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Mescaline quantification in live cacti using TLC

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An1cca

An1cca

Rising Star
Hello guys,

Before taking a break from psychedelic research, I wanted to post my notes concerning a promising technique of cactus analysis. I also added some notes on ethanolic extraction and purification.

As it was too much to post, I've put it all into an illustrated document. But don't worry, I gave it a Nexus-look&feel 8)

May it spark further research!
 

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Thanks! The descriptions is so detailed, I like the technique improvements and the purification method. Excellent job.

Downloaded instantly and will try this on fresh cacti.

Are there any chances this will make it into Wiki as an 'official' Tek?
 
blue.magic said:
Are there any chances this will make it into Wiki as an 'official' Tek?
Click to expand...

Hehe blue.magic, I'm glad you too think that this technique holds promise. However, I would wait with 'officializing' the technique until we have some more data. Especially, the data from 20M, 37M and 39M would be informative.

For personal reasons I need to take some time off, but I'm handing over my entire lab to the fellow-Nexian that is the father of all specimens except 'Arran' and who'll continue the research. So I think we'll getting more data the following weeks/months :thumb_up: .

Besides that, I hope anyone at the Nexus with access to live sacred cacti will take the step to TLC for personal or research purposes. That's what I'm doing anyway, having no ambition to drink cactus tea from specimen 30M :sick: ....
 
Thank you An1cca I'll dig into that document later, it seems very well presented at first sight. I've no cacti to work with right now but should lay hand on some because M is on top of the wish list.

You spoke of trying enzymes to take on with the slime but they did not help, what was that please? I had some hopes that pectinase enzymes would help.

I find your endeavoring mind set very inspiring and you brought a very positive light to the Nexus, would hate to see you leave even for short :oops:

:love:
 
Jees said:
Thank you An1cca I'll dig into that document later, it seems very well presented at first sight. I've no cacti to work with right now but should lay hand on some because M is on top of the wish list.
Click to expand...

I can only say: go for it! M is a joy for the senses and food for the soul. Depending on your climate, it may take a couple of years to grow enough for entheogenic purposes. So you could start a side-project already by sowing your own cacti. Hybridized high-quality Trichocereus seed is easily available these days.


Jees said:
You spoke of trying enzymes to take on with the slime but they did not help, what was that please? I had some hopes that pectinase enzymes would help.
Click to expand...

They might work. A pure pectinase however doesn't seem to do the trick. I used this one: http://www.erbsloeh.com/product_datasheets/en/PMB_TrenolinFrioDF_GB_001.pdf

Jees said:
I find your endeavoring mind set very inspiring and you brought a very positive light to the Nexus, would hate to see you leave even for short :oops:
Click to expand...

I think this mindset is the strength of the Nexus itself. And I'm not really leaving, just bringing more balance in my life. My problem is that I can't just do some research part of the time. When I'm into it, it's all over my mind, all the time. And that is hard to combine with a demanding job and family-life. So I'm picking up my meditation practice were I left it 10 years ago and hope to reintroduce my research-driven mind gradually and with balance in the following years. In the meantime, I will still follow the research that's being done with unsubsiding interest!:thumb_up:


I noticed that the document is hidden for non-members of the nexus. This might prevent interested visitors from reading, checking, validating or possibly debunking some of its content. Is there a way to make it accessible to guests?
 
An1cca, thanks a lot for sharing your work here. This is really cool stuff. I have a few comments and questions though. Please do not take them as a criticism of your work but more as thoughts to challenge :)

1.)
Did you try other solvents than the one provided by bunk police? My hunch would be that once should get good results when first adding HCl and then using a mixture of dH2O and IPA (9:1 perhaps) or dH20 and glacial acid (95:5). Personally I don't like Methanol as it is too toxic to deal with at home :(

2.)

I am curious why you went for the presented extraction method. After producing the the hydrochloride salts of the cactus' alkaloids I would have tried to clean this. Did you try just recrystallizing from dry IPA (you find this in the literature a lot) or from IPA/acetone?

Also, with the amount of effort you put into this, it would also be an option to perform a flash column chromatography on the extracts. I did this during my chemistry studies when extracting caffeine from tea and it worked very well (this was of cause caffeine freebase).

My feeling is that the hydrochloride salts of the different alkaloids have different solubilities in water and you can just run the thing with dH20 as a eluant. I observed this when using Ron69's tek using HCl and evaporating the liquid in a pot: The mescaline would crystallise first and the brownish alkaloids would stay in the dH2O much longer.

3.)

Did you try/thought about using the TLC method to quantify grasses for DMT/5-MeO-DMT? In this way we could find a way of using the bark of trees.

Again. Awesome work!
 
DrSeltsam, a critical attitude lies at the core of all science, so please be my guest :d

You're correct about methanol being too toxic to work with. That's why I chose basified ethanol as a solvent. However, for accessibility-reasons I chose to use Bunk Police's all-in-1 package, of which methanol is a part. Literature agrees that it is the solvent of choice for M and its salts (apart from water with its slimy properties :p )

Flash column chromatography, very interesting...must surely look into this the next time I engage myself in the process. As for the choice of salt: I fell in love with the sulfate-crystals, and isn't love blind sometimes :love: ? The fractional recrystallization you're talking about would make for an interesting purification method. Yet another thing worth digging into...

