burnt said:
you take that back!
I'd like to see you try to analyze cell lysates with an NMR.I'm hoping to have an orbitrap in a year or two. I do have a laser polarimeter now, inline with the DAD and MS
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DMT(s) Analysis Compilation [1H-/13C-NMR/HPLC/LC-MS]
- Thread starter Brennendes Wasser
- Start date
you take that back!
I'd like to see you try to analyze cell lysates with an NMR.I'm hoping to have an orbitrap in a year or two. I do have a laser polarimeter now, inline with the DAD and MS
benzyme said:burnt said:
you take that back!
I'm hoping to have an orbitrap in a year or two. I do have a laser polarimeter now, inline with the DAD and MS
yes its true, the orbitrap paired with a quadrupole mass filter and c-trap truly is the one and true god. I do submit. The NMR is succeeded.
sensitivity and routine analysis: ms > nmr
structure elucidation: nmr > ms
I'll submit though to high resolution MS combined with MS/MS to the N :twisted:
structure elucidation: nmr > ms
I'll submit though to high resolution MS combined with MS/MS to the N :twisted:
my next acquisition has got to be an Exactive. There are some on ebay, I'm just concerned about user-level servicing. My colleague/coworker suggested that instrument for what I usually test...small molecules, extracts, etc. He says it can effectively do structural elucidation rather effortlessly.
Apparently, Thermo became like Apple, and closed off alot of hardware troubleshooting in their software to end-users.
Apparently, Thermo became like Apple, and closed off alot of hardware troubleshooting in their software to end-users.
Tony6Strings
Rising Star
Yeah, reading these guys always makes me feel like an underachiever and a simpleton.
SpaceTraveller
Rising Star
Your heat treated red/Brown DMT - if exposed to sunlight in the same fashion as ordinary spice, does it still melt into a puddle and form N Oxide?
Just a hunch.
Also was the question of the effects of long term UV exposure ever answered?
Just a hunch.
Also was the question of the effects of long term UV exposure ever answered?
benzyme said:
Yeah, and they run the universities like a racket, best chance for you would be to find a freelancer, Thermo is far from consumer friendly
You mean an investigation if the melted Spice at 100 °C will also form some N-Oxide when exposed to sunlight?
Well I would definetly say yes, it is well homogenized and has a plain surface instead of being in crystaline chunks, so it should absorb UV-light very well. You expect an interesting outcome or why you have that idea ;D
Actually 2 days ago I had another idea, which may also be very interesting:
Place 1 sample Freebase and 1 sample Fumarates on a dish into UV-light and remove a little chunk every 2 days for GC/LC analysis. Then even without having a standard (and by assuming a linear dependency of signal intensity to mass concentration) one would be able to get some percentage values of
- the speed of the degradation is
- the efficiency of the protonation by FASA (and maybe even the anti-oxidative properties of fumaric acid) which should protect DMT compared to Freebase
It fell out of the kitchen window during a storm and broke into too many parts :x :? hahah ... then I placed a new sample from Christmas 2019 to Easter 2020 at the window - during winter to check how much 1 whole season of low-UV-weather would harm it. It looked just slightly degraded, totally fine compared to the 2-days-sample retrieved earlier here. Not sure if this is a direct hint, but based on appeareance I would say sun damage is minimal, even though it stood there for months. At Easter 2020 I replaced it with a new sample and it already looks quite degraded by now, but right now I just have a window facing North and this is definetly not causing the sample to get real UV-irradiation ...
Still I would have a longterm-sample with "winter exposure" and maybe I may move that other sample somewhere in direct sun. One could of course just irradiate a sample with laboratory equipment and calculate the equivalent dosage (time * intensity during summer), but I dont know if this would be comparable, as unusual degradation processes may occur when ramping up the intensity (and reducing time).
SpaceTraveller
Rising Star
Just a hunch
I feel like the heat treated form may prove more stable. I had a go at replicating your results but i started with a mixture of liquid alkaloids and crystal 50/50 from acacia acuminata, they ended up melting into a gorgeous golden mess (75 degrees C for about 5 hours in a small sealed jar) and now cooling at room temp the crystals are re-forming within the goop over a period of days. Will post results when complete.
When i have pure crystal in my hands again I'll be experimenting to see if I can replicate your mass increase result!
Seems weird. Have you ever tried again?
