As I went through this forum, I couldn't find yet anyone mentioning this patent (attached to this post), so I will shortly mention it and it's teks. If it has been already mentioned, please add the link and stop me! LOL


There are several teks in this patent, the simplest would certainly be the one using HCl.

First the water extraction:

1) Dried, ground to powder
2) 240g powder in 60 ounces distilled water at 45F (7C) for 24h (fridge)
3) Filtered

In the patent the water extract had a muscimol to ibotenic acid ratio from 0.29.

Conversion tek with HCl:

1) Prepare HCl in water 1:7 (someone might chime in how much that is)
2) Add to water extract until PH 2.6 is reached
3) Maintain at 195-212F (90-100C) for 3h

Result in the patent is a change of muscimol to ibotenic acid ratio from 0.29 to 53.89.


Conversion tek with (glutamate decarboxylase, and pyridoxal phosphate P-5-P)

Special items needed:

* Pyridoxal phosphate (P-5-P) can be bought easily as supplement online.
* I have no idea where one could get Glutamate decarboxylase, so probably this tek isn't an easy one in this respect

1) 14mg of glutamate decarboxylase per 2ml of water extract
2) then add 0.3 mg of pyridoxal phosphate (P-5-P) per 2ml water extract
3) Maintained at 37C (99F) for 4h
4) refrigerated

Result in the patent is a change of muscimol to ibotenic acid ratio from 0.29 to 92.77.


The third tek is IMHO especially interesting, as it uses lactobacillus cultures to create the needed glutamate decarboxylase, and otherwise is basically the same conversion as above.

I find this very interesting as the old northern preparation was putting it in beer. So maybe the original key point would be, to put it into the beer still in the fermenting process to get a similar result.

But back to the original "fermenting tek" in the patent:

Conversion tek with (lactobacillus cultures, and pyridoxal phosphate P-5-P)

Special items needed:

* lactobacillus showing glutamate decarboxylase (e.g. L. plantarum is widely available)
* Pyridoxal phosphate (P-5-P) can be bought easily as supplement online.
* Calcium carbonate
* Sugar

1) 10,000,000 CFU of Lactobacillus (showing glutamate decarboxylase), 0.4
gram of powdered calcium carbonate, 10 mg of pyridoxal-5-phosphate, and 4.5 teaspoons of table Sugar were added to the 60 ounces (1.8l) of water extract (from 60g dried Amanita).

2) This liquid was placed in a hermetic sealed container, stirred, and the contents agitated until thoroughly mixed. The initial pH of the combined solution was approximately 6.0.

3) Frozen until thoroughly solid.

4) Placed in an incubator at 103F (40C)

5) After 12h an additional 10,000,000 CFU of Lactobacillus is added

6) Wait 2.5 days
After 18 hours of fermentation, the pH dropped to approximately 3.8-4.0, and remained stable for the duration of fermentation.

It has been mentioned (probably a number of times) but I don't think there is a dedicated thread... until now?

I've attached a couple papers to supplement discussion of the acidic conversion technique. The paper by Tsunoda et al. (1993) is one of the patent's references and discusses the acid conversion tek, some of the more destructive and inefficient traditional teks and extraction through soaking. The paper by Neilsen et al. discusses the acid tek in greater detail (among other things).

The Tsunoda et al. (1993) paper is also available online, in case the PDF requires additional font packs on your computer: https://www.jstage.jst.go.jp/article/shokueishi1960/34/2/34_2_153/_pdf/-char/en

2 Strange things, why not just take the water extract and slowly add HCl solution until pH 2,6 is reached?

1M or 2M HCl should be fine to slowly get to this point.


Which makes this thing even stranger: 1:7 HCl to water - does this mean 1/7 weight in pure HCl-gas ? Probably not, but then as it should be HCl solution it's not written what molarity . So I don't see why not just taking the extract and simply adding some diluted HCl-solution until pH 2,6 is reached.


Ok and yes the 2nd TEK seems rather impossible, you would have to buy this enzyme and use it correctly, I guess that also would be overkill chemistry when you could just decarboxylate in boiling water with some HCl

And THX for sharing, indeed that's an easy and quick way to prepare Amanita. Wasn't there just someone asking?

@all:

Thanks a lot. As usually it seems I'm the last getting aware of anything...

HCL tek don't give a very high conversion rate.

How about this alternative to the HCl conversion tek?

1. Dry and grind the mushroom.
2. Put the powder in a pan, add vinegar and simmer for 3 hours.

Benefits:
1. A potentially more efficient extraction than the cold water soak.
2. Conversion using a safer and more kitchen-friendly acid (especially since simmering is involved).
3. Both of the above steps combined into one.

Or am I missing something?

Currently give this a go.

Have dried 100g, let it sit in cold distilled water for 3 days, filtered. Added p5p, caclicum carbonate, sugar and the bacteria as per tek. Currently freeze thawing. Will start fermentation soon. The bacteria strain as mentioned in the tek should ferment and produce GAD enzyme for conversion. Current pH 6 should see it drop while the bacteria does its Job over a day or 2.

Thinking about evaporating, using methanol to clean it up and leave the calcium carbonate behind, decant evaporate and the treat with hcl at 90c for 3 hours just to ensure close to full conversion then evaporating again for a crude (and hopefully fully convert) extract.

Will post results in a few days. Will be interesting to see the weight of the extract. 200mg of muscimol should be there (with decent conversion), 400mg of calcium carbonate (which I'll separate) and who knows what other carbs and proteins left behind.
Thinking of running the filtrate through activated charcoal too, as per patent to remove any unwanted non polar impurities still left.

Dried extract should be smokable.

Excited to finally be trying this. I originally floated an idea for a crude extract when I first joined the forum.

I successfully fermented the solution as ph dropped as per patent, followed by a carbon filtering (as noted as a option in the patent) which like the previous filtering took forever. (Should have taken a sample but it was late I was over it and I forgot).
Unfortunately some carbon found its way through the lab filter and couldn't be effectively removed.
Liquid was evaporated and 50g of sludge was left (presumably with activated carbon contamination).

Decided to throw the whole kitchen sink at this experiment and tried a HCL soak as well (as mentioned also in the patent). To ensure no ibotenic acid was left. I then evaporated. Then I really screwed up, got paranoid due to the sludge that HCL might have got trapped in it so I used ammonia to neutralize it and again evaporated this back to the sludge.

Split the sample in to 2:
One sample was pulled with methanol and evaporated (a small sample was taken for testing). A lot of the sludge was dissolved by the methanol as expected.


The other half was pulled with EA. EA left the sludge behind and when evaporated rendered 250mg of oil with some tiny crystal like formations in the oil.

It would be interesting to find out if the EA sample contains muscimol. Bioassay may follow.
Might be worth getting tested?