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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