Ethyl Acetate Approach for Harmala

[downwardsfromzero]

Love ya work DFZ,
For memory the EA stayed red after freezing precipitating the harmala. This wasn't crude harmala extracted EA. It was with already purified harmala that was then dissolved in EA to test freeze precipitation.

Thanks

I'll earmark a sample of freebase solution for temperature cycling tests. It seems the reversible colour change displayed by my initial test precipitate was a peculiar anomaly.

On further examination of the CASBA-CIBLO precipitation it appears that the primrose-yellow suspension aggregates to form dirty orange masses as the solution settles. Much as one might expect, settling is more rapid with the more concentrated fraction.

Speaking of settling, I just decanted the solution of re-acidified freebase that fell on the floor. Despite slow filtering through a generous cotton plug, there was a couple of milimetres thickness of black 'harmala crud'™ sat at the bottom of the jug. This kind of highlights the value of patience in harmala extractions as this has had over a week's settling time. I'll be leaving the decanted solution to settle for a few more days to help clean that up - something any home winemaker should understand.

PS - thought I'd best mention, the repaired microgram scales turned out to be way off in the calibration tests, not to mention lacking stability. I'm still restricted to centigram resolution until this gets sorted out.

 

[_Trip_]

I find when doing the tek the "citric EA" jug will crash out a little more over a few days as does the based harmala jug. (only a small amount)

I usually decant the citric EA and use the harmala citrate goo for the next run. As for the FB harmala that crashes over a few days in the left over base jug, instead of being a nice white-grey harmala it too is a almost black crude harmala. I have no idea how much of it would be usable (could be worth a LC-MS). I always figured it was mostly impurities?

The reason I wrote up the tek to be as quick as it is, is because the end result is very decent purity and color (IMO) and can be done quick with very decent yields. One can always do repeat runs on spent material as well for a better yield. I guess I'm just impatient. The other reason for the quick tek as well, is, if you live in a country where harmala is illegal you can do it in a couple hours rather than having 'manufacturing evidence' sitting around for weeks.

I always figured it was just impurities in the base jug that crashed out when left for a few days or a week. At the very least it looks impure by color. I guess ammonia wash could clean it up and more harmala could be recovered?

 

[Loveall]

_Trip_, thanks for your work on the TEK and the mass spec analysis.

It may be interesting to do your harmala TEK with calcium carbonate.

Reason is that the pH should be in the high ~9.5 to 10 range, right where harmalol becomes neutral if I understand this paper correctly.

At this pH range harmine and harmaline should be in freebase form amd neutral. Nothing new here. However, harmalol should be neutral (isoelectric) and could move into the EA (?).

The resulting goo from citric salting could have Harmine, harmaline, and harmalol (?).

Harmalol in rue may be substantial and >1%. We are currently throwing it all away with our current teks. It is difficult to get easily, but your TEK using calcium carbonate may be a path (?)

 

[_Trip_]

I like where your head is at.

Will calcium carbonate base well if it is not water soluble?

Thoughts on separation? Will it be left in the base water when base precipitating harmaline and harmine with sodium carbonate?

 

[Loveall]

I like where your head is at.

Will calcium carbonate base well if it is not water soluble?

Thoughts on separation? Will it be left in the base water when base precipitating harmaline and harmine with sodium carbonate?

I think it should dissolve in the acidic rue well, then stop dissolving as it becomes alkaline. Final pH should be in the right range which is what matters I think. I could be wrong.

Separating the different harmala citrates could be tricky. Harmalol is difficult to work with but open to ideas.

 

[downwardsfromzero]

Calcium carbonate does get used as a base in some organic synthesis reactions. Its lack of solubility need not be a problem.

I'm wondering now if harmalol is behind the orange to pink colours of harmala citrates that I've made. The crude freebase was precipitated with ammonia around pH 10, then dissolved in BuAc, and precipitated with citric. When my lablet is up and running again for autumn I'll be sticking some of this colourful stuff through a little chroma column.

I think the colour comes from oxidation. I would also not be at all surprised to find that harmalol was light-sensitive. Therefore, anyone who really wants to get to grips with this stuff should get up to speed with inert atmospheres and work under dim red light. Does make it look rather like an acid lab, though :lol:

 

[Loveall]

Good luck witj Butyl Acetate DWF.

I ran a test with 30g of rue following the general srcipt in _Trip_'s tek. Did not do microwave and therefore used less water. I did not heat up the EA since I'm just testing.

