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joze



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Cool to see your progress. You should try this reaction at many different pH values.
I would recommend getting some pH indicator strips and testing the pH of your mushroom tea aftering boiling, before you add the reagent. Different amounts of mushroom may change the pH slightly. Buffer your mushroom tea to a set of different pH (perhaps pH 3, 5, 7, 9) and then carry out the reaction and see if different pH values lead to significantly different color changes. You may find that this reaction is much more sensitive if you buffer the reaction precisely.
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27755191 - 04/28/22 02:18 PM (1 year, 8 months ago) |
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I mean really shouldn't the reagent react weather or not theirs water? Wouldn't it penetrate the material nonetheless? And I figure the pigment wouldn't effect the reagent, I guess because it would be non polar I would assume? So adding the water to the solution would inherently mix the two, but I could be very wrong to assume the reagents are non polar. Does this make since?
Aqueous reaction... Okay I looked it up, I guess that make since. So I guess that would suggest the reagents are non polar and contain no water?
I'm just trying to understand this.
And of it just costs a bottle of whiskey why shouldn't everyone do it?
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27755242 - 04/28/22 02:57 PM (1 year, 8 months ago) |
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Quote:
joze said: Cool to see your progress. You should try this reaction at many different pH values.
I would recommend getting some pH indicator strips and testing the pH of your mushroom tea aftering boiling, before you add the reagent. Different amounts of mushroom may change the pH slightly. Buffer your mushroom tea to a set of different pH (perhaps pH 3, 5, 7, 9) and then carry out the reaction and see if different pH values lead to significantly different color changes. You may find that this reaction is much more sensitive if you buffer the reaction precisely.
Funny that you mention that because this was exactly my line of thinking this morning after observing two more trials with different ratios of extract to reagent. I’d expected a stronger reaction after increasing the amount of extract to 2ml but, when the opposite happened, I got to thinking and I’m pretty sure the result was due to me unwittingly raising the pH too much with the increased extract volume. I got out my pH test paper and sure enough the mushroom extract was more basic than I’d assumed. Once I get back home tonight, my plan is to use drops of phosphoric acid to adjust the pH of the extract so that it is slightly acidic before adding it to the reagent bottle. Does that seem sensible or would you suggest a different buffer?
I may also experiment with using methanol instead of water for the extract but one thing at a time…
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27755260 - 04/28/22 03:18 PM (1 year, 8 months ago) |
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Quote:
QM33 said: I mean really shouldn't the reagent react weather or not theirs water? Wouldn't it penetrate the material nonetheless?
No, not really. It doesn’t necessarily have to be water — methanol, for instance, would also work — but both molecules, DMAB and psilocybin, need to be in the same solution for the reaction to happen.
Quote:
QM33 said: And I figure the pigment wouldn't effect the reagent, I guess because it would be non polar I would assume? So adding the water to the solution would inherently mix the two, but I could be very wrong to assume the reagents are non polar. Does this make since?
I’m not entirely sure what you’re getting at here but maybe it will help you understand to tell you that there are no non-polar solvents involved in this experiment. Psilocybin is polar-soluble and both methanol and water are polar solvents. The other part of the 85% phosphoric acid is also water, so there are only two solvents involved, water and methanol, and both are polar.
Quote:
QM33 said: And of it just costs a bottle of whiskey why shouldn't everyone do it?
I think everyone should do it But first someone’s gotta figure out how to mix it all up in a way that yields consistent and useful results, which is what I’m up to
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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QM33
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Okay my bad I kinda drew a blank on the methonol, I got focused on the phosphoric acid solution I guess. Which yes contains some water, and I guess what I was getting at as since it has some water in it, to go along with the aqueous solution thing, then you may not need to 'add more' for the reaction to occured. And since I was ting thinking about the methonol, I was wondering if the solution I suppose would pull the pigment if it were non polar. But ya methonol duh.
Totally, idk I thought you meant someone could use elrich for what your trying to put together. I see.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27755303 - 04/28/22 03:55 PM (1 year, 8 months ago) |
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Quote:
QM33 said: Okay my bad I kinda drew a blank on the methonol, I got focused on the phosphoric acid solution I guess. Which yes contains some water, and I guess what I was getting at as since it has some water in it, to go along with the aqueous solution thing, then you may not need to 'add more' for the reaction to occured. And since I was ting thinking about the methonol, I was wondering if the solution I suppose would pull the pigment if it were non polar. But ya methonol duh.
Totally, idk I thought you meant someone could use elrich for what your trying to put together. I see.
I am using Ehrlich’s reagent. DMAB = Ehrlich’s reagent. It can be prepared in a number of different ways with various acids and solvents
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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QM33
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Okayyyy. I thought elrich was dmab+85%phosacid for some reason. Idk if I read the begging of this thurough enough. But with the way things are looking I'll definetly go back and clearify some of this! Thank you!
-------------------- OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum... There are known knowns, there are known unknowns, there are also unknown unknowns. With great privilege comes great responsibility.
  Quantom Qups PROOF AND Soft Drops Turn your Swab to a Syringe and Syringe to Multiple Syringes! No Pours (QuantomStyal)Magic Fruit Leather DMT for IandI
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joze



Registered: 11/10/20
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Quote:
SpectreOfCommunism said:
Quote:
joze said: Cool to see your progress. You should try this reaction at many different pH values.
I would recommend getting some pH indicator strips and testing the pH of your mushroom tea aftering boiling, before you add the reagent. Different amounts of mushroom may change the pH slightly. Buffer your mushroom tea to a set of different pH (perhaps pH 3, 5, 7, 9) and then carry out the reaction and see if different pH values lead to significantly different color changes. You may find that this reaction is much more sensitive if you buffer the reaction precisely.
Funny that you mention that because this was exactly my line of thinking this morning after observing two more trials with different ratios of extract to reagent. I’d expected a stronger reaction after increasing the amount of extract to 2ml but, when the opposite happened, I got to thinking and I’m pretty sure the result was due to me unwittingly raising the pH too much with the increased extract volume. I got out my pH test paper and sure enough the mushroom extract was more basic than I’d assumed. Once I get back home tonight, my plan is to use drops of phosphoric acid to adjust the pH of the extract so that it is slightly acidic before adding it to the reagent bottle. Does that seem sensible or would you suggest a different buffer?
I may also experiment with using methanol instead of water for the extract but one thing at a time…
Yes a dilute phosphoric acid solution should work great. Adjust dropwise
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27755543 - 04/28/22 07:15 PM (1 year, 8 months ago) |
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Anyone trying this testing who doesn't have at least high school chemistry experience: read up on handling of strong acids, they can fuck you up, ruin your eyes, your lungs, your skin.
Handle with care, cover eyes, wear gloves, considering wearing a respirator and or doing the test outside or where you have good ventilation. Do not breathe the fumes. Don't do in presence of pets or kids.
Have a good base ready to neutralize in case of exposure. Solution of baking soda would work. Don't dump the used tests down the drain, especially if you have a septic system. Careful what you mix with acids, some common household substances can produce toxic fumes with common strong acids.
Also, methanol is very toxic, unlike the ISO we use all the time in shroom tek.
I seem to remember reading somewhere about an indole test that didn't involve acids. Was it Hoffman's Agent maybe?
One thing that might be fun would be to buy one of those 15 dollar Miraculix "PSILO-QTests", send it to a lab and find out what's in it. I don't see a patent statement on it too, so shouldn't be an issue.
This is interesting, a "narco check" cheap spot test for cops to test for shrooms, says doesn't contain acids, and returns pink for psilocybin and blue for psilocin. Weird. Seems fresh shrooms would return a mixture, maybe the brown in the PSILO-QTest? https://www.pharmadrugtest.com/identification-tests-for-solid-substances/80-id-test-identification-psilocin-psilocybin-hallucinogenic-fungi.html
This is an Idaho government link that talks about testing for shrooms, I wonder if this could be adapted for quantitative use. If so, thank you Idaho drug cops. I couldn't copy from this but read number 4, "color spot test" using "fast blue bb" (which is toxic):
https://isp.idaho.gov/forensics/wp-content/uploads/sites/10/documents/archivedAMs/before/Controlled%20Substances/08%20psilocyn%20sop%20rev%205.pdf
I'd love to adopt a test for growers that borrows from cop tek used by a state where even spores are illegal. haha.
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Edited by nektar61 (04/28/22 11:32 PM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 3
#27756013 - 04/29/22 02:08 AM (1 year, 8 months ago) |
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It’s late, so this won’t be a comprehensive update but I wanted to post a picture of the latest run and say that buffering / acidifying the sample solution does seem to be the correct thing to do because I am getting a much stronger color-reaction by adding a few drops of dilute phosphoric acid to the extraction solution:

And for comparison, here is the exact same procedure but with no acid added to the extract:

Tomorrow I will experiment with extracts of different pH values to find the parameters that yield the most robust reaction
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Alan Rockefeller
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27756048 - 04/29/22 02:56 AM (1 year, 8 months ago) |
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Quote:
nektar61 said: One thing that might be fun would be to buy one of those 15 dollar Miraculix "PSILO-QTests", send it to a lab and find out what's in it.
Unfortunately labs like that don't exist, and if they did, they would need to charge many thousands of dollars to reverse engineer the contents. Analytical chemistry isn't as simple as running a sample through a LCMS and having the computer spit out the names of the chemicals in the mixture.
Quote:
This is interesting, a "narco check" cheap spot test for cops to test for shrooms, says doesn't contain acids, and returns pink for psilocybin and blue for psilocin. Weird. Seems fresh shrooms would return a mixture, maybe the brown in the PSILO-QTest?
That test is Ehrlich's reagent. Miraculix tests don't use Ehrlich's - there's definitely a good reason why, but I haven't asked Felix what it is. He surely started with Ehrlich's and found it unsuitable for mushroom potency testing.
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: Alan Rockefeller] 1
#27756117 - 04/29/22 04:46 AM (1 year, 8 months ago) |
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What does miraculux use?
-------------------- OmManiPadmeHum,OmManiPadmeHum, OmManiPadMeHum... There are known knowns, there are known unknowns, there are also unknown unknowns. With great privilege comes great responsibility.
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joze



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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27756509 - 04/29/22 12:24 PM (1 year, 8 months ago) |
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Quote:
QM33 said: What does miraculux use?
It's proprietary, nobody really knows.
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joze



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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 2
#27756714 - 04/29/22 03:18 PM (1 year, 8 months ago) |
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Hey SpectreOfCommunism, can you do me a favor as you develop this reagent more 
I'm curious how many other indole derivatives are present in mushrooms that could interfere with this test's accuracy. Alongside a known active mushroom, can you dehydrate and test a known inactive mushroom? Maybe even an oyster or a portabello from the store? It would act as a good control.
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Re: DIY Psilocybin Content Percentage Detection [Re: Alan Rockefeller] 1
#27757022 - 04/29/22 08:47 PM (1 year, 8 months ago) |
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Quote:
Alan Rockefeller said: Miraculix tests don't use Ehrlich's - there's definitely a good reason why, but I haven't asked Felix what it is. He surely started with Ehrlich's and found it unsuitable for mushroom potency testing.
Another test we might consider trying to find the open source test is Hofmann's Reagent. Tests for a lot of Indole substances, turns blue for psilocybin and psilocin. https://www.smplest.eu/products/hofmann-test
Alan, have you used it? Do you know if it turns more blue with more of the substance present?
Interesting substance, turns different colors with different indoles. Yellow for DMT, for instance.
Can test 1/10 of a blotter of acid, so seems pretty sensitive.
Here is another supplier: https://testkitplus.com/product/hofmann-reagent
More info here: https://dosetest.com/product/hofmann-reagent-testkit/
"Named for Albert Hofmann, the creator of LSD, Hofmann reagent is a slight tweak on Ehrlich reagent which offers more specificity when testing for LSD or DMT.
Ehrlich reagent is something of a “binary” reagent, usually only responding with a pink/purple colour to a variety of substances (although there are always exceptions, please refer to results.dosetest.com for more information).
Hofmann reagent, meanwhile, clearly delineates between LSD, DMT and other substances which would give a positive Ehrlich result, as well as ruling out the possibility of 5-HTP being added to a sample to “fake” a reaction, as can be done with Ehrlich reagent."
"Hofmann contains sulfuric acid under 15% and in sufficient amount is strong enough to burn skin and clothing."
