Gramine

[pitubo]

We should figure out how to add another carbon thingy to said bridge then.

Oh come on people, chemistry is not playing with legos. You can't just pick up a molecule and arbitrarily add or remove parts. it does not work like that.

Can we please discuss the practical aspects of gramine? For example:

Is gramine really very toxic? I just can not find reliable information about it. There's a lot of assumptions going around, but surprisingly little hard data.

For instance, gramine is apparently present in the leaves of barley. Surely that gets eaten occasionally by cows, sheep and goats. Where is the "dead farm animal scare" in relation to barley leaves and gramine?

The LD50 for mice and rats is about 50 mg/kg body weight, which would translate to more than 2 gram for the average human being. Good luck smoalking 2 gram freebase unknown alkaloid extract in one go.

Another issue is the combination of gramine with maoi. It is not unreasonable to expect that gramine is digested and metabolized in similar ways as dmt. The oral LD50 of dmt is probably also way over 2 grams for the average human being. But I would not carelessly ingest 2 gram dmt while maoi inhibited. So what is the case with gramine and MAOIs?

 

[entheogenic-gnosis]

We should figure out how to add another carbon thingy to said bridge then.

It's called methylene insertion. This would be the reverse of part of the biosynthesis of gramine from tryptophan where a methylene bridge is removed - curiously, labelling studies show that the same nitrogen atom is retained in the molecule during this process even though it is attached to the carbon atom that gets removed.

So this is our side chain "ch2-ch2-nh2"

The carbon connected to the amine nitrogen (nh2) is the "alpha" carbon, the next is the "beta" carbon...

So your saying the alpha carbon is removed without effecting the amine nitrogen? Fascinating...

I'm only in my second year of organic chemistry and I am still learning, so sorry for all the clarification, I just want to be sure I'm understanding this correctly....


-------

For the person who Said " organic chemistry is not like playing with legos"

It kind of is, here is an example:

Row 7a of the periodic table (halogens) all have a single gap in their atomic shell, which they desperately want to fill...

Now say we take 2,5-dimethoxy-phenethylamine, the four position of this molecule is perfect for completing the gap of the atomic shells of halogens (as the hydrogen connected to the carbon at that position offers an electron)...

So kind of like legos, due to electrons and gaps in electrons, you can add to molecules.

You can subtract pieces of molecules as well though this process can be a bit different.

Regardless, it IS very possible to say "I think I want to add this too or subtract this from this molecule" and to be able to preform such a feat (after the notebook chemistry confirms its possible) in the real world...

Look at the benzene ring, August Kekulé was going crazy trying to figure out C6H6, there was no way he could complete the formula in a linear fashion...

Carbon has 4 gaps in its atomic shell, hydrogen has one, now poor August Kekulé was frustrated and goes to sleep, where he dreams of a snake eating it's own tail, then it all comes together, A RING! This was the answer!
Every carbon has a hydrogen on it, which leaves three gaps left in the atomic shell of every carbon, which are filled by single and double bonds to their neighboring carbon atoms, forming a six remembered ring.

...I don't know, I see the atoms as being like legos, the electrons could be represented as the "bumps" and the gaps could be represented by the "impressions"...

Molecules as well, if you know the "rules" of the molecule you can manipulate it, Albert Hoffman, Alexander shulgin, Daniel Trachsel, David e. Nichols, etc...actually focused a good deal of their careers on this sort of molecule manipulation.

This is just my thoughts on the matter though, and as i said before, I'm still early in my learning, and have a long way to go.


You were right about the gramine toxicity bit though, there's really no evidence that it's toxic in humans, though there is quite a bit of data in the animal realm involving this compound, a quick abstract and some links are below...

Some hematological and histopathological effects of the alkaloids gramine and hordenine on meadow voles (Microtus pennsylvanicus) - PubMed

Meadow voles (Microtus pennsylvanicus) were used to evaluate the relative toxicity of the alkaloids, gramine and hordenine, which are present in reed canarygrass (Phalaris arundinacea) and to assess their effects on the quality of this grass as a forage. One hundred and twenty meadow voles, 31...

www.ncbi.nlm.nih.gov

Some hematological and histopathological effects of the alkaloids gramine and hordenine on meadow voles (Microtus pennsylvanicus).
Goelz MF, Rothenbacher H, Wiggins JP, Kendall WA, Hershberger TV.
Abstract
Meadow voles (Microtus pennsylvanicus) were used to evaluate the relative toxicity of the alkaloids, gramine and hordenine, which are present in reed canarygrass (Phalaris arundinacea) and to assess their effects on the quality of this grass as a forage. One hundred and twenty meadow voles, 31 days of age, were fed gramine (0, 0.125, 0.25, or 0.5% of a nutritionally complete diet) or hordenine sulfate (0, 0.15, 0.31, or 0.62% of the diet) for 21 days. The effects of treatment on growth, mortality, hematology, blood chemistry, and histology of body organs were examined. Approximately one-third of the voles died when fed either 0.25 or 0.50% gramine. Voles that survived on gramine diets had kidney lesions, glycosuria, higher intakes (P less than 0.05), and lower weight gains (P less than 0.01) than control animals. Voles fed 0.25 and 0.50% gramine had increased circulating reticulocyte levels (P less than 0.01) and those fed 0.125% gramine had higher alkaline phosphatase activity (P less than 0.05) than the control voles. Hordenine did not affect vole diet intake, weight gain, or rate of mortality. Voles fed hordenine developed kidney lesions and glucose was detected in the urine of 62% of these animals. Gramine was more toxic than hordenine on a molar equivalent basis. Some hematological and histopathological effects of the alkaloids gramine and hordenine on meadow voles (Microtus pennsylvanicus) - PubMed

gramine toxicity - Search Results - PubMed

gramine toxicity - Search Results - PubMed

www.ncbi.nlm.nih.gov

I could not find anything anywhere confirming that gramine is toxic in humans....

------

-eg

 

[pitubo]

So kind of like legos, due to electrons and gaps in electrons, you can add to molecules.

Sorry to keep straying off topic, but no, it's not lego. With lego, you get to pick up the individual parts and choose where to plug on new parts. In chemistry, you cannot expect molecules to react magically where you want them to react.

Say you want to chlorinate the 3 carbon in your above example? How would you stop the action on the 4 carbon happening instead? What about the amine nitrogen? Isn't that going to want to react with halogens? What if side reactions cleave one of the methoxy groups?

Then there are endless subtleties about the conditions favorable to the intended reaction, such as catalysts, solvent effects, etc.

 

[downwardsfromzero]

"kind of like lego" =/= "is lego". But pitubo has a point. Internet forums like this one have a plethora of posts where people with inadequate to zero understanding of chemical synthesis (no offence!) post questions similar to steppa's, a particular example that keeps cropping up being the "how do we get the hydroxy group off of psilocin to make DMT?" one. I find this as irritating as I suspect pitubo does (although a small portion at the back of my brain is reserved for mulling this over without prejudice).

The gramine question is potentially more do-able, it would be nice to be able to avoid using cyanide though.

So you['re] saying the alpha carbon is removed without [a]ffecting the amine nitrogen? Fascinating...

The process is mediated by pyridoxal bound to the side-chain nitrogen. The tryptophan get decarboxylated, the alpha carbon gets hydroxylated and a shuffle of bonds occurs - which is where the pyridoxal shows off its niftiness in bouncing pi bonds around - so the beta carbon jumps onto the nitrogen to produce pyridoxal bound N'-formyl indolemethylamine. Hydrolysis, SAMe, yada yada. Hey presto, gramine with the same nitrogen atom as there was in the starting tryptophan.

However:
Biosynthesis of gramine

The biosynthesis of gramine was tested by growing the germinating barley in media containing pyruvic, 2-14C acid, W-benzyloxycarbonyl-tryptophan β-14C and tryptophan, β-14C. Iso-nicotinylhydrazide was also added in a series of experiments to inhibit the cleaving action of pyridoxal phosphate. The presence of the inhibitor reduces the total uptake of tryptophan and benzyloxycarbonyl-tryptophan by a factor of 15 - 20, while the uptake of pyruvic acid and the total production of gramine were reduced by a factor of only 0.5. This fact suggests an alternative route for the biosynthesis of gramine, when the path via tryptophan is blocked.

Isn't nature wonderful?

 

[pitubo]

I find this as irritating as I suspect pitubo does (although a small portion at the back of my brain is reserved for mulling this over without prejudice).

It is not really irritating to me, it is just wrong. It pleases me to weed out myths and misunderstandings that are sooner or later detrimental to us all. I am probably wrong myself regularly and I appreciate it if other people can point out any of my wrongs in a fair and straightforward manner. They give me a chance to learn and help me to stop polluting the common infospace.

With regard to conversion of gramine into dmt, I think that - even if the chemical transformation were easy, safe and cheap - it makes no sense to do so when it is so easy to extract dmt in quite pure form from reasonably well-obtainable plant sources, whereas the same can not be said for gramine.

 

[steppa]

pitubo,

In regard to

it would be useful to get some of the sharp minds here thinking about how we can get rid of potential gramine for those extracting from these plants....

The question of converting gramine into DMT is exactly on topic.

With regard to conversion of gramine into dmt, I think that - even if the chemical transformation were easy, safe and cheap - it makes no sense

It would make sense if the conversation could be done in presence of DMT as we wouldn't even need to worry about gramines toxicity or about how to get it out then. It would be the ultimate solution to the problem of gramine occuring in our extractions. Thus I think it should be allowed to ask without getting witty remarks. But that's just my opinion of course.

Of course I see, that this may be difficult...but hey...as asking a question doesn't cost a thing, I thought I might as well give it a shot.

