Hi ava, thanks for your well wishes. How terrible to hear of your poor morning glories! It really beggars belief the pettiness we can encounter sometimes. I managed to get a few seeds from my plants that were in the sunniest location, the others I think got planted out too late after a failure of some early seedlings. I also think there may be a temperature/day length issue at my latitude. But more of that in the right thread.

The 1kg rue extraction really is an undertaking (and I baulk at the thought of an 8kg one!) but it was in part inspired by your THH reports, ava. It would help to be able to press the seeds and, despite serendipitously having found just the right bottle jack, I haven't rigged anything up yet. Instead I'm on about the ninth boil and still getting alklaoids - the size of my PC limits my boils to 1.7L at a time so I think next time I'll stick with a half kilo instead. State of play is I have three gallons of based tea, one of which has got to the point of a 2nd A/B cycle for the harmine fraction. I think my alkaloid concentration has been too high, which impedes crystallisation. My small-scale test on the Magnesium reduction went a lot more smoothly because of the lower concentration at the base stage.

There's also the question of rue tea's perpetual ability to precipitate ultra-fine black crud alongside its incredible foaming abilities. Both of these make vacuum filtration far more of a challenge than I anticipated, and I'm still puzzling over exactly how I'll be cleaning up my fritted funnels. Filter papers seem both to clog and to let through the fine particulates. The trusty cotton plug gravity filter has already demonstrated its worth in cleaning up the tea as it comes out of the PC and we'll have to see what it can make of the filthy-looking supernatant that gets syphoned off the precipitated alkaloids.

My putative magnesium ammonium phosphate which precipitated between pH 5.5 & 6.2 has remained undissolved in 96% ethanol, which is a good sign. After pipetting off, the ethanol had been evaporating in a Petri dish so we'll see if any harmine crystals appear there. It's beyond question that the low-pH precipitate here was MgNH4PO4, or at least largely so, simply because of the quantity in comparison with the higher pH precipitates totalling about 150mg (it was a 5g rue tea directly reduced with magnesium ribbon as a test so this is not entirely disappointing). The low pH precipitate amounts to some 2.02 grams - plus another amount which I've set aside from the very first ammonia diffusion precipitation (and which reminds me that they need weighing).

The question is whether my 150mg precipitate is harmaline or THH. I need to get a new pH probe so I've been relying on narrow-range pH papers for this and I've established that the two high pH (10-12/12-14) range papers have stopped working, presumably due to old age. It's therefore at present unclear whether my ammonia was getting to a high enough pH to precipitate THH, but there was a clear gap between the low pH precipitate and the high pH one. I was hoping to have titrated this but managed to break the tap key for my burette. Don't try cleaning your equipment when you're tired!

Because there was no clear harmine precipitate we are faced with at least a couple of possibilities - the harmine has perhaps become trapped in the MgNH4PO4, or maybe magnesium is capable of reducing harmine (which would turn out to be rather interesting). I'm pretty sure the seeds started out with some level of harmine present but this wasn't expressly tested so we can't exclude the possibility that there's simply no, or very little, harmine there although that seems rather unlikely. As Mark Twain once said (and Shulgin quoted him in this), "I love how science gives such an abundant return in speculation for such a trifling investment of fact."

Where did the harmine go? Has THH been precipitated? Will I ever be able to afford some decent laboratory equipment? The experimentation continues...

Another update: I've found the harmine! Here's what happened...

After redissolving in phosphoric acid the solids that were precipitated from the initial reaction by ammonia diffusion (described elsewhere on the forum), magnesium ammonium phosphate ('MAP') was precipitated in three fractions by dropwise addition of aqueous ammonia (specific gravity 910 when fresh, which it wasn't). The initial solubility test was carried out on a small sample from the 0.64 grams of dirty off-white precipitate which had started to form at pH5.5. [The typical harmala crud had had time to settle slowly on top of it while it stood overnight.] Evaporation of this solution left but the faintest oily smear on the evaporation dish. Near UV light (from a 'spy pen' toy) produced no visible fluorescence on the dish. The residue when redissolved in ethanol gave only a faint line of yellowish-green fluorescence along the light beam, indicative of the inevitable presence of harmaline contamination due to incomplete washing.

The second fraction, comprising 1.22 grams of clean, white precipitate on a par with titanium dioxide in appearance, formed between pH 5.5 and 6.0. It settled out quickly, giving an indication of its higher density. This was a large hint that it was more dense than the organic fractions could be. An ethanol solubility test help confirm this - the white solid remained undissolved as far as could be discerned and the supernatant produced no noticeable fluorescence with the spy pen except for the unavoidable harmaline traces. The powder also felt noticeably gritty in comparison with many organic substances.

The third fraction formed at around pH 6.2. The 0.16 grams of beige powder settled more slowly and was noticeably brown compared to the whiteness of the previous fraction as it formed. It became much paler on drying. The ethanol solubility test cleaned up the white powder a little and the solution gave a faint deep blue fluorescence which could be considered characteristic of the tiny amount of harmine present.

***So, yes - there's the harmine, it seems.***

There was also an insoluble fraction left over from the initial ammonia diffusion precipitation. This seems to be basic magnesium carbonate, as there was an effervescence on washing it with phosphoric acid. I'll be doing a few more tests on this material later today.

One more thing to bear in mind when working with ammonia and phosphoric acid is that ammonium phosphates will exert a buffering effect. This, combined with my apparently lousy narrow-range pH papers, makes the pH values quoted above maybe less than reliable. The fact that the harmine apparently precipitated at pH 6.2 could be down to co-precipitation effects with MAP that will have been present as a fine suspension of crystallisation nuclei, or could be that the pH papers couldn't cope with the conditions of buffering, strongly coloured solution, and the evaporation of ammonia from the papers at PH >7 which really hampers measurement of higher pH using this method.

Good news is I'll be trying out a self-build pH probe to test with my ancient pH meter. More on that elsewhere, when it happens.

I've unilaterally decided to sticky this thread as it's pretty useful and tends to disappear down the list as time goes by.

Hopefully it'll act as a reminder on some of my science projects that got sidelined last summer.

Nice research you're doing here, downwardsfromzero!

A cheap microscope will already give you a lot of useful information on the identity of harmala alkaloids. See elsewhere in this monster-thread...

Best wishes also for a revealing 2023

How did I miss this? Thanks, An1cca - hope you have a fascinating year too!

I was searching again for my THH conversion notes that I got from somewhere on the forums and they tell to just stirr Harmalin in Zinc + Acetic Acid for 1,5 h.

Then I went back to the VDS Protocols and they stirr for some hours and then to continue to stirr for 6 hours. Quite a difference

So what is the concens of how long to stirr? I used that 1,5 h back in the days and my melting point was pretty exact. Still is there a short finalized protocol everybody uses, that is hidden inside those 30+ pages?

That will depend upon your DHH concentration and the surface area of the zinc. If you're in a hurry, you can check under uv-light for the blue fluorescence of THH that is easily overpowered by the green of DHH. Or do a recrystallization in-situ on a microscopic plate of a drop of solution and verify the absence of DHH-like crystals.

I read that fluorescence info many times, but thought if it's a mix of both I would still not know about how much I have and/or still need to convert.

So that means even traces of DHH will make it green and absence of green will indicate conversion must be complete?

If it's blue, it can't hurt to let it react for another hour. If it's green, you're sure you're not yet there. If you don't have a microscope, I've seen an interesting technique that points a common laser-pointer at a hanging drop of solution. The image is magnified hugely on the wall behind it. You could try this with your solution and please report back if it works... https://youtu.be/bYEuo3j2Ly0
Good luck!