Hey DFZ,
how did your experiments go?




Although the Tek has delivered good results it is still in developmental stage. I believe the lab results speak for themselves so I have (struggled to) add a wiki page. Feel free to add to it.

I have also fixed some photos and hope to add more as many observers of the forum are visual learners.

Hi _Trip_, this is great work, thank you 🙏

I'm finding a potential application where the EA extract is done later. Start as usual with the water/vinegar extracts and base precipitation Sakkadelic style.

Then, dry and extract the crude harmalas with boiling EA. It leaves a bunch of gunk behind and seems very selective. Indeed, upon cooling white free base harmala candidate begins to precipitate. It is sparkly and white I don't even need to salt. I think freezer temp will Xtalize a lot of the FB. The used EA should be usable for the next batch too.

So instead of:

- Acidic water extraction, base, decant, multiple water washes, manske, dissolve and base again, rinse, etc, etc, dry.

Another possible apporach could be simply:

- Acidic water extraction, base, decant, dry, EA re-X

Also, as FYI, I re-Xed the precipitated water pulls separately. The first pull had a lot more junk. For what it's worth, the ratio of junk/product is highest in the first pull. I think Sakkadelic mentioned doing a quick water wash before extracting to eliminate the junk up front without affecting product. That could make sense, junk solubility in water must be super high based on the re-x gunk I get from the 1st pull.

That is interesting, I never got any precipitation early on with room temp EA (during tail end of winter in the shed) however the EA may not have been cold enough, may have been a EA saturation issue or too many impurities.

Out of curiosity what percentages and time have you got your modified Tek down to?

I've found with this approach more water in the dissolving stage before basing seems to really effect color of freebase extract yet it doesn't represent purity in contrast with lab results (B vs C). You can get quiet a whitish precipitation with the standard tek, however, extra clean up steps make it more tan (but produce a clearer signal in the LCMS). I've also have not bothered with sodium carb or ammonia washes at the end with the last filter which could produce a even whiter product.

But not crystal white, which is intriguing Loveall.

It does seem EA is quite selective, I have had runs when harmala citrate has crystalized but this was with a CaCl2 dry before salting. Makes me think if an optimal ratio of water to paste was worked out citrate crystals would form over goo.
This was tested (sample C) a chemical dry and mini A/B at the end was performed, however lab results did not vary enough to be considered an issue (and as previously mentioned came out more impure by look but produced a cleaner signal). Which I found strange.

I wonder if a straight to base with calcium hydroxide, boiling EA pull and freeze precipitate could work??? Harmala cruystals straight up? then a quick ammonia or sodium carb wash???
Could be the quickest Tek to date. Your thoughts?

Either case solid work Loveall. Look forward to hearing what comes from it.

I unfortunately have had no time to experiment and contribute at the moment.

I guess the hot STB and freeze could work. Definetly a good idea worth a try.

I've been working my way through 4 gallons of rue tea, somewhat on the periphery. Settling took ages so I ended up just leaving it be for a couple of weeks. In the next few days I'll have separated and washed the sediment and was planning to dissolve a decent-sized portion of crude harmala freebase in butyl acetate once it has dried off. Then I'll be replicating the citric precipitation at this larger scale using a citric acid solution. Updates when it's happened.

I can't get FB harmala to crash out of EA with freezer temps. Are you sure that white precipitation is harmala? And I've never seen harmala crash out of EA upon cooling unless there is an ideal saturation ratio criteria I'm not meeting.

Interestingly, the EA turned from yellow to red upon freezing it overnight. I would like to know the chemical reaction happening there.

Ive been experimenting with clean up steps since i dont have ammonia available. Sodium carbonate washes dont seem to clean it up. I've alos learnt Harmala is also soluble in iso and MEK.

I'm pretty sure it was harmalas crashing, but now I'm doubting myself.

This was very concentrated and already relatively pure harmine/harmaline from a water base crash.

It may have less plant compounds than your example. Besides lower concentration, sometimes, plant compounds from raw plant extract can complex with the product and enhance solubility.

My sample was from FB harmala already extracted then re-dissolved in EA.
So then maybe it was likely a concentration issue?

Edit: It is indeed a saturation issue. Harmala does crash out of EA if minimal EA is present. However, mine was not white but that's expected as the Tek used to obtain the harmala I used is different than the Tek you're using Loveall.

Quick update I have done multiple tests on this now. The key in this Tek to getting 'whitish' harmala is the amount of water used in the dissolving of citrate step.

