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Invisiblejoze
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Quantitative psilocybin/psilocin reagent
    #27559526 - 11/27/21 12:37 PM (2 years, 3 months ago)

Hey all. I know that Miraculix makes a quantitative test for the determination of psilocybin, but unfortunately they only sell single-use kits. I'd love to start testing the psilocybin content of my fruits and performing small experiments, but I wish I could figure out what their reagents are, and make it in bulk. I can't find any research on this topic, but I was hoping one of y'all might have an idea where to look.


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InvisibleAssyrian
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27559548 - 11/27/21 01:01 PM (2 years, 3 months ago)

Quote:

joze said:
Hey all. I know that Miraculix makes a quantitative test for the determination of psilocybin, but unfortunately they only sell single-use kits. I'd love to start testing the psilocybin content of my fruits and performing small experiments, but I wish I could figure out what their reagents are, and make it in bulk. I can't find any research on this topic, but I was hoping one of y'all might have an idea where to look.




It'd be just Ehrlich reagent with a chart calibrated for different concentrations.

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Invisiblejoze
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Re: Quantitative psilocybin/psilocin reagent [Re: Assyrian]
    #27561845 - 11/29/21 10:55 AM (2 years, 3 months ago)

Quote:

Assyrian said:
Quote:

joze said:
Hey all. I know that Miraculix makes a quantitative test for the determination of psilocybin, but unfortunately they only sell single-use kits. I'd love to start testing the psilocybin content of my fruits and performing small experiments, but I wish I could figure out what their reagents are, and make it in bulk. I can't find any research on this topic, but I was hoping one of y'all might have an idea where to look.




It'd be just Ehrlich reagent with a chart calibrated for different concentrations.




Is it really just Ehrlich reagent? The reaction looks like a slightly different color to me.


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Offlinem0n0id
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27561857 - 11/29/21 11:14 AM (2 years, 3 months ago)

Interesting, looks like there's plenty of suppliers out there with Ehrlich's reagent for relatively cheap. And it is definitely good for psilocybin/psilocin testing.

I suppose if one was looking for comparative potency testing program for genetics selection you wouldn't even need to know how much actives there are quantitatively, just proportionally to other strains.

And if you did want quantitative results you could use the Miraculix kit to calibrate on the other high quantity Eherlich reagent.

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Invisiblejoze
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Re: Quantitative psilocybin/psilocin reagent [Re: m0n0id]
    #27561926 - 11/29/21 12:33 PM (2 years, 3 months ago)

From the reading I've done, it seems like Ehrlich's reagent is typically used to detect psilocybin, but isn't precise enough to be used to reliably measure psilocybin concentrations. However I've peeked at a few papers that have modified the reagent to improve the precision and accuracy, so maybe I'll look into this some more. Now my best guess is the Miraculix reagent is a modified Ehrlich's reagent.


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Offlinem0n0id
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27562025 - 11/29/21 02:25 PM (2 years, 3 months ago)

I'm by no means an expert but here's what I'd do. (I did pass a few basic chemistry classes haha, but never did organic chem.)

What I'm hypothesizing here is that the color resulting from the reaction can be controlled across tests by controlling all variables besides color.

Variables / Rough Steps:
1. Quantity of mushrooms (by weight)
2. Fineness/consistency of powedered mushrooms (powdered for uniform/max surface area)
3. Quantity of water (or other solvent) and mushrooms mixed to solve psilocybin / psilocin / other tryptamines from mushroom.
4. Time and tempreature allowed for tryptamines to disolve into solvent.
5. Quantity of Ehrlich's and solved tryptamines mixed
6. Time and tempreature for Ehrlich's to react with tryptamines.

Assuming all these variables kept the same for each test, you should be able to get a quantitative measure of tryptamines present. At that point all you would need to do is calibrate the process against a test with known quantitative read-out like the PSILO-QTest, then you'd be set to do as many tests as you desired.

Seems like a fun set of experiments to do! This is way low on my priorities, but would be cool if someone developed a tek.

