culture not because it has a higher alkaloid concentration, it's actually considerably lower, but because it is that much easier.
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