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nerdintheshell



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Re: DIY Psilocybin Content Percentage Detection [Re: nerdintheshell] 3
#27702993 - 03/21/22 12:01 AM (1 year, 10 months ago) |
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The video I saw in case anyone is interested: link
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nerdintheshell] 2
#27703006 - 03/21/22 12:36 AM (1 year, 10 months ago) |
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Here is a better chart if anyone is considering TLC.
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shr00med
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Registered: 03/10/21
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Quote:
justanotherhuman said:
And it seems like the OP is probably more interested in chromatographic approaches to this than I β personally, Iβd like to see a widely accessible DIY method become available to the average Shroomerite, which is why Iβm more fixated on the gradient-scale approach.
I'm definitely interested in creating a reliable tek for citizen scientists not only to follow, but also as something that can hopefully lead to more advanced DIY discoveries. It's a shame we had to go political with things, but I'm thankful this thread is active and for the encouragement from justanotherhuman to proceed with what would've been just another personal pondering.
My plan of action will be as follows: Test different weighing dried material with reagent, seeing if the chromogenic results differ with what should be different potencies. If the reagent proves to only show the presence of psilocybin, and not the quantity (which is my hypothesis), then I will resort to making my own TLC plates and go from there.
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SingularFusion


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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27704172 - 03/21/22 11:36 PM (1 year, 10 months ago) |
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the tlc plates sounds doable... I seen guys over at the nexus some years ago already testing various potential DMT plant candidates on tlc plates. Seems fairly accurate if you are able to interpret the results properly, probably takes a touch of practice but surely can be learned
I would love some open source colour gradient thing, that would be ideal. I would be willing to pay for miraculix but the idea of a box of tests arriving through customs addressed to a private individual pretty much closes the deal for me... perhaps my paranoia has grown to be too large, but I will be paranoid and safe lol
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QM33
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Registered: 04/09/20
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Loc: Oregon
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Re: DIY Psilocybin Content Percentage Detection [Re: SingularFusion] 1
#27706536 - 03/24/22 07:01 AM (1 year, 9 months ago) |
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I'm going to sub just in case one of you cracks that code
-------------------- 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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nerdintheshell



Registered: 09/11/21
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Re: DIY Psilocybin Content Percentage Detection [Re: SingularFusion] 3
#27708958 - 03/26/22 12:59 AM (1 year, 9 months ago) |
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Just made my first batch of diy tlc plates. Ran a test with a sharpie and it came out pretty good. Hopefully I can try with psilocybin soon. I don't have a vacuum pump but could probably use the one at a local maker space.
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nerdintheshell] 1
#27709411 - 03/26/22 11:51 AM (1 year, 9 months ago) |
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Quote:
nerdintheshell said: Just made my first batch of diy tlc plates. Ran a test with a sharpie and it came out pretty good. Hopefully I can try with psilocybin soon. I don't have a vacuum pump but could probably use the one at a local maker space.

Nice job!

I'm not so sure you really need to use a vacuum pump to reduce the extraction, do you? I don't see a reason why a different/simpler extraction method couldn't work, like the 1% (0.01g) concentration mix of mush powder into 3cc methanol, talked about in this thread. Unless I'm overlooking/missing something? Perhaps there's reduced stability of psilocin in HEET vs the acetic acid, or maybe it would be harder to accurately dilute? https://www.shroomery.org/forums/showflat.php/Number/9042075#9042075
-------------------- ποΈ π why you are empty. Hunt for the habitat not the mushroom.
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QM33
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Registered: 04/09/20
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 2
#27709487 - 03/26/22 01:12 PM (1 year, 9 months ago) |
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Okay so idk if this was mentioned.
But why couldn't you get a bottle of the miraculous, other sterile vials, serperate the reagent in say even drop or 100-300 microliter portions, and introduce samples at the relative weight. Assuming the reagent would be hard or nearly impossible to see for comparisons sake, maybe you could, after reaction, put the reagent on a more appropriate container for viewer, and or use with either a stereoscope or maybe even a microscope that could be "calibrated " to the color scale in some manner, assuming the colors would view differently under a microscope....
Any thoughts? About any of it?
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Icyurmt
Strange