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Offlinem0n0id
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Re: Quantitative psilocybin/psilocin reagent [Re: m0n0id]
    #27562027 - 11/29/21 02:27 PM (2 years, 3 months ago)

Joze, curious what other reagents you've found that might be more promising. Please share names/links when you have some time.

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OfflineEclipse3130
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27562166 - 11/29/21 05:01 PM (2 years, 3 months ago)

Just buy more of them.. they're really not expensive for a test result. Much cheaper than sending each fruit to a lab


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Invisiblejoze
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Re: Quantitative psilocybin/psilocin reagent [Re: Eclipse3130]
    #27562948 - 11/30/21 10:27 AM (2 years, 3 months ago)

Quote:

m0n0id said:
Joze, curious what other reagents you've found that might be more promising. Please share names/links when you have some time.




Here's a very old paper from 1960 that uses a modified Ehrlich's reagent to obtain better precision with low concentration ranges of indoles. Could be applicable.



Quote:

Eclipse3130 said:
Just buy more of them.. they're really not expensive for a test result. Much cheaper than sending each fruit to a lab




Lucky for me I work in a lab.


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InvisibleQM33
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27564213 - 12/01/21 06:31 AM (2 years, 3 months ago)

Keep us posted Jose.

Start looking into the makeup on the reagent, maybe see if it can be synthesized easily. And the quantitative test must just a a percentage to solution ratio, although I know the reagents can be corrosive ide be interested to see how "concentrated" or dangerous they are at a better level of understanding.


Interesting from wiki
Ehrlich's reagent or Ehrlich reagent is a reagent containing p-dimethylaminobenzaldehyde (DMAB) and thus can act as an indicator to presumptively identify indoles and urobilinogen. Several Ehrlich tests use the reagent in a medical test; some are drug tests and others contribute to diagnosis of various diseases or adverse drug reactions. It is named after Nobel Prize winner Paul Ehrlich who used it to distinguish typhoid from simple diarrhoea


Then

para-Dimethylaminobenzaldehyde is an organic compound containing amine and aldehyde moieties which is used in Ehrlich's reagent and Kovac's reagent to test for indoles. The carbonyl group typically reacts with the electron rich 2-position of the indole but may also react at the C-3 or N-1 positions.[1] It may also be used for determination of hydrazine.


And a number of sellers across the web..
https://www.scbt.com/p/4-dimethylamino-benzaldehyde-100-10-7?lang=en&cur=usd&gclid=Cj0KCQiA15yNBhDTARIsAGnwe0WOqzFlcLGPzun4h8nD6li8RK-NrsJZMeg6CDchKP0sfpCxOisK_JoaAiu9EALw_wcB

https://www.grainger.com/product/9FK87?ef_id=Cj0KCQiA15yNBhDTARIsAGnwe0XyTl_Q6dXN5nLGV6QQEF70_ftB4dQbLhCe9dqVaY7KnBKRBwtvOqUaAqyqEALw_wcB:G:s&s_kwcid=AL!2966!3!496359972172!!!g!437519823839!&gucid=N:N:PS:Paid:GGL:CSM-2295:4P7A1P:20501231&gclid=Cj0KCQiA15yNBhDTARIsAGnwe0XyTl_Q6dXN5nLGV6QQEF70_ftB4dQbLhCe9dqVaY7KnBKRBwtvOqUaAqyqEALw_wcB&gclsrc=aw.ds


So it seems as tho Elrich is just this chemical... And it would probably be safe to assume that the miraculux test kits are solutions of this chemical,

I wonder if the Hoffman test is also this chemical...

But maybe if you could find a way to dilute the chemical without destroying it, anything to thin it out really... So that I could be more easily viewed. Or maybe even if you could use a single drop and then put it next to a color wheel...

But you would also need to make tests to calibrate your own legend, meaning you would need to send samples to a lab, have them test them, use the same products on your own test and creat a scale based on what you know and how your test works....