Below are two examples, same batch of seeds, same EA pulls, same run, same everything except the one on the left used 4 times more water before basing. If using 100g of seed and 1lt of hot EA when pulling then 850ml should give a relatively clean harmala result. 1.25lt of water before basing will give an even whiter looking harmala.

The downside, although not tested could be that the yield decreases. Currently however, results indicate this may not be the case.

Although I must stress lab results between 2 of my samples indicated color did not equate to purity.


On a side note phosphoric acid appears to act very similarly to citric acid in preliminary tests. I'll have to do a full run to confirm if the end results are any different.

Had about 1g of zinc converted "cleanish" harmalas.

Decided to try an EA re-X. Used 200ml of EA. Most dissolved upon boiling (but not all). Not sure if what remains is contamination or if the boiling EA was saturated.

Filtered the re-x after cooling (do not handle hot solvents, they can spontaneously ignite). Gunk ended in the filter, so that was nice.

Freeze precipitation caused the solution to crash xtaline material (R1) and to change color from orange to a deep red.

What is the red stuff? I evaporated the EA, and washed what was scraped up with alkaline water (R2). See image of the resulting red powder below.

Doing manske on R1 and R2 is showing needles on both sides. The R1 manske is is the less yellow I have ever seen.

I wonder what R2 is? It is only 100mg, but I think it has been in all my harmala extractions since it won't manske out and precipitates in mild alkaline water. Only way it separates for me is with EA re-X.

It does appear when (harmala) EA freezes the EA goes red. So I wonder what reaction is happening there.

When I used already extracted harmala and freeze precipitated it in EA the harmala remained more or less the same color maybe slightly more red?

Anyway my thoughts are and I could be totally wrong, is that it is likely a saturation issue. I would bet R2 is mostly still harmala just with few more impurities. Perhaps the harmala crashes first and impurities don't crash as easily. However, if there's a saturation issue present as well, I would guess harmala crashes first and then impurities and left over harmala remain in the EA.
I base this on absolutely no facts.

Cool looking red crystals though. Sorry Loveall, when you say zinc converted do you mean THH?



I've had success with cleaner harmala simply by using more water in dissolving of citrate step. And I don't wash with ammonia. Although I'm thinking of purifying sudsy ammonia so I can see how clean I can get it in the final step. Sodium carbonate doesn't seem to work the best.

But I love your work with the freeze precipitation. It would be interesting to do a base, pull then freeze run. Just to see what comes out of it. If I have time this week I may give that a quick go.

When I say zinc conversion I meant converted from harmine/harmaline rue mix to harmine/THH. I didn't make that clear, thanks for asking.

DFZ,
just out of curiosity what is the latest with the BuOAc?

I've been too busy gardening The BuOAc is sat in the can next to my jar of crude alks and I just keep nibbling the alks

There was also a huge backlog of other stuff due to me being me plus some medical issues. And being a parent, of course.

I did repair my microgram scales, finally, so I'm hoping to try out a few solubility tests this evening. It would be great if BuOAc turns out to be a viable alternative solvent for quick cleaning of harmalas.

If it turns out well, I'll slate a test for extraction from the seeds as the PC boils took ages, mostly due to the cooling phase of the process. The good thing about the cooks is that the alks can be reacted directly with ascorbic acid - as I've mentioned elsewhere.

Ascorbic+PC-cooked alks are way smoother than the equivalent amount of crude rue tea would be, and the effects from a dose of over 400mg are clearly distinct from those that might be expected from a mixture containing 50% harmaline. And perhaps the other relevant point here is that I didn't want to be playing with stinky solvent while going through the weeks-long process of ramping up my doses or harmala alks!

By the way, to make sure I hadn't simply developed a tolerance to the side-effects of harmaline I did make sure to take a week off before trying a further 400+mg dose. TLC results will help a lot here but I still need to find my gypsum stash so I can make myself a few plates. If that fails to show up I'll probably do some column chroma instead.

OK, so I've had a little play around with 2.1 grams of decidedly damp freebase harmalas from the PC cooks with ascorbic acid based with ammonia. They were still damp after washes with 5% ammonia.

8 pulls with c. 20 mL BuOAc each time making a total of 150 mL of a yellow-coloured extract which fluoresces blue! All the pulls from start to finish fluoresced the same shade of blue, and the remaining solids have lost their reddish cast, going from kind of somewhat reddish cigar brown to a much darker shade. The fluorescence contrasts markedly with the green of an aqueous solution of the mixed harmala ascorbate.

It got to the point where I had to stop work for the night so it still remains to be seen whether the remaining solids will dissolve if further fractions of BuOAc are used. Also on the cards is checking what colour fluorescence the residual solid might produce when dissolved in either more solvent and/or in acid.

