Harmine, harmaline and THH from Syrian Rue. Verification and finetuning of the VDS-protocols

[An1cca]

Rather than to make a 'static' PDF, I would like to create a dynamic information source that can be updated as new experiments bring on data. Like in the OP of this thread? I will first try that. Later, I can always copy-paste this into a wiki I suppose.

Sorry for the newbie question, but how do I upload a picture on the Nexus-server? Can I use the address that appears when hovering over a picture in this thread for example? And if so, how can I upload one without having to upload it first into a thread?

Thanks guys! And may the Vine reward you with transforming journeys for the analysis of the samples, Endlessness:thumb_up: !

Edit: Thanks to Intezam, I now know how to upload pictures...

 

[Jees]

Any picture you have uploaded there in the posts, can be re-uploaded in another post, like here for example:

I went to your post, set pointer over a pic of choice, click right button, choose COPY IMAGE LOCATION and that clipboard content can be image-pasted anywhere (but I suspect it has to stay within the nexus realm, is that right?)

An1cca you have done so much for the community already you deserve a break from vessels :thumb_up: . I would like to see that MAO-saurus one day

Next week I expect my microscope and I hope to catch up with your works. Thank you for knowing what exactly to look for.

...I don't think there's any unreacted DHH left as microscopic examination of the mother liquor and redissolving-reprecipitation of the residue give identical results where no DHH can be recognized. Also, Endlessness et al. confirmed this by LC/MS...

I never considered non-complete reactions, but only thinking of the mass balance before-after. My 50% yield wondered me why so much of mass was not showing up as a precipitate after basing. Then asking myself: if they don't show up precipitating then where the hell are they? No matter reduced or un-reduced, I expected them (near) all to precipitate. Not so, hence my wondering why. Stirring and waiting might indeed help as you say, makes sense actually, if we think of crystallizing as a process that takes time then better not harvest right after basing but wait .... wait for the crystals to grow.
Thank you for pimping the yield up to 77%, we might get better and better :thumb_up:

 

[Jees]

Whatever VDS stream followed, the base is a very clean starting point that goes further than most of us ever did. TBH I've never did 4 to 5 successive manskes before, I just did 2 because never caring much of potential % of other-plant-stuff. It did "the job" so easy was good enough. But here, I think we have to shape ourselves a notch better and follow VDS recommendations to really start with the best we can.

First time in my life I'm doing 5 manskes wondering if it's not overkill, well one sees the product really getting purer and crystal growing getting faster as you go, not the first 2 manskes but after that things go 'super'. My yields have never been so low but more pure. I think this is mandatory when going for VDS style of works. It should be highlighted that there is no cutting corners here like jumping into the latest module for THH with in-a-hurry derived crude extracts.

If we come to a tek, the first module should be the seeds --> {harmine+harmaline} the best we can. We've been extracting rue for so long with little concern, because it simply always worked, that now a new attitude should be advocated when one wants to go VDS way.

For example there are situations you are simply not going to see the pH depressions at all, where you are not able to harvest decent amount harmine at 7 - 7.2 . These are situations in large volumes where crystal grow rate is heavily compromised (lesser purity), and then thing suddenly seem to answer more the good ole 8.75 cut point separation technique as proposed by Infundibulum!
I second VDS his claim that at 8.5 all harmaline can be retrieved, but I am a bit on a limp out here (for now) to suggest a condition for such to happen. Conditions that are strongly promoting crystal grow rate ('pure' materials in smaller volumes). But if you take conditions that mute or slow crystal grow rate, VDS claims are simply not valid, I've found so far.

So we need a stamp "VDS approved" or something before embarking further VDS routes.

Luckily a manske has become a no brainer thanks to VDS hands on. In a 250 ml vessel, poor your alkaloids, top in 50 ml vinegar (7 to 8%) + 50ml water, hot-bath to melt it all. Then heat 150 ml of water + 50 gr salt, dissolve all salt, and poor that into the beker, swirl, stand, cool off, coffee filter, no need to dry just get on with wet alks to the next manske. Repeat until 2 filtrates look the same. I used to make such a fuzz of it in 1 liter volumes carefully pH-ing etc.
Thank you VDS and An1cca to bring it in here :thumb_up:
BTW An1cca how did you stumble on that paper?

 

[An1cca]

BTW An1cca how did you stumble on that paper?

I must admit it was not that difficult: it's still the third hit on Google when looking for "harmine isolation" . Since it was very recent and I had been lurking the Nexus' forums too long, it was my time to do my share...

I made a stirrer based upon this concept: http://www.instructables...table-Magnetic-Stirrer/ It works like a charm!

We'd all love to see what you've come up with, I'm sure!

Well downwardsfromzero, I attached some pics and a little movie from my improvised magnetic stirrer. As I said, the handle of the potentiometer got very hot so I had to run it at intervals. I'm sure someone a little more into electronics would come up with a smarter circuit...