I feel like the heat treated form may prove more stable. I had a go at replicating your results but i started with a mixture of liquid alkaloids and crystal 50/50 from acacia acuminata, they ended up melting into a gorgeous golden mess (75 degrees C for about 5 hours in a small sealed jar) and now cooling at room temp the crystals are re-forming within the goop over a period of days. Will post results when complete.
When i have pure crystal in my hands again I'll be experimenting to see if I can replicate your mass increase result!
Seems weird. Have you ever tried again?
Thermo indeed is pretty bad customer service and has made it difficult to use their equipment outside of them. Q-exactive is pretty amazing instrument but the claim that it can do structure elucidation effortlessly is not a claim I'd buy. Sure its compound discover software is powerful but it can't solve new structures. if a compound is in a compatible database it can tentatively id. Anyway Thermo has market on these instruments for now would love for a competitor to come out with something similar bring down price etc.
The Q-exactive is an instrument ive used regularly for academic research, it is indeed powerful and no doubt the most powerful for analysis of small molecules within plant extracts. orbitrap resolution is usually down to 4-5 decimal places, although unfortunately even after a few years age they need frequent recalibration.
The MS-MS mode is also a very powerful tool for structure elucidation especially when it might pertain to less abundant trace alkaloids / compounds that one might want to identify within an extract. The 'collision power' within the quadrupole can be tuned by increasing or decreasing the voltage for fragmentation of stronger or weaker bonds, this is my favorite tool as it lets you generate multiple fragmentation patterns of a single ion/ionized compound. If one has a good idea of the parent structure, this is more than sufficient for structural elucidation.
Of course, if the parent structure is completely unknown or of high complexity, then complete characterization with NMR or x-ray crystallography is necessary. However the drawback with these methods is the necessity of the purified isolate of the unknown compound in question, which in the case of trace compounds requires mass scale extraction and separation or via total synthesis or semi-synthesis.
Whereas the amount of data you can generate with just milligrams of plant material using the q-exactive is pretty mind blowing.
The MS-MS mode is also a very powerful tool for structure elucidation especially when it might pertain to less abundant trace alkaloids / compounds that one might want to identify within an extract. The 'collision power' within the quadrupole can be tuned by increasing or decreasing the voltage for fragmentation of stronger or weaker bonds, this is my favorite tool as it lets you generate multiple fragmentation patterns of a single ion/ionized compound. If one has a good idea of the parent structure, this is more than sufficient for structural elucidation.
Of course, if the parent structure is completely unknown or of high complexity, then complete characterization with NMR or x-ray crystallography is necessary. However the drawback with these methods is the necessity of the purified isolate of the unknown compound in question, which in the case of trace compounds requires mass scale extraction and separation or via total synthesis or semi-synthesis.
Whereas the amount of data you can generate with just milligrams of plant material using the q-exactive is pretty mind blowing.
Well still nothing new since ages, but I actually had a sample of what I believed to be the pure jungle ready. But upon measuring I saw that it still contained mostly DMT ... So I discarded to post anything from that here, but the GC at least showed that there are 4 minor peaks besides DMT, that come in roughly the same quantity.
Gonna prepare another sample soon, which definetly totally spice-exhausted Jungle, prepared with Xylene extraction.
But meanwhile I got an UV-Vis of DMT ready and an IR-spectrum. No idea why anyone would use that, but its data nevertheless, so I attached it to "7.) Pure Spice".
Gonna prepare another sample soon, which definetly totally spice-exhausted Jungle, prepared with Xylene extraction.
But meanwhile I got an UV-Vis of DMT ready and an IR-spectrum. No idea why anyone would use that, but its data nevertheless, so I attached it to "7.) Pure Spice".
Brennendes Wasser said:
Pretty much what we've been trying to tell people for years but some people don't seem to accept it.
Brennendes Wasser said:
You have the spectra of those minor peaks ? Was the signal good enough to get a clear reading of that spectra? Wondering how it compares with what I found here. Or was it just GC but no coupled MS ?
Brennendes Wasser said:
How are you planning to prepare this sample? Excited to hear the results
Sadly it was not with MS, so there's not so much so say about that data. What I did was depleting a basic soup with hot Naphtha many times and that's why I thought I will just pull pure Jungle afterwards with Toluene. Seemingly there is still some Spice to be dragged out ...
Next time I will just directly use Xylene (no Toluene available, but wont change much) on the basic soup and then after evaporating in vacuo I will just throw boiling Naphtha on this. That should deplete it much better, as I'm then now extracting from an oily alkaloid mixture and not from a solution, which carries a dozen other things. I guess this way the Spice will come out of the other Alks pretty easily.
Next time I will just directly use Xylene (no Toluene available, but wont change much) on the basic soup and then after evaporating in vacuo I will just throw boiling Naphtha on this. That should deplete it much better, as I'm then now extracting from an oily alkaloid mixture and not from a solution, which carries a dozen other things. I guess this way the Spice will come out of the other Alks pretty easily.
Do several naphtha pulls on that extract, I've tested doing one pull and it was still mostly DMT.
Other than that, sounds like a great plan and I'm excited to see what you find out!
Other than that, sounds like a great plan and I'm excited to see what you find out!
JesusTheEntheogene
Jesus-Iesus-Zeus
I came up with a Yellow, orange Xylene extract following Shaggy d Entheogenist's jungle tek and am looking into obtaining a spectrometer I was curious if you have any suggestions on what type will give me the most accurate results. Second I would love to have it analyzed I have no idea what I am doing I quit school in 3rd grade and just research myself but I have a microscope, I can say there is clear differences in the *Jungle spice variant compared to the previous Pure white extracts I obtained following Cyb's tek much more purging effects throughout the body but I feel like I may have a nice sample with a very good stability, as It consisted almost nearly identical to some pure gold thc consistency into what was a distillate honey oil and hardened into a yellow orangish hard rocky oil substance and seemed evidently stronger lasts longer and smokes much more. That being said I also took pictures throughout the whole process periodically to record the color and physical changes and add various techniques. I was also wondering what would happen if one were to get say a Kilo of full spectrum procuct and place it within a contained zone such a lab glass box and vaporized it within a flash contained it and let it settle if this would be able to derive various significant blotches of separationendlessness said:
So finally something new added ... labelled as 8.) 1H-NMR of Harmala Alkaloids (No Manske) (but need to scroll down a litle bit). Stuff derived just from an acidic boil and then precipitated with NaOH. The Alkaloids were washed until water was neutral.
In short:
The ratio of Harmalin/Harmin seems practically exact 1:1, with an maximum possible excess of 9 % in regards to Harmalin.
Also no Vasicin could be observed. Possible reason would may be excessive washing with water as Vasicin may be more water soluble than the other 2 alks. So basically a Manske step can be skipped (can be skipped anyways as Vasicin is just unhealthy to pregnants' which should also take no Drux), as excessive washing of Alkaloids may also just remove it and be way faster than Manske.
So you would be wondering if evaporation and sequential solidification of a full spectrum pull would possibly cause a separation?
I think its pretty unlikely, a lot of these components start to turn brown while heating, indicating at least a partial decomposition while being heated. Also having a wild mixture will maybe also increase chances of reactions while getting hotter and hotter. If you could very precisely cool your vessel down, you would then maybe still achieve a fractional solidification, but again, a wild mixture will not fully evaporate and leave behind a lot of black tar. So any analysis on its content would be better by doing any sort of chromatographic separation with sequential Mass Analysis by Mass Spectrometry.
In short:
The ratio of Harmalin/Harmin seems practically exact 1:1, with an maximum possible excess of 9 % in regards to Harmalin.
Also no Vasicin could be observed. Possible reason would may be excessive washing with water as Vasicin may be more water soluble than the other 2 alks. So basically a Manske step can be skipped (can be skipped anyways as Vasicin is just unhealthy to pregnants' which should also take no Drux), as excessive washing of Alkaloids may also just remove it and be way faster than Manske.
JesusTheEntheogene said:
So you would be wondering if evaporation and sequential solidification of a full spectrum pull would possibly cause a separation?
I think its pretty unlikely, a lot of these components start to turn brown while heating, indicating at least a partial decomposition while being heated. Also having a wild mixture will maybe also increase chances of reactions while getting hotter and hotter. If you could very precisely cool your vessel down, you would then maybe still achieve a fractional solidification, but again, a wild mixture will not fully evaporate and leave behind a lot of black tar. So any analysis on its content would be better by doing any sort of chromatographic separation with sequential Mass Analysis by Mass Spectrometry.
The lack of vasicine and related compounds confirms my own findings too.. I think potentially they dont even precipitate in the first place and stay behind, dissolved in water, during crystallization of harmalas. This should be relatively simple to confirm.
Thanks for sharing!
Thanks for sharing!
I would like to add EA may not pick up vasicine even when using the EA dry tek approach. Although it could be more likely it doesn't form a citrate.