There was no time to try out a citric acid precipitation yet. Firstly, the final pull of the wash is still settling; the remaining solid had become completely powdery after starting out as a paste. This indicates that the butyl acetate does incrementally dissolve out the water in small amounts. The solvent was dried by swirling over sodium chloride crystals before use to assist this process of drying out the paste, although initially the solvent was used as it came and instead the first 3 pulls were removed and dried half way through the process and then returned to the paste.

One observation with the first few (~4) pulls was that the paste became more liquid as the more soluble harmala base dissolved out more quickly than the water, leaving less solid in the paste. By the sixth pull, the paste started to become crumbly and on the seventh it was a clumpy powder. Separation was quick and easy with all these pulls. Only on the eighth pull had the solids become so dry that fine powder dispersed into the solvent, hence requiring an overnight settling period.

Instead of doing a preliminary citric acid precipitation test, I clumsily dropped the beaker containing the remaining 80+ grams of harmala paste onto the stone floor. After a lengthy string of expletives, I was greatly relieved that I hadn't bothered to dry out the paste as this meant that most of it remained in one large blob in a bigger fragment of beaker and the rest of it was confined to a few smaller pieces which were easy to recover with minimal losses. All of the blobs, shards and fragments were placed into a larger beaker and doused with dilute acetic acid.

The harmala acetates solution will be filtered and processed in due course. This was something I'd been meaning to do for several weeks but it was all too easy just to keep chipping away at the nice big blob of yummy alkaloids, so now my hand has been forced and this whole mishap may be considered to have progress as a silver lining

A further update on the freebase harmala/butyl acetate solubility test:

The powdery solid was transferred to a Pasteur pipette containing a small cotton plug as a filtering device. After spending several hours topping up the solvent with another pipette, I finally rigged up a syphon system using a suitably bent piece of glass tubing and a beaker. This 'automatically' keeps a layer of BuOAc topped up in the filter pipette. The solvent is percolating at a rate of about 4½ drops per minute, which equates to roughly 270 drops an hour. That's somewhat less than 7mL per hour for these particular pipettes, meaning 50mL should get through the night unattended.

One interesting observation so far is that the powder, which had already lost a slight reddish tinge in the paste form to become a dark olive colour, has since had further pale colour leached from it in a layer starting from the top. This is of course where the greatest amount of contact time with fresh solvent has occurred. This dark chocolate coloured layer has been expanding downwards over the past 48 hours and I would anticipate the lighter colour to have been leached completely in another couple of days. (The colour leaching was slower at first before the syphon system was made as it kept running dry while unattended.)

While the appearance and fluorescence of the leachate are the same as those of the initial stirred/mashed batch, I'll keep the fractions separate on account of the changes in the colour and consistency of the solids. Wouldn't it be handy if a simple solvent percolation method could separate the different alkaloids? It's kind of a lazy form of not-quite chromatography.

One thing this solvent won't be winning prizes for is speed (at least with my methods) and it looks as though 300mL will be needed for a mere 2 grams of alkaloids so I'm not sure how favourably that would compare with the EtOAc (EA) in a rue seed extraction.

DFZ,
I wouldn't say its slow per say, you could speed up the filtering if using coffee filters or lab grade vacuum pump filter setup, easily enough.



Are you saying the ascorbic acid and pressure cooker potentially produced THH (blue glow)? or the BuOAc just pulled harmine? (which I believe also glows blue).

I'm suggesting there's a strong possibility that at least some THH may have been produced by the ascorbic PC cooks. This is not only based on the blue (with a hint of turquoise) fluorescence I've been seeing here. [The fluorescence from harmine that I've seen is a deeper blue and the pictures of THH fluorescence tend to be much more in the direction of turquoise.] The colour of the fluorescence is a fickle indication but the smoothness of my subjective experiences with the crude alkaloid precipitate also hints that the harmaline, if any, is present only at a greatly diminished concentration.