Luckily a manske has become a no brainer thanks to VDS hands on. In a 250 ml vessel, poor your alkaloids, top in 50 ml vinegar (7 to 8%), hot-bath to melt it all. Then heat 150 ml of water + 50 gr salt, dissolve all salt, and poor that into the beker, swirl, stand, cool off, coffee filter, no need to dry just get on with wet alks to the next manske. Repeat until 2 filtrates look the same.

Perfect Jees, this is already a part of the first "seed to mixed extract"-route. Have you written down your recent extraction steps as well? Were they comparable to VDS's? If they were, we could also make these steps 'Nexus-checked'. And BTW, what do you mean by "Repeat until 2 filtrates look the same"?

 

[An1cca]

I attached a little movie of THH-precipitation by 'bombing' it out of solution (4x speed). Because we now suspect this might considerably reduce yields, it is better to look at a movie of it than to actually do it :d

 

[downwardsfromzero]

I attached a little movie

Thank you! The sound tracks were quite pleasing too

 

[DreaMTripper]

Beautiful

 

[Jees]

That is nice tinkering An1cca :thumb_up:

* * *

...what do you mean by "Repeat until 2 filtrates look the same"?...

In the picture is the filtrate water of manske 4 and 5, or was it 5 and 6? It's the first time the waters look alike, the little mist in n+1 doesn't matter, then they're the same. I remember the color of n-1 to be a tad dirtier than n. So no more cleaning up can be reached by repetitive manskes. I chose semi-logic/semi-intuitively the color gradient over the different manskes as a measure. IIRC VDS used basing of the filtrate as a measure, if he could see fall out when basing the filtrate, he kept repeating manske, and found that 5 is enough. Well I'm about there too at 5, quite convenient, 5 seems the magic number 8)

BTW am checking out on my microscope, I get one eye bigger :surprised

* * *

For rue, I've never had a fixed workflow, it was never two times the same I'm afraid, in a manner of speaking. For tek ideas I think of as kitchen friendly as possible, like vinegar, whole seeds for easiest filtering and squeezing them, and an easy base like sodcarb. That should do I guess. I better start trying one out just like that.

 

[Jees]

I've suggested to do the pH depressions on cleaned products, but 2 tests have indicated lye to be sabotaging the pH depressions. Even on clean products(!) even when lye is well diluted --> the harmine/DHH fell out at the very same pH levels but without a pH depression. :surprised

It looks like one has to use a "weak" base, one that does not dissolves completely so that there is an equilibrium between it's dissolved and un-dissolved states.
So a weak base can "accordion" in and out of it's dissolved state-ness, lye cannot do that as it always dissolves completely being a strong base.

For ammonia as a weak base: NH3 + H2O <--> NH4(+) + OH(-)
<--> being a 2 way volatile (vulnerable) equilibrium point.

I'm temped to suggest that crystal grow rate has an influence on the weak base's accordion-able equilibrium point, and this being the reason for the pH depressions. I think that during the pH depression the crystal growing will force the weak base a notch back into it's undissolved state hence pH drops.

Since the pH depressions for harmine are bigger than for DHH, perhaps harmine's crystallization has a larger impact on the weak base equilibrium point. On top of that at somewhat higher pH levels (like for DHH at 8.0 ) the accordion-ability of the weak base might be different (smaller/lesser) than at 7.

I redissolved the same batch harmine+DHH and based with ammonia and now very crisp pH depression occurred at 7.2 to 6.2 for the harmine (wow 100% on top of VDS this time), and a smaller one 8.37 to 8.13 for DHH.

I've tried to fit some explanation to the witnessing, all comments welcome, please correct me when you think so. Chemists?

 

[Jees]

I've also thought about another explanation for the pH depressions in relation to crystal growth:

Like when the molecules are able to "grow" on a crystal raster, their energetic state encourages to do that due the lower energy state seeking (like a ball wants to fall once it can). So when a Hydrochlorided molecule (salted form) find it's way to attach to a crystal raster, then by FB'ing it kicks it's H+ ion in the soup hence lowering the pH.

Explanation could fit (if I'm not producing BS) but then this mechanism would also work on lye, which it doesn't [1]. So I'm left with the weak-base explanation for real pH depressions of former post.

Cheers, by the way.

[1] I could see a very tiny horizontal pH plateau for harmine, not for DHH, but no depression at all. So the H+ kickout by crystallizing/FB'ing could have been that.

 

[An1cca]

Very interesting Jees! Do you still use vinegar to aid in dissolving the post-Manske hydrochloride salts? I ask because you speak of hydrochloride salts instead of acetate salts...

So VDS was confirmed again... I'm waiting 'till the LC/MS results are in before compiling all this interesting research into verified teks. Thanks a lot Jees:thumb_up:

 

[Sakkadelic]

Very interesting Jees! Do you still use vinegar to aid in dissolving the post-Manske hydrochloride salts? I ask because you speak of hydrochloride salts instead of acetate salts...