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27757120 - 04/29/22 11:31 PM (1 year, 8 months ago) |
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Quote:
nektar61 said:
Quote:
Alan Rockefeller said: Miraculix tests don't use Ehrlich's - there's definitely a good reason why, but I haven't asked Felix what it is. He surely started with Ehrlich's and found it unsuitable for mushroom potency testing.
Another test we might consider trying to find the open source test is Hofmann's Reagent. Tests for a lot of Indole substances, turns blue for psilocybin and psilocin. https://www.smplest.eu/products/hofmann-test
Alan, have you used it? Do you know if it turns more blue with more of the substance present?
Interesting substance, turns different colors with different indoles. Yellow for DMT, for instance.
Can test 1/10 of a blotter of acid, so seems pretty sensitive.
Here is another supplier: https://testkitplus.com/product/hofmann-reagent
More info here: https://dosetest.com/product/hofmann-reagent-testkit/
"Named for Albert Hofmann, the creator of LSD, Hofmann reagent is a slight tweak on Ehrlich reagent which offers more specificity when testing for LSD or DMT.
Ehrlich reagent is something of a “binary” reagent, usually only responding with a pink/purple colour to a variety of substances (although there are always exceptions, please refer to results.dosetest.com for more information).
Hofmann reagent, meanwhile, clearly delineates between LSD, DMT and other substances which would give a positive Ehrlich result, as well as ruling out the possibility of 5-HTP being added to a sample to “fake” a reaction, as can be done with Ehrlich reagent."
"Hofmann contains sulfuric acid under 15% and in sufficient amount is strong enough to burn skin and clothing."
Hofmann reagent (also known as Van Urk reagent) is made with DMAB, sulfuric acid, and ferric chloride. If my memory serves me correctly, ferric chloride is what is typically used for etching PCB circuit boards and so it should be available online — I haven’t used it in ages because I find cupric chloride to be a far better etchant for copper clad boards. Sulfuric acid can be bought from some hardware stores as “professional” grade drain opener.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nektar61
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Quote:
SpectreOfCommunism said: Hofmann reagent (also known as Van Urk reagent) is made with DMAB, sulfuric acid, and ferric chloride. If my memory serves me correctly, ferric chloride is what is typically used for etching PCB circuit boards and so it should be available online — I haven’t used it in ages because I find cupric chloride to be a far better etchant for copper clad boards. Sulfuric acid can be bought from some hardware stores as “professional” grade drain opener.
Your update looks good, looks like progress.
Keep in mind that ground shrooms degrade psilocin with time, especially if in solution. Might want to make a fresh batch of shroom material each day of testing. That will better simulate the way it will be used anyway.
My uneducated guess is that Miraculix PSILO-QTest is Hoffmann's with a substitution or an addition.
FWIW:
"There are two copper chlorides: copper (I) chloride https://en.wikipedia.org/wiki/Copper(I)_chloride CuCl
and copper (II) chloride. CuCl2 https://en.wikipedia.org/wiki/Copper(II)_chloride
"The former is white (if pure); the latter is already green, in it usual hydrated state, although anhydrous cupric chloride is a light brown. In no case would I describe any of the possible candidates as ‘colorless’ - in chemistry at least that term is used to describe clear solids, not white ones.
"Copper (II) chloride generally gives a greenish-blue solution in water, tending more towards blue with greater dilution.
"Solutions of copper (I) chloride are clear, although a concentrated solution in hydrochloric acid can be brown; what complexes are responsible for that I don’t remember."
from https://www.quora.com/Does-colourless-copper-chloride-change-to-green-and-brown-when-reacting-with-water?share=1
Ferric Chloride has two forms: https://en.wikipedia.org/wiki/Iron(II)_chloride FeCl2
and https://en.wikipedia.org/wiki/Iron(III)_chloride FeCl3
-------------------- -NEW? Start here.
Edited by nektar61 (04/30/22 12:09 AM)
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Alan Rockefeller
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27757137 - 04/29/22 11:57 PM (1 year, 8 months ago) |
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Quote:
nektar61 said: Alan, have you used it? Do you know if it turns more blue with more of the substance present?
No.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: Alan Rockefeller] 1
#27757150 - 04/30/22 12:19 AM (1 year, 8 months ago) |
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Alan,
I've thought a lot about the fact that shrooms turn blue when bruised or cut. (And some turn green, I've had B+ and Z-Strain variety cubes that turned green.) I'm thinking about this bruising in conjunction with these testing ideas.
