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

[An1cca]

I suggest doing a recrystallization on a microscopic slide. DHH crystals are very typical...

The UV pictures make THH an improbable candidate.

Let's call it Nexine and apply for the Nobel-prize

Nice research, Jees!

 

[Jees]

...Let's call it Nexine...

Love that word on first sight. Nexine: a considered substance that, at least for the time being, is enigmatic.


* * *

...I think the 10% rule you describe will be very practial indeed. For me, it was difficult to make the difference between precipitating Zn-salts and excess bicarbonate, but as you say, the bicarbonate falls to the bottom rather quickly, while the zinc salts stay floating for a longer time...

Yes one must read bottom layer thickness after 60 to 90 seconds, then its like the picture.
Note that the bottom layer thickness is like what is on top of original level 100. One could as well set a level mark at begin + a second mark above it like 10% of height. Then add sodbicarb till level reaches the +10% mark. That level is independent of settling time.

* * *

...One thing we have to consider with this method is the fact that THH-rich mother liquor can stay behind in the 10% sedimentation at the bottom. How will we 'wash' the bicarbonate without getting the zinc salts? Perhaps we should just add some water (10% ?) to the sediment that remains after the mother liquor was poured through the filter. Then we stir until this water is saturated with bicarbonate as well before we pour it over our filter. Just a suggestion...

I was thinking of having a bottle at hand of wash water that has a bottom layer of undissolved sodbicarb. That should flush well I suppose.

* * *

...the only thing that was not reproducible is the clean-up of the harmine-fraction with sodium carbonate as stated in his protocol 4...

this relates to:

clean up harmine- and THH-fraction separately by redissolving them in very dilute vinegar and reprecipitating them with a minimum amount of base (Na2CO3, ammonia or other)

On harmine levels: "Minimal amount of base" means not making ZnOH/ZnCO3 into a soluble zincate, this goes for ammonia too. So yes I agree it's not the best of clean-ups achievable.
Basing higher with sodcarb say 11, or with ammonia say 9, will make soluble zincate. Filter and zincate fall trough. Then wash harmine. Can't see much of another way, any other ideas for good harmine cleanup? <--- EDIT how could I forget: manske :thumb_up: if the ZnStuff stays liquid there, still to be tested.

Still catching up with studying your posts, you have been busy lately

 

[Jees]

...Next time, when I do A/B's on syrian rue, I'll limit the highest pH to 9.00 to cut out this portion of whatever it is.

So new rue was boiling and based the fist wash, euhh strangely no pH depression? In a mix of so much non alkaloids things behave differently? I saw that after pH 9 a lot more turned milky, I would not take a chance in these phases and still base to 12. After cleaning up a lot then incorporating a max pH 8.5 or 9 would be enough to eliminate the 'nexine'.

This was on my mind lately: these absolute pH numbers mean not so much. For example An1cca had to base to 10 with ammonia to see the ZnOH/ZnCO3 go liquid again. Then I was reducing deems n-oxide, filtered Zn off, based with ammonia:
- at our very expected pH 6 the ZnStuff became appearing;
- this increased untill pH 8 where it started to really go liquid again;
- above 8 all was water clear again;
- at pH 9 there was clouding again because of the deems FB'ing;
- this remained and did not disappear by raising pH;

So the point where ammonia turns ZnOH/ZnCO3 (EDIT for ammonia only ZnOH) into soluble zincate is 8 or 10? It's both depending on circumstances. Something to live with I suppose.

This is the reason I will keep basing my rue first washes full of 'contaminations' to 12, and stop trying to fine-tune nexine out in those phases. Once things are cleaned up and pH depressions become visible, then a max pH 8.5 to 9 would be a better moment to avoid the nexine.

:want:

 

[DreaMTripper]

Could nexine be harmalol? Also yes pH levels required will no doubt be affected by the many other compounds and molecules present in the organic soup that are all pushing and pulling the OH and H ions.
Was the same amount of water used in the two experiments?
I've actually lost track now! Are you talking about Sodium carbonate then ammonia or just Zn then ammonia?
Its getting late I'm confused will leave this here...

"Characteristic reactions of Zn2+:

Zinc(II) ion forms complex ions readily.

Aqueous Ammonia:

Zinc(II) ion reacts with aqueous ammonia to precipitate white gelatinous Zn(OH)2:

Zn2+(aq) + 2NH3(aq) + 2H2O(l) <==> Zn(OH)2(s) + 2NH4+(aq)

The zinc(II) hydroxide precipitate dissolves in excess ammonia:

Zn(OH)2(s) + 4NH3(aq) <==> [Zn(NH3)4]2+(aq) + 2OH-(aq) "

Zinc

 

[ijahdan]

Just a thought, if thh survives a manske, forming thh hcl crystals, as an1cca has shown, couldnt this be used to separate it from the zinc compounds, or would they form zinc hcl?

 

[downwardsfromzero]

So the point where ammonia turns ZnOH/ZnCO3 into soluble zincate is 8 or 10? It's both depending on circumstances. Something to live with I suppose.