- Mixed 1 part water and 0.25 parts calcium carbonate with ground seeds. Verified pH was alkaline by rubbing a sample of the paste on pH paper. pH is between 9 and 10, may be a little lower than our target of high 9 for harmalol, or maybe it is perfectly OK, IDK.

- Pulled with EA at room temperature. Looks redder than the tek, I think that is consistent with Harmalol. It could be the same red we throw away after the first basing in the standard water based extraction. It glows green under a cheap UV light.

- EA currently resting in the fridge to standarize and lower the water ammount before salting.

Will decant/filter and salt with citric. Hoping to get a goo rich in Harmine, Harmaline, and Harmalol citrates. Harmaline (and Harmalol?) could be converted to THH and tetrahydroharmal (that is potentially 6 different harmalas).

 

[Loveall]

Update:

* No water formed in the fridge, this EA from rue calcium carbonate is a lot dryer than the ones from CIELO
* Instead of water I got what looks like harmal FB precipitating. Makes sense since _Trip_ has reported how important temperature is.
* After 18h the extract is noticably redder (see image below). Could this me harmalol oxydation? We may need to salt right after extracting?
* Adding a tartaric acid salting test bases on _Trip_'s result

 

[_Trip_]

I always grind my acid to a powder, it seems to help the reaction. It will be interesting to see what you get I have a feeling EA straight from rue and salted with tartaric could give goo but let us know.

 

[downwardsfromzero]

Loveall - looks promising with the calcium carbonate there! I can't wait to see the results of adding an acid.

I have decent amount of magnesium carbonate so I'll give that variation a try over the coming days. After that, maybe I'll have a go using the strontium oxide I've mentioned from time to time for the past however many years :lol:

If this works out with calcium carbonate, it's another - yet another - revolutionary step for plant extractions. Perversely perhaps, if for whatever reason calcium carbonate might be unavailable it can still be made by combining sodium carbonate or bicarbonate with calcium chloride.

So here's another idea off the top of my head - the seeds could be treated with sodium carbonate solution [IIRC either benzyme or mindlusion mentioned doing this as part of some other kind of extraction scheme] and then mixed with a calcium chloride solution. Perhaps it would then be necessary to wash out the resulting sodium chloride - but that defies _Trip_'s point of making the process as quick as possible so there would need to be some other kind of discernible benefit to this spur of the moment idea.

 

[Loveall]

The citric salted quickly (1/2 of the 1st room temp pull from 30g of rue seeds). Recovered 250mg of red powder that was easy to decant (see image). This may change to a goo over the next few days, will monitor. I'm surprised I did not get a good directly. What is in the red powder? Don't know but hoping for relatively pure harmine/harmaline/harmalol citrates.

Tartaric is settling as a mix of white powder and goo. I can see both in the jar. It is salting more slowly.

 

[_Trip_]

Tartaric is settling as a mix of white powder and goo. I can see both in the jar. It is salting more slowly.

I found tartaric isn't anywhere near as soluble as citric is there a chance the white powder is excess.

 

[downwardsfromzero]

That extra hydroxy group probably doesn't help. Malic is more the way to go, I'd think (and always did - tartaric acid is a metabolic poison, really).

 

[_Trip_]

Planning to test malic. DFZ, are you saying when heated it creates a toxic metabolite? or it is toxic in general?

Tartaric acid is a naturally occurring acid in fruits I thought it had a good safety profile?

 

[downwardsfromzero]

Obviously, tartaric acid is far less toxic than, say, fluoroacetate - otherwise it wouldn't be approved as a food ingredient (at least so one might hope). I'm slightly stumped for a precise reference about this piece of biochemical trivia, in fact. The main thing is that humans don't absorb dietary tartrate particularly much, but if we did it could (IIRC) interfere with some aspects of the Krebs metabolic cycle.

Metabolism of L(+)-and D(-)-tartaric acids in different animal species - PubMed

L(+)-Tartaric acid, which is the naturally occurring form, is used as a food additive in a variety of foodstuffs. Recently, there has been an increasing interest in using synthetic (D,L)-tartaric acid as a substitute. Several toxicological studies have been published on L(+)-tartaric acid, but...

pubmed.ncbi.nlm.nih.gov

Re‐evaluation of l(+)‐tartaric acid (E 334), sodium tartrates (E 335), potassium tartrates (E 336), potassium sodium tartrate (E 337) and calcium tartrate (E 354) as food additives

The Panel on Food Additives and Flavourings () provides a scientific opinion on tartaric acid‐tartrates (E 334‐337, 354) when used as food additives. The Scientific Committee for Food () in 1990 established an acceptable ...

www.ncbi.nlm.nih.gov

It's likely that tartaric acid is poisonous to dogs:

What causes grape toxicity in dogs? Playdough might have led to a breakthrough

We’ve known for years that grapes are toxic to dogs. But we didn’t quite know why. Now we might, and homemade playdough is the reason.

www.aaha.org

 

[Loveall]

The citric salted quickly (1/2 of the 1st room temp pull from 30g of rue seeds). Recovered 250mg of red powder that was easy to decant (see image). This may change to a goo over the next few days, will monitor. I'm surprised I did not get a good directly. What is in the red powder? Don't know but hoping for relatively pure harmine/harmaline/harmalol citrates.

