lets start w the article:
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 mothballs (06/18/16 05:22 AM)
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so, what does that say????? 100 g corn sugar 200g succinic acid (ebay) 20 g potasium phosphate. (americanspice.com) 100g epsom salt 100g yeast extract 600mg vitamin B1 500mg green vitroile (naturalpigment.com) 100mg blue vitriole (amazon) 10mg amonium molybendate(ebay) 7mg manganeese chloride (ebay) 6mg white vitrole (naturalpigments.com) 100L water
this will make an lc specifically designed by albert hoffman to make a particulary potent mycelium and is most likely the most scientific data out there on the nutes required for potency. as far as the extraction goes,.... 75 l of meoh for >1 kilo of dryed matter.....?????? WTF, mr.hoffman, phytochemistry wasnt your strong suit. put it on a meoh reflux in a 3L soxholet system to yeild perfect crystals lickity split!
Edited by mothballs (06/18/16 05:20 AM)
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