It07:Psilocybin

Psilocybin
IUPAC Systematic Name
4-Phosphoryloxy-N,N-dimethyl-tryptamine
Identification
CAS Number	520-52-5
Beilstein Registry No.	273158
PubChem	10624
Chemical Data
Molecular Formula	C12H17N2O4P
Mol. Mass	284.25 g/mol
SMILES	C[NH+](C)CCC1C[NH]C2CCCC(OP(=O)([O-])O)C12
Physical Data
Melting Point	493-501K
Density	1.34gcm-3
Legal Status
Prohibited (S9)(AU) Schedule III(CA) Class A(UK) Schedule I(US)

Psilocybin Mushrooms

Psilocybin (a.k.a. psilocybine) is a psychedelic alkaloid (naturally occurring amine) of the tryptamine family, found in psilocybin (magic mushrooms). It is a psychoactive substance and used to suppliment types of 'transcendence' such as meditation and psychedelic psychotherapy.
It is present in many species of fungi such as those of the genus Psilocybe, such as Psilocybe cubensis and Psilocybe semilanceata but is found in many other genera. The intensity/duration of the trip varies depending on the species of mushroom, quanitity consumed and individual physiology.

Chemistry

Psilocybin is a drug administered in an inactive form that is then converted to the active compound psilocin in the body by the removal of phosphate groups. The reaction occurs under acidic conditions or by phosphatases in the body. It is a zwitterion that is soluble in water, quite soluble in methanol and ethanol and insoluble in most organic solvents.

Albert Hofmann of the University of Delaware was the first to recognize the chemical structure of psilocybin and psilocin. Hofmann's colleagues, however, were unsuccessful in their attempts to isolate the active compound.

Biology

Psilocybin occurs naturally and high concentrations can be found in some species of Psilocybe and Panaeolus (known collectively as psilocybin or psilocybian mushrooms), and lower concentrations are present in some species of Agaricales. The potency varies significantly between species and even between specimens of one species in the same batch. Smaller, younger mushrooms are higher in alkaloids and are of a milder taster than larger mushrooms. Mature mycelium also contains some psilocybin which can be extracted with an acidic solution (typically citric acid or ascorbic acid is used). Young mycelium does not contain very many alkaloids. Numerous types of psilocybin mushroom bruise a blue colour when handled/damaged due to the oxidation of active compounds, however, bruising is not a conclusive way of establishing the potency of a mushroom.

Pharmacology

Psilocybin is dephosphorylated in the body at a rapid rate to psilocin which acts as a partial agonist at the 5-HT_2A serotonin receptor within the brain where it mimics the effects of serotonin (5-HT)_1A and 5-HT_2A/2C agonist.

Applications

The mycelium of any psychoactive psilocybin mushrooms can typically be cultivated without huge difficulty and can be grown in ordinary jars in 10-12 days under typical conditions. It is possible to create a large scale psilocybin factory in a small room (such as that of the average university student) that can produce approximately 5000 doses of psilocybin per week.

Finding the mushroom

Only few spores from one mushroom are required to develop a culture that will produce large quantities of psilocybin for a number of years. The common Ysidro mushroom, psilocybe cubensis 'Singer' is most easily obtainable, readily cultivated, most resistant to disease and psychoactively the strongest. Although the sale of magic mushrooms/growing kits is formally illegal within the UK, additional information can be found on a number of internet websites. Psilocybe cubensis is normally found growing on/near cow dung in pastures between February and November. NOTE: there are many species of toxic mushrooms which could be mistaken for some other species of psilocybin fungi. It is reccommended that literature be consulted before searching for psilocybin mushrooms.

