After these tests, here is the next strategy I'm thinking of pursuing.

First a bulk test to confirm that we can use the Na+ cation resin to catch and relese psilocin as follows:

1) Loosely follow Casale's enzymatic psilocin extraction into dilute vinegar. Will likely use fresh mushrooms though (should not matter since this is an aqueous extract).

-Room temp dilute acedic acid extract pH~5.5 (psilocin and psilocybin)

-Heat cycle up to 70C to convert psilocybin to psilocin.

-Pull acidic water with Na+ cation resin

-Rinse resin with neutral water

-Pull resin with bicarbonate water (or alcohol) and neutralize with an organic acid

This should give a potent extract that is relatively clean. A lot of poteins would still be present though. At this point a bioassay would be done to confirm things are as expected.

To create a cleaner extreact:

2) To reduce protein content, add a protein removal step

- Cold acidic water extract (pH ~ 5.5)

- Cation resin protein removal. This will remove any psilocin present at this time also, but it should be a low amount. We are hoping that the dephosphorilating enzymes remain though.

- Heat cycle up to 70 C. If we did not lose the enzymes, psilocybin will convert to psilocin.

- Pull extract with cation resin. Only the new psilocin should be pulled.

- Rinse resin with neutral water

- Pull resin with a slightly basic solution and try to isolate and stabilize psilocin which should be pure. A vacuum evaporation may be needed at this point.

The choice of pH 5.5 is higher than Casale's (which was pH4). Hope is that it still works roughly the same. This pH is chosen as a guess since most proteins and enzymes could be near their isoelectric point (see post #5) and not interact with the resin. This improves the chances that the dephosphorilating enzymes are left in solution with strategy 2).

For reference, attached are a couple papers on the dilute acetic extraction and conversion to psilocin.