need help, testing Acacias with TLC

[Endeavor]

I want to test some suspect acacias for DMT, all the information i could find on TLC and colorimetric testing was for testing pure substances, like pills and such to see if they were legit. So I'm not sure how i do a TLC on a tree...

I suppose its just grind up bark/leaves and make a solution with that, put that solution on the paper and then the paper in the jar with the other solution? :?


So is this pretty much how it goes, what solvent(s) do i use (for the bark and the paper)? Will a solution just from the bark contain enough alkaloid to show up in the test?

 

[endlessness]

Check these links and let me know if it didnt answer some question you might have had..

How do analytical methods work? (TLC, UV-Vis Spectrophotometry, GC-MS, LC-MS, etc) - Plant Analysis and Substance Testing - Welcome to the DMT-Nexus

Colorimetric test results (Marquis, Mecke, Ehrlich, etc) for different alkaloids - Plant Analysis and Substance Testing - Welcome to the DMT-Nexus

Rf for different alkaloids (TLC/Paper/Column chromatography) - Plant Analysis and Substance Testing - Welcome to the DMT-Nexus

Solvent choice depends on what is available to you and what according to literature separate well the compounds that might be there. Ive been doing some acacia TLC with methanol:ammonia at 100:1.5 and it works good at separating acacia compounds:

The Acacia Analysis Thread - Plant Analysis and Substance Testing - Welcome to the DMT-Nexus

I guess ethanol or ipa:ammonia would also work fine

Also, notice that bark and leaves often have different alkaloid content so its worth it to test them separately. TLC is usually quite sensitive, small amounts are enough to see the spots. A cold methanolic soak of any plant and then putting a few microliters in the plate works fine.

 

[nen888]

..i posted the following in entheogenic effects of NMT p.3

'M'(2002) subjected a number of acacias to reagent testing, following Roveli(1967)'s recommendation for alkaloid field testing..
2-4 grams of fresh plant material was ground in a mortar and pestle with sand..this was mixed with DCM (or hexane), the sand and goo filtered,
and the solution (approx. 10-20ml) put in a test tube..
..to each test tube, the reagent was added (parallel tests were also done with Mayer's reagent, but the results were slightly less clear)..solutions with large amounts of alkaloid would become thickly cloudy, whereas trace amounts would result in very mildly cloudy but still mainly clear solutions.. (the + to ++++ strength scale)..

some of the color changes observed (with Silicotungustic reagent) were:

DMT (P. viridis, A. phlebophylla leaf) - salmon pink>orange

and in acacia info thread p.12#228:

.as for the above mentioned TLC method, i wish i had a video of JJ's workshop at EGA to send you, but the basics are this: mix ammonium sulfate with a small amount of NaOH in a damp bottle..run a hose from the bottle to a jar containing acetone or MEK..the NaOH will release the ammonium as a gas which will start bubbling into the acetone..buy a standard TLC plate..mix sample in a tiny amount methanol or methylated spirits..by glass capilliary tube or other method place a small spot of sample near the bottom of the TLC plate..place TLC plate upright in beaker with acetone/ammonium..wait 15 minutes..
..now, after drying the plate reagents such as Ehrlich's can be applied..or, bake/heat the plate (don't know optimum temperature) ..tryptamines, after heat treatment will fluoresce under UV (don't have the actual wavelengths handy) ..ß-carbolines will show up UV without heat treatment..
if looking for dmt/nmt, even in a very crude gummy quick extrcact the smell should be an indicator..

 

[Endeavor]

..i posted the following in entheogenic effects of NMT p.3

'M'(2002) subjected a number of acacias to reagent testing, following Roveli(1967)'s recommendation for alkaloid field testing..
2-4 grams of fresh plant material was ground in a mortar and pestle with sand..this was mixed with DCM (or hexane), the sand and goo filtered,
and the solution (approx. 10-20ml) put in a test tube..
..to each test tube, the reagent was added (parallel tests were also done with Mayer's reagent, but the results were slightly less clear)..solutions with large amounts of alkaloid would become thickly cloudy, whereas trace amounts would result in very mildly cloudy but still mainly clear solutions.. (the + to ++++ strength scale)..

some of the color changes observed (with Silicotungustic reagent) were:

DMT (P. viridis, A. phlebophylla leaf) - salmon pink>orange

and in acacia info thread p.12#228:

.as for the above mentioned TLC method, i wish i had a video of JJ's workshop at EGA to send you, but the basics are this: mix ammonium sulfate with a small amount of NaOH in a damp bottle..run a hose from the bottle to a jar containing acetone or MEK..the NaOH will release the ammonium as a gas which will start bubbling into the acetone..buy a standard TLC plate..mix sample in a tiny amount methanol or methylated spirits..by glass capilliary tube or other method place a small spot of sample near the bottom of the TLC plate..place TLC plate upright in beaker with acetone/ammonium..wait 15 minutes..
..now, after drying the plate reagents such as Ehrlich's can be applied..or, bake/heat the plate (don't know optimum temperature) ..tryptamines, after heat treatment will fluoresce under UV (don't have the actual wavelengths handy) ..ß-carbolines will show up UV without heat treatment..
if looking for dmt/nmt, even in a very crude gummy quick extrcact the smell should be an indicator..

Thanks, very helpful of both of you. a Few questions though


I guess I'll do the; methylated spirits/plant solution with the ammonia sulfate/NaOH/acetone. Seems Simple

Endlessness, you seem the expert... what temp and how long do i heat the plate? is there any particular type of UV light i should get?

is beta-carbolines and tryptomines really the only two things that will show up florescent?

sorry for being frustratingly clueless :lol:

 

[AkashicFields]

I think someone was doing this and gave a talk at the recent Entheogenesis conference in Melbourne. Sorry can't remember who maybe this guy EnTheo.net is for sale | HugeDomains You may be able to find out more via their web-site or facebook page....

 

[endlessness]

Im not sure about the baking temperatures because Im working with Merck Silica Gel 60 F254 plates, which are pre-treated with some reagent that make all spots shine under UV at 254nm, no need to heat or anything.

They are more expensive but if you use the 20x20cm plates, you can cut them in half and its already enough run distance to separate well most substances, and your costs are halved. With these plates I use, no need to play with ammonium sulfate or NaOH or whatever, just put a very small amount of the solution with alkaloid in the plate, let it run on methanol:ammonia, let it air dry and look under UV... Compare heights/Rf and drop some reagent like marquis/pdmab to confirm color change is as expected for the identified substance.

 

[Endeavor]

I'm still a little unclear :? what do you use as the alkaloid solvent when using methanol:ammonia as the dip?

 

[endlessness]

Just so we understand ourselves better, by "dip", do you mean eluent, as in, the solvent that will be inside your box and that the plate will soak up to separate the substances, yes?

And by alkaloid solvent, you mean the one that the alkaloid or or plant mixture will have been dissolved in, and that you will add a small amount to the bottom of the plate/paper so that later it is taken upwards by the eluent, yes?

Ok so, as for alkaloid solvent, it doesnt matter what it is, as long as it dissolves the alkaloid in the first place.. I say this because, once you get a tiny amount of this alkaloid solvent and put it in the plate, you have to wait for this small part of the plate to dry before putting your plate inside the box with the eluent. We use methanol because we also play around a lot with alkaloid salts, and if it was something like, say, acetone, it wouldnt dissolve the salts.. But as I mentioned, as long as its a solution that has your alkaloids and dries up before letting it run, its all good, it could even be a bit of (acidic) water with your alkaloids.

 

[Endeavor]

Awesome :d thanks heaps (and for the promotion)

I'll get right onto the testing, damn TLC plates are expensive though