Information on preparing cultures

Many varieties of bacteria and fungal spores could overrun the mycelial cultures if careful measures are not followed: The room should be cleaned, tidied and dusted prior to working (this may take some time for the average student). The desk should be cleared of all work/lecture materials and sprayed with disinfectant. Arms, hands, nails and most of the body should also be rinsed with disinfectant/soap, whilst the mouth should be washed with antiseptic mouthwash. Clean clothes should then be applied and the mouth covered with a face mask (or clean scarf). The head should then be shaved, or if hair is valued, a hairnet can be worn. The gaps around windows and doors should be covered with rolled towels to exclude drafts, and the number of people entering the room should be severely limited (particularly anyone incapacitated by alcohol consumpion). If possible a hood should be constructed around the desk and no-one should be allowed to enter whilst work is in progress.

All utensils should be sterilised: glassware boiled in water for 30 mins and metals heated under a flame (or putting them in a clean oven at a high temperature will suffice)

Making a spore print

A spore print is a collection of spores on a flat surface that can serve a number of purposes. Spore prints are the standard method of collecting spores for later germination on agar media. To make a spore print take a mushroom with the cap fully open and gills exposed. With a sharp sterile knife cut the stem off as close to the gills as possible. Place the cap gills-down on a clean white sheet of paper. Cover the cap with a clean inverted bowl. Let this stand for 24 hours and if after this time a good spore print has not formed, tap the cap lightly with the flat side of a spatula. Fold the paper several times to ensure the print is well contained.

Preparing the media

PDA (Potato Dextrose Yeast Agar) can be prepared as follows: Wash 250g of unpeeled potatoes and slice them into 2-3ml slices. Wash them several times until the rinsings are clear, drain in a collinder and rinse with distilled water. Cook in distilled water until soft, strain the cooking liquid through a flannel shirt (lookout for people wearing these in lectures) and collect this liquid in a flask. Rinse the boiled potatoes further with distilled water and add this liquid to the liquid in the flask and eat the potatoes. Make the liquid in the flask up to 1 litre with distilled water, bring to the boil and add 15g of agar, 10g dextrose and 1.5g yeast extract. The agar should be added slowly to prevent over-boiling. While the liquid is hot, pour it into a petri dish (or other such culture container e.g. boiling tubes 1/4 full), half-filling the dish.

MEA (Malt Extract Agar): To one litre of gently boiling distilled water add 20g of malt extract, 20g agar (slowly), 100mg of potassium phosphate dibasic and 100mg calcium carbonate. While still hot, pour the liquid into the culture dishes.

Starting the culture

Having obtained psilocybin mushrooms, any part of the fungus can be used for inoculation. To develop a culture from the spores: The spores should be scraped from the spore print into 10ml of sterilised water (in a jam jar). This should be well shaken, a further 90ml of sterile water added, and shaken further. A number of petri dishes or other containers containing sterilised agar media should be at hand. The lid should be lifter slightly from each container and with a sterilised pipette/syringe drops of spore water should be placed on four different parts of the agar surface, covering the container immediately afterwards. Allow the dish to stand at room temperature for 3-5 days and radial growths of monokaryotic mycelium will occur at each point. When any two mycelium have grown such that they make contact with each other, mating has occurred and within a few days these mycelia will have become dikaryotic organisms. Any portion of this tissue can now be used to seed other cultures.

Raising crop cultures

The most vigorous appearing mycelia should be selected. These can be identified as the largest and fastest growing specimens that are not contaminated. Mycelia are pure white fibrous mats that sometimes have a light bluish tinge. Any deviation from this appearance signifies the presence of contaminants. The selected mycelia should be transferred from the agar based stock cultures to the liquid broth cultivation jars. The jars should have been prepared and sterilised three days before transferring and allowed to stand for several days to make sure that they are sterilised properly. The uncontaminated jars can now be inoculated. The room should be re-cleaned at this stage. The lid of the stock dish is lifted just enough to pick up a fragment of mycelium with an inoculation loop that has been flame sterilised. Place the mycelium in the broth and immediately cover the jar. Repeat this such that all the jars have been inoculated and refrigerate unused stock cultures. The jar covers should be tightened and the jars should be shaken well to disperse the inoculum through the broth. This process also aerates the medium since the mycelium requires oxygen for life support and growth. The lids should then be loosened slightly and placed on a 'growing' shelf for 10-12 days at room temperature. The maximum growth and highest percentage of psilocybin occurs about four days after all the broth's sugar content has been used up, and the mycelium should be harvested at this time.

