didn't realize I started this thread.

looking around on e.bay, I found this

I have one, it is an Omni International GLH (general laboratory homogenizer) 750W rotor-stator homogenizer, which works wonders for breaking down dried materials and cells. Very useful for what we're trying to do here. Rotor-stators and ultrasonic homogenizers (even the baths), are great for getting the alkaloids into methanol or your solvent of choice; quicker than a soxhlet, but you could even do that afterwards.

I honestly think some column work is essential for this project. I know there exists a solvent-partitioning method, pioneered by Hoffman, but it seems like more of a hassle. Column work with a ternary (or quat) mobile-phase readily separates out psilocybin, and psilocin, using acetic acid. I'll look into simplifying it, as I've already shown the separation occurs using HPLC.

benz - how does that differ from a sonicator?

It’s essentially a high-speed blender, up to 35,000 rpm. the probe has a sawtooth stator, and a rotating shaft with sawteeth as well. the bubbles it creates also cause cavitation, because of the high speed rotation.

I just realized my post a couple posts ago in this thread was a little difficult to read through, as I simply laid out my various experimental results without compiling into a set of instructions. Digging through the notebook now....



20g of dried mushroom is crushed but not powdered, then added to 250ml distilled water. Glacial acetic acid is added until pH 4 (only takes a few drops). Mixture is placed in a beaker on a lab stand and the beaker is suspended in boiling water until the internal temperature reaches 75C for 2-5 minutes. While heating, a funnel is plugged with a cotton ball and loaded with several ice cubes. Upon removing the solution from heat, it is poured first through a coffee filter to facilitate straining, then into the iced funnel. This water-acid extraction process is repeated for improved yield.

As soon as it's fitered, it should be basified slowly with addition of a saturated sodium bicarbonate solution. Once effervescence ceases, add a bit more bicarbonate solution, followed by a splash of concentrated sodium hydroxide solution, keeping pH between 9.5 and 10. Addition of hydroxide solution causes a noticeable darkening of color, though nothing dramatic. Immediately add 50-75mL chloroform, swirl, separate, and repeat for a total of 100-150mL chloroform.

Place chloroform in freezer overnight to precipitate dissolved water (there is quite a bit!), quickly filter through a cooled cotton filter in a cooled funnel to yield a very transparent light yellow solution. Extract alkaloids into aqueous solution via titration using tartaric or citric acid. Tartaric is the one I've had the best results from. Back-extract the aqueous solution with naphtha or similar to remove dissolved chloroform, which becomes a hazard when open-evaporating the aqueous solution.

Evaporate the aqueous solution using a plate, a fan, and a hot plate, but keep the temperature low (around 50C). This yields a light-colored solid compound, for which you'll need to use your imagination to determine a use.

I tend to dry aggressively, then store in an air-tight container with a desiccant (I use Na2SO4). When I want to consume the alkaloid extract, I make a volumetric solution using 1 drop = 1mg ratio, and 50:50 ethanol:water as the solvent. I typically use this solution for abortion of migraine attacks, though it is also very effective as a psychedelic in dosages from 15-30mg.

Great post lysurgeon, do you have any pictures on what you did

Thanks for all the details lysurgeon. Did you use P. Cubensis? Since we think psilocybin is converted to psilocin during the warm acid extraction for P. Cubensis, this should give psilocin tartarate with excess tartaric acid, right? Did you ever send it out for analysis? How is the shelf-life? Any info on yields?

Great contribution Lysurgeon.

How do the effects / side effects compare with whole mushrooms?
Apart from the obvious satisfaction of scientific pursuit and experimentation involved in producing it, does the extract have any benefits over existing methods of consumption?

Update: the emulsion is eliminated by drying the mushroom/Ca(OH)2 paste in the oven dry at low temp (170F). Not sure why it works, one possibility is that proteins are denatured.

Of the many tests done, this is surfacing as most promising:

- Grind fresh mushrooms in a splash of water and excess Ca(OH)2.
- Spread the resulting paste in a shallow Pyrex pan and dry in the oven at ~170F. Note we are using a part of the bufo extraction.
- Pulverize the dry paste and add white vinegar with 30% ammonium sulphate amiming for pH~4 (I got 4.8, near the isoelectric point of psilocybin). Note that we are using a part of the DMT max ion extraction.
- Psylocybin (hopefully) is now at its Isoelectric point. Pull with Xylene to get it out of the ammonium sulphate Ionic water Extract (PIXIE). Xylene as a solvent for Psylocybin was proposed years ago on the Nexus.
- Use FASA on the Xylene to get (hopefully) Psilocybin Fumarate (loose white powder one can call PIXIE dust). Once again, we are using an idea that is already on the Nexus, this time from the DMT FASA approach.

Stayed tuned for moar updates to see how far we get with this PIXIE process. If it works credit to the Nexus as a whole, each step comes from ideas and established procedures used elsewhere.

Below are some pictures of the process. I got 240mg of PIXIE dust from about 25g of fresh mushrooms. Yield is too high to be psilocybin alone, so the fluffy power is maybe 10% psilocybin at best. Will bioassay and send for analysis (powder sent earlier was pulled at pH of 9, not as near to the Isoelectric point as this powder at pH 4.8 ). Also added a slide from the zwitterions thread where we asked for some help.

