Carrots to reduce harmaline to THH?

[modern]

Old digital camera picked up the colors better than my phone you can see the color and tones. Also there are clear separations here. I think it is possible the thh value may be too high and the tailing may be due to excess amount? I'll try an even more diluted spotting to test next time.

 

[modern]

So just to get a better handle on TLC work I continued to develop these plates.
I think I may be doing the vanillin stain work because only pot I found was the bottom line as a very faint brown mark. (phenol?)
The potassium permanganate only works properly on thicker plates but due to destructive nature will be a last resort.
Still waiting on my ninhydrin and will make some dragendorff reagent as well for when I start attempting dmt spotting and my different psychotria.

EDIT: more research... maybe the thin line that vanillin is showing is harmalol or harmol which are both phenol.

 

[modern]

So after cleaning and having fairly white precipitate from doing a few a/b I decided to start titration to separate the different harmalas. But strangely I got 3 precipitates.
At 7.5ish range I started to get a some brown dots then lots of brown precipitate when allowed to rest. I filtered then added little more lye and around 8ph range I got lots of white light precipitate which is most likely the harmine. Then I kept adding lye and no preipitate formed but after allowing it to sit for a while I got some precipitate which from my past experience is harmaline and thh. What is the first precipitate? harmol?

1st pic is the 14ph
2nd pic is ph around 8+
3rd is brown precipitate around 7.5 ph

I will repeat the a/b once more to see if I can clean up a little better but these were distinct and each required a lot of lye between each step not like 1ml more caused precipitate. (after swirling)

 

[Transform]

Following the order of the pictures, what's to say they're not THH, then harmaline , then harmine?

Did you start with caapi? (I've forgotten…)

 

[modern]

Following the order of the pictures, what's to say they're not THH, then harmaline , then harmine?

Did you start with caapi? (I've forgotten…)

Yea caapi

I've never been able to precipitate THH on its own always with harmaline. But it could be I guess HOWEVER by the amount of precipitate of the middle powder that one HAS to be harmine since harmaline is rarely found in high amounts in caapi particularly in relation to harmine. Harmine is always the dominate harmala.

I'll do another A/B regardless to see if I can clean up more. I'll also be trying to follow the TLC procedure found here to compare the rf values to have more certainty.

Maybe just dirty harmine that precipitated with impurities.

 

[Transform]

Yea caapi

I've never been able to precipitate THH on its own always with harmaline. But it could be I guess HOWEVER by the amount of precipitate of the middle powder that one HAS to be harmine since harmaline is rarely found in high amounts in caapi particularly in relation to harmine. Harmine is always the dominate harmala.

I'll do another A/B regardless to see if I can clean up more. I'll also be trying to follow the TLC procedure found here to compare the rf values to have more certainty.

Maybe just dirty harmine that precipitated with impurities.

Oh, in which case you could be right about the harmol. A couple of presumptive tests would be solubility in sodium hydroxide solⁿ. and insolubility in NaHCO₃ solⁿ., and forming a blue to purple precipitate with iron III chloride solⁿ. This latter test might not run cleanly because of the possibility of oxidation of the harmol. I get the impression that there's a bit of a paucity of information on harmol.

 

[modern]

Oh, in which case you could be right about the harmol. A couple of presumptive tests would be solubility in sodium hydroxide solⁿ. and insolubility in NaHCO₃ solⁿ., and forming a blue to purple precipitate with iron III chloride solⁿ. This latter test might not run cleanly because of the possibility of oxidation of the harmol. I get the impression that there's a bit of a paucity of information on harmol.

Well I precipitated by slowly adding saturated lye solution so it is insoluble in NaOH. What I can test is adding vanillin tlc stain on some dry powder and heat to see if it changes colors since it is a phenol it should react.

Ionization processes of some harmala alkaloids.