It's easy to imagine how the conversion pH will be different as Zn(OH)2 and ZnCO3 are different compounds. And depending on the exact conditions Zn(OH)2 and ZnCO3 will have co-precipitated in different proportions. These conditions include temperature, air pressure and CO2 concentration, reagent concentrations and rate of addition... etc...

The more you look...



I'm loving this thread, btw!

Highly instructive, I'm currently looking into using a digital (Kids') microscope - Intel Play - for looking at crystals and sorting out open source drivers for the thing is a bit of a nightmare. Any suggestions (apart from "use something else"!) would be most welcome.

Just a thought, if thh survives a manske, forming thh hcl crystals, as an1cca has shown, couldnt this be used to separate it from the zinc compounds, or would they form zinc hcl?

Zinc chloride is VERY soluble at slightly acidic pH.

 

[Jees]

Good news that Zn chloride will not manske, thanks.
And thanks DreaMTripper for hinting ammonia only forms ZnOH, I edited my post, should have known better but this ZnOH/ZnCO3 combo got a notch too hard imprinted LOL.

When writing things, some strange elemental errors keep surviving an re-read, it happens to everyone, and it's a good thing others help finding those pesky thinko/typo lapses :thumb_up:

And yes I must get myself a microscope, and when browsing I get tempted to even spend a nickle more than absolutely needed, is it the snob in me I dunno but I get attracted to something "better" easily.
I have no clue about OMAX quality, but the -60% is really tempting, even if I have to pay for 100 euro import taxes to EU.

OMAX 40X-2500X LED Digital Trinocular Lab Compound Microscope with 5MP Camera and Mechanical Stage for sale online | eBay

Find many great new & used options and get the best deals for OMAX 40X-2500X LED Digital Trinocular Lab Compound Microscope with 5MP Camera and Mechanical Stage at the best online prices at eBay! Free shipping for many products!

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I know its more than necessary and maybe chinese under quality, but it's so tempting. Any remarks welcome. Otherwise that Bresser BioDiscover is a load cheaper, doubts, doubts LOL, the luxury to be able to choose

 

[Intezam]

Good news that Zn chloride will not manske, thanks.
And thanks DreaMTripper for hinting ammonia only forms ZnOH, I edited my post, should have known better but this ZnOH/ZnCO3 combo got a notch too hard imprinted LOL.

When writing things, some strange elemental errors keep surviving an re-read, it happens to everyone, and it's a good thing others help finding those pesky thinko/typo lapses :thumb_up:

And yes I must get myself a microscope, and when browsing I get tempted to even spend a nickle more than absolutely needed, is it the snob in me I dunno but I get attracted to something "better" easily.
I have no clue about OMAX quality, but the -60% is really tempting, even if I have to pay for 100 euro import taxes to EU.

OMAX 40X-2500X LED Digital Trinocular Lab Compound Microscope with 5MP Camera and Mechanical Stage for sale online | eBay

Find many great new & used options and get the best deals for OMAX 40X-2500X LED Digital Trinocular Lab Compound Microscope with 5MP Camera and Mechanical Stage at the best online prices at eBay! Free shipping for many products!

www.ebay.com

I know its more than necessary and maybe chinese under quality, but it's so tempting. Any remarks welcome. Otherwise that Bresser BioDiscover is a load cheaper, doubts, doubts LOL, the luxury to be able to choose

Well, we'd first purr-chase one of these kids microscopes. They should work just fine. Then later One (1) could upgrade the equipment? If need be....

500 $ -- that's a years supply of bird food (jerky type/precioussss) for the lot of four (4) tribes (nations) :shock:

http://www.ebay.fr/itm/172367951171

http://www.ebay.com/itm/172292998087

http://www.ebay.com/itm/291920317826

 

[downwardsfromzero]

Microscopes: I was lucky, a friend found me one at the dump for free. It was fun to refurbish it as best I could; a job lot of eyepieces and objective lenses from a popular auction site proved very inexpensive. It's even possible to take out the eyepiece and poke a digital compact camera lens in the hole for photographs. Now, what to do with all these spare lenses?

I need help compiling the drivers for the intel play qx3 USB kids' microscope and the associated cpia whatnots as I am almost completely clueless when it comes to coding. It would just be nice to get a USB microscope up and running on my zero budget. :thumb_up:

 

[endlessness]

This thread is epic




(by the way, save all your products, for example this unknown precipitated product, and I'll get it lc-ms tested )

 

[Jees]

Thank you Intezam for landing my feet back on the ground
That bracket for fixing a smartphone on the scope is witty :thumb_up:
And this is even more flabbergasting:

Also gonna check a second hand site by the way.

* * *

Had removed Zn today from a 2gr DHH reduction. So to precipitate the ZnCO3 and traces of harmine I wanted 7.5 but that was not possible with only 10% of undissolved sodbicarb at the bottom, it was like 20% as in the picture.
They don't grant us a simple visual clue it seems, put some loids in and the game changes.
So back to measuring, put a probe in it. Forget the 10% rule, was worth a try

 

[Jees]

Then that goes trough a cotton ball in high tech over pressure
Strange thing that filtering changes pH a bit.
That is a one way air-valve from an aquarium shop, hardly an euro.

 

[Jees]

Blue enough ? It's almost my avatar

 

[An1cca]

Nice work Jees! Perhaps you might consider something like this: https://www.pelletlab.com/filtering_kit (with the money you didn't spend on the expensive microscope:wink: )

Anyway, I think the pH is dependent upon multiple factors, that's why I think it's important to follow a standard protocol. Like the one used in VDS's article.

This is one I followed now to make a microscopic study of DHH to THH conversion:

3010mg DHH + 3020mg Zn-powder in 75ml of acetic acid 7% (pic01). The solution was brown-red (pic02). Only 15 minutes after dropping in the zinc, the solution's color became much lighter (pic03). The color didn't change much in the following 6 hours (pic04). After treating the solution with activated charcoal (60mg + 10 min of boiling), the solution's color was very pale (pic05). After addition of ammonia, THH precipitated as a white solid (pic6 and pic7). The other half of this experiment, where the ammonia-free route was followed after reduction, posed some serious problems. I'll elaborate on that later on. Now I'll post the microscopic evolution separately...

 

[An1cca]

And now for the microscopic evolution of the above DHH to THH reduction. It seems that after 3-6 hours, the reaction is complete. Apparently, VDS noticed this as well when UV-visualising the solution (I didn't check this for now).

All samples were recrystallized from the mother liquor on the microscopic slide using ammonia 12%.

It seems that relatively quickly, the specific 2-dimensional fern-like DHH crystals disappear and consecutively form a three-dimensional slender-crystal radial cluster and eventually, these crystals get smaller and more boulder-like after 2 hours. Sublimated THH showed the same crystallisation form (see previous posts). After cleaning with activated charcoal, the THH-solution recrystallized as (almost) perfect spheres composed of radial THH-crystal clusters. This form was also published in VDS's article. I also put in some pictures of 'fractured' clusters to show individual crystals. I labelled the pictures with their respective reactions times.

 

[Jees]

Microscopes rule :thumb_up: Wonderful to nail the necessary time, splendid
Then we could also find out what impact heat has on the outcome, I feel so slow in here :lol:

 

[An1cca]

Strange thing that filtering changes pH a bit.

Yep, I noticed this as well. Actually, I think it might account for the clouding that re-appeared in the filtrate during my last ammonia-free precipitation. I used the other half of post #134's reduced and charcoal-cleaned solution (pic01). To this I added sodium carbonate until 10% remained undissolved at the bottom underneath the zinc salts (pic02). After filtering, the filtrate spontaneously started clouding again (pic03). When washing with saturated sodium bicarbonate solution, there appeared a light-brown precipitate and clouding kept on occuring. I refiltered the filtrate through the residue a few times to get rid of it, but it seemed to get worse (pic04-05). Then, when the filtrate was more or less clear, I started adding sodium carbonate powder. It immediately clumped and a sticky yellow-brown goo formed (pic06). In considerable panic:shock: , I dissolved everything (previous residue as well) in about 200ml of acetic acid (went perfectly) and added 100ml of ammonia 12%: NOTHING happened:surprised . After standing for many hours, some THH collected on the bottom, but almost all was lost....

So, although I can not explain this fully, DO NOT WASH YOUR PRECIPITATES WITH SATURATED BICARBONATE SOLUTION. I might try this again without washing and see if sodium bicarbonate would still cause this strange precipitation (and use dissolved sodium carbonate in the first place (20g/100ml)). For now however, I have quite a lot of THH in the freezer, so perhaps another Nexian might re-check this route. Jees, will you basify your filtrate with sodium carbonate or ammonia?

On a side note, it seems my yields are lower now. The ammonia-precipitation gave about 58% yield. I'm not sure if it's caused by the activated charcoal (I don't think so) or by varying concentrations of reagents. Van Der Sypt used quite dilute solutions and got high yields, so perhaps my theory of using concentrated solutions is counterproductive. Something else to check...

I also want to repeat my call for using a standardised protocol so that we can compare our findings. VDS's protocols are detailed and apparently researched, perhaps we should stick more closely to them?

 

[An1cca]

Wonderful to nail the necessary time, splendid

Well Jees, it would have been nice to also have pictures at t=240 and t=300 min, but there's more to life than crystal porn:grin:

Actually, perhaps it is better to NOT let the reagents over-react. Perhaps we should stop after 6h? Just trying to explain the differences in yield...VDS let the mixture react for only 9h.

 

[endlessness]

Why do you suppose AC can't make you lose yields?

AC is even used in some cases of alkaloid poisonining. It has been documented that quite some alkaloids can be absorbed by AC (im trying to find the original reference since my link in this post is dead).

 

[An1cca]

Why do you suppose AC can't make you lose yields?

Of course it will reduce yields, but it seems improbable to me that 60mg could absorb more than 150mg of THH. Also, as I showed in post #113, a charcoal-cleaning and reprecipitation of post-Manske THH had a yield of 87%. Reprecipitation will always give a yield like that (or even less!), even without AC...