Registered: 04/02/20
Posts: 1,625
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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27709513 - 03/26/22 01:47 PM (1 year, 9 months ago) |
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Quote:
QM33 said: Okay so idk if this was mentioned.
But why couldn't you get a bottle of the miraculous, other sterile vials, serperate the reagent in say even drop or 100-300 microliter portions, and introduce samples at the relative weight. Assuming the reagent would be hard or nearly impossible to see for comparisons sake, maybe you could, after reaction, put the reagent on a more appropriate container for viewer, and or use with either a stereoscope or maybe even a microscope that could be "calibrated " to the color scale in some manner, assuming the colors would view differently under a microscope....
Any thoughts? About any of it?
Alan had mentioned the possibility of splitting them up into smaller portions here. I can't see a reason why it wouldn't work to some degree. https://www.shroomery.org/forums/showflat.php/Number/27576775#27576775
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.
-------------------- ποΈ π why you are empty. Hunt for the habitat not the mushroom.
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nerdintheshell



Registered: 09/11/21
Posts: 74
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27709608 - 03/26/22 03:15 PM (1 year, 9 months ago) |
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So I could probably just use elrich's reagent as the solvent in the steps workman provided and skip the spraying step? Then to find the concentration I just dilute the methanol solution until spots don't show up? Then once the spots don't show up anymore I know what concentration? So if I dilute the 1% solution to 0.5% and the spots don't show up anymore I know the concentration?
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Icyurmt
Strange


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Posts: 1,625
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Re: DIY Psilocybin Content Percentage Detection [Re: nerdintheshell] 1
#27709729 - 03/26/22 05:38 PM (1 year, 9 months ago) |
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Quote:
So I could probably just use elrich's reagent as the solvent in the steps workman provided and skip the spraying step?
I'm not quite sure, I have to go back and reread workman's post to understand what you are referring to. I believe you would want to use one of the solvents from the chart above with an already established rf value so you know what you are looking at though. I think elrich is only used as a developer enabling you to see the spots but? I haven't played with TLC plates for about 20 years now, hopefully somebody more familiar/knowledgeable than myself will chime in soon.
Quote:
Then to find the concentration I just dilute the methanol solution until spots don't show up? Then once the spots don't show up anymore I know what concentration? So if I dilute the 1% solution to 0.5% and the spots don't show up anymore I know the concentration?
If you just wanted to compare 2 samples to one another or to a known reference, you would only have to extract and then spot them side by side, run it in the solvent, and then once developed with elrich; the size and intensity of the spots that form at the corresponding RF should give a quick indication as to which one is more potent.
For a semi-quantitative test, I believe you would dilute it down until spots no longer show up visible at the corresponding RF for the specific alkaloid you are testing. Then using published minimum detectable amounts for whatever developer you used (I think it's 20ng/mL for psilocybin with p-DMAB, but you should double check) It should now possible to do the math backwards for your dilutions and calculate the amount in your original extraction/sample if I'm understanding things correctly.
-------------------- ποΈ π why you are empty. Hunt for the habitat not the mushroom.
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QM33
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Registered: 04/09/20
Posts: 4,739
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27709735 - 03/26/22 05:45 PM (1 year, 9 months ago) |
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Quote:
Icyurmt said:
Quote:
QM33 said: Okay so idk if this was mentioned.
But why couldn't you get a bottle of the miraculous, other sterile vials, serperate the reagent in say even drop or 100-300 microliter portions, and introduce samples at the relative weight. Assuming the reagent would be hard or nearly impossible to see for comparisons sake, maybe you could, after reaction, put the reagent on a more appropriate container for viewer, and or use with either a stereoscope or maybe even a microscope that could be "calibrated " to the color scale in some manner, assuming the colors would view differently under a microscope....
Any thoughts? About any of it?
Alan had mentioned the possibility of splitting them up into smaller portions here. I can't see a reason why it wouldn't work to some degree. https://www.shroomery.org/forums/showflat.php/Number/27576775#27576775
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.
Cool thanks, I thought so I just didn't know if this was the same thread. Idk I'll post in there maybe, but ide like to expand on that also..
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27709758 - 03/26/22 06:09 PM (1 year, 9 months ago) |
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Quote:
Cool thanks, I thought so I just didn't know if this was the same thread. Idk I'll post in there maybe, but ide like to expand on that also..
By all means, please expand on it here, I'm certainly interested. I linked it just to merge these two as I remembered Alan bringing it up before and thought it was a great idea.
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27710141 - 03/27/22 05:21 AM (1 year, 9 months ago) |
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I mean the first step would be reacting the reagent in a full bott and smaller sample and comparing it to a full vial on the scale and then comparing smaller samples, even in different container sizes, to the same scale and see how close the same reaction looks in different sizes, and containers, know the container is going to change the way the sample looks and percieves it's colors.
Then you would need to decide how close it is and if you would need to make a new scale, which I think you most likely would, probably a "lighter" one relatively, but probably in the same shades.
Uhhg and I just looked, they don't tell you how much you get, of course. If I had to guess though it looks like a milliliter or a CC about, so maybe 1000uL?
Idk and I'm cheap haha, and I would need alot of these, so idk you would need to get it down to 2-250uL just to get 4 or 5 probably...
Also I noticed they have a 10% deviation from HPLC.... So idk if that's even worth it in the long run... Maybe. Probably maybe.
But idk if you can even have ideas after that, until trying stuff like that, comparing smaller reacted portions to larger ones, containers and figuring out the original scales relavence then it's just guessing beyond what could stop you right away.
Maybe I'll get some in the future. Idk magic myco and dudes with access to HPLC are already moving along.
Foodfurthought
-------------------- 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
Posts: 928
Loc: PNW
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Dividing Miraculix kits into smaller aliquots and doing psilocin/psilocyin detection on a smaller scale would work, but the problem you will run into is a higher variability the smaller your samples get. If they're already reporting 10% deviation from HPLC, you start doing it on a smaller scale and your deviation would probably increase.
Quote:
justanotherhuman said: I'd point out, however, that this is accomplishing the same goal using the same principle as what I'm talking about -- the main difference being that semi-quantitative TLC is plotting a concentration curve using however many dilutions whereas what I'm talking about is plotting a concentration curve to a colorimetric scale based on the intensity of the reagent's color reaction. The Miraculix test serves as a proof of concept.
An advantage to a Miraculix-style approach is that it necessarily requires a very small sample (~0.1g) near the threshold for the color reaction in a relatively small given quantity of reagent. Another advantage is that, after having initially determined the colorimetric scale, tests will yield results in a matter of minutes and without further effort. The trade-off, of course, is accuracy -- properly executed TLC will certainly be more accurate. But I'd argue that -- for most DIY-minded Shroomery members -- the accuracy trade-off is minimal enough to justify avoiding chromatography.
I'd love to see a DIY colorimetric reagent come to fruition. I would suggest that when you establish a concentration curve using colorimetric data, you repeat the process multiple times with reagent that is fresh, 1 day old, 3 days old, 1 week old, etc. Certain reagents deteriorate quickly and this can significantly affect your results.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 3
#27712129 - 03/28/22 09:25 PM (1 year, 9 months ago) |
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Update: I have ordered some p-DMAB so that I can mix up some Ehrlichβs to begin running tests to see if I can DIY a reasonably accurate colorimetric reagent test. Ideally, the result will be a tek that anyone can follow to mix their own reagent for determining psilocybin potency Weβll see what happens
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Alan Rockefeller
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 2
#27712338 - 03/28/22 11:59 PM (1 year, 9 months ago) |
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Quote:
joze said: Dividing Miraculix kits into smaller aliquots and doing psilocin/psilocyin detection on a smaller scale would work, but the problem you will run into is a higher variability the smaller your samples get. If they're already reporting 10% deviation from HPLC, you start doing it on a smaller scale and your deviation would probably increase.
The other issue is that with smaller samples, having an extremely accurate scale would be much more important.
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joze



