Incubation of psilocybin or psilocin with a crude enzyme extract resulted in the formation of a.blue color usually after 5-15 minutes incubation and gave a linear rated over 180-minute-period (Figure 10). The addition of NaCN was effective in delaying the blue color formation, but did not influence the phosphatase activity'.
The dephosphorylation of PDP by a crude enzyme extract proceeded at a rapid rate as shown the liberation of phenolphthalein (30-120 seconds for enzyme extract A and 5-25 minutes for enzyme extract B). The dephosphorylation of psilocybin was rapid for both enzyme extracts. The blueing phenomena occurred in 3-5 minutes incubation in enzyme extract A and after 15 minutes in enzyme B. The liberation of psilocin reached a maximum just before the reaction mixture turned blue (Figure 11). When the psilocin was used as substrate, its concentration remained
unchanged before it turned blue.
The use of citric acid could possibly work as a preservative, and could be easily tested(if it does not turn blue).
What are your thoughts on adding antioxidants into an already blued solution, which makes the blue go away?
It is not quite as simple as that, the anti-oxidants are useful before the reaction as they can prevent the it from happening in the first place. Once it has been done, it is not as simple to put the toothpaste back into the tube so to speak. There has been a little research into the blue product, and they have not had much success finding out exactly what is was. There are theories to what it is, but at the end of the they are just theories......
Your last question I am sure you will be glad you asked;
In 1985 in the Journal of Forensic Science some researchers realized that because of the phosphoryloxy group that psilocybin has it makes it very poorly soluble in non-polar solvents (like ether, xylene ect). So they converted their samples into psilocin and used a classic A/B extraction and got great yields of very clean psilocin.
So to answer your first question;
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.
Basically putting dry sample into vinegar making sure pH is 4, standing for 1 hour, place in a boiling water bath(saucepan with boiling water in it) until it gets to 70*C. Cool and filter. Then.....
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.
I have a paper that uses (I think, I have to double check this) bicarb soda to adjust the pH to 8 and said it was much gentler on the psilocin molecule which is vulnerable at higher pH.
Hope this helps....