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shr00med
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DIY Psilocybin Content Percentage Detection 5
#27682139 - 03/04/22 07:48 AM (1 year, 10 months ago) |
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I've viewed a couple posts regarding methods of HPLC and GC using professional equipment, and I'm interested if I could figure out a way to DIY it. This is a new topic for me as of this morning, so apologies if this makes absolutely no sense. Because technically all kinds of chromatography have the same principles of separating components, would it be possible to use Column Chromatography to detect percentages of psilocybin/ psilocin instead of Liquid Chromatography? If so, what would be used as the mobile phase? Maybe another method could be something like this, but I don't know what chemical would be the "detection" solution, or how I would gauge different colored results.
Edited by shr00med (03/04/22 08:04 AM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27683058 - 03/04/22 08:04 PM (1 year, 10 months ago) |
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I have been looking into the second option you listed. Chromatography obviously would work but that’s a bit outside the usual DIY skills and materials availability. I believe that company is using an Erlich reagent which basically just changes color in the presence of psilocybin (or any indole alkaloid, I think) and I believe it’s is the same reagent that pigs use in their field test kits.
I am currently looking into whether there is a good DIY way to reliably make the reagent and, if there is an easy and cheap enough way to do so, the next step would be to “calibrate” the process by determining an appropriate amount of mushroom material (say 1/10th of a gram) and reagent to combine and then observing the spectrum of color change in response to various dilutions of material. I might for example take some powdered mushroom material of average potency, measuring out first 1/10 of a gram, than 1/20, 1/30, etc and mixing them all in turn with the same amount of reagent to establish a rough color chart. This would have to be repeated with many different samples known to be more potent and known to be weaker and then ultimately we would have something similar to a pH test strip With a reasonably accurate scale to compare relative potency. But it wouldn’t give us an exact mg of psilocybin per g of mushroom :/
Anyway, that’s what I’ve got so far — that would be cool if you figured out a more precise way though
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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SpectreOfCommunism
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Okay, so basically to make Ehrlich's reagent, you need p-dimethylaminobenzaldehyde -- or DMAB, for short -- and some ethanol that is at least 95% pure and some hydrochloric acid.
The ethanol can be purchased at most liquor stores and I've always sourced my hydrochloric acid from hardware stores where it's sold as "muriatic acid" for cleaning driveways. And I've found that 100g of DMAB can be purchased on Amazon for $35: https://www.amazon.com/dp/B00DYO8J2A/
So for about $60, an interested person could totally make a decent amount of the reagent at home and, using the procedure I outlined above, could design a test that could measure the relative potency of a given quantity of powdered mushroom material -- and, I suppose, if the published research on the range of % by weight of psilocybin in cubensis is accurate, one could "map" the lower and higher ends of that range to the reagent's color-scale to approximate the % by weight of psilocybin using the test.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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shr00med
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I appreciate your response, looks like I got some experimenting to do! I’ll make sure to keep the tread updated with my findings and failures.
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Alan Rockefeller
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Quote:
justanotherhuman said: I believe that company is using an Erlich reagent which basically just changes color in the presence of psilocybin (or any indole alkaloid, I think) and I believe it’s is the same reagent that pigs use in their field test kits.
Wouldn't Erlich's reagent give a pink color in the presence of tryptamines? The Miraculix kits turn brown.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: Alan Rockefeller] 1
#27694454 - 03/13/22 11:24 PM (1 year, 10 months ago) |
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Quote:
Alan Rockefeller said:
Quote:
justanotherhuman said: I believe that company is using an Erlich reagent which basically just changes color in the presence of psilocybin (or any indole alkaloid, I think) and I believe it’s is the same reagent that pigs use in their field test kits.
Wouldn't Erlich's reagent give a pink color in the presence of tryptamines? The Miraculix kits turn brown.

Ehrlich’s reagent will turn pink to dark purple in the presence of indole alkaloids like psilocybin and yes the miraculix color gradient does seem to be different — I hadn’t seen their chart yet, thanks for the new info. I’ve read about several different “modified” Erlich’s reagents, which i assume would result in “modified” gradients as well, and so it’s possible they’re using a proprietary version (maybe so they could patent an otherwise well-known test?).
In any case, it’s the same principle and so I think a similar test could be DIY’d using Ehrlich’s reagent but I will do some research tonight to see if I can narrow down what particular reagent they use and whether there may be any advantage to it
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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fossilshark
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You are just assuming its the same principle based on the fact that you can't figure it out.
An Ehrlich reagent will be set off by plenty of other alkaloids in mush, it is not accurate for testing psilocybin potency. If you just want to confirm the presence of indoles, sure an Ehrlich will work. But then try to prove to someone its not being tripped by something else.
The miraculix would not be able to clame any sort of accuracy if it was a simple modified Ehrlich in my opinion.
I think the person who developed miraculix deserves the compensation required for selling it seeing as its pretty revolutionary and definitely an original idea, and trying to bootleg it is kinda scummy.
-------------------- LITFA LITFA LITFA
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: fossilshark] 1
#27695122 - 03/14/22 02:48 PM (1 year, 10 months ago) |
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Quote:
fossilshark said: You are just assuming its the same principle based on the fact that you can't figure it out.
I am “assuming” so based on multiple observations and sound inferences, such as (1) the fact that Miraculix itself refers to its proprietary test-solution as a “reagent” and (2) the fact that said reagent indicates indole alkaloids are present by a change in color which is proportional to their concentration in the solution, all of which is also true of Ehrlich’s and other DMAB-based reagents; moreover, we know that (3) a number of modified forms of Ehrlich’s reagent exist (e.g. Hoffman’s reagent) which indicate the presence of indole alkaloids with different colors, that (4) law enforcement agencies use Ehrlich-type reagents in psilocybin field tests, and that (5) the chromogenic (aka color-changing) response of such reagents varies in intensity depending on the concentration of the target compounds.
From these facts, we can deduce that the observed color-change in given amounts of such reagents will vary with the concentration of indoles in given amounts of a material, which means we can use a colorimetric scale to determine a sample’s alkaloid content with reasonable accuracy. Now, we can either conclude that Miraculix is telling the truth and that they use such a reagent and colorimetric scale in their psilocybin test or, on the other hand, we could speculate wildly that the Miraculix team has discovered some utterly new and secret process which just happens to look identical to the methods we already know about and understand.
Quote:
fossilshark said: An Ehrlich reagent will be set off by plenty of other alkaloids in mush, it is not accurate for testing psilocybin potency. If you just want to confirm the presence of indoles, sure an Ehrlich will work. But then try to prove to someone its not being tripped by something else.
The miraculix would not be able to clame any sort of accuracy if it was a simple modified Ehrlich in my opinion.
Can you name some of these “plenty of other alkaloids” that would be present (in any significant amount) in any of the commonly cultivated psilocybe species? There are small amounts of indoles like baeocystin, norpsilocin, etc but these only appear in pretty minute amounts and, in any case, they are chemical analogs of the indoles we’re looking for. I'm aware that some chemicals (e.g. pyridoxine) can lead to false positives in Ehrlich tests but none of them would be present in homegrown samples of cubensis.
Quote:
fossilshark said: I think the person who developed miraculix deserves the compensation required for selling it seeing as its pretty revolutionary and definitely an original idea, and trying to bootleg it is kinda scummy.
I'm not sure a color-changing reagent for detecting drugs is a particularly "revolutionary" or "original idea -- I mean reagent color-tests are kinda the standard of drug detection worldwide and have been for awhile. What Miraculix has done is to add a colorimetric scale to such reagent tests, which is cool and everything but hardly revolutionary. And no one said they shouldn’t be paid for their product — I simply pointed out that, as is common practice throughout the whole rotten capitalist pharmaceutical industry, it’s likely that this company opted to develop a proprietary version of the known psilocybin-indicating reagents because patented commodities are more profitable.
That’s just the internal logic of capital, my dude
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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WhiteThumbScience
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27701959 - 03/19/22 10:56 PM (1 year, 10 months ago) |
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This sounds like a good idea but in practice this method will not be accurate just due to the bio pathway related indole/tryptamine components in mushrooms that do vary greatly between just maturity levels alone in the very same strain/variety!!!
-------------------- Just say NO to police searches!!! -d-DEX-25 Mathematics knows no races or geographic boundaries; for mathematics, the cultural world is one country. -David Hilbert
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WhiteThumbScience
Full Rick


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I notice your very anti capitalist but let me ask you if companies cannot profit from the billions it takes to develop these new technologies then who is gonna develop them and why?
Sounds like you want to live in the world of no new products or inventions because there will be zero incentive for anyone to do anyone unless "forced" by a command and control economy?
Remind me what economic system(capitalism is just the result of a free society doing trade and thus is default remember) brought us the modern age?
Oh right it was the communists right??
-------------------- Just say NO to police searches!!! -d-DEX-25 Mathematics knows no races or geographic boundaries; for mathematics, the cultural world is one country. -David Hilbert
Edited by WhiteThumbScience (03/19/22 11:01 PM)
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SpectreOfCommunism
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Quote:
WhiteThumbScience said: I notice your very anti capitalist but let me ask you if companies cannot profit from the billions it takes to develop these new technologies then who is gonna develop them and why?
Workers will make everything in the future just as they do now and just as they have always done.
Quote:
WhiteThumbScience said: Sounds like you want to live in the world of no new products or inventions because there will be zero incentive for anyone to do anyone unless "forced" by a command and control economy?
This is where you demonstrate that you are debating in bad faith. See, if you really believe that my position is that absurd, then you are an even bigger fool than I am to debate it -- and, on the other hand, if you don't really believe my position is that absurd, then you are deceitful for implying so. In either case, it is a strawman and a bad-faith argument. Do better.
Quote:
WhiteThumbScience said: Remind me what economic system(capitalism is just the result of a free society doing trade and thus is default remember) brought us the modern age?
Oh right it was the communists right??
If you'd ever read Marx (which, of course, you haven't), you'd know that communists regard capitalism as a necessary and, initially, progressive stage of social development which brings about a great rise in productivity, innovation, etc and that the development of socialism is only possible because of and within the context of capitalism.
Anyway, you're new here and so I'll just gently let you know that this is off-topic and that there is a strict no-flaming policy here in the Shroomery. If you're interested in actually learning what a communist perspective really is (and, if you'd like, to debate it), you're welcome to send me a private message. This thread, however, was created to discuss a different topic
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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WhiteThumbScience
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I was responding directly to your statements first which included statements about my countries economic/political structure and tied that into the OPs post!
Second I am simply debating something with you and I will apologize it did come off in sort of bad faith and I do apologize for that I did not expand enough and instead reverted to immediate criticism of the position I know you clearly hold!
Third I have read the works of Marx and by later Lenin and both are radically different! Both do rely on late stage capitalist concentration(a real danger in an unregulated or severely corrupt system that no other alternative legal remedy is available but workers revolt and possibly complete revolution!).
I will agree true communism has never been attempted because it requires late stage toxic capitalism which is not even close to were we are at? If that developed then we can talk the benefit to cost ratios of command economics ECONOMICALLY but what we can't ever say is that such a system includes freedom because inherent in its principles is the control of all goods and services which cannot be free by definition!!!
Really want the government to have control of your life in at the very least regards to all financial decisions beyond one's own credit given as privilege instead of by one's own merit!
I'll agree this is more off topic but I feel I had to clarify and apologize. Though I even agree on some things as I believe not enough people have studied ALL sides and ALL opinions something more people need the ability to do including myself despite beliefs I formed sometimes I shut out the other side before they even give their opinion 😥
-------------------- Just say NO to police searches!!! -d-DEX-25 Mathematics knows no races or geographic boundaries; for mathematics, the cultural world is one country. -David Hilbert
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Icyurmt
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Quote:
Can you name some of these “plenty of other alkaloids” that would be present (in any significant amount) in any of the commonly cultivated psilocybe species?
Getting back to the topic, wouldn't serotonin, tryptophan, and 5-htp, give false positives with ehrlich? Those are not only found in active panaeolus but also the inactive ones like foenisecii etc.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27702718 - 03/20/22 06:38 PM (1 year, 10 months ago) |
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Quote:
Icyurmt said:
Quote:
Can you name some of these “plenty of other alkaloids” that would be present (in any significant amount) in any of the commonly cultivated psilocybe species?
Getting back to the topic, wouldn't serotonin, tryptophan, and 5-htp, give false positives with ehrlich? Those are not only found in active panaeolus but also the inactive ones like foenisecii etc.
Yes, tryptophan will do it (unsure about the others) and that is why I included the qualifier “in any significant amount.” I understand that some precursors like tryptophan are present in trace amounts but, from what I’ve been led to believe by the limited research available on the topic, such chemicals don’t make up anywhere near the 1-2% psilocybin/psilocin content by weight in cubensis and therefore wouldn’t impact the results significantly. In fact, I believe that many of the studies wherein they determined the ~1-2% range are actually referring to total indole content. Correct me if I’m wrong.
And as to your other point, I will clarify that I am not suggesting that the reagent-based method we’re discussing would be useful for detecting indole content in anything but psilocybe cubensis and very similar mushrooms. And for that matter, I am also not suggesting that it wouldn’t be useful — just that, for the moment, I am only considering cubensis because they are by far the most cultivated magic mushroom and ya gotta start somewhere
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Icyurmt
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What about using semi-quantitative TLC?
Quote:
WillSolvem said: The easiest method that does not utilise a mass spectrometer:
Extract from 1g mushroom powder using 10ml vinegar, heat to 70*c cool and filter. then reduce to 50% of volume under vacuum then just spot and run tlc plates (available on ebay) and develop with Ehrlich's reagent (ingredients available on ebay), then using known dilutions you spot and run plates until there are no signs of a colorimetric reaction in the rf value responsible for psilocin. then calculate and plot a concentration curve and you have a quantitative value for your original 1g sample.
Rf values
Psilocybin 0.40 Baeocystin 0.51 Bufotenine 0.54 Psilocin 0.55 Serotonin 0.65 Tryptophan 0.68
using a 7:3 methanol/vinegar eluent over silica
http://en.wikipedia.org/wiki/Ehrlich's_reagent
https://www.shroomery.org/forums/showflat.php/Number/19917169#19917169
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27702904 - 03/20/22 09:38 PM (1 year, 10 months ago) |
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Quote:
Icyurmt said: What about using semi-quantitative TLC?
Quote:
WillSolvem said: The easiest method that does not utilise a mass spectrometer:
Extract from 1g mushroom powder using 10ml vinegar, heat to 70*c cool and filter. then reduce to 50% of volume under vacuum then just spot and run tlc plates (available on ebay) and develop with Ehrlich's reagent (ingredients available on ebay), then using known dilutions you spot and run plates until there are no signs of a colorimetric reaction in the rf value responsible for psilocin. then calculate and plot a concentration curve and you have a quantitative value for your original 1g sample.
Rf values
Psilocybin 0.40 Baeocystin 0.51 Bufotenine 0.54 Psilocin 0.55 Serotonin 0.65 Tryptophan 0.68
using a 7:3 methanol/vinegar eluent over silica
http://en.wikipedia.org/wiki/Ehrlich's_reagent
https://www.shroomery.org/forums/showflat.php/Number/19917169#19917169
That would certainly work -- one disadvantage would be the cost of thin-layer chromatography plates.
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.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Icyurmt
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You can buy quite alot of TLC plates (~.60¢ each) for the same price as 4 Miraculix tests and with known rf values already published, one could also start testing right away, without the need to figure out what reagent may work best or fiddling with calibrating a colorimetric scale. True it may not have the same ease of use and smaller sample size as with the miraculix, but it is an accurate, already available, relatively inexpensive, DIY way of determining potency regardless of species.
Interested to see what you eventually come up with.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
Edited by Icyurmt (03/20/22 10:53 PM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 2
#27702962 - 03/20/22 11:11 PM (1 year, 10 months ago) |
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Quote:
Icyurmt said: You can buy quite alot of TLC plates (~.60¢ each) for the same price as 4 Miraculix tests and with known rf values already published, one could also start testing right away, without the need to figure out what reagent may work best or fiddling with calibrating a colorimetric scale. True it may not have the same ease of use and smaller sample size as with the miraculix, but it is an accurate, already available, relatively inexpensive, DIY way of determining potency regardless of species.
Interested to see what you eventually come up with.