The ionization and UV-visible spectral properties of some harmala alkaloids have been investigated spectrophotometrically. Harmaline and harmine were found to have pKa values of 9.55 +/- 0.04 and 7.45 +/- 0.03, respectively. The ionization of harmalol was characterized by two processes which...

molpharm.aspetjournals.org

"Harmaline and harmine were found to have pKa values of 9.55 +/- 0.04 and 7.45 +/- 0.03, respectively. The ionization of harmalol was characterized by two processes which could be spectrophotometrically isolated from one another, allowing pKa values of 8.62 +/- 0.15 and 11.30 +/- 0.23 to be determined"
"For harmol, pKa values of 7.90 and 9.47, reported at 21 degrees by Perrin [N.Z.J. Sci. Technol. 388:688-694 (1957)]"

IDK what pKaH is I'll do more research to understand better.
pKa1
9.51
pKaH1
7.86

 

[Transform]

Well I precipitated by slowly adding saturated lye solution so it is insoluble in NaOH. What I can test is adding vanillin tlc stain on some dry powder and heat to see if it changes colors since it is a phenol it should react.

Did you filter off the precipitate before adding more NaOH though? You'd need to check the precipitate's actual solubility in excess dilute NaOH.

You might want to use a drop of a (methanol?) solution of the unknown compound, maybe on the unused corner of a tlc plate, for testing with vanillin, since there's a chance that any colour reaction might not show up on bulk ("bulk") powder.

EDIT: I think pKₐH¹ refers to the removal of the proton from the pyrido- nitrogen, with an equilibrium of 50% protonation at pH 7.86.

The data also seems to indicate that harmol is rather more acidic than harmalol.

 

[modern]

Did you filter off the precipitate before adding more NaOH though? You'd need to check the precipitate's actual solubility in excess dilute NaOH.

You might want to use a drop of a (methanol?) solution of the unknown compound, maybe on the unused corner of a tlc plate, for testing with vanillin, since there's a chance that any colour reaction might not show up on bulk ("bulk") powder.

EDIT: I think pKₐH¹ refers to the removal of the proton from the pyrido- nitrogen, with an equilibrium of 50% protonation at pH 7.86.

The data also seems to indicate that harmol is rather more acidic than harmalol.

I had the concentrated acidic harmala solution after washing excess lye and compor impurities off. Then added acid. I would then slowly add lye solution to which there is a precipitate that forms but when swirled dissolves so keep adding slowly until precipitate remains. Then settle and filter off before adding more lye to get more precipitate.

I’ll make some tiny solutions of 1mg/1mL and run TLC on them using same mobile solvent phase as my first attempts.

Did you filter off the precipitate before adding more NaOH though? You'd need to check the precipitate's actual solubility in excess dilute NaOH.

You might want to use a drop of a (methanol?) solution of the unknown compound, maybe on the unused corner of a tlc plate, for testing with vanillin, since there's a chance that any colour reaction might not show up on bulk ("bulk") powder.

EDIT: I think pKₐH¹ refers to the removal of the proton from the pyrido- nitrogen, with an equilibrium of 50% protonation at pH 7.86.

The data also seems to indicate that harmol is rather more acidic than harmalol.

you may be right I weighted the precipitates and brown was more than second one. Volume wise seems like much less but was denser.

Maybe just dirty harmine

 

[Transform]

I'd say dump a bit of the brown powder into dilute NaOH and see if any dissolves.

Doing another TLC run seems a rather good idea as well.

The data also seems to indicate that harmol is rather more acidic than harmalol.

Just to clarify further, this is the acidity of the phenolic proton that is being referred to. Harmol would almost definitely dissolve in dilute NaOH if the pKₐ figures are reliable.

 

[modern]

So after a clean up doing a/b brown didn’t lighten up but was able to seperate now a brown almost crystalline and tan precipitate after adding a little more lye solution.

Something new that I’ve not read anywhere is I got precipiatate similar to manske but just dissolved the middle batch ph 8.5 range in near saturated citric acid solution to dissolve. It dissolved well and I filtered and left overnight and woke up to crystals that are identical to the hcl crystal from salt brine in vinegar.
I could be wrong but does pkb come into play here? Since ph was around 2-3?
Regardless I filtered and collected and then added little more lye to water and have some precipitate at the 8.5 ph range but almost all crashed out as citrate? I thought that harmalas were messed with enough to not have any surprises now a days

First pictures are of the citric solution which the harmalas crashed out of overnight. Visually very similar to the hcl salt from manske. The last picture is of the brown suspected harmol which I’ll now test adding to lye solution for solubility test. Much cleaner and crystalline but not any lighter. I did get better desperation with remaining solution showing cleaner precipitate.