Hmm lol


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Invisiblejoze
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Re: Quantitative psilocybin/psilocin reagent [Re: QM33]
    #27564667 - 12/01/21 12:16 PM (2 years, 3 months ago)

Yeah Ehrlich's reagent is very simple and Hofmann reagent is just a modified version. From wiki: The reagent is prepared by dissolving 0.5–2.0 g of p–dimethylaminobenzaldehyde (DMAB) in 50 mL of 95% ethanol and 50 mL of concentrated hydrochloric acid and is best used when fresh. ... The van Urk reagent, which uses 0.125 g of p-DMAB, 0.2 mL of ferric chloride solution (25 g/mL) in a solution of 65% sulfuric acid. This is sometimes referred to as the Hofmann reagent or p-DMAB-TS (Test Solution) and gives slightly different colours with different indoles.

I think the hardest step would be getting a reference standard. Hard to get 99.9% pure psilocybin.


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InvisibleQM33
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27564702 - 12/01/21 12:40 PM (2 years, 3 months ago)

I wonder if 4 aco Dmt would react similarly..? Or even regular Dmt, might be able to base it off that......?

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OfflineAlan RockefellerM
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27576775 - 12/11/21 12:31 AM (2 years, 3 months ago)

Quote:

joze said:
Hey all. I know that Miraculix makes a quantitative test for the determination of psilocybin, but unfortunately they only sell single-use kits. I'd love to start testing the psilocybin content of my fruits and performing small experiments, but I wish I could figure out what their reagents are, and make it in bulk. I can't find any research on this topic, but I was hoping one of y'all might have an idea where to look.




I speak to Felix regularly, who invented the Miraculix kits, and I am pretty sure it's not Ehrlich's.  I asked him if it was a secret what was used, and he said it was.  He would probably tell me if I asked directly, but I don't like to pry.  He already shares so much cool information with me without having to ask pointed questions.

The kits have a relatively large volume of solution, I imagine you could split them up into lots of little tubes (like 200 uL PCR tubes) and do lots of tests with one kit.  You'd need a pretty accurate scale.

Another way to do it is with Ehrlich's reagent + TLC.

I know he spent quite a while getting the psilocybin extraction part working in a way that is repeatable.

The best way to test for psilocybin is HPLC, but it's expensive, and developing a good method isn't something a beginner would likely have good luck doing.

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Invisiblejoze
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Re: Quantitative psilocybin/psilocin reagent [Re: Alan Rockefeller]
    #27577207 - 12/11/21 11:10 AM (2 years, 3 months ago)

Quote:

Alan Rockefeller said:
Quote:

joze said:
Hey all. I know that Miraculix makes a quantitative test for the determination of psilocybin, but unfortunately they only sell single-use kits. I'd love to start testing the psilocybin content of my fruits and performing small experiments, but I wish I could figure out what their reagents are, and make it in bulk. I can't find any research on this topic, but I was hoping one of y'all might have an idea where to look.




I speak to Felix regularly, who invented the Miraculix kits, and I am pretty sure it's not Ehrlich's.  I asked him if it was a secret what was used, and he said it was.  He would probably tell me if I asked directly, but I don't like to pry.  He already shares so much cool information with me without having to ask pointed questions.

The kits have a relatively large volume of solution, I imagine you could split them up into lots of little tubes (like 200 uL PCR tubes) and do lots of tests with one kit.  You'd need a pretty accurate scale.

Another way to do it is with Ehrlich's reagent + TLC.

I know he spent quite a while getting the psilocybin extraction part working in a way that is repeatable.

The best way to test for psilocybin is HPLC, but it's expensive, and developing a good method isn't something a beginner would likely have good luck doing.



Thanks for weighing in Alan. I had a feeling it was a proprietary reagent different from Ehrlich's.

Your idea of splitting up the test solution is a good idea, maybe I could figure something out with 96 well plates. My lab is working towards getting an HPLC and ultimately an LC-MS, so hopefully some day I'll have access to that kind of technology.


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OfflineAlan RockefellerM
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Re: Quantitative psilocybin/psilocin reagent [Re: joze]
    #27579136 - 12/12/21 07:24 PM (2 years, 3 months ago)

If you could use 96 well plates and a plate reader that could be pretty accurate.

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