Pictures in order are:

-Fresh mushroom / Ca(OH)2 / splash of water paste
-Dry flaky material after drying the paste in the oven at 170F
-Paste rehydrated and brought to pH of 4.8 using vinegar with 30 % ammonium sulfate (for increased Ionic strenght at the Isoelectric point, the two I's in PIXIE).
-Xylene pull clouding up after adding some FASA drops (does not cloud up if pulling without dropping the pH)
-Resulting PIXIE dust from FASA reaction. It is truly "dusty": loose and fluffy and can be blown away easily. Smeels like Fairy feet.
-Plot showing the partition coefficient (logD) between water and octanol at different ionic strength (fixed pH 4 at the Isoelectric point) and another plot showing lodD of psilocybin vs. pH (fixed 0.1M ionic strength). These are cross-sections of a 3D plot, with their common point highlighed.

excellent work.
really great stuff.

I'm having issues (I usually do, but it's particularly difficult now) establishing communication. I suspect it's the network card on the computer, it's an old 3Com ethernet card with a BNC connector, on WinXP.

if for some reason I can't establish comm, I'm still going to do FTIR and HPLC on the sample.

*edit* nevermind, I got it.

That is some stellar work Loveall. I am very very impressed regardless of the outcome of assay. This is the spirit of scientific curiosity at its finest. Take a bow sir.

I look forward to this thread everyday

thanks Endlessness. this is the data I got. Loveall, thank you for your contribution, and elaboration on this challenging project. You have indeed advanced a difficult extraction, and it is most certainly appreciated. I'll be following and contributing to the endeavor, as it is quite relevant and important to the mental health of society as a whole. Thank you, sir.

2nd that! Great to see the recent progress here.

meant to post this, Loveall. the HPLC chromatogram using the 10:90:1 v/v MeOH:H2O:AcOH mobile phase, 286 nm.
I need to adjust the noise threshold, but there are no other peaks. what this means is only the fluorophore for the analyte was present. The mass chromatogram would show impurities regardless of wavelength.

Many thanks benzyme. So we think this peak must be psilocin, correct? It is a couple au high based on the post #73, is that an absolute calibration thing? One question that came up in chat last night (from Mindlusion) is where is the fumaric/fumarate? Another question is can you estimate the % of psilocin in the powder?

I've left the powder (calling it PIXIE dust) uncovered and in air for a few weeks and no changes in color or appearence have occured so far. I redisolved it in water to remove any excess fumaric acid that came out during the Xylene residue dry. After drying the water the resulting powder is very sparkly, it really looks like what one would call PIXIE dust

The other sample with the Xylene pull at pH 4.8 (near Psylocybin's isoelectric point of 4.1) should get to you soon. Based on logD (partition coefficient - image below for low ionic strength of 0.1M) Psylocybin should be more difficult to pull into a solvent in general compared to psilocin, (and I'm not sure about the solubility of psilocybin in Xylene compared to psilocin). However, since we saw (nor) baeocystin in an earlier test at high ionic strength, we may get it if it survived the extraction process. Based on mass/yield I don't think we can get more than 10% psilocybin in this last sample (if there is any).

it is off by 2-4 m.u., and that is a great question..where is the fumarate? psilo fumarate would appear as a peak @ 319-322.
I honestly forgot to look for fumarate. It readily dissolved in the mobile phase (10:90:1, MeOH:H2O:AcOH)
FTIR didn't provide any useful info..toluene and some polymer. I'll look at it again.
I'll also whip up some Ehrlich's to test

Update:

Our hero benzyme has been testing the pH4.8 pulls. To review:

-Fresh mushrooms (100g) where turned into a paste with a little water and ~excess Ca(OH)2
-Paste was dried in the oven at low heat (170C) to denature proteins/enzymes
-30% Ammonium sulfate water was added to the paste to rehydrate it (anmonoa smell results).
-Vinegar with 30% Ammonium sulfate added until pH got to 4.8 (near Psilocybin's isoelectric point of 4.1).
-Xylene pulls done, followed by FASA. A lot of white precipitate collected, roughly 1.5g (dry mushrooms should weigh 10g and have ~100mg of actives.

First conclusion is that a lot of other stuff got pulled by the Xylene while at pH4.8 and precipitated during FASA. In earlier tests when pulling at pH 12, nothing crashes out. This maybe means that we are pulling something with an -OH group such as proteins.

After a methanol extraction on the PIXIE dust by benzyme, psylocybin and psilocin peaks appear. The psilocin peak is somewhat surprising since we were not near it's isoelectric point (however the solution was at high ionic strength).

To make this extraction practical and more pure we need a way to separate out this extra material that crashes out during FASA. I'll do some testing at 60% ammonium sulfate to see if something crashes out of the water so it can be separated before the Xylene pull (of course it could be the actives themselves that could crash out first, but the heavier proteins may have a better chance of crashing out sooner and ammonium sulfate salting is a standard way to precipitate proteins). Another test could be to extract the pixie dust with denatured alcohol and see what we get.

Work continues, ideas and feedback welcome.

Have you tasted the pixie dust yet? I'm very curious to see how this goes.