Registered: 11/10/20
Posts: 928
Loc: PNW
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Re: DIY Psilocybin Content Percentage Detection [Re: Alan Rockefeller] 1
#27712781 - 03/29/22 11:39 AM (1 year, 9 months ago) |
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Quote:
Alan Rockefeller said:
Quote:
joze said: Dividing Miraculix kits into smaller aliquots and doing psilocin/psilocyin detection on a smaller scale would work, but the problem you will run into is a higher variability the smaller your samples get. If they're already reporting 10% deviation from HPLC, you start doing it on a smaller scale and your deviation would probably increase.
The other issue is that with smaller samples, having an extremely accurate scale would be much more important.
Yes, this is absolutely true as well.
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shr00med
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27716152 - 04/01/22 07:16 AM (1 year, 9 months ago) |
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I did invest in a scale; this one had me skeptical because of the price but it's been preforming wonderfully. I'm going to try making the reagent today with muriatic acid instead of hydrochloric just cause I already own it...
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joze



Registered: 11/10/20
Posts: 928
Loc: PNW
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27716397 - 04/01/22 11:18 AM (1 year, 9 months ago) |
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Quote:
shr00med said: I did invest in a scale; this one had me skeptical because of the price but it's been preforming wonderfully. I'm going to try making the reagent today with muriatic acid instead of hydrochloric just cause I already own it...
Lucky for you, muriatic acid is just dilute HCl. I'm not sure what the pH range of Ehrlich's reagent is but you may need to adjust it.
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