 

[modern]

@Transform didn’t dissolve but also doesn’t “clean up” when diluted in water.

Water around 7ph, lye solution around 9-10ph and 14ph lye solution.

I may do a citric acid precipitate or hcl salt brine to see if it will clean up but I was able to separate the brown from a tan precipitate.

Finishing clean up to run TLC on the different fractions I got.

 

[Transform]

@Transform didn’t dissolve but also doesn’t “clean up” when diluted in water.

Water around 7ph, lye solution around 9-10ph and 14ph lye solution.

I may do a citric acid precipitate or hcl salt brine to see if it will clean up but I was able to separate the brown from a tan precipitate.

Finishing clean up to run TLC on the different fractions I got.

It's looking like an impure harmine fraction then, probably! Did you check for fluorescence in the aqueous layer? Looking forward to see what the TLC turns up, in any case.

Also, I may have to have a play with this possibility of Manske-style harmala citrate precipitation. Did you get a positive confirmation that an appreciable amount of alkaloid was recovered, rather than, say, sodium citrate or something like basic calcium or magnesium citrate? The situation brings the crystalline precipitation of magnesium ammonium phosphate to mind. What if THH was forming a complex salt with citrate and a metal ion, for example?

 

[modern]

It's looking like an impure harmine fraction then, probably! Did you check for fluorescence in the aqueous layer? Looking forward to see what the TLC turns up, in any case.

Also, I may have to have a play with this possibility of Manske-style harmala citrate precipitation. Did you get a positive confirmation that an appreciable amount of alkaloid was recovered, rather than, say, sodium citrate or something like basic calcium or magnesium citrate? The situation brings the crystalline precipitation of magnesium ammonium phosphate to mind. What if THH was forming a complex salt with citrate and a metal ion, for example?

Im gonna save each fraction separately. The citrate seems most likely to be a harmala since very small amount was recovered from making the solution more alkaline. I’ll weigh it soon and see if it matches appropriate expected weight with citrate ion.

I’ll save a portion as the citrate precipitate and then try another cleanup.

I’ll probably get around to doing TLC this weekend.

 

[modern]

Só Im gonna assume my second precipitate is harmaline despite being weird ph 8.5 range.
So the citrate precipitate should have been around 550mg staring from 300ish mg freebase. I got 450ish mg but also had more precipitate in solution afterwards so fairly certain it is some harmala.

I took 200mg and placed in 50mL water and it didn’t dissolve completely with ph around 5-6. I decided to add more vinegar to dissolve it all and added magnesium strip in excess around 1:1 ratio harmala. I’ll allow it to react overnight and and add extra vinegar if needed to dissolve the rest of the magnesium.

I’ll precipitate this with mansake since I know it precipitates clean. Without the magnesium.

Once complete I’ll do TLC and save a portion for future TLC references and experiments.

 

[modern]

Since I made the ninhydrin solution I decided to just spray the harmala tlc plates I did of the extracts. The citric acid precipitate seems to have only been harmine and harmaline precipitates fairly clean when added a base afterwords. The harmaline produced after the citric acid salting produced a faint yellow stain when ninhydrin was sprayed without the need for heat. It is a secondary amine which aligns with "Secondary amines stains light yellow which is difficult to see." TLC stains

I made solutions for tlc of 10mg per 2 ml of alcohol and had different colors for them all. The magnesium treated harmaline has the marked blue stain and precipitated with manske salt brine leaving the magnesium in solution for anyone interested. I also made a more dilute 10ml solution adding water but ended up not needing it so dilute. 5mg/ml seems to be a good point spotting amount for a single application with most... some needed more concentrated since very weak uv light reaction

First picture is THH manske precipitate.
Second and third picture is the solutions of the different precipitates in a plastic egg carton. and final 2 are spots before running the TLC.

 

[modern]

Hard to see in the first photo but zooming in you notice a slight yellow stain that is from the ninhydrin spray and it aligns perfectly with the harmaline in the second photo under uv light. 3rd picture is just to better show the staining.

So something I noticed is NONE of the staining included the harmol/harmolol that the tea showed as a very faint thing line and also showed yellow staining with the vanillin stain which was the only phenol on the plates. I guess the harmol or harmolol might change due to the extreme ph conditions? The red/brown precipitate was just harmine that didn't want to clean up for whatever reason.