Harvesting and drying

The medium of each jar should be filtered through a clean flannel shirt and the mycelial material should be collected from the shirt and placed in a pyrex baking dish. The same should be done for each jar until each baking dish is about one third full with mycelia. These are then heated in an oven at no more than 200 degrees fahrenheit. The baking dishes should be checked periodically and when the material first appears to be dry the oven should be turned off and allowed to cool. Each jar should give roughly 5-10g of dry material.

Extraction

The dried mycelial material should be crumbled and ground. 0.1g of material should be combined woth 10ml of methanol in a flask and placed in a hot water bath for four hours. The liquids are filtered under suction, with the filtrate being saved. Extract the solid 3 more times in methanol.

Rotary evaporate off the liquids. Each 100g should contain at least 0.5g psilocybin/psilocin, or about fifty 10mg doses. Psilocin is very susceptible to oxidation and the extracted material should be kept in an air-tight container in a refrigerator. (a sack of silica-gel can be placed in the container to ensure the material remains dry.

Dosage

The standard dose for an average male is 6-20mg, with the average dose being taken as 10mg. A brown crystalline powder of roughly 25% purity is produced via the above method, with each jam jar containing about 25-30mg of psilocybin/psilocin which is 2-3 hits.

Medicine

Psilocybin has been studied in the treatment of a number of disorders. In 1961, Timothy Leary and Richard Alpert ran the Harvard Psilocybin Project and carried out a number of experiments regarding the use of psilocybin in the treatment of several personality disorders.

In the US, an FDA-approved study suppored by Multidisciplinary Association for Psychedelic Studies (MAPS) began in 2001 to study the effects of psilocybin on patients with obsessive compulsive disorder. In 2006, the MAPS study found psilocybin effective in relieving symptoms of obsessive compulsive disorder symptoms, in some cases for more than a few days.

Toxicity

The toxicity of psilocybin is relatively low. In rats the oral LD₅₀ (median lethal dose) is 280mg/kg which is almost 1.5 times that of caffeine. Death from psilocybin alone is unheard of at recreational/medicinal levels.

Effects

Psilocybin is absorbed through the linings of the mouth and stomach. The effects begin 10-40mins after consumption if held in the mouth for 20-60 minutes or if swallowed on an empty stomach and last for 2-6 hours depending on dose, species consumed and individual metabolism. Typical recreational dosage is from 10-50mg psilocybin.

The effects of psilocybin are typically pleasant and often ecstatic including a deep sense of connection to others along with confusion and hilarity. Bad trips can occur when psychedelic compounds are taken in a negative environment, in unexpectedly high doses or when difficuly areas of the psyche are triggered.

At low doses, hallucinations may occur such as walls that appear to breathe, a vivid enhancement of colours and the animation of organic objects. At high doses the experience is less social and intense spiritual experiences are often catalysed.

A small number of people are unusually senstitive to the effects of psilocybin where doses of as little as 0.25g of dried Psilocybe cubensis mushrooms can result in the effects associated with medium and high doses and vice versa. Both mental and physical tolerance to psilocybin builds and dissipates quickly. Taking psilocybin more than three to four times a week can result in diminished effects. Since tolerance dissipates after a few days, frequent users often keep doses spaced five days to a week apart.

Adverse Effects

People that have relatives with schizophrenia should be particularly careful when consuming any hallucinogenic drug due to the risk of triggering psychosis. In very rare cases the use of hallucinogens can initiate a malady known as Hallucinogen persisting perception disorder. (HPPD).