I've commonly heard, and noticed on my own, that the amount/ speed of bluing isn't always directly proportionate to the strength of the shrooms. Sometimes is an indicator, but not as a 1:1 proof. If it did, we would't need a test, the test would be inbuilt.
Alan, is that because the bluing is do to psilocin, not psilocybin, and a shroom could be strong but not blue much if it's got a lot of psilocybin but not a lot of psilocin?
(fell down some wormholes, here's an explanation involving enzymes that doesn't entirely answer that question); https://doubleblindmag.com/blue-bruising-mushrooms/ --- all: Maybe an even better test than Miraculix PSILO-QTest could be made....with more variation or even different colors for different amounts of alkaloid.
Here's a thought: that Miraculix test turns darker shades of brown with higher psilocybin / psilocin. Brown, in pigments (as opposed to beams of light) doesn't really exist. Brown is just very dark yellow or orange. So maybe it's a test that turns yellow or orange in presence of shroom alkaloids, with an additional substance to amplify the effect by a magnitude or two. -- What about Keller's reagent?
"Psilocin is relatively unstable in solution due to its phenolic hydroxy (-OH) group. In the presence of oxygen it readily forms bluish and dark black degradation products. Similar products are also formed under acidic conditions in the presence of oxygen and Fe3+ ions (Keller's reagent)." https://en.wikipedia.org/wiki/Psilocin#Chemistry -- "In organic chemistry, Keller's reagent is a mixture of anhydrous (glacial) acetic acid, concentrated sulfuric acid, and small amounts of ferric chloride, used to detect alkaloids. Keller's reagent can also be used to detect other kinds of alkaloids via reactions in which it produces products with a wide range of colors." https://en.wikipedia.org/wiki/Keller%27s_reagent
-------------------- -NEW? Start here.
Edited by nektar61 (04/30/22 03:09 AM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27757798 - 04/30/22 03:51 PM (1 year, 8 months ago) |
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Quote:
nektar61 said: Alan,
I've thought a lot about the fact that shrooms turn blue when bruised or cut. (And some turn green, I've had B+ and Z-Strain variety cubes that turned green.) I'm thinking about this bruising in conjunction with these testing ideas.
I've commonly heard, and noticed on my own, that the amount/ speed of bluing isn't always directly proportionate to the strength of the shrooms. Sometimes is an indicator, but not as a 1:1 proof. If it did, we would't need a test, the test would be inbuilt.
Alan, is that because the bluing is do to psilocin, not psilocybin, and a shroom could be strong but not blue much if it's got a lot of psilocybin but not a lot of psilocin?
(fell down some wormholes, here's an explanation involving enzymes that doesn't entirely answer that question); https://doubleblindmag.com/blue-bruising-mushrooms/ --- all: Maybe an even better test than Miraculix PSILO-QTest could be made....with more variation or even different colors for different amounts of alkaloid.
Here's a thought: that Miraculix test turns darker shades of brown with higher psilocybin / psilocin. Brown, in pigments (as opposed to beams of light) doesn't really exist. Brown is just very dark yellow or orange. So maybe it's a test that turns yellow or orange in presence of shroom alkaloids, with an additional substance to amplify the effect by a magnitude or two. -- What about Keller's reagent?
"Psilocin is relatively unstable in solution due to its phenolic hydroxy (-OH) group. In the presence of oxygen it readily forms bluish and dark black degradation products. Similar products are also formed under acidic conditions in the presence of oxygen and Fe3+ ions (Keller's reagent)." https://en.wikipedia.org/wiki/Psilocin#Chemistry -- "In organic chemistry, Keller's reagent is a mixture of anhydrous (glacial) acetic acid, concentrated sulfuric acid, and small amounts of ferric chloride, used to detect alkaloids. Keller's reagent can also be used to detect other kinds of alkaloids via reactions in which it produces products with a wide range of colors." https://en.wikipedia.org/wiki/Keller%27s_reagent
The bluing reaction is a result of psilocin oxidizing but it’s not that simple because bruising also causes the dephosphorylation of psilocybin to psilocin which in turn oxidizes resulting in the bluing reaction Sauce: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7004109/
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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