After the manske, your harmalas will be in the form of hydrochloride salts. While you are correct that adding vinegar to freebase harmaloids will cause them to dissolve as an acetate, the use of NaCl to saturate the aqueous solution causes chloride ions to supplant the acetate radical (owing to their difference in electronegativity), thus forming the HCl salt, which precipitates when cooled.

So you were right the first time. Remember, "the path of the Tao is not always the easy one, but it is always the right one."

I hope that clears things up.

 

[Jees]

Sakkadelic that 2nd quote was on my mind indeed but I'm wondering if it is relevant in our case of non-solid in-liquid situation right before basing. An1cca had a good point reconsidering this.

While we start from a full on manske product (H-Cl form) as soon as we dissolve it again (to base it later to get a freebase) then usually I add water only and maybe some heat also. To answer An1cca: even without adding vinegar or any other acid, it dissolves in the water and pH is usually between 3.5 and 4. But what condition is the alkaloid in at that stage? A non-precipated H-Cl form? This is what I thought but now I'm unsure :lol:

Anyhow, this all concerned a failing theory about the nature of pH depressions and not much worth further dabbing.

My bet is on the weak-base theory to explain the pH depressions, it's not just ideas-wanking but hands on important to choose your base and why. VDS never went in depth about this. I thought he never mentioned lye because he was all over harm reduction, but there seem to be even more pressing reasons, like lye doesn't create pH depressions (in my wee attempts).

 

[DreaMTripper]

The alkaloids are in an ionic state when dissolved. Binding with the hydrogen and oxygen is more attractive than the chlorides. So you have dissociated cl- ions and the others that I do not know the formula of!
More here Molecular Dipole Moments

 

[Jees]

Thanks Dreamtripper I'll dig into that.

Meanwhile have been playing with my new microscope toy. The included Bresser MicroOcular II usb camera is worth next to nothing, resolution too low and focus sucks. The software didn't work too LOL. The gave it with the set for no extra money, I can guess why. So I took pics out of hand with a pocket camera.

First pic HARMINE
Sec pic HARMALINE
3rd pic THH

All on same magnification factor 100x so you can compare crystal size.
It looks like darkfield microscopy but it's not, I used the overhead lightening instead of light from below.
Crystallization not in situ, but just like it forms in the vessel.
Crystal porn :shock:

 

[An1cca]

I wonder what recrystallized DHH looks like when viewed in this manner. Frozen jungle?

 

[Jees]

Ah you mean the in situ DHH ferns, I'm still working how to get my in situ's right as they are quite a wild result for now, how exactly do you do them please? Perhaps I lack some basics on microscopy.

Clearly microscopy takes more than pile-and-look, it's all new to me, many hours and trials were needed to get the above pics. It's very easy to get unsatisfying visual results, a bit like a photo-shoot.

 

[Jees]

Meanwhile, have a look at Nexine, the enigmatic precipitation that easily survives all manskes, and crashes out at higher pH's after the harmaline does.

It looks hard but it's a fluffy dusty powder, gel-like in suspension, takes more time to settle and hard to filter as some goes straight trough the coffee filter, does not reduce under zinc as DHH does.
There is some color difference between 2 harvests, but that is just due more or lesser cleaning up steps. It's the same 'stuff'.

It looks like it crystallizes but in no definite shape like the other ones, there's no real structure it seems. This is the same magnification as the pics a post back. If it does have structure then serious magnification is needed to recognize it, but even my heaviest factor cannot really show a pattern of growth. It looks more randomly solidifying. What the heck is this bird?

For now I have no idea if it comes out of the seeds, or that I'm creating it by ph 12 treatments or so, like demolishing DHH into Nexine? My next batch will have max pH of 10.5 on the raw seeds soup.

 

[An1cca]

I'm so curious about he looks of your Nexine when recrystallized on the slide. I dissolve a speck of alkaloid in a small drop of vinegar on the slide and then put a drop of ammonia 12% right next to it. I use a syringe with needle to do this. Then I shake the slide so the 2 drops mix. It instantly turns milky. Under the microscope, you can see how trillions of floating particles reorganize themselves in the course of a few minutes into the typical crystal formations we've seen in the above posts. And these can differ a lot from the ones that were obtained by just precipitating the alkaloids from solutions with a lot of other ions present. In fact, this recrystallization is in my experience the most reliable way of identification. I've seen harmine crystallize very much like harmaline and THH very much like harmine. Recrystallized on the slide however, they show their true nature...

I would have sworn Nexine to be DHH, but since it is not readily converted to THH, I don't have a clue anymore. Frodo, sorry I mean Jees, you are the only one that can walk the path to wisdom and liberate us from the dark abysses of ignorance :lol: ....

 

[Intezam]

:lol: How much nexine quantity was there compared to harmine/DHH/THH? Could it be harmane? Harmol? Harmalol? It looks loike sannnnnnnd......:thumb_up: