Hey all. A very close friend of mine asked me to report on a 'presumed' successful THH "synthesis". (Simple reduction) There were some bumps along the way, but it all worked out well. This is awful laboratory technique.

Used many peganum harmala seeds, salted out ala Manske, then performed an A/B on the Manske Product.

The resulting 4.8 g tannish powder was dissolved in enough 10% HCl to completely dissolve it and put on a hot plate with a reflux condenser. The mix was brought to a boil. Zinc powder was slowly added to the round bottom flask, and given time to boil and fizzle before more was added. Zinc continued to be added over the course of an hour until fizzing stopped. ~50 mL 10% HCl was added and more zinc was slowly added. Anytime one of the reagents was clearly expended, more of it was added (very scientific of him, eh?). Eventually, the dark red colour of the mix turned yellow, then clear with just the faintest hint of pale baby yellow. This was left to continue fizzing away for an hour or so. When it became obvious that the compound was not getting any lighter in colour, a few scoops NH4Cl was added and the whole mix was filtered through celite (...you don't have celite and a fritted funnel? Then a coffee filter lol) and the alkaloids were extracted with six additions of ~75 mL Ethyl acetate. The ethyl acetate was rotovapped. (What, you don't have a rotovap? Food Dehydrator'ed then.... lol). Dirty brown goo was left behind. 10% HCl was added and this was shaken (awful technique) this resulted in a disgusting coloured nasty chunky liquid. This was rotovapped. To aid the the drying process, blasts of DCM were added to the round bottom periodically and shaken.

This refused to dry (due to the awful technique of acidifying with aqueous) and so the brown glob had ~75 mls of ethyl acetate and 75 mls DCM added. It was shaken vigorously. Everything but the good stuff went into solution leaving brown liquid and white crystals. These were filtered and washed with DCM. These crystals were taken up in 150 mLs of isopropanol/methanol in a 50:50 ratio. This was refluxed and once it began boiling, water was slowly added dropwise and given time to resume boiling until everything was dissolved and the solution was clear. The hotplate was turned off and almost immediately, beautiful fractally snowflake crystals began to form. The flask was allowed to cool to room temp and then put in the freezer. 150 mls of isopropanol were also put in the freezer.

When everything was cold, the crystals of the flask were filtered out and washed with the cold iso. Beautiful shimmering perfectly white crystals resulted. These were weighed and found to have a mass of ~3.5 grams. Thin Layer Chromatography revealed that the compound was indeed distinct from either harmine or harmaline. No NMR was run. Bioassay revealed good times and despite the huge amounts of coffee my friend had been drinking, no hypertension was noted, which most definitely would not be the case had it been harmine/harmaline.

A much more effective and much faster technique involved taking the ethyl acetate fractions and adding methanolic Hcl (made from methanol + acetyl chloride). The compound fell out of solution immediately, in similar yield and was filtered and recrystallized as outlined above.

Hello,

Thank you for the tests! What eluent did you use for the TLC ? What was the Rf compared to harmine/harmaline (or more simply, was the spot between both ?) Did you use UV or any reagent to confirm?

What if after filtering from the zinc/NH4CL, you just precipitated the base out of the acqueous solution, to see how much of a mixture you actually have?

Thanks for your tests as well!

I was under the impression that harmine cannot be reduced using zinc; zinc treatment on (impure) peganum harmala extracts does have an effect (since it can reduce harmaline). As for colour changes, hell knows what they mean, but they are often unreliable for judging said conversions.

I'm dying to see pics of your plates with controls and all!

It is possible that harmine was not reduced but simply did not crystallize out. This would mean that the harmaline was definitely the major component of the seeds. I am well aware that colour changes are unreliable for judging the conversion of a compound. But my yields would suggest that the harmine may have been reduced.

I ran the TLC plates in 5% methanol, 95% DCM if memory serves me correctly (you'll have to forgive me, I run a lot of plates and a lot of columns in a given day.

Hmm, I didn't actually measure Rf, but I run reactants, products and a co-spot on the same plate. The reactant had two distinct spots large spots (and a million little ones because it was impure as all hell), the product had one spot and the co-spot had three primary spots. Again, what the TLC plate even looked like I can't begin to try to recall; I run many TLC plates every day.

Unfortunately, Infunfibulum, it would have brought unnecessary attention to myself if I began to take pictures of my TLC plates while at work. Haha! I didn't have the foresight to take the TLC plates. I had no intention of posting anything, it was merely for personal consumption, but after getting home from work I decided 'what the hell?'.

TLCs were examined using a UV light source.

There was definitely no substantial quantity of harmine or harmaline in my final product as confirmed by the TLC plate. The reduction has been reported to produce THH from harmaline, and what I have is definitely active.

I feel safe in saying that what I have is several grams of pure white THH.

Whether the harmine reduced, didn't crystallize, or didn't move over in one of my solvent extractions, I can't say. TLC analysis of each fraction could do this.

Perhaps six months from now I can figure that out for you

Hey! Thanks for the further info!

I could try doing a zinc reduction with HCl (or with acetic acid? At least its known to reduce amine oxides) on mixed harmalas, and test out with TLC, compare with control product. I would not do all this over complication with solvents, I would just try to precipitate the bases, filter and test it out, then later think about separation through solvents if I have time and material

I have no THH standard to compare, but we do for harmine and harmaline. The standard eluent we've been using for trypt/phens/b-carbolines is methanol:ammonium hydroxide 100:2.5 , while it does work well for most substances, some dont separate well (for example 5-MeO-DMT stays about same height as DMT). Maybe a dual or triple solvent mix would be better.

Not so long ago I TLCed some "caapi copy" from a vendor who's known for having sold fake THH in the past. Here's what came up:

http://imageshack.us/pho...mages/836/dsc02483d.jpg/

So you see the harmala columns, the left one are the harmine/harmaline standards dropped together, the middle one is the caapi copy (the right one was some 4:1 caapi extract from some other vendor, seems to only have harmine). You can see just under harmaline on the middle column there is a darker spot that is clearly different than harmaline. I wondered if that might be THH, but according to it's polarity, it should theoretically be above harmaline and below harmine. I asked Entropymancer his thoughts on this, he said:

"On the TLC: You're right, it seems strange that THH would be very slightly below harmaline; reducing the double-bond would make the ring nitrogen available for some H-bonding action, which ought to increase affinity for the methanol and lead to a higher spot. Then again, the sigma-bonding character of alkanes gives them greater affinity for silica than alkenes; maybe this increase in affinity for the stationary phase compensates for the increase in affinity for the eluent."

Thats why I was wondering if your THH was between both others, clearly. How did it look under UV, anything like that? (thats at 254nm)

How much zinc did you use, more or less?

Hmm, well, I think it is definitely safe to safe that caapi copy is very very low in THH, if it does possess any. That spot is pathetic lol. But I'd be more inclined to believe it is a related harmala alkaloid that was present in the rue it was undoubtably extracted from. I definitely showed a nice, distinct separation. Caapi would show a whole lot more THH, would it not?

I have never heard of anyone using ammonium hydroxide as an eluent lol. But I've always used hexanes:ethyl acetate and if that doesn't do it by the time I work up to 100% EtoAc, DCM and methanol, and haven't had a need to look elsewhere. Seems strange to me to be utilizing a 'solvent' that is going to be buggering with your protons lol.

For caapi:
Harmine, 0.31-8.43%
Harmaline, 0.03-0.83%
Tetrahydroharmine, 0.05-2.94%

From Callaway JC, Brito GS & Neves ES (2005). Phytochemical analyses of Banisteriopsis caapi and Psychotria viridis Journal of Psychoactive Drugs 37(2): 145-150.

I would think you would show a much more pronounced THH spot. And a less pronounced harmaline spot.

For rue:
Harmane, 0.16%
Harmine, 0.44[31]–1.84%
Harmaline, 0.25%–5.6%
Harmalol, 0.6%–3.90%
Tetrahydroharmine, 0.1%

My bet is Harmine, Harmaline and Harmalol. But... ugh.. the exposed hydroxyl group would probably bind the plate more firmly than that small separation would indicate.

I really wish I would have kept those plates. I'll tell you what. I'll bring some of each into the lab and run it again. I hope I don't get fired lol.

That'll setlle this.

About ammonium hydroxide in eluent mixture: It is commonly used as minor part of eluent systems.

It's been mentioned in publications several times. Just off the top of my head because I was checking out these papers a couple of days ago, you can see it.

Virginia Department of forensic sciences, controlled substances manual

Fish et al (1955) Piptadenia alkaloids

Bussey, R. (1974) Thin layer chromatographic differentiation of amphetamine from other primary-amine drugs in urine

The methanol:ammonia is what we use as standard mixture for different tryptamines and phens, because in our experience it works well (and cheap ), except for the few exceptions as I mentioned, which we use other solvent systems.

I will see if I can get that caapi copy GC-MSed... After the bad history of this vendor I dont doubt it wouldnt even have THH.

So your THH was indeed between harmine and harmaline?

What about the zinc, how much of it did you use?

Did you use any colorimetric reagent on the spots for confirmation?

Like... half a 7 dram vial filled with loosely packed Zn shavings. No reason to fear excess when someone else is paying for it

I don't remember where my THH sat. I will confirm that with my buddy sometime over the next few days.

My buddy always uses UV light followed by permanganate+heat.

I believe that Caapi Copy is just a mixture of alkaloids extracted/processed from rue, so if FV’s THH didn’t actually contain THH, then it’s reasonable to assume that Caapi Copy wouldn’t either, since Caapi Copy is just a blend of their harmine, harmaline, and “THH” products in a particular ratio meant to mimic caapi.

Also for anyone planning to do this, I must give the obvious warning H2 gas is evolved. Which is of course highly flammable. Don't do this indoors on a big scale near any sparking electronics, water heaters, stove-tops, etc.

First of all congratulations on the success and of course for trying!

While you are at it, what do you think of Magnesium as a reduction agent it this reaction?

It can be easily acquired if the lab does not stock it from firestarter kits : the ones with the metal bar you scrape (magnesium) and you start it of with a spark. The reason for it is that Zinc has a reduction potential of -0.76 while magnesium has -2.37 (lower is stronger in this case). Aluminium also has a "better" reduction potential than zinc, with it being -1.66 (for those who cannot find magnesium or zinc, aluminium foil is...aluminium!). So, i wonder if aluminium or even better magnesium could give better results.

Another question would be if mixtures of those metals could give good results, for example aluminium + zinc, or magnesium +aluminium. I wonder if when one of the metals reduces the organic molecule (metal getting oxidised) the other metal with the stronger reduction potential could "regenerate" it by reducing it back...

Hm, am i missing something in all of the above?

Have you run across an eluent that separates DMT and 5-MeO better?


-D.

according to literature, 2N acetic acid should work good for separating DMT (Rf 0.5) and 5-MeO-DMT (Rf 0.6), though I havent tried it yet.

With methanol:ammonia, DMT has slightly higher Rf, and differentiation is possible with some reagent. We use PDMAB-TS, which makes DMT turn yellow, while 5-MeO-DMT starts at yellow but then goes green. I wanna try to drop a dmt+5-meo standard together and see how it appears, I would post back once i do.