TLC has been used for the quantification of DMT and gramine in phalaris: Mimosa hostilis and extract analysis thread (FASA, FASW, Naphtha, Limo, "jungle spice" etc - Plant Analysis and Substance Testing - Welcome to the DMT-Nexus. Endlessness already did a great job investigating this:thumb_up:
 
An1cca, beautiful work :) thank you so much for all the work and sharing it!

Im on the phone now, going on memory from what i remeber of your process but... did you consider simply taking a picture of the plats under uv with a camera in a stand instead of using the reagent to visualize the spots and scan? It could work well enough that a person could do this without having to purchase anything extra (though it does seem to work beautifully the way you did it)
 
An1cca, beautiful work :) thank you so much for all the work and sharing it!

Im on the phone now, going on memory from what i remeber of your process but... did you consider simply taking a picture of the plates under uv with a camera in a stand instead of using the reagent to visualize the spots and scan? It could work well enough that a person could do this without having to purchase anything extra (though it does seem to work beautifully the way you did it)
 
Indeed Endlessness, visualization and analysis under UV is certainly possible. However, in my experience this only works for extracts. When using the 'Direct'-sampling method, there is simply too much UV-blocking material ('smear' ) present to allow for M-quantification. That's why I had to switch to the ninhydrin-method (page 3). An added benefit of the latter is the fact that you can do all visualization/picture-taking under daylight, much easier.

Ninhydrin is available as fingerprint-spray e.g. Forensics Lab 8.3: Revealing Latent Fingerprints Using Ninhydrin. I bought 25g of the powder for about 38 euro's. This makes litres of solution and you only need 0,3ml for every plate. Perhaps Bunk Police could add 0,5g and a bottle of ethanol to their kits? Fiji-software is free and easy to use. So this addition would open up a whole world of (semi-)quantification...

Urtica, I only bought Marquis-reagent from Bunk Police. I used it twice, then stopped using it because it had no added value to TLC-visualization. It is destructive as well, so the plates cannot be compared later on...
 
Great, thank you for that link.

It is a bummer that the Marquis destroys the TLC plate. It would be nice to hold onto them in physical form.
 
Whoa,

An1cca, your approach is exceedingly impressive. I tip my hat to you!

I am sticky-ing this thread because all of what you describe in the attached document are not only directly applicable all sorts of plants and fungi, but also because, for the very first time, your work uprovides a means for (semi)quantitation of chemicals of importance in plants and fungi!

Apart from the quantitative innovations you bring, i.e. Fiji / ImageJ, I particularly liked the inventivenes e.g. using a drill for a home centrifuge.

I am so amazed to have found your post, it really deserves to be on the spotlight.
 
You're making me blush, guys :oops: .

It all isn't that hard to do, it just takes a systematic approach and some persistence.

A (big) part of me really wants to continue this work, but a wiser part tells me to do some research inside first. Vipassana (with psychedelic boosters) requires persistence and a systematic approach as well ;).

I really hope someone will peer review these techniques. There are many interesting loose ends to look into!
 
Yes I also applaud this write up again!

I am thinking that I will investigate this process once things calm down in my life enough for me to resume research.

Now if only we had some reference standards and could figure out what some of the other alkaloids are in those cacti. I still really want to wrap my head around people's subjective feelings that the different spp's have different effects & see if there is a clear difference in chemical make up that would account for this. I have a sneaking suspicion that it is placebo but would love to dig deeper.

Also, maybe someday we as a community can figure out why the PC clone is active. My own analysis and a few others that I have seen make it look like there are only a couple milligrams of M per kilogram of fresh cactus, but my own bioassays have shown that there is distinct entheogenic activity from that clone once you get above 3 kg or so...

Cheers again, An1cca! This is the kind of research that I love to see on this forum.
 
An1cca said:
I really hope someone will peer review these techniques. There are many interesting loose ends to look into!
Click to expand...

I am going to reproduce the extraction method. I have already got most of the utensils and chemicals needed so I will test it in few weeks on 25 grams of powdered T. Peruvianus.

TLC part will come next. I have an expertise in digital image processing so I am very interested in possible improvements/automation of the measurement process.
 
An1cca said:
I really hope someone will peer review these techniques. There are many interesting loose ends to look into!
Click to expand...

What certaintly needs doing is a standard dilution curve. Your approach An1cca, as it stands right now, estimates alkaloid concentrations in relation to a set reference, but what we do not know is whether there is a linear relationship between e.g. mescaline amount and mescaline-ninhydrin stain.

You sound like a science-leaning person so I take it that you know what I mean by a dilution curve. Since as you state ninhydrin overstaining saturates the signal and causes intense backround staining.

On another note, for peak volume quantification, have you thought or tried rolling ball background subtraction? That approach automates the guesswork of where a peak starts or ends.

Again on another note, if you do try a standard dilution curve, could you also use UV (preferable a wavelength that is easily obtainabel by hobbyists, such as for quick satitisation) and examine whether there is a different relationship spot volume vs mescaline concentration when compared to ninhydrin staining?
 
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