There are numerous caveats though.
Firstly, the first three cooks were precipitated separately and have not been analysed yet as they have so much more of the rue crud that processing was a total nightmare, even with vacuum filtration. So that's where the harmaline might be.
Secondly , I may just have been incredibly lucky and got a rare batch of rue seeds that were unusually high in THH. This has not been controlled for and all those seeds have been used up, although I did go and buy another kilo - fortunately with the same packaging - from the same shop.
Thirdly, I now have four fractions from the test sample post-butyl-acetate. While all three liquid fractions have the same pale blue-turquoise fluorescence, I don't know if this is simply a solvent-related phenomenon. Crashing with citric and dissolving the citrates in water could change everything, in the sense that there might be a change in the fluorescence colour at least, both because of the different solvent and because this is currently the fluorescence of the freebase which is being observed rather than that of the protonated cation. Then there's the solid that remains in the pipette. This needs to be quantified and examined for solubility in acidic water along with any resulting fluorescence that may arise. It does mostly look like rue crud but maybe there's still an alkaloid or two in there.
Fourthly, coming back to the rue crud/solid remnant - if this turns out to be a significant quantity of pharmacologically inert material it would force me to reassess my interpretation if my subjective experiences and drialnk(?) moar... My subjective tests do still hint that I've been tasting a mixture that was predominantly harmine + THH, though.
Fifthly, I want to re-iterate the unreliability of making assumptions based on subjectively observed fluorescence. And I should apologise for still not having found my gypsum for making TLC plates

Here's some reading - for my benefit as much as anyone else's!
Solvent Effects on Fluorescence Emission
Photophysics of harmaline in solvent mixtures
Experimental and computational study of solvent effects on one- and two-photon absorption spectra of chlorinated harmines
It would be very helpful if anybody knew of specific, pre-existing data regarding solutions of harmala alkaloids in butyl acetate - I still find it a little hard to believe that I might actually be doing novel research.

Specifically, the BuOAc extraction process has taken about five days, although it could have been made quicker by starting with dry freebase and putting it straight into a syphon-fed percolation column. The observation of how the BuOAc dries the paste is of limited use in the practical execution of preparing successive fractions of harmala alkaloid solution.

If the goal was to get mixed harmala freebase into solution as quickly as possible I would think warming it up would help considerably. I maybe could have used vacuum filtration on the rue cooks although a kilo of seeds still would have taken ten or more batches going through my largest Buchner funnel. This kind of brings me to the other point about my approach here - it's at odds with the spirit of the alkyl acetate approach that you've outlined here because I've used water cooks for the ascorbic acid investigation and then used precipitated freebase for a somewhat incidental solubility experiment. I guess that should be my sixth caveat.

And to round off, a little update - the three fractions of BuOAc extract were progressively lighter yellow in colour, the first being morning piss colour, and the third being the equivalent after a pint of water and five cups of tea, with the second showing a shade somewhere between. This is also matched by the volumes of each fraction - 150, 100 and 50 mL respectively. The fluorescence follows a similar pattern, which brings me onto the final point worthy of note. If you have a point source beam of UV for examining your solutions, it's very easy to see that the beam will penetrate to a depth that inversely correlates with the concentration of the fluorescent solute. More molecules = more absorption. Thus, the spy pen light beam only gets a couple of centimetres into the first fraction, 4cm into the second, and passes all the way (4cm) through the third fraction and can then penetrate 1cm of the first fraction as well. This might give some indication of the relative concentrations in each of the fractions.

Citric acid precipitation should proceed this coming morning.

EDIT: Maybe even more tangential but there is this: Pilot study on the uptake and modification of harmaline in acceptor plants: An innovative approach to visualize the interspecific transfer of natural products. https://doi.org/10.1016/j.phytochem.2020.112362 Pretty awesome paper!

As anticipated from preliminary test tube experiments, the fractions are all giving nice, primrose-yellow precipitates on addition of citric acid saturated BuOAc (CASBA!). On aggregation the precipitate appears more orange, although this could be a yellow harmala gunk that has coagulated. The precipitate is very fine at the moment and takes quite some time to settle.

Because the most concentrated fraction was also the largest (150mL), I'll be testing 25mL portions of it with a number of different methods. CASBA was of course quite effective, but the precipitate is always very fine. Next up, medium-coarse (from the packet) citric acid crystals; to be followed by finely ground CA. Also available are tartaric, malic, and fumaric acids but I'd like to get hold of some succinic acid too if at all possible. I'm hoping at least one of these options gives a result which settles more quickly.



One thought on why, in the earlier tests, the precipitate turned pink and then went back to yellow: it could be that the particles became coated with a reddish impurity that is insoluble below about 16°C but which redissolves when the temperature rises again. Or there is a phase transition between different crystal forms at this temperature. This seems to have been mirrored in the reports of a similar colour change upon refrigeration of the EA precipitate.

Does this colour change reverse on warming if the precipitate is kept in contact with the EA solution? What happens when the precipitate is filtered off cold then allowed to warm up once dry? And what happens if the precipitate is cooled with a portion of fresh EA?

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.