For sure, both approaches have merit methinks — just a matter of which mix of advantages/disadvantages one wants to take on And it seems like the OP is probably more interested in chromatographic approaches to this than I — personally, I’d like to see a widely accessible DIY method become available to the average Shroomerite, which is why I’m more fixated on the gradient-scale approach. The initial labor would be greater than TLC but, after someone does it, the scale and amounts of reagent and mushroom material could be shared here in a public forum, after which it would be just a matter of purchasing DMAB and hydrochloride acid and then following the tek Kinda like an open-source Miraculix
I like your “hunt the habitat“ signature btw — it’s the most important insight I’ve gleaned in all my years of foraging
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nerdintheshell



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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 3
#27702978 - 03/20/22 11:34 PM (1 year, 10 months ago) |
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Quote:
You can buy quite alot of TLC plates (~.60¢ each) for the same price as 4 Miraculix tests
On YouTube there's also videos on making your own. One video I watched from the channel "TheHomeScientist" says you can even use corn starch as the stationary. If so, the costs per plate could go down immensely and open this method up to pretty much anyone with a vacuum setup and the reagents.
Maybe you could boil down the solution also? It would definitely mess with results a bit but if you're just trying to look for the more potent culture it would still give you a definitive answer of which one is more potent without requiring expensive Miraculix tests.
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Icyurmt
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Quote:
personally, I’d like to see a widely accessible DIY method become available to the average Shroomerite,
Same here, I fully agree 👍 I'm all for open source science.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
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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
Stranger
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 — personally, I’d like to see a widely accessible DIY method become available to the average Shroomerite, which is why I’m 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
(NOT A PUPPET!) ❤❤❤❤❤



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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



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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
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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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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.
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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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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.
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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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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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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
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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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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 Ehrlich’s 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 We’ll see what happens
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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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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



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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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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27716466 - 04/01/22 12:12 PM (1 year, 9 months ago) |
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I'm also curious: what are y'all planning on using as a reference? The Miraculix kit has a scale they've developed themselves with their specific reagent. You're talking about determining the potency of the mushrooms, which is essentially the same as talking about the concentration of psilocybin/psilocin. Okay, so you can test your mushrooms with this reagent and measure the color, but what are you going to compare the value to? Unless you are able to source a relatively pure sample of psilocybin or psilocin in a known concentration, you will be limited to comparing relative potencies of mushrooms rather than measuring the actual concentrations.
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Edited by joze (04/01/22 12:13 PM)
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27716484 - 04/01/22 12:20 PM (1 year, 9 months ago) |
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But I mean shit, that could have its own value in a way if it was significantly cheaper or quicker maybe..?
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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27716496 - 04/01/22 12:30 PM (1 year, 9 months ago) |
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Quote:
QM33 said: But I mean shit, that could have its own value in a way if it was significantly cheaper or quicker maybe..?
Oh, I absolutely agree. I just think it's important for everyone to be aware of what they're actually measuring.
A quick, inexpensive method to compare relative potency of different cultures would be borderline revolutionary for the hobby grower. Being able to say okay, these golden teachers are more potent than the B+ I grew, that would be awesome. But at the same time, it would be difficult to compare potencies with other growers if you don't have that true measurement of potency.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27716502 - 04/01/22 12:33 PM (1 year, 9 months ago) |
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Totally, but you might have about all you need to guarantee your isolating you most potent genetics, and maybe once you got to a certain point you could take it for further analysis, rather than testing multiple genetics with more costly analysis...
What's going on here?! I want some
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27716506 - 04/01/22 12:36 PM (1 year, 9 months ago) |
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Agreed! Looking forward to seeing where this thread goes!
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 3
#27717225 - 04/01/22 10:42 PM (1 year, 9 months ago) |
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Quote:
joze said: I'm also curious: what are y'all planning on using as a reference? The Miraculix kit has a scale they've developed themselves with their specific reagent. You're talking about determining the potency of the mushrooms, which is essentially the same as talking about the concentration of psilocybin/psilocin. Okay, so you can test your mushrooms with this reagent and measure the color, but what are you going to compare the value to? Unless you are able to source a relatively pure sample of psilocybin or psilocin in a known concentration, you will be limited to comparing relative potencies of mushrooms rather than measuring the actual concentrations.
Firstly, I'd say -- for the purposes of most of us anyway -- even being able to consistently tell whether this sample is relatively low-potency, this one average, and this one high-potency would be a pretty heckin' useful test.
And secondly, concerning a reference, I can think of a few approaches that don't involve already having any known quantities of psilocybin.
One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder. Because this powder is a randomized mixture composed of thousands of individual fruits representing thousands of cubensis strains, we can be all but certain that samples taken therefrom will be very close to the average potency -- and if our analysis shows the same results after multiple small samples (e.g. 150mg), we can be 99.99...% certain.
So that procedure will give us the middle or average potency and that alone would be enough of a starting point to create a scale of "low, middle, high" potency through repeated trials.
Another starting point would be to use the known molar masses of the substances in question -- in this case, psilocybin/psilocin and p-DMAB -- and the fact that the reagent's color-reaction is the result of each molecule of p-DMAB binding to two indole molecules (psilocybin/psilocin) to work out the upper and lower limits of the reaction with given weights of both materials.
I think the statistical approach will be more practical -- but we'll see Currently I have p-DMAB, as well as a few different acids to make different forms of the reagent, but I am waiting on an online order of phosphoric acid because my research leads me to believe that using phosphoric acid and methanol will give me the most sensitive reagent to work with. Once that arrives, I plan to proceed along the statistical approach I outlined above.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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joze



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Quote:
SpectreOfCommunism said:
Quote:
joze said: I'm also curious: what are y'all planning on using as a reference? The Miraculix kit has a scale they've developed themselves with their specific reagent. You're talking about determining the potency of the mushrooms, which is essentially the same as talking about the concentration of psilocybin/psilocin. Okay, so you can test your mushrooms with this reagent and measure the color, but what are you going to compare the value to? Unless you are able to source a relatively pure sample of psilocybin or psilocin in a known concentration, you will be limited to comparing relative potencies of mushrooms rather than measuring the actual concentrations.
Firstly, I'd say -- for the purposes of most of us anyway -- even being able to consistently tell whether this sample is relatively low-potency, this one average, and this one high-potency would be a pretty heckin' useful test.
And secondly, concerning a reference, I can think of a few approaches that don't involve already having any known quantities of psilocybin.
One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder. Because this powder is a randomized mixture composed of thousands of individual fruits representing thousands of cubensis strains, we can be all but certain that samples taken therefrom will be very close to the average potency -- and if our analysis shows the same results after multiple small samples (e.g. 150mg), we can be 99.99...% certain.
So that procedure will give us the middle or average potency and that alone would be enough of a starting point to create a scale of "low, middle, high" potency through repeated trials.
Another starting point would be to use the known molar masses of the substances in question -- in this case, psilocybin/psilocin and p-DMAB -- and the fact that the reagent's color-reaction is the result of each molecule of p-DMAB binding to two indole molecules (psilocybin/psilocin) to work out the upper and lower limits of the reaction with given weights of both materials.
I think the statistical approach will be more practical -- but we'll see Currently I have p-DMAB, as well as a few different acids to make different forms of the reagent, but I am waiting on an online order of phosphoric acid because my research leads me to believe that using phosphoric acid and methanol will give me the most sensitive reagent to work with. Once that arrives, I plan to proceed along the statistical approach I outlined above.
That would be a good method of defining your average potency and then testing cultures to see whether they are above-average or below-average. I'm curious to see how your trials turn out. Another question, how are you all planning to measure your data? Do you have a colorimeter or something? Or are you just eyeballing shit?
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27721860 - 04/05/22 08:38 AM (1 year, 9 months ago) |
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I was planning on eyeballing it, definitely using my camera as well. Reagents tend to change color and/or darken with time, so timing the reaction would be essential. Colorimeter would be ideal, but this *future* tek is targeted for those with limited resources. Also- I don't see why you couldn't take a sample with an averaged, known value (for example Pan Cyanenscens has 2.5% psilocybin and 1.194% psilocin so ~3.69% [with ~10% variability] alkaloid content) and test the same fruit yourself, recording the color.
Edited by shr00med (04/05/22 08:49 AM)
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joze



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Quote:
shr00med said: I was planning on eyeballing it, definitely using my camera as well. Reagents tend to change color and/or darken with time, so timing the reaction would be essential. Colorimeter would be ideal, but this *future* tek is targeted for those with limited resources. Also- I don't see why you couldn't take a sample with an averaged, known value (for example Pan Cyanenscens has 2.5% psilocybin and 1.194% psilocin so ~3.69% [with ~10% variability] alkaloid content) and test the same fruit yourself, recording the color.
Do you not see the problem with what you're saying?
You're wanting to use this reagent to accurately measure alkaloid content, because psychedelic mushrooms can vary in their potency. But for your reference, you want to use a psychedelic mushroom, because you think you can assume its alkaloid content... If you're so confident that each species has <10% variability, then what's the point in testing?
I will go to my death bed arguing that if you want to use 1) a DIY reagent with slightly variable concentrations and 2) eyeball your results, you will need a better reference sample to obtain accurate results.
Quote:
SpectreOfCommunism said: One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder. Because this powder is a randomized mixture composed of thousands of individual fruits representing thousands of cubensis strains, we can be all but certain that samples taken therefrom will be very close to the average potency -- and if our analysis shows the same results after multiple small samples (e.g. 150mg), we can be 99.99...% certain.
One possibility for obtaining a better reference actually comes from what SpectreofCommunism said above. If you made a homogenized batch of a bunch of powdered mushroom, you could then store that as a "reference sample" jar as long as you store it in a way that the alkaloids don't significantly degrade over time. If you got a Miraculix test kit (since we actually know these ARE precise), you could measure the alkaloid content of the reference sample with both Miraculix as well as your DIY reagent multiple times. If your DIY reagent gave you relatively consistent results, you could correlate that color change to the specific concentration that the Miraculix results gave you.
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 2
#27722219 - 04/05/22 03:39 PM (1 year, 9 months ago) |
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Quote:
shr00med said: Also- I don't see why you couldn't take a sample with an averaged, known value (for example Pan Cyanenscens has 2.5% psilocybin and 1.194% psilocin so ~3.69% [with ~10% variability] alkaloid content) and test the same fruit yourself, recording the color.
The content varies a whole lot more than that - every species varies by at least 50%, maybe more.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27722515 - 04/05/22 08:00 PM (1 year, 9 months ago) |
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joze said:
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SpectreOfCommunism said:
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joze said: I'm also curious: what are y'all planning on using as a reference? The Miraculix kit has a scale they've developed themselves with their specific reagent. You're talking about determining the potency of the mushrooms, which is essentially the same as talking about the concentration of psilocybin/psilocin. Okay, so you can test your mushrooms with this reagent and measure the color, but what are you going to compare the value to? Unless you are able to source a relatively pure sample of psilocybin or psilocin in a known concentration, you will be limited to comparing relative potencies of mushrooms rather than measuring the actual concentrations.
Firstly, I'd say -- for the purposes of most of us anyway -- even being able to consistently tell whether this sample is relatively low-potency, this one average, and this one high-potency would be a pretty heckin' useful test.
And secondly, concerning a reference, I can think of a few approaches that don't involve already having any known quantities of psilocybin.
One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder. Because this powder is a randomized mixture composed of thousands of individual fruits representing thousands of cubensis strains, we can be all but certain that samples taken therefrom will be very close to the average potency -- and if our analysis shows the same results after multiple small samples (e.g. 150mg), we can be 99.99...% certain.
So that procedure will give us the middle or average potency and that alone would be enough of a starting point to create a scale of "low, middle, high" potency through repeated trials.
Another starting point would be to use the known molar masses of the substances in question -- in this case, psilocybin/psilocin and p-DMAB -- and the fact that the reagent's color-reaction is the result of each molecule of p-DMAB binding to two indole molecules (psilocybin/psilocin) to work out the upper and lower limits of the reaction with given weights of both materials.
I think the statistical approach will be more practical -- but we'll see Currently I have p-DMAB, as well as a few different acids to make different forms of the reagent, but I am waiting on an online order of phosphoric acid because my research leads me to believe that using phosphoric acid and methanol will give me the most sensitive reagent to work with. Once that arrives, I plan to proceed along the statistical approach I outlined above.
That would be a good method of defining your average potency and then testing cultures to see whether they are above-average or below-average. I'm curious to see how your trials turn out. Another question, how are you all planning to measure your data? Do you have a colorimeter or something? Or are you just eyeballing shit?
My hope is that, once all is said and done, my hypothetical test will be eyeball-able but, during my work of calibrating the test, I plan to use a combination of taking very consistent photos against the same background and photo-editing software that will allow me to sample the colors in a digitally consistent way in order to devise a colorimetric scale
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Alan Rockefeller
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Quote:
SpectreOfCommunism said: My hope is that, once all is said and done, my hypothetical test will be eyeball-able but, during my work of calibrating the test, I plan to use a combination of taking very consistent photos against the same background and photo-editing software that will allow me to sample the colors in a digitally consistent way in order to devise a colorimetric scale
Make sure you use manual mode on the camera, with the camera and sample in the exact same place and with the exact same lighting, otherwise the automatic exposure and auto white balance will affect the colors.
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Re: DIY Psilocybin Content Percentage Detection [Re: Alan Rockefeller] 4
#27753993 - 04/27/22 08:01 PM (1 year, 8 months ago) |
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I've got an update for you all I've officially begun experimenting to find a procedure like the one I've outlined earlier in this thread and my initial results are very promising. To recap for anyone who hasn't followed closely, my goal is to design a DIY colorimetric test that will allow the average Shroomerite to determine the potency of their magic mushrooms using a reagent (p-DMAB) and a color-scale much like those little pH test-strips people use for pools and fish-tanks.
Anyway, without further ado, here is a quick write-up of my procedure so far:
For my starting material, I prepared 78g of fine powder by grinding a gallon-sized ziplock bag of cracker-dry cubensis specimens representing hundreds (if not thousands) of different strains grown from MS syringes of 4 well-known varieties* under the same conditions on brown rice-flour substrate. As I've explained earlier in this thread, this procedure ensures that the starting material is very close to the overall average potency of cubensis.
For my reagent solution, I dissolved 5g of p-dimethylaminobenzaldehyde (p-DMAB) into 100ml of concentrated phosphoric acid (85%) which was added slowly** to 100ml methanol (99.9%) to achieve a 1:1 solution. The reagent solution was then stored in an amber bottle at room temperature.
For the initial run, I carefully weighed out 200, 100, 50, 25, and 12.5mg samples in five small glass jars and 5ml of boiling water was added to each of them. The jars were then placed into a hot water bath for 15 minutes to extract the alkaloids. Meanwhile, a pipette was used to dispense 3ml of reagent solution to five standard-size 2-dram vials. After 15 minutes, a 1ml syringe was used to draw 1ml of each extract through a cotton ball in order to filter out the powdered mushroom material*** before being added to the reagent vials.
Results: After about 1 hour, a color reaction developed so that there is a clear gradient-scale from vial #1 (200mg) to #2 (100mg) to #3 (50mg). Vials #4 and #5 (25mg and 12.5mg) show little to no evidence of a reaction.

Although further trials are obviously needed in order to "calibrate" the test so that a useful gradient-scale can be devised, these results show that the concept is likely workable.
In order to become a useful test, the next challenges to overcome are (1) to increase the intensity and optimize the visibility of the color-reaction, (2) to establish the upper and lower limits of the color-reaction so that a gradient-scale can be devised, and (3) to establish the most useful amounts of both the reagent and extraction solutions so that samples of average potency will result in a color corresponding to the middle of the gradient-scale.
Regarding (1), there are a number of possible solutions to the problem — for example, the amount of extract added to the reagent could be increased, i.e. using 2 or 3ml rather than 1ml, without needing to use greater amounts of mushroom material. Or alternatively, a more efficient solvent like methanol could be used. Another avenue to explore would be to react the two solutions in a hot-water bath instead of at room-temperature.
As to (2) and (3), simple trial and error should be enough to overcome these problems assuming (1) has first been solved.
And lastly, I'd love to hear from you all if you have any constructive criticism or suggestions about how to proceed from here 
Notes: *Mazatapec, B+, Huautla, and Treasure Coast **This is an important step because adding a strong acid to methanol results in an exothermic reaction that produces a lot of heat ***Syringe filters would be ideal, but hey ya gotta use what you got
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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SpectreOfCommunism
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Addendum: I want to clarify that the intended goal of this first trial run was not to have a fully functional and useful quantitative test but rather to establish some starting values to work with in subsequent trials and particularly to establish a lower limit of sensitivity for the reagent solution. (Which this has accomplished.)
I also want to add that the whole bill for the materials to do this experiment and many more similar experiments has only been about $75 so far. Ehrlich‘s reagent is available on Amazon for about $30, a liter of concentrated phosphoric acid is available for about $20, and methanol (although it is pretty hard to track down) is only about $5 per gallon. The 2-dram vials I used are about 25$ for a 100-pack. And these materials are enough to do thousands of these types of tests.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Addendum Part II: The Reckoning: Tonight I plan to run two more experiments and, in order to isolate variables effectively, I will change only one parameter at a time. For the first run, I will be repeating the procedure except that I will add 2ml (rather than 1ml) out of the 5ml of extract to the reagent solution. My expectation is that the color-reaction will be stronger than in the initial experiment. And for the second run, I will repeat the initial procedure except that I will react the sample mixture in a hot-water bath by bringing water to a boil and then shutting off the burner before nesting the vials in the pan. I expect that this will result in a faster reaction and possibly increase the intensity of the final color.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27754393 - 04/28/22 12:12 AM (1 year, 8 months ago) |
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Good ideas here. (except the new guy who thought that developing a DIY testing process is harming some random dude's IP, haha.)
If you want to try to make a color test chart that can be used by others, as A-Rock said, turn off the auto functions on camera. Also should white balance camera, and mention make and model of camera, plus color temp of light. At least what the light is sold as.
Also would be good to include a photo using that camera / settings/ lighting of a standard color checker card like this https://www.ebay.com/itm/224566675111?hash=item34493786a7:g:jMQAAOSwJFhhEhzw
I'll get some Erlich's when I get paid in a couple weeks. I certainly have some strong clone shroom and some weaker MS shroom I could start by comparing.
I do predict that the price of milligram-accurate lab tests will go down, especially with the Oregon therapeutic protocols requiring testing (potency, plus microbials) of every batch not over 5 pounds of shrooms.
If someone developed an accurate mail-in TLC or other lab test that's 10 dollars instead of 100 dollars, they'd be flooded with customers from day one until forever.
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Edited by nektar61 (05/01/22 12:32 AM)
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27754400 - 04/28/22 12:29 AM (1 year, 8 months ago) |
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nektar61 said: Good ideas here. (except the new guy who thought that developing a DIY testing process is harming some random dude's IP, haha.
If you want to try to make a color test chart that can be used by others, as A-Rock said, turn off the auto functions on camera. Also should white balance camera, and mention make and model of camera, plus color temp of light. At least what the light is sold as.
Also would be good to include a photo using that camera / settings/ lighting of a standard color checker card like this https://www.ebay.com/itm/224566675111?hash=item34493786a7:g:jMQAAOSwJFhhEhzw
All good suggestions and I fully intend to be rigorous and transparent about camera settings, light, etc once I reach the endgame-stage where I'm devising a gradient-scale for the use of everyone -- I didn't bother with that rigor this time around because this was just the initial stage where I chose a somewhat arbitrary starting point to start gathering data. Once I've trial'd and error'd enough to determine the most useful ratios of reagents, solvents, etc, to position my averaged sample at the center of the colorimetric reaction, then the last phase (hopefully inshallah) will be to take some very scientifically-minded photos lol -- but these photos are just "process" photos to let those following the thread in on what's happening over here
As I write this sentence, I am in the process of observing round #2 and #3 I'll probably have another round of photos before the night is out
Thanks for your engagement and thoughtful response
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nektar61
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Quote:
SpectreOfCommunism said: ...but these photos are just "process" photos to let those following the thread in on what's happening over here
...Thanks for your engagement and thoughtful response 
One suggestion I'd have is maybe double the amount of test shrooms or double regent perhaps. (or both?)
I think brighter results would have more meaning and be easier to discern differences. At quick glance the one on the far left is the only one that looks like much of a reaction in your tests.
Maybe double the amount of DMAB in your reagent but keep the amount of ethanol and HCL same as you've been using?
Does anyone here know if filtering out the powdered mushroom material is needed, is it used in commercial kits? I doubt it's needed in the field tests cops use for LSD, which only indicate presence, and maybe not amount, but would work on a much smaller amount, micrograms of indole compound compared to the milligrams with shrooms.
Thank you for jumping in feet first on this. I was happy to see this thread, I was wondering the same thing recently.
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Edited by nektar61 (04/28/22 02:17 AM)
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27754542 - 04/28/22 05:34 AM (1 year, 8 months ago) |
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Ya I'm confused as to why you essentially made a tea prior to the test. Imo your pulling out pigmentation that is going to variabloze it most likely. Seems as though the reactions are even somewhat tea colored..
And actually idk why you would even do that, because what your doing is adding a bunch of water, 33-25% depending on how your looking at it, the mushroom material could have been added powdered..?
This should keep the reagent more colorful, and idk, not a chemistry guy, but adding that much water to a solution may make it generally less effective.
I don't think filtering is necessary.
Lol nektar, double the shrooms or reagent or both? Do both your right back where you started haha.
And that color card looks like it belongs on the third episode of Barney.
Your probably going to need to see what color is your primary color and see what kind of shades your getting, and probably much more intentionally make your own scale.
I think your onto something.
And why would you need or want elrich for this? I thought the whole point was to make your own reagent for much cheaper and then make your own scale that would be completely different than elrich?
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Re: DIY Psilocybin Content Percentage Detection [Re: QM33] 1
#27755098 - 04/28/22 01:11 PM (1 year, 8 months ago) |
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QM33 said: Ya I'm confused as to why you essentially made a tea prior to the test. Imo your pulling out pigmentation that is going to variabloze it most likely. Seems as though the reactions are even somewhat tea colored..
And actually idk why you would even do that, because what your doing is adding a bunch of water, 33-25% depending on how your looking at it, the mushroom material could have been added powdered..?
I’m doing so for several reasons — firstly, because we know that psilocybin and it’s relatives are polar and water-soluble and so a water-extraction contains the only thing we’re concerned with testing. Secondly, the DMAB color-reaction is an aqueous reaction and so the alkaloids need to be dissolved one way or another — doing so in a known quantity of a good solvent for a controlled duration is a good approach which is repeatable while also removing >95% of the non-alkaloid content.
And lastly, because the Miraculix kit also works by making an extract of the mushroom powder with boiling water and, since Miraculix is the only example I have of a functioning colorimetric indole-test, that seems like a good place to start.
I don’t think pigments are going to be an issue and I’m not sure how a water-extract would add more of them than adding the powdered mushroom directly as you suggest — why would the extract have more pigment than the thing it was extracted from itself?
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QM33 said: I don't think filtering is necessary.
I think it is. Without filtering, there would be different proportions of rehydrated mushroom-mush to water-extract in every test which would be impossible to know or control. Also, as you already pointed out, this would add a lot of confounding colors to the reagent which would muddy the reaction. Plus, the mushroom material will clog the syringe without the cotton ball.
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QM33 said: And why would you need or want elrich for this? I thought the whole point was to make your own reagent for much cheaper and then make your own scale that would be completely different than elrich?
My goal is to design the cheapest, most effective, most accessible DIY colorimetric test possible. And since 100g of DMAB (enough to make about 8 liters of concentrated reagent) only costs as much as an OK bottle of whisky, the cost and effort to synthesize the reagent at home doesn’t seem justifiable
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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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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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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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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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.
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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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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!
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joze



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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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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?
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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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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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nektar61
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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."
-------------------- -NEW? Start here.
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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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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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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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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27757822 - 04/30/22 04:16 PM (1 year, 8 months ago) |
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Quote:
nektar61 said: 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.
Not sure where you’re getting your info on dry mushroom powder degrading with time — I see no reason why powdered dried mushrooms would have any difference in shelf-life from non-powdered dried mushrooms. You’re correct about wet mushrooms or wet mushroom powder — those will be inactive before long.
I explained earlier in this thread why it’s necessary to prepare a large batch of mushroom powder:
Quote:
One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder. Because this powder is a randomized mixture composed of thousands of individual fruits representing thousands of cubensis strains, we can be all but certain that samples taken therefrom will be very close to the average potency -- and if our analysis shows the same results after multiple small samples (e.g. 150mg), we can be 99.99...% certain.
So that procedure will give us the middle or average potency and that alone would be enough of a starting point to create a scale of "low, middle, high" potency through repeated trials.
In short, the whole purpose of powdering several ounces of genetically diverse cubensis material at once is to create a sample of very precisely average potency without which all these experiments would proceed by groping along in the dark. So grinding small samples each day on the scale of milligrams would not just be more tedious but also rob the experiments of their value altogether
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nektar61
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Quote:
SpectreOfCommunism said: I explained earlier in this thread why it’s necessary to prepare a large batch of mushroom powder: .... One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder.
I'd just use a clone to keep more even consistency.
FWIW, my comments here aren't directed at only you, but are more thinking out loud with the readers of the thread. I'm spitballing for anyone doing this, and am taking notes with what people bring to it.
As for grinding reducing potency, I don't have proof of this, I don't have a test for potency...(that's why I'm on this thread)
But I've heard over and over that grinding increases surface area which increases oxidation which increases psilocin in shrooms degrading to non-actives.
Interesting aside (and maybe somehow a hint toward finding the perfect DIY test, and maybe speaking to my thought of making a fresh sample each time), the bluing in bruised shrooms involves as you said, (an enzyme) turning psilocybin into psilocin, and another enzyme turning psilocin into a bluing...but the bluing is a longer chain that is close chemically to the chemical in Indigo dye that produces the bright blue.
If you make an ethanol (Everclear) extract of powdered shrooms, then a water extract, mix them, let it dehydrate a bit with a fan on it, it is BRIGHT blue, almost magical looking. For a little while. Within an hour or two, it's brown, and the potency goes down. I've done it.
Would be a nice party trick to do that and serve the blue liquid, maybe from a chalice if someone were into that. But even in paper cups, it's stunning looking.
I wonder if the test we seek is inbuilt...maybe making a slurry of shrooms and comparing the immediate blue is the answer, or part of the answer. Maybe especially if the shrooms are fresh. (Though a test needs to be able to test dried shrooms.)
Or maybe the proper test would use something that would imitate the enzymes in the shrooms, but have more of them than shrooms have.
-------------------- -NEW? Start here.
Edited by nektar61 (04/30/22 05:03 PM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27757878 - 04/30/22 04:57 PM (1 year, 8 months ago) |
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Quote:
nektar61 said:
Quote:
SpectreOfCommunism said: I explained earlier in this thread why it’s necessary to prepare a large batch of mushroom powder: .... One possibility (and the one I'm currently favoring) is to use the statistical principle of large numbers -- say I take a few ounces of dried samples grown from multispore inoculates of multiple varieties and grind it all to a uniformly mixed powder.
I'd just use a clone to keep consistency more even.
As for grinding reducing potency, I don't have proof of this, I don't have a test for potency...(that's why I'm on this thread)
But I've heard over and over that grinding increases surface area which increases oxidation which increases psilocin in shrooms degrading to non-actives.
Interesting aside (and maybe somehow a hint toward finding the perfect DIY test, and maybe speaking to my thought of making a fresh sample each time), the bluing in bruised shrooms involves as you said, (an enzyme) turning psilocybin into psilocin, and another enzyme turning psilocin into a bluing...but the bluing is a longer chain that is close chemically to the chemical in Indago that produces the bright blue.
If you make an ethanol (Everclear) extract of powdered shrooms, then a water extract, mix them, let it dehydrate a bit with a fan on it, it is BRIGHT blue, almost magical looking. For a few hours. within 8 hours it's brown, and the potency goes down. I've done it.
Would be a nice party trick to do that and serve the blue liquid, maybe from a chalice if someone were into that. But even in paper cups, it's stunning looking.
Yes, in any aqueous solution, psilocybin will be rapidly converted to psilocin which is very unstable and will oxidize your to pretty blue inactive substance — but again, that requires water and there is no more water in a powdered dried mushroom than in a non-powdered dried mushroom. The enzymatic dephosphorylation of psilocybin requires water to occur which is why we dry our mushrooms as fast as possible — the faster you dry them, the fewer actives are converted to inactives.
Anyway, i’d be happy to take a look at any evidence to the contrary but, as someone who has been hunting and growing and eating these mushrooms for decades, I can tell you with 100% certainty that I have never seen any evidence that powder dried mushrooms are any less potent than non-powdered dried mushrooms. And from a chemistry perspective, I can’t think of any reason why psilocybin co tent would be impacted by grinding the dried material. I mean think about it – the mortar and pestle is basically the symbol of herbalism and alchemy throughout history. If grinding up herbs and mushrooms harmed the medicines locked in their cells, then I would think the mortar and pestle would be fairly anathema within the chemical sciences by now
And concerning the clone idea, yes that would be consistent but here would be no way to know how potent it was relative to the average potency. The measurements would be consistent but they would only be consistent relative to themselves and not to the overall range of potencies observable in cubensis — we’d be right back where we started, with unknown (albeit consistently unknown) sample potencies. With the statistical approach of using large sample sizes, we can be certain that we are measuring average samples which gives us a starting point that we can use to construct other non-average samples
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Icyurmt
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 The chart above is for mushroom powder; unless it is well protected from oxygen and light, it can lose a lot of potency very quickly. This is the study thats from, it looks at tryptamine loss in mushrooms under various conditions as well as the effects of drying. PDF Stability of psilocybin and its four analogs in the biomass of the psychotropic mushroom Psilocybe cubensis
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
Edited by Icyurmt (04/30/22 05:09 PM)
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27757898 - 04/30/22 05:11 PM (1 year, 8 months ago) |
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Quote:
Icyurmt said: The chart above is for mushroom powder; unless it is well protected from oxygen and light, it can lose a lot of potency very quickly.
Well there we go.
It's possible that SpectreOfCommunism's contrary results can be human vs chemical (makes sense, if human brain was a good test, we wouldn't need a chemical test.)
Or SpectreOfCommunism's shrooms had a lot of psilocybin and not much psilocin, so grinding didn't degrade much.
-- All:
According to: https://www.swgdrug.org/Monographs/PSILOCIN.pdf (use a VPN, that's a cop site, "Scientific Working Group for the Analysis of Seized Drugs" haha):
Psilocin is only slightly soluble in water. Psilocybin is soluble in water.
and Psilocin is soluble in methanol. Psilocybin is only slightly soluble in methanol.
So maybe methanol isn't a good solution to use. Or am I missing something and the acid takes care of that or something with commercial tests and with what SpectreOfCommunism is trying? This thread is getting harder to navigate, which is good, means it's getting a lot of response. I really think this is a "throw the crowd at it" problem.
It's weird that most commercial tests and cop tests seem to be using methanol, but that might mean the mainly end up only testing the Psilocin and are not even testing much of the Psilocybin in a given sample.
I remember the reason I used ethanol and then water for my "magic blue chalice liquid" was somewhere I found that Psilocin dissolves well in one and Psilocybin dissolves well in the other.
There is some info in that link above about color change results for yet more reagents with Psilocybin and Psilocin that might help with all this:

I'm thinking maybe the PsiloQ test may be mostly Marquis reagent (simple to make: "a mixture of formaldehyde and concentrated sulfuric acid, which is dripped onto the substance being tested.") https://en.wikipedia.org/wiki/Marquis_reagent
It turns greenish-brown for Psilocin, and dull orange for Psilocybin. Both together would likely be brown
"The United States Department of Justice method for producing Marquis reagent is the addition of 100 mL of concentrated (95–98%) sulfuric acid to 5 mL of 40% formaldehyde.[1]: 12 Different compounds produce different color reactions. Methanol may be added to slow down the reaction process to allow better observation of the colour change."
One advantage we have for the test we're trying to make is it only has to test for two things. It doesn't have to test for adulterants or other drugs like a street drug test.
-------------------- -NEW? Start here.
Edited by nektar61 (04/30/22 05:31 PM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 3
#27757940 - 04/30/22 05:43 PM (1 year, 8 months ago) |
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Methinks you guys are misinterpreting that study. From the study:
Quote:
“We found that the dried fungal fruiting bodies had a better yield from a fungal powder than from whole pieces. Conversely, in the case of fresh fungal fruiting bodies, the extraction of larger unprocessed pieces of fresh fruiting bodies was found to be more effective than chopping these mushrooms into small pieces”
The researchers say that for dried specimens they got “a better yield from a fungal powder than from whole pieces” and that the opposite was the case for fresh (i.e. water-containing) mushrooms, which agrees exactly with what I’ve said.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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SpectreOfCommunism
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Also from that study:
Quote:
In summary, the yields of the analytes from the dried mushroom powder in comparison with the unprocessed dried mushrooms were as follows: 16% increase in psilocybin, 13% increase in psilocin, 74% increase in baeocystin, and 40% increase in aeruginascin.
And as for the degradation of tryptamines in powder form, they’re talking about this degradation over the course of 15 months, particularly when left in the light in the presence of oxygen. Which, yeah, that’s a silly way to store them.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Icyurmt
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Quote:
Methinks you guys are misinterpreting that study. From the study: Quote:
Quote:
The researchers say that for dried specimens they got “a better yield from a fungal powder than from whole pieces” and that the opposite was the case for fresh (i.e. water-containing) mushrooms, which agrees exactly with what I’ve said.
That's from the section 3.1 that was looking specifically at extraction efficiency, not storage effects. That says that it's more efficient to extract from powdered than it is from whole dry pieces, which I don't think anybody was disagreeing with. The storage of fresh mushrooms is covered in section 3.6.
Quote:
And as for the degradation of tryptamines in powder form, they’re talking about this degradation over the course of 15 months, particularly when left in the light in the presence of oxygen.
And as shown on the chart above and in section 3.7, approximately half the initial potency was lost within only one month when the powered mushrooms were stored, even for those in a bag in the dark.
] From the conclusion.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
Edited by Icyurmt (04/30/22 07:36 PM)
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27758067 - 04/30/22 08:17 PM (1 year, 8 months ago) |
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Either way, it’s an interesting paper — it’s unfortunate that they don’t include a comparison of potency loss over time between whole dried mushrooms and powdered dried mushrooms. Like, I recognize that powder dried mushrooms tend to lose potency over time just like every other form of mushrooms but drastic potency loss after more than a year seems to be normal compared to whole dried mushrooms. A few searches on here and on the internet at large seems to indicate that the general consensus is that properly dried and stored mushrooms will last about a year — although my own experience is that they last longer. But I vacuum pack mine, so maybe the lack of oxygen in the packaging has served me well idk — I know I’ve eaten three year old cyanescens that seemed as potent as the day I dried them
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 2
#27758075 - 04/30/22 08:25 PM (1 year, 8 months ago) |
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Quote:
Icyurmt said: And as shown on the chart above and in section 3.7, approximately half the initial potency was lost within only one month when the powered mushrooms were stored, even for those in a bag in the dark.
right.
There is some first-hand (non-scientific) info on Shroomery of someone storing non-ground shrooms in a hot garage for 10 years and still having them be very strong.
Forrester's quote here, in the vacuum pack and long-term storage thread: https://www.shroomery.org/forums/showflat.php/Number/27660424#27660424
Seems to imply that a warm garage is less damaging to potency than air is. I'll ask him to clarify. (He did not vacuum pack, he stored in a jar, but did not grind the shrooms.)
I've probably seen 30 people on here recommend against grinding shrooms for storage....all say it degrades potency quicker.
I'm going to do my testing using a clone (x7x plin clone I use for everything, it's the main thing I grow now). And only stems or only caps, since stems are supposed to be a little stronger than caps. I'll leave it un-ground until I test.
I think clones make sense over grinding, not just because grinding degrades, but because we're testing tiny amounts, and even a ground mix of wildly varying potency ms shrooms could have noticeably different amounts of actives a few cm apart in the same bag. --
Quote:
SpectreOfCommunism said: although my own experience is that they last longer. But I vacuum pack mine, so maybe the lack of oxygen in the packaging has served me well idk.. 
Yes. I also vacuum pack and have started a ten-year study (with clones). Vacuum packing is about the opposite of grinding.
Think about activated charcoal. It's ground fine to increase surface area, to increase absorption. When you grind shrooms you're vastly increasing the amount of surface area. =-- Intersting that article mentions shrooms losing potency in freezer. But they meniton -80 c. Who the fuck has a freezer that goes to that (-112 f). I guess labs do.
I store live cultures in a dedicated dorm fridge for 6 plus months (and counting) at 2 c (about 35 f), I assume you could store dried shrooms at that.
I think the problem with freezing or even cold is that you draw water in.
I'm intrigued by the "inert gas" comment, I was thinking that the other day and wondering about buying a tank of argon. Is about 300 bucks.
There isn't a lot of info on long term storage of shrooms, but from what I've heard, best bet is maybe vacuum sealed, in the dark, at room temp.
-------------------- -NEW? Start here.
Edited by nektar61 (04/30/22 09:10 PM)
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27758118 - 04/30/22 09:06 PM (1 year, 8 months ago) |
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Quote:
it’s unfortunate that they don’t include a comparison of potency loss over time between whole dried mushrooms and powdered dried mushrooms.
They did look at mostly whole mushrooms stored for 3 months in the dark at room temp in section 3.6. I believe. Those apparently showed little to no loss during that time.
I think you would probably still want to grind/homogenize before testing with clones just because of the variation that can occur in different parts like cap and stripe. Doing so closer to the time of testing though would make the most sense imo.
Quote:
I'm intrigued by the "inert gas" comment, I was thinking that the other day and wondering about buying a tank of argon. Is about 300 bucks.
I think either buying of making something like this purgeable storage box, out of an otterbox or pelican case, would be the most ideal for long-term storage. Tig welders will often have argon, so if you know anyone who welds you can probably convince them to fill up a few balloons for you.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27758122 - 04/30/22 09:09 PM (1 year, 8 months ago) |
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Nektar, what are you going to compare your clone material to? How do you get from “reagent plus mushroom results in color-reaction” to something you can quantify? My whole point with the grinding of many, many different mushrooms with different genetics is that, by doing so, probability theory and the law of large numbers guarantees that such a sample will contain the mathematical average quantity of alkaloids. This then allows us to extrapolate that 1/2 of a given sample will give us mushroom material that is 0.5x average potency, 2/1 of the same sample will give us mushroom material that is 2.0x the average potency, etc and so on, which gives us a way to devise a quantitative scale that can be re-used.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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SpectreOfCommunism
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Icyurmt, 3.6 deals with “fresh fruiting bodies” not whole dried mushrooms. I’ve read the thing a few times now and I can’t find any part where they compare potency loss between whole dried and powdered dried mushrooms. Additionally, all the comparison tables seem to indicate that they’re only comparing the storage of dry powder and fresh fruiting bodies under various temperature and lighting conditions
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nektar61
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Quote:
SpectreOfCommunism said: Nektar, what are you going to compare your clone material to? How do you get from “reagent plus mushroom results in color-reaction” to something you can quantify? My whole point with the grinding of many, many different mushrooms with different genetics is that, by doing so, probability theory and the law of large numbers guarantees that such a sample will contain the mathematical average quantity of alkaloids. This then allows us to extrapolate that 1/2 of a given sample will give us mushroom material that is 0.5x average potency, 2/1 of the same sample will give us mushroom material that is 2.0x the average potency, etc and so on, which gives us a way to devise a quantitative scale that can be re-used.
A lab would order a small known sample of each chemical. We can't do that, so....
I haven't gotten to that point yet. I've ordered a couple reagents. I will test using a dried clone harvested in the past month or two, to try to get rough quantitative color variation results, then test again to confirm it's repeatable.
I'd consider it a win if I can get a different visible color comparison result between testing 150 mg and testing 300 mg dried of the same clone. Something that does that consistently would allow a profile to be formed. (unless doing that throws off the test and gives different results from the same amount of two samples where one is twice the potency.)
If I get really into it, maybe I'll send a sample of it off to a lab (like the one A-Rock) runs, pay the 100 dollars+ to find the actual percent quantity of that. That could get expensive quick though. I still don't think you'd have to test 6 or 7, I think you could test 1, and use 6 or 7 varying amounts of that shroom to test. If you knew the percent in the clone, that might work.
Or maybe ask some people in the Psilo Cup to send samples of their clones, since those have been tested with HPLC and the amounts are known.
So far I will just be looking for something that gives a reasonable color difference between say, a tiny bit, a little bit, a bit, and a lot. From there it can be fine tuned.
How do we know if the PsiloQ / Miraculix test even did the last step of taking the color chart they made, having HPLC lab tests on all the samples, and putting the correct percentages on the color chart?
I'm not saying they're scammy, it looks legit, but I think the calibration could be done with one shroom clone sample that has gone to a lab for tests.
They don't claim on the site that they tested, they only claim within 10% accuracy of HPLC lab tests. If I made and sold that test, and had done the lab tests to prove that amount, I'd provide the data in the marketing materials. Again, not saying they didn't, but why not?
Though maybe they did the tests, since Alan called the guy who runs it by his first name on here. haha. If you're on a first-name basis with A-Rock, you might just run a lab.
I'd be happy to just be able to tell weak from strong from super strong shrooms at home. That would be useful to me. And I bet to a lot of other people.
I'm using a clone because it makes sense. Using a mix of ground shrooms doesn't make sense to me. And I have seen enough people on this site post about doing an "experiment" then getting shot down by TCs (with good reason) because they didn't use a clone. Search "experiment" in advanced search and read the results.
I'm mainly interested in this for me to have the test. But I will share any of my results with the community. I don't want to produce data that the community will dismiss outright. -- Here's some of the info from what they claim, might be useful to review for anyone interested in working on a test:
Quote:
This test is performed in under 1/2 hour, and requires only 150mg of starting material.
--Fast evaluation after 30 minutes. --Detection range between 0.1 - 2 % active ingredient content. --Only 150 mg starting material is required. --10 % deviation from HPLC analysis. --Evaluation of the test kits with the naked eye and evaluation scale. --The PSILO-QTest for concentration determination is a single-use test.
INSTRUCTIONS
--Preparation: Crush dried mushroom material. Weigh out approx 150 mg of starting material. --Extraction: Add weighed starting material to the supplied extraction liquid (yellowish liquid in translucent plastic bottle) --Detection: Use the sterile syringe/needle to take up extraction solution, and add it to the detection solution (the clear liquid in glass bottle) --Evaluation: Allow color complex to develop (over about 1/2 hour) and evaluate with evaluation scale.
-------------------- -NEW? Start here.
Edited by nektar61 (04/30/22 10:12 PM)
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27758150 - 04/30/22 09:53 PM (1 year, 8 months ago) |
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Quote:
SpectreOfCommunism said: Icyurmt, 3.6 deals with “fresh fruiting bodies” not whole dried mushrooms. I’ve read the thing a few times now and I can’t find any part where they compare potency loss between whole dried and powdered dried mushrooms. Additionally, all the comparison tables seem to indicate that they’re only comparing the storage of dry powder and fresh fruiting bodies under various temperature and lighting conditions
Section 3.6 was looking for the optimal drying and storage conditions to maintain tryptamine content. The fresh fruiting bodies in group 2, were left at room temperature in the dark for 3 months (taken along with the comment in that section about the confirmation of drying not reduce the tryptamine content, I believe that implies they were allowed to naturally dry, as fresh fruits would not last 3 months without doing so on their own).
All of the comparison charts they give appear to be specific to each section they are from. The study does not do a direct comparison with the samples from section 3.6 and the dried fungal powder in 3.7, but they do report on the various concentrations/% found, so it's possible to compare those results with the concentrations/losses found with the powder from section 3.7.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
Edited by Icyurmt (04/30/22 10:38 PM)
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 2
#27758175 - 04/30/22 10:37 PM (1 year, 8 months ago) |
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If the PsiloQ test were a lot cheaper, I'd just buy it and wouldn't even be on this thread.
15 bucks for a one-use visual test with 90% accuracy seems expensive. It's probably 1 or 2 reagents mixed, maybe with some other common chemical. I'd wager it's a lot cheaper to make.
If it were the price of Reagents, like 30 bucks for 20 tests, (where the sellers still make a profit), I'd just buy it.
Kudos to the guy for cracking the code, but I think he's overpricing it.
Though with the Shroom-Boom ramping up I'm sure there are yuppies who'd buy crates full at his price.
-------------------- -NEW? Start here.
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27758194 - 04/30/22 10:58 PM (1 year, 8 months ago) |
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If not marquis perhaps it's mecke? Does anyone know what color mecke reagent turns when reacted with psilocybin/psilocin? I can't find any info when I looked but when reacting with DMT and lsd it looks very similar to the PsiloQ tests color. I know mecke also reacts with sugars so you would likely want to use an extract solvent that accounted for that if possible. Anyone know how soluble psilocybin/psilocin is in something like hexane or perhaps something better?
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27758198 - 04/30/22 11:00 PM (1 year, 8 months ago) |
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Quote:
Icyurmt said:
Quote:
SpectreOfCommunism said: Icyurmt, 3.6 deals with “fresh fruiting bodies” not whole dried mushrooms. I’ve read the thing a few times now and I can’t find any part where they compare potency loss between whole dried and powdered dried mushrooms. Additionally, all the comparison tables seem to indicate that they’re only comparing the storage of dry powder and fresh fruiting bodies under various temperature and lighting conditions
Section 3.6 was looking for the optimal drying and storage conditions to maintain tryptamine content. The fresh fruiting bodies in group 2, were left at room temperature in the dark for 3 months (taken along with the comment in that section, I believe that implies naturally dried, as fresh fruits would not last 3 months without doing so).
All of the comparison charts they give appear to be specific to each section they are from. The study does not do a direct comparison with the samples from section 3.6 and the dried fungal powder in 3.7, but they do report on the various concentrations/% found, so it's possible to compare those results with the concentrations/losses found with the powder from section 3.7.
I dunno, I'm not seeing/reading anything in there to suggest that they are comparing dried to dried and powdered mushrooms -- in the conclusion, they only mention dried-powdered versus fresh and, in all the comparison tables, they indicate dried-powdered versus fresh. There are a few passages where the wording is vague and so, out of context, it seems like they could be referring to dried whole mushrooms -- but it would be really weird if they left out 1/3 of the subjects they're comparing in all the tables and then go on to not even mention the 3rd thing being compared in the conclusion.
Unless they're referring to dried mushrooms as "fresh mushrooms" idk -- but then up is down and left is right and words have no meaning lol
-------------------- 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: Icyurmt] 2
#27758212 - 04/30/22 11:17 PM (1 year, 8 months ago) |
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Quote:
Icyurmt said: If not marquis perhaps it's mecke? Does anyone know what color mecke reagent turns when reacted with psilocybin/psilocin? I can't find any info when I looked but when reacting with DMT and lsd it looks very similar to the PsiloQ tests color. I know mecke also reacts with sugars so you would likely want to use an extract solvent that accounted for that if possible. Anyone know how soluble psilocybin/psilocin is in something like hexane or perhaps something better?
Mecke is actually my best guess for what Miraculix is doing, as well -- but I also can't find any info on Mecke and psilocybin. However, because it's reaction with other indoles like DMT seems to fit the color profile, it seems worth investigating
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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Icyurmt
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Quote:
Unless they're referring to dried mushrooms as "fresh mushrooms" idk -- but then up is down and left is right and words have no meaning lol
I think fresh mushrooms left out at room temperature for 3 months would soon become dried mushrooms all on their own, they all start out fresh and drying was a part of the "processing" they were looking at with the tests in that section. It also helps explain why they cite that test of group 2, as further support for their conclusion that drying in the dark at room temp does not cause a reduction of tryptamine content. The "preparation" of the fresh mushrooms is described at the beginning of that section and in section 2.2.
They report on the %/levels of various tryptamines found in those samples after 3 months of storage. The tests on the power did not look specifically after 3 months but instead after 1week, 1month, 2months, and 15months(different sections so different set of tests). Even so, a comparison of the levels found after one month as power vs 3 months mostly whole in the dark, can still be done from the data they provide.
-------------------- 👁️ 🌊 why you are empty. Hunt for the habitat not the mushroom.
Edited by Icyurmt (04/30/22 11:58 PM)
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt] 1
#27758239 - 04/30/22 11:48 PM (1 year, 8 months ago) |
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Quote:
Icyurmt said: If not marquis perhaps it's mecke?
I think it's Marquis. Might even be that and nothing else.
I ordered some marquis along with some Ehrlich.
I don't know if you saw the chart I posted on the previous page, but the two actives in shrooms, one turns Marquis brown and one turns yellow. Dark yellow liquids look brown.
--
Quote:
Icyurmt said: I know mecke also reacts with sugars so you would likely want to use an extract solvent that accounted for that if possible.?
One of the cop drug test links I posted earlier on this thread mentioned extracting with methanol then washing with acetone to remove sugars, but that was prepping for GC. https://www.swgdrug.org/Monographs/PSILOCIN.pdf
That page also mentions using Froehde’s Reagent. Turns yellow (eventually) for both alkeloids. Again, dark yellow liquids look brown.
The PsiloQ test produces brown.
"The Froehde reagent is used as a simple spot-test to presumptively identify alkaloids, especially opioids, as well as other compounds. It is composed of a mixture of molybdic acid or a molybdate salt dissolved in hot, concentrated sulfuric acid, which is then dripped onto the substance being tested.
"The United States Department of Justice method for producing the reagent is the addition of 100 ml of hot, concentrated (95–98%) sulfuric acid to 0.5 g of sodium molybdate or molybdic acid."
"The Virginia Department of Forensic Science method uses 0.5 g ammonium molybdate per 100 ml H2SO4 (conc.)[2]
"Unheated sulfuric acid can be used to prepare the reagent in a less dangerous manner, but 2–4 hours must be allowed for the molybdate to dissolve." https://en.wikipedia.org/wiki/Froehde_reagent
That's an interesting one. Turns a lot of different colors, can test presence of a lot of types of drugs. Look at that link on the wikipedia link.
I'm going by the guess that anything that can change color to detect presence of something might change color more in the presence of more of that thing. Some reagents maybe more than others.
-------------------- -NEW? Start here.
Edited by nektar61 (05/01/22 12:01 AM)
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Forrester
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27758372 - 05/01/22 04:54 AM (1 year, 8 months ago) |
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Quote:
nektar61 said: There is some first-hand (non-scientific) info on Shroomery of someone storing non-ground shrooms in a hot garage for 10 years and still having them be very strong.
Forrester's quote here, in the vacuum pack and long-term storage thread: https://www.shroomery.org/forums/showflat.php/Number/27660424#27660424
Seems to imply that a warm garage is less damaging to potency than air is. I'll ask him to clarify. (He did not vacuum pack, he stored in a jar, but did not grind the shrooms.)
I've probably seen 30 people on here recommend against grinding shrooms for storage....all say it degrades potency quicker.
Yeah I think it's pretty well accepted that it's oxidation that degrades the actives over time, not temperature. My experiment was very anecdotal of course, but during that ten years they were stored, the jars were moved several times between hot garages and basements (I lived in 3 different places over the period) so the temperature was far from stable. I think the chitin cell walls of un-ground fruit bodies keep the actives pretty well protected from oxidation, and wide swings in temperature didn't seem to cause any issues. Vacuum packing would obviously be an even better way to do it, to get the oxygen out of there, but I'm not sure how much difference it makes over the long term as long as the fruit bodies are intact.
If you're interested in the "purging with an inert gas" idea, you can always just buy one of these:
wine preserver
But I wouldn't waste the money for storing mushrooms. This stuff was recommended by a few on here years ago for storing other powders (RC's mostly) to keep them protected from oxidation. It would be interesting to see if it worked to help extend the shelf life of DMT freebase which is pretty short.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: Forrester] 1
#27760386 - 05/02/22 06:30 PM (1 year, 8 months ago) |
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I confirmed with Forrester via email, the shrooms that lasted 10 years in varying amounts of heat with no noticeable degradation of potency were NOT ground (as I suspected), they were whole.
They were not PE, probably B+. Someone else said PE on the long-term storage thread.
-------------------- -NEW? Start here.
Edited by nektar61 (05/02/22 08:18 PM)
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27760569 - 05/02/22 08:19 PM (1 year, 8 months ago) |
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Does anyone (A-Rock?) know if psilocin is soluble in Isopropyl alcohol?
I'm thinking 70% ISO would be better extraction solvent for testing shrooms, especially since growers have it on hand.
Methanol is VERY toxic, not something people who don't know chemistry should play with....at least not the wide group of people we'd be aiming for with an open-source grower's test.
The 30% water in 70% ISO would dissolve the Psilocybin, and I'm hoping the ISO would dissolve the psilocin.
Most chemistry sites that have profiles of chemicals list solubility in everything else: water, methanol, ethanol, acetone, ether. Not ISO.
Web search mostly points to Shroomery extractions. haha. This extraction said it made a strong goo, but could have just been https://www.shroomery.org/forums/showflat.php/Number/15982455
Most tests use methanol. According to this: https://www.swgdrug.org/Monographs/PSILOCIN.pdf
psilocin dissolves in methanol, but psilocybin is only slightly soluble in methanol. If that's the case, common tests are mostly only testing the psilocybin, not much of the psilocin.
I'm sure we want to test both.
Of course Everclear (ethanol and water) would work, and work well. But Iso alcohol 70% is what most of us already have on hand.
-------------------- -NEW? Start here.
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SpectreOfCommunism
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27761942 - 05/03/22 08:11 PM (1 year, 8 months ago) |
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Quote:
nektar61 said: Does anyone (A-Rock?) know if psilocin is soluble in Isopropyl alcohol?
I'm thinking 70% ISO would be better extraction solvent for testing shrooms, especially since growers have it on hand.
Methanol is VERY toxic, not something people who don't know chemistry should play with....at least not the wide group of people we'd be aiming for with an open-source grower's test.
The 30% water in 70% ISO would dissolve the Psilocybin, and I'm hoping the ISO would dissolve the psilocin.
Most chemistry sites that have profiles of chemicals list solubility in everything else: water, methanol, ethanol, acetone, ether. Not ISO.
Web search mostly points to Shroomery extractions. haha. This extraction said it made a strong goo, but could have just been https://www.shroomery.org/forums/showflat.php/Number/15982455
Most tests use methanol. According to this: https://www.swgdrug.org/Monographs/PSILOCIN.pdf
psilocin dissolves in methanol, but psilocybin is only slightly soluble in methanol. If that's the case, common tests are mostly only testing the psilocybin, not much of the psilocin.
I'm sure we want to test both.
Of course Everclear (ethanol and water) would work, and work well. But Iso alcohol 70% is what most of us already have on hand.
Psilocybin and psilocin are BOTH soluble in water and methanol. Both are strongly polar molecules that will dissolve in most polar solvents and will not dissolve in nonpolar solvents like petroleum ether, naphtha, etc. Think about it — if psilocin wasn’t soluble in water, then mushroom tea made from fresh mushrooms would be no more potent than mushroom tea made with 1/10 of that weight of dried mushrooms because fresh mushrooms are more potent primarily because of their psilocin content which is largely destroyed by the drying process. Also you can literally watch psilocin oxidize in an aqueous solution because that aqueous solution will turn blue. Psilocin is slightly MORE soluble than psilocybin in methanol and psilocybin is slightly MORE soluble than psilocin in water but they are both soluble in polar solvents in general.
Also, isopropyl alcohol is toxic too btw and ordinary people handle methanol all the time whenever they put gas-line antifreeze and water-removers like HEET into their gas tank. Or whenever people put winter-season wiper fluid in their cars. Or whenever people use denatured alcohol as a household solvent or paint cleaner. Just use gloves and don’t drink the stuff. Simple as that.
-------------------- Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus
 
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 3
#27764174 - 05/05/22 10:03 AM (1 year, 8 months ago) |
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Here's results of my first test. I'm saving these on my Journal. ----
First test, May 5, 2022, Ehrlich's Reagent
Crushed up 1.28 grams of penis envy. Was harvested 3 months ago, was dried in dehydrator for 24 hours. Is strong, per my eating a gram a while back. Is not clone, is multispore, will be using clone for further tests.
Crushed with spoon on a plate (I've ordered an electric grinder). Put in test tube, added 20 ml Isopropyl alcohol 70%. (measured with syringe.)
Let sit at room temp for 6 hours. Shook 10 shakes each every 2 hours. Used a coffee glass as a test tube holder:
Filtered with sieve and funnel. Result is mostly clear straw-colored liquid.
Poured some into 4 small vials. Shook test tube 5 shakes between each pour.
Vial 1 got .5 ml solution. vial 2 got 1 ml solution. vial 3 got 2 ml solution. vial 4 got 3 ml solution. (all measured with syringe.)
Topped each off with 70% ISO so they were all 3 ml.
Unless I'm missing something, I think this step is important, otherwise you'll get different dilutions of the purple color reaction. Shook each one 10 shakes.
Added 10 drops Ehrlich's Reagent to each vial.
Shook vials all together 10 shakes at start, and every 15 min, including at end.
After one hour, result is inconclusive / paradoxical:
Doesn't seem to be much difference in purple tint, if anything, seems stronger 1, 2, and 3, which all had less material than 4.
Somehow maybe that IS the result? I don't know.
Will retry this test again at some point with 10 ml ISO instead of 20, to get a stronger solution to see color better.
Next, I will try Marquis reagent. I have higher hopes for that one, and did even before this test, based on reading.
(The remaining shroom / ISO extract in the test tube got thrown out. I'm going to mix fresh for each test.)
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Edited by nektar61 (05/05/22 10:28 AM)
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27764314 - 05/05/22 12:20 PM (1 year, 8 months ago) |
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4 almost looks foggy no?
Maybe you maxed out the reagent.?
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27764456 - 05/05/22 01:51 PM (1 year, 8 months ago) |
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Quote:
nektar61 said: After one hour, result is inconclusive / paradoxical:
Doesn't seem to be much difference in purple tint, if anything, seems stronger 1, 2, and 3, which all had less material than 4.
Somehow maybe that IS the result? I don't know.
Will retry this test again at some point with 10 ml ISO instead of 20, to get a stronger solution to see color better.
Next, I will try Marquis reagent. I have higher hopes for that one, and did even before this test, based on reading.
(The remaining shroom / ISO extract in the test tube got thrown out. I'm going to mix fresh for each test.)
You're probably experiencing difficulties due to the alkalinity of the mushrooms. See SpectreOfCommunism's post I quoted below.
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…
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27765058 - 05/05/22 08:11 PM (1 year, 8 months ago) |
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Quote:
joze said:
You're probably experiencing difficulties due to the alkalinity of the mushrooms. See SpectreOfCommunism's post I quoted below.
Thank you both.
So my shrooms need to take some acid?
All I have handy is vinegar. That good? Maybe 1 ml per 5 ml of solvent?
Is this going to make toxic gas? I think HCL and isopropanol make chlorine. I guess I already did that, since there is HCL in Erlich's. I did all this in a bathroom with the window open and a strong ceiling fan to vent.
Do you think I'll need the acid with other reagents? I have some Marquis now. That's formaldehyde and concentrated sulfuric acid.
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Edited by nektar61 (05/05/22 08:17 PM)
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27765145 - 05/05/22 09:08 PM (1 year, 8 months ago) |
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If you're experiencing the same effect, you'll likely need to buffer the pH regardless of the reagent you're using. I don't really know though, different reagents may be less sensitive to pH.
White vinegar (acetic acid) will work well, you could also make a dilute solution of HCl.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27765193 - 05/05/22 09:58 PM (1 year, 8 months ago) |
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Quote:
joze said: If you're experiencing the same effect, you'll likely need to buffer the pH regardless of the reagent you're using. I don't really know though, different reagents may be less sensitive to pH.
White vinegar (acetic acid) will work well, you could also make a dilute solution of HCl.
Thank you.
I'll try white vinegar. I like to use what's on hand. Also, if I do anything worth sharing with other growers, I'd like to have as much of it as possible safer things from the supermarket.
Some people are reluctant to order chemicals, can get you on a list. I also still maintain that ISO is less toxic than methanol, despite SpectreOfCommunism saying it didn't matter.
Methanol is fine for them, for me, since I have a heart condition, the less toxic the better.
ISO is considered "relatively non-toxic."
On the other hand, "Ingesting as little as 10 mL of pure methanol can cause permanent blindness by destruction of the optic nerve."
I know he said "don't drink it", but someone will accidentally ingest (or even just inhale it), so I think for the common worker bee average shroom grower, less toxic is better, and less likely to get you on a list is better.
White vinegar is a better chemical for the average bear to be using than concentrated HCL (where diluting it could cause a spill or breathing fumes).
Most people have no idea how to safely work with common lab chemicals.
I know there are acids in the reagents, but we're not using much of the reagent, and we're diluting it, dropwise, into a larger amount of alcohol.
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27765873 - 05/06/22 11:53 AM (1 year, 8 months ago) |
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Quote:
nektar61 said:
Quote:
joze said:
You're probably experiencing difficulties due to the alkalinity of the mushrooms. See SpectreOfCommunism's post I quoted below.
Thank you both.
So my shrooms need to take some acid?
All I have handy is vinegar. That good? Maybe 1 ml per 5 ml of solvent?
Is this going to make toxic gas? I think HCL and isopropanol make chlorine. I guess I already did that, since there is HCL in Erlich's. I did all this in a bathroom with the window open and a strong ceiling fan to vent.
Do you think I'll need the acid with other reagents? I have some Marquis now. That's formaldehyde and concentrated sulfuric acid.
I just realized I didn't directly answer all of these questions.
In terms of toxicity, you shouldn't be generating anything seriously harmful. Having a window open and being mindful should keep you plenty safe. However, please avoid dumping your waste down a sink. DMAB is a pollutant. Keep a bucket of waste or something and find a waste disposal center.
For how much acid you should be using: I think it will take some trial and error to find the right amount to use, but you should be able to find out a constant pH that gives the strongest reaction with DMAB. You should get some pH test strips and see how much the mushroom tissue affects the pH. I have outlined a general procedure that I think would work well for you, based on your above procedure.
- Add sample and isopropyl (or whichever solvent you're using) to test tube. Let sample sit for 6 hours.
- Filter sample. Test sample pH and record.
- Use syringe to measure sample into test tube(s).
- Adjust sample pH dropwise with white vinegar. Re-test pH and buffer to ideal range (perhaps start with pH 6 or 7).
- Use syringe to adjust sample volume with isopropyl (if necessary).
- Add 10 drops of Ehrlich's reagent, shake, wait one hour.
- Record color change.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27766368 - 05/06/22 06:15 PM (1 year, 8 months ago) |
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Quote:
joze said:
I just realized I didn't directly answer all of these questions....... [/LIST]
Thank you.
My next tests will be with Marquis agent and no extra acid, then I'll try both reagents again with extra acid, buffered to neutral. I have some pH strips.
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 2
#27766934 - 05/07/22 06:44 AM (1 year, 8 months ago) |
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Test 2, Marquis Reagent
Ground dry strong cube clone (x7x) to fine with new grinder. Zeroed scale with paper before adding powder.

Added 20 mil ISO alcohol 70%
in 4 oz jar. Shook every hour for 6 hours:
Strained result, then filtered with coffee filter.
Lost some of it during filtering, so didn't have enough to do all planned tests. Just did 1 test, plus a part of another.
Vial 1: .5 ml shroom solution Vial 1: 1 ml shroom solution Vial 1: 2 ml shroom solution Vial 1: 3 ml shroom solution
On first 3, used more ISO to make up volume to 3 ml like vial 4.
Vial 5; 1.5 ml shroom solution + 1.5 ml white vinegar.
added 10 drops Marquis reagent to each vial:
Was an exothermic reactions. Vials got very warm to the touch soon after adding the reagent.
Shook every 15 min for 90 min. Result doesn't look useful. 1-4 are about the same color as before reagent. Maybe 4 is a little more yellow (which is one known reaction to psilocybin). But not enough to call it useful. Vial 5 looks milky and inconclusive.
I'm posting these because even non-results should be included, to avoid cherry picking results.

Next time I'll make it a point to photograph the before and after on the same background to make any differences more clear.
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Edited by nektar61 (05/07/22 07:16 AM)
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med] 1
#27768195 - 05/08/22 06:32 AM (1 year, 8 months ago) |
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Test 3, different things, and some promising results:
- I decided the thing to do was to try a few things, and look for ANY bold result...either brighter colors or very dark reaction.
Then do further tests next time to see if that can be tweaked to be quantitative.
First thing different from my previous tests: Instead of an alcohol, I tried a solution of an acid in water as the extractor. Something somewhere was telling me to do that (without the alcohol).
I had white vinegar on hand, I use it to clean my water distiller. It says on the label that it's adjusted to 5% acetic acid. The rest is mostly water.
Second new thing: I did a shorter extraction of my powdered shrooms, I did 2 hours instead of 6 hours. A test like this shouldn't take all day.
Third, fourth, and fifth new thing: Added heat. Also used more shrooms, and more extractor solution, so I'd have more to work with. Once I soaked the shrooms (3.24 grams of BBM clone) in 40 ml of white vinegar for 2 hours, I put the test tube in a glass of warm (90 f) water. Put it back there between pouring vials.
The extraction was very slightly blue, likely just shroom bluing reaction:
I used vials numbered from the last tests that I ended up not having enough extract to use last time. Vial number 7 is missing and was not used. This is the vials after adding reagent to only the first one (#6), but not the rest:
This is after adding different things. Got some dark results and some colorful results in 10 minutes.
They all have 1 ml of extraction, except the first, one, Vial 6, has 3 ml. (Once I started pouring, I realized I didn't have enough to put 3 ml in each one): LEGEND: Vial 6: 15 drops Ehrlich's Vial 8: 15 drops Marquis Vial 9: 5 drops Marquis Vial 10: 15 drops Ehrlich's Vial 11: 7 drops Ehrlich's, 7 drops Marquis, 7 drops Isopropyl alcohol 70% (water 30%) Vial 12: CONTROL. Nothing added.
This is at 15 minutes. Some color starting to fade.

This is at 1 hour. Purple on #10 and #11 mostly faded. 
I think Vial 10 (15 drops Ehrlich's) looks most promising with its purple result.
Though Vial 6 (15 drops Ehrlich's in 3x as much extract) and Vial 11 (7 drops Ehrlich's, 7 drops Marquis, 7 drops Isopropyl alcohol 70%) could be promising.
Interesting that the same amount of Ehrlich's in 3x as much solution turned brown, where in the 1 ml it looks purple.
Next test's goal would be to have stronger reaction of that purple or brown, then try with different amounts to see if can be tweaked to be quantitative.
Anyone who wants to jump in and try some, I'd love to see it, especially with stronger acids I don't feel comfortable using.
Might also add: -Heat. Warm water bath for the reaction.
--Check and photograph each minute for the first 15 minutes after adding reagent(s).
--Stronger extract? Counting loss from absorption into coffee filter used for filtering, and loss of absorption into shroom powder, I'd guess I ended up with something like 100 mg of shroom material per ml. Could double that.
--Stronger acid? I'm not going to mess with that, but some other people on here might.
--Try the extraction for 1 hour, not 2. I think most of the material might be extracted in 1 hour. Maybe not, but worth a try. The less this test costs, in time and money, the more useful it might be.
Note the Ehrlich's gives off heat when added to the extract. Also did when added to just ISO alcohol in previous tests. Might be reaction with the shroom components, not sure.
I'm saving used vials to neutralize outside with baking soda before sealing. Good idea? Will wear eye protection and good mask. Used both plus fan and open window for all my tests.
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Edited by nektar61 (05/08/22 12:44 PM)
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27768544 - 05/08/22 11:24 AM (1 year, 8 months ago) |
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Awesome! Great progress nektar. I appreciate how well you're documenting your process.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze]
#27768615 - 05/08/22 12:11 PM (1 year, 8 months ago) |
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Quote:
joze said: Awesome! Great progress nektar. I appreciate how well you're documenting your process.
Thank you. I try. What I lack in schooling I make up for in that my day job includes writing weekly reports on the whole warehouse, so organization is second nature. Though I did actually pay attention in high school chemistry, and I'm old enough that they used to teach chemistry in high school when I was there.
I'm wearing gloves, eye protection, a good mask and have a fan blowing everything out the window (or work outside when it's warm enough). Also keeping some baking soda solution handy in case any acid (like both of these reagents, or even the vinegar) gets on skin or in eyes. I got some of this vinegar on a paper cut a while back, hurt like the devil, and it's much weaker than the acids in the reagents.
Don't smoke or have flame around this. A little bit of hydrogen is being given off in that exothermic reaction.
For anyone wanting to jump in on these experiments and throw chemicals against the wall and see what sticks (hopefully based on some reading more than just a hunch), here's a possibly useful post I found on the Hive, chemicals you should not mix for one reason or another (usually because they result in either immediate fire, or immediate toxic gas): -----
Incompatible Chemicals By Boris
A wide variety of chemicals react dangerously when mixed with certain other materials. Some of the more widely-used incompatible chemicals are given below, but the absence of a chemical from this list should not be taken to indicate that it is safe to mix it with any other chemical!
acetic acid: chromic acid, ethylene glycol, nitric acid, hydroxyl compounds, perchloric acid, peroxides, permanganates acetone: concentrated sulphuric and nitric acid mixtures acetylene: chlorine, bromine, copper, fluorine, silver, mercury alkali and alkaline earth metals: water, chlorinated hydrocarbons, carbon dioxide, halogens, alcohols, aldehydes, ketones, acids aluminium (powdered): chlorinated hydrocarbons, halogens, carbon dioxide, organic acids. anhydrous ammonia: mercury, chlorine, calcium hypochlorite, iodine, bromine, hydrofluoric acid ammonium nitrate: acids, metal powders, flammable liquids, chlorates, nitrites, sulphur, finely divided organic combustible materials aniline: nitric acid, hydrogen peroxide arsenic compounds: reducing agents azides: acids bromine: ammonia, acetylene, butadiene, hydrocarbons, hydrogen, sodium, finely-divided metals, turpentine, other hydrocarbons calcium carbide: water, alcohol calcium oxide: water carbon, activated: calcium hypochlorite, oxidizing agents chlorates: ammonium salts, acids, metal powders, sulphur, finely divided organic or combustible materials chromic acid: acetic acid, naphthalene, camphor, glycerin, turpentine, alcohols, flammable liquids in general chlorine: see bromine chlorine dioxide: ammonia, methane, phosphine, hydrogen sulphide copper: acetylene, hydrogen peroxide cumene hydroperoxide: acids, organic or inorganic cyanides: acids flammable liquids: ammonium nitrate, chromic acid, hydrogen peroxide, nitric acid, sodium peroxide, halogens hydrocarbons: fluorine, chlorine, bromine, chromic acid, sodium peroxide hydrocyanic acid: nitric acid, alkali hydrofluoric acid: aqueous or anhydrous ammonia hydrogen peroxide: copper, chromium, iron, most metals or their salts, alcohols, acetone, organic materials, aniline, nitromethane, flammable liquids, oxidizing gases hydrogen sulphide: fuming nitric acid, oxidizing gases hypochlorites: acids, activated carbon iodine: acetylene, ammonia (aqueous or anhydrous), hydrogen mercury: acetylene, fulminic acid, ammonia mercuric oxide: sulphur nitrates: sulphuric acid nitric acid (conc.): acetic acid, aniline, chromic acid, hydrocyanic acid, hydrogen sulphide, flammable liquids, flammable gases oxalic acid: silver, mercury perchloric acid: acetic anhydride, bismuth and its alloys, ethanol, paper, wood peroxides (organic): acids, avoid friction or shock phosphorus (white): air, alkalies, reducing agents, oxygen potassium: carbon tetrachloride, carbon dioxide, water potassium chlorate: acids potassium perchlorate: acids potassium permanganate: glycerin, ethylene glycol, benzaldehyde, sulphuric acid selenides: reducing agents silver: acetylene, oxalic acid, tartaric acid, ammonium compounds, fulminic acid sodium: carbon tetrachloride, carbon dioxide, water sodium nitrate: ammonium salts sodium peroxide: ethanol, methanol, glacial acetic acid, acetic anhydride, benzaldehyde, carbon disulphide, glycerin, ethylene glycol, ethyl acetate, methyl acetate, furfural sulphides: acids sulphuric acid: potassium chlorate, potassium perchlorate, potassium permanganate (or compounds with similar light metals, such as sodium, lithium, etc.) tellurides: reducing agents zinc powder: sulphur Swio would have just posted the link, but it contains info on bomb making etc. and swio thinks that there is enough of these things going off around the world!
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Edited by nektar61 (05/08/22 12:34 PM)
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Re: DIY Psilocybin Content Percentage Detection [Re: shr00med]
#27769476 - 05/09/22 06:53 AM (1 year, 8 months ago) |
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I was wondering why I had two Erlich's tubes (different amounts of liquid / shroom alkaloids) where one with more was brownish and one with less was lavender.
I found something similar on an article called Quantitative Analysis of Psilocybin and Psilocin in Psilocybe Baeocytis byHigh-Performance Liquid Chromatography and by Thin-Layer Chromatography. Is about TLC but would these correspond to the simpler type we're doing here? If not, may help with those wanting to do home TLC:
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27769908 - 05/09/22 02:53 PM (1 year, 8 months ago) |
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Hey nektar.
I've been doing some more reading on this topic, and I think you may actually have gone too acidic now, if you're using undiluted white vinegar to extract the mushroom tissue. In the paper "A Simplified Method for the Analysis of Hydroxyproline in Biological Tissues," (1996), the authors write that "A pH of the acetate-citrate buffer between 6.0 to 6.5 yielded maximal absorbance values for all samples. At pH values below 5, the formation of chromophore was found to be negligible; however, alkaline pH of the buffer did not influence the absorbance of standard Hyp except for those at pH 8.0."
This basically means that the DMAB reagent did not produce much of a color change when the sample was buffered below pH 5, and was optimal between pH 6 and 6.5. White vinegar is around pH 2.5, according to a quick Google search. I would mess around with using a couple of dilutions of white vinegar, maybe 1:1, 1:3, and 1:5 or something like that, and see how that effects the color change reaction. The same may be true for the Marquis reagent as well.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze]
#27770678 - 05/10/22 05:01 AM (1 year, 8 months ago) |
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Okay I must have missed something.
Why is the marquis test even an option? Is Marquis and mecke for md(m)a?
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Re: DIY Psilocybin Content Percentage Detection [Re: joze]
#27770685 - 05/10/22 05:18 AM (1 year, 8 months ago) |
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Quote:
joze said: Hey nektar.
I've been doing some more reading on this topic, and I think you may actually have gone too acidic now, if you're using undiluted white vinegar to extract the mushroom tissue.
Thank you. Next time I'll try white vinegar:distilled water 1:1 as my extractor, and I'll do a pH test on it.
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61]
#27770698 - 05/10/22 05:42 AM (1 year, 8 months ago) |
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That image you cited from the 1975 article talking about marquise reacting to psilocybin doesn't seem to exist anymore and only seems to be referenced on the "cops", page. Actually it's a book, and it looks creepy haha. Some old times medicine shit. I think your wasting your time on that one.
This is kinda funny tho, elrich showing a positive result on a button mushroom because of tryptophan https://www.protestkit.eu/how-to-test-psilocybin-magic-mushrooms/
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27774468 - 05/12/22 06:35 PM (1 year, 8 months ago) |
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Quote:
nektar61 said: Test 2, Marquis Reagent
Ground dry strong cube clone (x7x) to fine with new grinder. Zeroed scale with paper before adding powder.
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I just noticed this thread while looking for something completely different. Following with interest.
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 2
#27775768 - 05/13/22 03:45 PM (1 year, 8 months ago) |
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Quote:
joze said: I've been doing some more reading on this topic, and I think you may actually have gone too acidic now....
Test 4, using white vinegar 50/50 with distilled water as extractor. (all my tests archived on my Journal, here.)
First I tested the pH of my 5% acetic acid white vinegar. Looks like 3. Wow. Vinegar is fucking stronger acid than I thought. No wonder it hurt like satan when I got it on a paper cut recently:
pH of my vinager diluted 50/50 with distilled water. Looks almost the same, but it's a little different. I think it's pH 4:
Since it's an acid, and the reagent I'm using (Ehrlich's) is even stronger (it's about half concentrated hydrochloric acid), I made some eye wash solution just in case. This is 30 grams of baking soda into 300 ml of distilled water. Not all of it dissolved, but it's good to have on hand, and can save eyes or skin if you get acid on them. Also I can use it to neutralize my tests (outside) after the experiment is over and before I throw them in the trash:
This is the pH of the eye wash solution. Looks like an 8, mildly alkaline:
I used the 50/50 vinegar/water solution to extract from 5 grams of BBM dried shrooms. I ground them to a powder right before the tests. Mixed 25 ml distilled water with 25 ml white vinegar. Soaked in solution for 2 hours, this time kept the jar in a water bath at 115 f.
I strained it then filtered it. Lose some in filtering. Came out of bluish. Is likely shroom blue oxidation reaction. This is bluer than previous test extractions, I surmise that the warm water bath added this time increased either the amount of psilocin / psilocybin extracted, or the enzymes that make the bluing, or both.
This is the pH of the extract. Looks like a 4, or a 4.5. Same as or slightly less acidic than the extraction solution before the extraction. (I took the solution out to test with a pipette, I did not put the litmus paper in the solution.)
I added these these amounts of extract added to fresh vials, then brought them up to 3 ml with distilled water (all measured with syringe): Vial 1 got .5 ml solution. Vial 2 got 1 ml solution. Vial 3 got 2 ml solution. Vial 4 got 3 ml solution. Vial 5 got 3 ml solution (almost, I ran out).
I added 15 drops of Ehrlich's reagent to vials 1, 2, 3, and 4. Vial 5 is a control, I added no reagent to it.
This is 5 minutes after adding reagent:

This is 10 minutes after adding reagent:
This is 20 minutes after adding reagent:

This is 30 minutes after adding reagent:
This is 40 minutes after adding reagent:
This is 50 minutes after adding reagent:

When done, I took the vials outside, dumped them into a plastic bag, and poured some of the eye wash solution on them. When it stopped fizzing, I sealed it up, doubled bagged it, and threw it out.
I wore a good filtered mask, gloves, and eye protection for this and every step of the tests. I also had a window open, and a fan blowing out the window, and a ceiling fan on.
CONCLUSION: Not much. I think that diluting the acid was a step back. I think the last test, test 3, with straight 5% acetic acid white vinegar extractor had the strongest color reaction between vials of any of the 4 tests I've done.
I promise you, aqueous acid solution is the extractor we want to use. Must have water to dissolve the psilocybin, and must have acid to make a salt of the psilocin so it dissolves well in water.
According to: https://www.swgdrug.org/Monographs/PSILOCIN.pdf (use a VPN, that's a cop site, "Scientific Working Group for the Analysis of Seized Drugs" haha):
Psilocin is only slightly soluble in water. Psilocybin is soluble in water.
Same is confirmed elsewhere online with chemical library sites.
I think we want to try MORE acidic than vinegar, but I'm not going to do it. pH 3 is my limit. But if someone who knows how to use it safely wants to try a solution of an acid that's pH 2, I think you'll get the results we seek, then can make a color chart. Even stronger acetic acid might do it, but could also probably use hydrochloric, sulfuric, or phosphoric. But I'm not fucking with any of that.
If that works better, but isn't perfect, you might want to try again adding a small amount of ferric chloride, to get the effect of Keller's Reagent too. That reacts with the enzymes we're testing for.
So I've done 4 tests, I think I've found some useful info, and I'm passing the torch. -- EDIT: after looking again at the results, I think the tests at 20 minutes may have been useful. It's hard to tell because there is just the bluing with the dilution, but at 20 min it looks like the ones with more drugs in it looks more dark brown. Might be on to something here. What do you think?
If someone has some weak shrooms and some strong shrooms and wants to repeat this test with two extraction solutions, that might be good. I'm getting burnt out on breathing acid fumes.
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Edited by nektar61 (05/14/22 11:39 AM)
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nektar61
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Quote:
SpectreOfCommunism said: ...Hofmann reagent ...is made with DMAB, sulfuric acid, and ferric chloride.
Anyone know how much ferric chloride to how much of the other ingredients? Can't find it anywhere.
A lot of people sell Hofmann's, but no one seems to want to give up the recipe for the secret sauce. (not surprising)
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: joze] 1
#27787387 - 05/21/22 07:55 PM (1 year, 8 months ago) |
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TEST 5. BRING ON THE HEAVY METAL (Iron that is.... ferric chloride solution 40% - printed circuit board etchant) -- I think this one is showing promise. Just need to tweak the amounts. Added ferric chloride 40% to Ehrlich's reagent, sort of made an Ehrlich's / Hoffman cross.
I ground 2.4 grams of BBM clone shrooms (dry). Soaked in 40 ml white vinegar. (comes as 5% acid, 95% water).
Soaked for 80 minutes. Stirred vigorously every 20 min, plus at start and end.
Strained, then filtered. Got this:
This is a weaker solution than I made last time, which I think is good. Last time was so strong it had more bluing, which could interfere with reading color-based test results.
Here's what went into each vial:
Vial 1: 3 ml shroom extract. Vial 2: 3 ml shroom extract. Vial 3: 3 ml shroom extract. Vial 4: 3 ml shroom extract.
(This is pic before adding some different amounts of shroom extract to each of the vials after #4)
Added 10 ml of Ehrlich's reagent to each vial in Vials 1-4. Not much reaction.
Added tiny amounts of ferric chloride solution to Vials 1-4.
Vial 1: Added .2 ml ferric chloride solution. Vial 2: Added .4 ml ferric chloride solution. Vial 3: Added .75 ml ferric chloride solution. Vial 4: Added 1 ml ferric chloride solution.
Got INSTANT reaction of dark green on all, could not tell them apart:

Wanted to see if it was just a reaction to the vinegar. Poured shroom extract from Vial 5 into Vial 6 (which then had 6 ml extract).
Washed out Vial 5 with distilled water. Filled it with just 3 ml of white vinegar. Added ferric chloride solution, .25 ml. Did not turn green (shown in above photo). Turned straw / urine yellowish.
For Vials 6-9, adjusted to these amounts of shroom extract:
Vial 6: 6 ml shroom extract. Vial 7: .5 ml shroom extract, adjusted with distilled water up to 3 ml. Vial 8: 1 ml shroom extract, adjusted with distilled water up to 3 ml. Vial 9: 2 ml shroom extract, adjusted with distilled water up to 3 ml.
Then added the Iron: Vial 6: Added 3 drops ferric chloride solution. Vial 7: Added 1 drop ferric chloride solution. Vial 8: Added 5 drops ferric chloride solution. Vial 9: Added 2 drops ferric chloride solution.
I was trying different things to try to get SOME visible variation.
It's very hard to see, but I think that the ones with less had slightly lighter reaction. Hard to tell but under bright light, could see it. Couldn't photograph well though. Below pic is, left to right, Vials 6-9.
As you can see if you look closely, Vial 7 is slightly lighter green than the others. It had the least shroom extract and the least ferric chloride:

CONCLUSION: the VERY fast VERY abrupt reaction is going in the right direction. I think next step is to make Ehrlich's reagent with much smaller amounts of ferric chloride 40% mixed in. Maybe start with 1 drop per 3 ml and see, then go down or up from there to get see if we can get a quantitative reaction.
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Edited by nektar61 (05/21/22 08:03 PM)
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7Suns
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27792925 - 05/25/22 08:40 PM (1 year, 7 months ago) |
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Not sure if this is anything useful but have you guys also tried fluorescing the different concentrations under black light ? I’ve seen something about using black light to check tartaric powder extracts posted by achuma I believe
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: 7Suns]
#27793022 - 05/25/22 09:50 PM (1 year, 7 months ago) |
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Quote:
7Suns said: Not sure if this is anything useful but have you guys also tried fluorescing the different concentrations under black light ? I’ve seen something about using black light to check tartaric powder extracts posted by achuma I believe
I tried both long UV (black light) and short (mineral light, the kind that requires goggles) on the last batch I did, no florescence, but I did it after I added the reagents. Will try before on next.
A lot of things can respond to UV though.
When you say "tartaric powder extracts" do you mean LSD tartrate, or ergotamine tartrate? Or something else.
I've heard of UV light being used to separate out levo and dextro LSD in a chromo column. Not sure if that applies here.
One thing I do plan to do if I, or someone else here, gets something that works for quantitative visual reagent tests: test it on non-psychedelic shrooms to make sure we're not getting a hit on something other than the psilo alkeloids.
Thank you.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: 7Suns]
#27793044 - 05/25/22 10:11 PM (1 year, 7 months ago) |
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Quote:
7Suns said: I’ve seen something about using black light to check tartaric powder extracts posted by achuma I believe
OK, you mean this, right?: https://www.shroomery.org/forums/showflat.php/Number/22960162
I don't think that's florescence. That said, keep them coming. I'm interested in ANYTHING that will help figure this out.
Real florescence is almost shocking, and gets your attention even at low levels. I think what they were seeing is reflection of the visible component of their UV light. They also don't even say what type of UV it is. So probably black light, not what they probably want for this. I tried that and a "real" UV light, and didn't see anything. As I said, was after tests, and will repeat again before tests. (but then not test that tube, as UV can degrade chemicals.)
I don't think achuma got pure psilo alkaloids. They say it's 1/10 the weight of dried shrooms. Should be more in the magnitude of 1/100th.
They say it's active, but probably also contains a lot of sugars, proteins, and other non-actives that will mess with the results. I'm still going though the thread, but I think I agree with this: https://www.shroomery.org/forums/showflat.php/Number/22964630#22964630
Quote:
Regarding your UV "fluorescence" I believe what your observing is mere reflection of your blacklight. Psilocybin and psilocin lacks the conjugated double bond found in lysergic acid derivatives that emit fluorescence (opposed to just reflecting it).
This is what milk looks like under blacklight, it's not glowing, just reflecting.
Also see this thread for more fluorescence of compounds that contain conjugated double bonds (again more lysergic acid derivatives)
Edit: you have demonstrated that light can be reflected, you still lack proof of actual fluorescence
and this; https://www.shroomery.org/forums/showflat.php/Number/22965121#22965121
Quote:
What is being stated in that article is if you excite psilocybin with a 267nM light it will fluoresce at 335nM (which cannot be detected by the human eye) and if psilocin is excited with a 260nM light it will fluoresce at 312nM (which cannot be detected by the human eye). The human eye can only detect wavelengths from 390 to 700 nM
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7Suns
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#27793308 - 05/26/22 06:34 AM (1 year, 7 months ago) |
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Yes that’s the link, yea I wasn’t sure if it was useful or not
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: 7Suns] 1
#27797129 - 05/28/22 11:48 PM (1 year, 7 months ago) |
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TEST 6 modified Hofmann's Reagent, plus UV tests
I ground fine 2.5 grams of dried BBM clone. Added 40 ml 5% acid white vinegar. Shook at start and every 15 min and at end for one hour. Strained, then filtered twice with coffee filter. Result:
Here's the Ehrlich's Reagent I'm using:
Here's a fresh 3 ml bottle after adding 1/10 of 1 ml of ferric chloride, showing the slight color change (just dilution). I'm making a very modified variation (almost a distant cousin) of Hofmann's reagent:
Here's the solution I added, and a syringe showing trying to measure just .1 ml. Was hard, air bubbles, and end of syringe neck, but I think I got it after this to just 1/10 ml and added to the fresh bottle above.
Below, Vial #7, shows 3 ml shroom extract plus 1 drop ferric chloride solution only, no Ehrlich's Reagent added. Has immediate dark green reaction like test 5 with ferric / Ehrlich's. I believe now that the tests last time are just showing a reaction with something in the shrooms plus iron only. No reagent needed, so test is not conclusive.
It may even be measuring proteins or sugars. Did not show last time with just ferric and vinegar, is something in shrooms.
NOTE: To anyone who does these tests, if you get something you think is conclusive, then also test on non-psychedelic mushrooms to rule that out.
The other vials here are 2 ml shroom / vinegar extract with nothing added:
Other tests I did as long as I had everything out.
2 ml shroom / vinegar extract. 3 drops modified Hofmann's: 
2 ml shroom / vinegar extract. 5 drops modified Hofmann's: 
2 ml shroom / vinegar extract. 2 drops modified Hofmann's:

.5 ml shroom / vinegar extract. 1.5 ml distilled water. 2 drops modified Hofmann's:
I'm pretty sure these are just showing dilution of the reaction of vinegar and ferric. So this probably isn't detecting active alkaloids.
UV LIGHT: I had a little leftover vinegar / shroom solution so I tried UV light.
These are the lights I used. Cheap long wave black light flashlight on the left, and a better shortwave UV light for rock collecting on the right. If you do these tests, the shortwave requires eye protection.

Here's the unused shroom / vinegar extract illuminated in the dark with cheap long wave black light;
Here's the solution illuminated in the dark with better short wave UV light;
Neither of these are fluorescing. There is a little reaction with the glass on the long wave one. Other than that the liquid is just reflecting a little bit of the viable component of these UV lights
CONCLUSION: No conclusion, but science includes showing what doesn't work. I did show some things that might not need to be tried again, or someone can repeat them to show they're repeatable. But they seemed pretty conclusively non-reactive to me.
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QM33
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 2
#27798661 - 05/30/22 06:07 AM (1 year, 7 months ago) |
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“I have not failed. I've just found 10,000 ways that won't work.”
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Icyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: nerdintheshell] 1
#27861451 - 07/13/22 04:56 PM (1 year, 6 months ago) |
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Old thread, but if anyone is still interested in TLC there was a post recently describing relatively simple process for comparison testing that might be worth checking out.https://www.shroomery.org/forums/showflat.php/Number/27830910#27830910
Quote:
Notes on sample preparation and procedure for the above shown TLC]
100mg of carefully weighed, blended/homogenized mushroom powder from each sample was added to a small beaker, 5ml of methanol was added over top while also washing down the sides of the beaker. The beakers were covered with plastic wrap and secured with rubber bands then transferred to an ultrasonic bath in hot tap water, sonicated for 8 minutes with heat, 5 minute rest, followed by another 8 minute sonication with heat. Samples were allowed to settle for 10 minutes but no filtration, decanting, or centrifuging was done. A calibrated pipette was used to transfer 10µg of each sample to the chromatography sheet. Sheet was developed in a sealed glass chamber saturated with eluent consisting of 100 parts methanol: 1.5 parts ammonia. After development the sheet was allowed to dry while undergoing ultraviolet examination, then freshly mixed Ehrlich's reagent was applied by spray bottle until the sheet was saturated, followed by drying with a heat gun. Sheet was periodically photographed every 5 minutes until colors reached full development (maximum brightness).

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MariaTruthSeeker
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Re: DIY Psilocybin Content Percentage Detection [Re: Icyurmt]
#27976801 - 10/01/22 03:14 PM (1 year, 3 months ago) |
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Quote:
Icyurmt said: Old thread, but if anyone is still interested in TLC there was a post recently describing relatively simple process for comparison testing that might be worth checking out.https://www.shroomery.org/forums/showflat.php/Number/27830910#27830910
Quote:
Notes on sample preparation and procedure for the above shown TLC]
100mg of carefully weighed, blended/homogenized mushroom powder from each sample was added to a small beaker, 5ml of methanol was added over top while also washing down the sides of the beaker. The beakers were covered with plastic wrap and secured with rubber bands then transferred to an ultrasonic bath in hot tap water, sonicated for 8 minutes with heat, 5 minute rest, followed by another 8 minute sonication with heat. Samples were allowed to settle for 10 minutes but no filtration, decanting, or centrifuging was done. A calibrated pipette was used to transfer 10µg of each sample to the chromatography sheet. Sheet was developed in a sealed glass chamber saturated with eluent consisting of 100 parts methanol: 1.5 parts ammonia. After development the sheet was allowed to dry while undergoing ultraviolet examination, then freshly mixed Ehrlich's reagent was applied by spray bottle until the sheet was saturated, followed by drying with a heat gun. Sheet was periodically photographed every 5 minutes until colors reached full development (maximum brightness).

Good reference. I have been wondering which would be better, using TLC or a Miraculux like solution as described in this thread. I think the TLC solution described above could be simplified a little more, a single TLC sheet could be cut into several small sheets and developed in a standard 16oz mason jar. As you can see from the picture in that above thread, visualization is quite good with Ehrlichs and you can easily see both psilocybin and psilocin separately which is a big advantage over the Miraculux/reagent solution. Psilocin is significantly more potent by weight and the Miraculux type solutions may not be accurate for high psilocin species (like Pan Cyans). If someone came up with an exact procedure others could replicate, then you could measure the area of the spots using Gimp or other photo editing software and build out a table/graph of spot sizes to actual percentage for each alkaloid. I'm not sure how doable that would really be though, as spot size alone may not be adequate, maybe the darkness/color matters too? The other problem is everyone's extraction would have to be the same, while sonication is probably the best known extraction method, everyone would have to do it the same way with the same wattage, etc, and not many would own a sonicator so it wouldn't be cheap/easy for many people. The standard solution would probably have to use a free extraction method (maybe just soak for 24 hours in solvent with periodic shaking?). Likewise, to be reproducible, everyone would need a proper calibrated pipette, they aren't that expensive but still this is another tool people would have to buy and another barrier to accessibility.
I suppose just doing "comparisons" like was done between the Penis Envy and Pan Cyan to get a good sense of the potency difference is probably adequate for many people. You could use this to compare old stored samples vs. freshly grown/dried samples to see how much potency has been lost, or to compare one species to another, etc.
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Suckatshrooms
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Re: DIY Psilocybin Content Percentage Detection [Re: nektar61] 1
#28009921 - 10/21/22 08:15 PM (1 year, 3 months ago) |
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HCL is nasty. There’s almost always a better solution. Sulfuric acid is great. It’s cheap, easy to find locally and stays in solution so it’s much safer. HCL is a gas so respirators are needed, much like it’s basic brother, ammonium hydroxide. HCL outside a fume hood will rust everything in your lab as well.
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hjalmar
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Re: DIY Psilocybin Content Percentage Detection [Re: Suckatshrooms]
#28036538 - 11/06/22 01:33 PM (1 year, 2 months ago) |
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Quote:
Suckatshrooms said: Sulfuric acid is great. It’s cheap, easy to find locally and stays in solution so it’s much safer.
Depends where you live. In the EU since a few year ago the sale of sulfuric acid over 15% to private individuals has been banned. They're selling 10% H2SO4 now as drain cleaner, I doubt if it will even do that, only licensed professionals are allowed to handle to concentrated stuff now (i.e. you pay 150 euros instead of 15).
Ayway shouldn't be a problem to handle muriatic acid (the 25% HCl the hardware store stocks) as long as you take the necessary precautions.
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nektar61
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Re: DIY Psilocybin Content Percentage Detection [Re: hjalmar]
#28039162 - 11/07/22 08:13 PM (1 year, 2 months ago) |
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Quote:
hjalmar said:
Quote:
Suckatshrooms said: Sulfuric acid is great. It’s cheap, easy to find locally and stays in solution so it’s much safer.
Depends where you live. In the EU since a few year ago the sale of sulfuric acid over 15% to private individuals has been banned. They're selling 10% H2SO4 now as drain cleaner, I doubt if it will even do that, only licensed professionals are allowed to handle to concentrated stuff now (i.e. you pay 150 euros instead of 15).
Ayway shouldn't be a problem to handle muriatic acid (the 25% HCl the hardware store stocks) as long as you take the necessary precautions.
5% acetic acid vinegar, can you get that there? It's pretty low Ph, and I was trying to use it for my extractions here.
I think it might work.
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