_Trip_ said:
Are you saying the ascorbic acid and pressure cooker potentially produced THH (blue glow)? or the BuOAc just pulled harmine? (which I believe also glows blue).
I'm suggesting there's a strong possibility that at least some THH may have been produced by the ascorbic PC cooks. This is not only based on the blue (with a hint of turquoise) fluorescence I've been seeing here. [The fluorescence from harmine that I've seen is a deeper blue and the pictures of THH fluorescence tend to be much more in the direction of turquoise.] The colour of the fluorescence is a fickle indication but the smoothness of my subjective experiences with the crude alkaloid precipitate also hints that the harmaline, if any, is present only at a greatly diminished concentration.
There are numerous caveats though.
Firstly, the first three cooks were precipitated separately and have not been analysed yet as they have so much more of the rue crud that processing was a total nightmare, even with vacuum filtration. So that's where the harmaline might be.
Secondly , I may just have been incredibly lucky and got a rare batch of rue seeds that were unusually high in THH. This has not been controlled for and all those seeds have been used up, although I did go and buy another kilo - fortunately with the same packaging - from the same shop.
Thirdly, I now have four fractions from the test sample post-butyl-acetate. While all three liquid fractions have the same pale blue-turquoise fluorescence, I don't know if this is simply a solvent-related phenomenon. Crashing with citric and dissolving the citrates in water could change everything, in the sense that there might be a change in the fluorescence colour at least, both because of the different solvent and because this is currently the fluorescence of the freebase which is being observed rather than that of the protonated cation. Then there's the solid that remains in the pipette. This needs to be quantified and examined for solubility in acidic water along with any resulting fluorescence that may arise. It does mostly look like rue crud but maybe there's still an alkaloid or two in there.
Fourthly, coming back to the rue crud/solid remnant - if this turns out to be a significant quantity of pharmacologically inert material it would force me to reassess my interpretation if my subjective experiences and drialnk(?) moar... My subjective tests do still hint that I've been tasting a mixture that was predominantly harmine + THH, though.
Fifthly, I want to re-iterate the unreliability of making assumptions based on subjectively observed fluorescence. And I should apologise for still not having found my gypsum for making TLC plates :lol:
Here's some reading - for my benefit as much as anyone else's!
Solvent Effects on Fluorescence Emission
Photophysics of harmaline in solvent mixtures
Experimental and computational study of solvent effects on one- and two-photon absorption spectra of chlorinated harmines
It would be very helpful if anybody knew of specific, pre-existing data regarding solutions of harmala alkaloids in butyl acetate - I still find it a little hard to believe that I might actually be doing novel research.
you could speed up the filtering if using coffee filters or lab grade vacuum pump filter setup, easily enough
Specifically, the BuOAc extraction process has taken about five days, although it could have been made quicker by starting with dry freebase and putting it straight into a syphon-fed percolation column. The observation of how the BuOAc dries the paste is of limited use in the practical execution of preparing successive fractions of harmala alkaloid solution.
If the goal was to get mixed harmala freebase into solution as quickly as possible I would think warming it up would help considerably. I maybe could have used vacuum filtration on the rue cooks although a kilo of seeds still would have taken ten or more batches going through my largest Buchner funnel. This kind of brings me to the other point about my approach here - it's at odds with the spirit of the alkyl acetate approach that you've outlined here because I've used water cooks for the ascorbic acid investigation and then used precipitated freebase for a somewhat incidental solubility experiment. I guess that should be my sixth caveat.
And to round off, a little update - the three fractions of BuOAc extract were progressively lighter yellow in colour, the first being morning piss colour, and the third being the equivalent after a pint of water and five cups of tea, with the second showing a shade somewhere between. This is also matched by the volumes of each fraction - 150, 100 and 50 mL respectively. The fluorescence follows a similar pattern, which brings me onto the final point worthy of note. If you have a point source beam of UV for examining your solutions, it's very easy to see that the beam will penetrate to a depth that inversely correlates with the concentration of the fluorescent solute. More molecules = more absorption. Thus, the spy pen light beam only gets a couple of centimetres into the first fraction, 4cm into the second, and passes all the way (4cm) through the third fraction and can then penetrate 1cm of the first fraction as well. This might give some indication of the relative concentrations in each of the fractions.
Citric acid precipitation should proceed this coming morning.
EDIT: Maybe even more tangential but there is this:
Pilot study on the uptake and modification of harmaline in acceptor plants: An innovative approach to visualize the interspecific transfer of natural products.
Redirecting Pretty awesome paper!
The BuOAc is sat in the can next to my jar of crude alks and I just keep nibbling the alks