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Invisibleshr00med
Stranger
Registered: 03/10/21
Posts: 29
DIY Psilocybin Content Percentage Detection * 5
    #27682139 - 03/04/22 07:48 AM (1 year, 10 months ago)

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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OfflineSpectreOfCommunism
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Registered: 10/17/12
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Loc: Willamette Valley, Oregon
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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)

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


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Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus



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OfflineSpectreOfCommunism
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Registered: 10/17/12
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 2
    #27683112 - 03/04/22 08:53 PM (1 year, 10 months ago)

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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Invisibleshr00med
Stranger
Registered: 03/10/21
Posts: 29
Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 1
    #27683744 - 03/05/22 11:16 AM (1 year, 10 months ago)

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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OfflineAlan RockefellerM
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 1
    #27694088 - 03/13/22 05:26 PM (1 year, 10 months ago)

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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OfflineSpectreOfCommunism
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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)

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


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Found Species: Ps. Azurescens, Ps. Cyanescens, Ps. Baeocystis, Ps. Semilanceata, Pan. Cinctulus



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Offlinefossilshark
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 3
    #27694824 - 03/14/22 10:09 AM (1 year, 10 months ago)

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.


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LITFA LITFA LITFA


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OfflineSpectreOfCommunism
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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)

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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OfflineWhiteThumbScience
Full Rick
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Registered: 03/15/22
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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)

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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OfflineWhiteThumbScience
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 1
    #27701960 - 03/19/22 11:01 PM (1 year, 10 months ago)

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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OfflineSpectreOfCommunism
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Registered: 10/17/12
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Re: DIY Psilocybin Content Percentage Detection [Re: WhiteThumbScience] * 1
    #27701981 - 03/19/22 11:43 PM (1 year, 10 months ago)

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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OfflineWhiteThumbScience
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 1
    #27702066 - 03/20/22 03:48 AM (1 year, 10 months ago)

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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InvisibleIcyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: WhiteThumbScience] * 2
    #27702463 - 03/20/22 01:42 PM (1 year, 10 months ago)

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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OfflineSpectreOfCommunism
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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)

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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InvisibleIcyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 1
    #27702876 - 03/20/22 09:02 PM (1 year, 10 months ago)

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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OfflineSpectreOfCommunism
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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)

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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InvisibleIcyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 2
    #27702958 - 03/20/22 10:46 PM (1 year, 10 months ago)

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.
:popcorn:


--------------------
👁️ 🌊 why you are empty.

Hunt for the habitat not the mushroom.


Edited by Icyurmt (03/20/22 10:53 PM)


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OfflineSpectreOfCommunism
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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)

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.
:popcorn:



For sure, both approaches have merit methinks — just a matter of which mix of advantages/disadvantages one wants to take on :smile: 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 :smile: 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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Offlinenerdintheshell
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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)

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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InvisibleIcyurmt
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Re: DIY Psilocybin Content Percentage Detection [Re: SpectreOfCommunism] * 2
    #27702985 - 03/20/22 11:44 PM (1 year, 10 months ago)

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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