Research Sceletium alkaloid extraction

[Woolmer]

I performed a CIELO-type extraction on 43 g of Sceletium tortuousm with 80 ml water and 13 g CaOH. It was a bit too much water as a tiny amount of water came out in the first pull when I squeezed it with the coffee plunger. The water did not seem to pass through the coffee filter though.

I dumped the citric acid without any stirring and it resulted in a very thick goo. Unfortunately, there was plastic that came loose from the lid of my jar and some probably dissolved into the ethyl acetate. I had to redissolve the product back in water and try to filter the plastic out.

Yield 1.37% mesembrine (and related alkaloids) citrate

I tried to make the benzoate salt by doing a mini A/B with NaOH but struggled to redissolve the freebase back into ethyl acetate or acetone. The bit that did dissolve I salted with by dropping the benzoic acid directly and it made a less waxy but still not crystalline product.

I might try to dissolve some in pg/vg to vape it, though I am a bit wary of the plastic from the lid.

The second picture below is next to a mescaline extraction that I was doing at the same time, right after having added the citric acid. The last picture is the citrate salt.

 

[Woolmer]

I repeated a second extraction using 50 g S. tortuosum.

75 g water, 13 g CaOH, 5 x EA pulls totaling about 420 g. The amount of water was perfect and none came out when squeezing hard with the French press.

This time I did a fridge rest before salting. Something started precipitating in the fridge so I placed the EA in the freezer. About 30 mg worth of some white precipitate formed. I vaporized a small amount (1~3 mg?) but could not feel effects past placebo. It seems to have a very low boiling point as it vaporizes very quickly on my convection e-mesh setup. It is likely not mesembrine as the literature states mesembrine freebase being a brown oil.

The EA was then salted with citric acid, yielding 550 mg (1.1%) of the alkaloid salt. This time I redissolved the citrate alkaloids into water and then evaporated the water on a plate in the oven at around 60 °C. Interestingly, the resulting extract is very hard to scrape and when scraped small shiny crystals can be seen. Shortly after scraping the extract seems to have picked up water and gone back to a thick syrup texture.

Since the yield came out lower than the first extraction I decided to combine it with the leftover sceletium from the first extraction and do more pulls. I was able to (so far) recover 170 mg more.


I tried playing around with other salts (benzoate and fumarate) but struggled to redissolve the freebase (which I converted using NaOH) into either ethyl acetate or limonene. **I found out recently that NaOH and ethyl acetate should not be combined due to hydrolysis. I have only been able to recover 100 mg of freebase by evaporating the lye solutions and pulling with acetone so I am not sure what has happened to the rest.

 

[Transform]

Nice work! Have you tried the product? Can you run a TLC with it to see how many components get pulled with this method?

 

[Woolmer]

I have not gotten around to trying it properly yet. Only vaporized a tiny bit of the crystalline material on e-mesh during the comedown of a bufotenine experience so I could not quite tell the effects .

I will try run some TLC plates.

 

[Woolmer]

I have vaporized a bit more and put small amounts under my tongue. The effect so far has just been a subtle mood boost, but perhaps I just need to up the dose. It's hard to weigh given the stickiness.

Attached is a TLC plate I ran with DCM:MeOH 3:1. The sample was dissolved in MeOH. The two columns are just two replicates.

 

[aizoaceous]

I'm not sure what to make of that TLC. Can you get a commercial standardized extract to compare? If you haven't found Srinivas Patnala's thesis already, then it also has a chapter on TLC (and it's generally excellent).

And I assume that's purchased milled powder? I tried two commercial sources myself, and both yielded around 0.2% mesembrine, by HPLC with commercial extract as my standard. I think that's potentially a significant underestimate, since my standard is ">3%" and public analyses for it show up to double, and I'm giving freebase and you're giving citrate, and mesembrine isn't the only alkaloid. Your yield still seems suspiciously high to me (though such yields are certainly possible; the powders I've bought are weak, but my best plants are well above that.)

I've been extracting twice into acidified water, centrifuging or settling for a few days, filtering, making basic with ammonia, then extracting twice with EtOAc (again with centrifuging or long waiting). I then evaporate off the solvent at 50 C, yielding a yellow oil or wax. I redissolve that in 0.5 M citrate buffer at pH 5.0, around 10 mg/mL mesembrine. I dose 100-200 uL under my tongue, and the euphoria is unmistakable. I'm not particularly happy with this method, since the emulsions are unmanageable without either a centrifuge or extreme patience. Patnala extracted initially with alcohol then evaporated that and redissolved in acidified water, and others also report that solves the problem. I guess the CIELO approach solves that too, but that also pulls the chlorophyll (but if you redissolve in water after evaporating off the solvent then maybe that's fine?).

 

[Transform]

Attached is a TLC plate I ran with DCM:MeOH 3:1. The sample was dissolved in MeOH. The two columns are just two replicates.

It's a good idea to include a standard spot, using something like caffeine, alongside your unknowns. This helps with cross-referencing between different runs, where variations in performance can occur because of variations in real-life conditions [temperature, humidity, ventilation, solvent composition etc.]

 

[Loveall]

Woolmer, have you tried to salt the EA directly with fumaric acid?

 

[Woolmer]

Woolmer, have you tried to salt the EA directly with fumaric acid?

I've not tried to salt with fumaric acid. Will try if I get a chance.

I'm not sure what to make of that TLC. Can you get a commercial standardized extract to compare? If you haven't found Srinivas Patnala's thesis already, then it also has a chapter on TLC (and it's generally excellent).

And I assume that's purchased milled powder? Your yield still seems suspiciously high to me (though such yields are certainly possible; the powders I've bought are weak, but my best plants are well above that.)

I've now gotten a commercial extract with known quantities of mesembrine, mesembrenone, and delta-7-mesembrenone that I'll use as a reference. I've seen Patnala's thesis but haven't looked into it enough. Yes, it is commercial milled powder.

I am also skeptical of my yield as I've yet to feel anything that's definitely past placebo even when dosing in the low milligrams.

I guess the CIELO approach solves that too, but that also pulls the chlorophyll (but if you redissolve in water after evaporating off the solvent then maybe that's fine?).

The chlorophyll does not crash out when salting the EtOAc anyway though, right?

 

[aizoaceous]

The chlorophyll does not crash out when salting the EtOAc anyway though, right?

I mean if you evaporate the solvent off to recover the alkaloids as freebase, instead of trying to precipitate them as salts. All the methods I've seen in the literature evaporate off, since that pretty much always just works. Precipitation is better when possible since it's selective, faster, etc., but some effort is often required to find a method that yields a cleanly-separable precipitate.

Evaporation is otherwise strictly worse, since it's not selective. But I was thinking that you could extract alkaloids into EtOAc with a CIELO-style method, evaporate off the solvent, then redissolve into acidified water (or buffer) and filter, settle, centrifuge, extract with naphtha, etc. to remove the fats. That said:

• The solution wouldn't be directly suitable for vaporizing. I believe sublingual or intransal is much more common, though.
• You might need to dry the EtOAc (like with anhydrous magnesium sulfate) before evaporating to avoid carrying over lime dissolved in the water dissolved in the solvent. I don't usually dry, but I'm using ammonia so excess evaporates. I guess I do carry over some ionic stuff from the plant, but not visibly much.
• There'd be no opportunity to weigh the product, so you'd be depending entirely on your TLC to judge the dose.

It might be worth the experiment, though. Sceletium is very rewarding to extract, since a small amount of material yields many active doses. The chemotaxonomy is also complex (per my thread), and I believe could be usefully studied by TLC; so I'd highly recommend it to anyone with an interest in plant drugs.