Tartaric is settling as a mix of white powder and goo. I can see both in the jar. It is salting more slowly.

Update:
The red powder did not become goo. It is stable. Candidate for harmine/harmaline/harmalol citrates.

The tartrate goo disappeared over time. It is all powder now. Will collect and report on it. Candidate for harmine/harmaline/harmalol tartrate.

 

[_Trip_]

The tartrate goo disappeared over time. It is all powder now. Will collect and report on it. Candidate for harmine/harmaline/harmalol tartrate.

I've seen citrate goo do the same thing and crystalise. Might be worth testing the exact solubility of tartaric acid in EA too.

 

[Loveall]

The tartrate goo disappeared over time. It is all powder now. Will collect and report on it. Candidate for harmine/harmaline/harmalol tartrate.

I've seen citrate goo do the same thing and crystalise. Might be worth testing the exact solubility of tartaric acid in EA too.

Here is the candidate harmine/harmalime/harmalol tartrate powder after catching in a filter, washing with fresh EA, and drying the solvent. 200mg collected. Beautiful fluorescence (second image). Will repeat the PG solubility test that _Trip_ did.

 

[downwardsfromzero]

Here we have the long awaited pic of rue harmala citrates crashed from butyl acetate! (Summer happened and the test ideas got greatly simplified.)

Beaker on the left has relatively light fluffy stuff, yellow in suspension and orange-brown in bulk when still moist with solvent. On the right, a denser, olive-brown, clumpy material. Both display a similar fluorescence as seen in Loveall's photo.

This material was prepared by drip-leaching rue bases (precipitated from rue tea using ammonia) with butyl acetate through a cotton wool filter plug in a glass pipette. Black, insoluble material remained trapped in the filter. A differential solubility was observed (although whether this was noted down is another matter entirely... [EDIT: here they are!]) and this is what gave rise to the two fractions as seen in the photograph. One of them dissolved first, it doesn't entirely matter which, but it could at least be observed by the relative colour of the material in the extraction tube.

To the two resulting batches of orange-yellow solution was added a saturated solution of citric acid in moistened butyl acetate ('CASBA').

Moist butyl acetate was prepared by repeatedly shaking 100mL butyl acetate, fresh from the tin, in a stoppered measuring cylinder with ca. 2mL of distilled water several times for about ten seconds each time, followed by a short pause to allow droplets to settle. After this became too boring and it looked like no more water was being absorbed, a saturated citric acid solution was prepared by dripping the moist butyl acetate through another glass filter column with a cotton wool filter plug, this time filled with citric acid. The packet claimed this was anhydrous citric acid but that seems a tad spurious; it may well have been the monohydrate. Its hydration level was not assessed.

It is possible to be reasonably confident about the solution being saturated because the unused CASBA deposited crystals of citric acid quite readily owing to some evaporation of the solvent during storage, as well as the preparation method being a reasonable guarantee of producing a saturated solution.

The material in the photo is what remains after allowing the precipitates to settle and removing the supernatant.

Rinsing with fresh butyl acetate should remove any excess citric acid from solvent residues and evaporation thereof. If the solubility of citric acid is insufficient for this cleanup, a rinse with anhydrous acetone will be attempted, where there is a better solubility, but the solubility of harmala citrates in acetone is yet to be determined (as far as I'm aware).

More data to follow. Time for a pull from the seeds with BA, and I ought to get that chroma sorted too. Little (tiny) column is ready and waiting to be packed.

 

[_Trip_]

It will be interesting to do a side by side comparison on which solvent holds freebase better. I've been wondering if there is a better solvent to use than EA.
As it stands 3.6% white grey harmala is common for a (hot) EA run. (I seem to get a fairly consistent 3.6% per run for some reason). Thus a re-pull on spent seeds is required to get over 4.5-5%. Both runs can be done within 4-5 hours and the first in 2.5-3 if one is being quick. It would be great to find a solvent that'll pull 5% straight up.