 

[pitubo]

Endlessness' original question was about the options for gramine removal. Usually that means some kind of separation process. Technically you are right that a hypothetical conversion of gramine would also effect a removal, but it just isn't practical or even realistic in this case.

The key to my argument is that a separation can be performed on a mixture of compounds, but a chemical transformation is rarely performed on a mixture of compounds.

Like I wrote earlier, you just cannot pick and choose the molecules that will undergo a reaction. The molecules "have a mind of their own" with respect to reacting. For this very reason, you'll notice that if you study multi-step chemical synthesis procedures, there is usually a work-up and purification step between each part of the larger synthesis scheme.

Applying these considerations to your proposed solution for the presence of gramine in plant extracts, it implies that in order to perform any transformation on gramine, the gramine would have to be separated and purified beforehand. But once that has been done, why bother anymore with the transformation? It is probably more efficient to extract more plant material for dmt than to process the separated gramine.

So I remain with my earlier assessment: converting gramine is not realistic, because we don't have a proven separation and purification procedure, and it is not relevant, because it is not economical and/or the chemical transformations are not kitchen-level. In fact I even wonder if it would not be "easier" to synthesize dmt from basic reagents than to turn gramine into dmt.

The talk about having enzymes do the work for us is IMHO no more than prototypical pie-in-the-sky stoner talk. If that route were realistic, why not have the enzymes make dmt in the first place? Can I has my gene-spliced-lsd-making-enzymes enhanced potato plants too?

 

[downwardsfromzero]

In fact I even wonder if it would not be "easier" to synthesize dmt from basic reagents than to turn gramine into dmt.

"If" I was to consider synthesizing DMT, gramine would be a reasonably good starting material. But there's more ways than one to dose skin a cat.

Can I has my gene-spliced-lsd-making-enzymes enhanced potato plants too?

I'll get my fairy to send you one. (Solanidine looks a bit like LSD, doesn't it? )

Perfecting a gramine extraction procedure would point towards effective ways of removing this largely unwanted alkaloid, although if it's insoluble in naphtha as suggested then we've sort of got that one nailed anyhow.

 

[pitubo]

Perfecting a gramine extraction procedure would point towards effective ways of removing this largely unwanted alkaloid

Agreed 100%

although if it's insoluble in naphtha as suggested then we've sort of got that one nailed anyhow.

How much more than a suggestion is it, really? IIRC endlessness is the source of this information (can't find it now, it's not in this thread). Can we verify/replicate this?

 

[downwardsfromzero]

Ironically, one might say, the simplest way to obtain gramine is by synthesis, rather than extraction. If you can get hold of indole you're virtually there. Then the solubility/extraction from matrix/possible separation from DMT tests could easily be carried out.

With a modicum of purification and daring/stupidity, human toxicity tests could even be performed. Hopefully not LD50 :? I'll say - just in case - really don't do this.

 

[1ce]

Is there any substantial evidence supporting gramine for being moderate-high risk for nuerotoxicity? Links I found stated it as inconclusive or probably not a factor in tests where phalaris aquatica was fed to sheep.

How does this scale in comparison to say.. methylmercury in tuna?

 

[downwardsfromzero]

(Human) toxicity will probably come through its adrenergic properties. Maybe gramine neurotoxicity is aimed more at insects?

Late hour addled thoughts off top of head.

 

[1ce]

I recall reading that gramine is used in industry to prepare other tryptamines. I don't think anything down that route would be more simplistic than chromatography. I'd aim to pinpoint a safe/negligible dose of gramine and go from there.

DMT-acetate being soluble in chloroform may be of interest, I wonder if that could be exploited to separate it from gramine-acetate.

 

[downwardsfromzero]

There's an evens chance at least that the gramine acetate would dissolve as well, it's all down to the experimentals at this stage. Good suggestion, you tempt me to locate some gramine as a toy

 

[1ce]

There's an evens chance at least that the gramine acetate would dissolve as well, it's all down to the experimentals at this stage. Good suggestion, you tempt me to locate some gramine as a toy

Yeah it seems easy to come by, maybe a bisolvent crystallization could separate the freebases. But you're right, the only way to be certain would be to try.

 

[Leithen]

Once the grasses are back in season I will be collecting as much as I can and doing as many tests as possible. This is once of the easiest source plants for a lot of North Americans to obtain and I think getting a well tested tek is crucial! :thumb_up:

Is there any substantial evidence supporting gramine for being moderate-high risk for nuerotoxicity? Links I found stated it as inconclusive or probably not a factor in tests where phalaris aquatica was fed to sheep.

I do remember reading in TiHKAL that there was no proof these toxins effected humans. There was also no proof that they did not but Shulgin did not sound entirely convinced of this.

 

[pitubo]

Wikipedia, in its lemma on gramine, states that it is used as a starting material for the synthesis of tryptophan, but gives no reference for that claim. The tryptophan lemma in turn states that it is mostly produced by means of fermentation by selected bacterial strains.

The TIHKAL entry for tryptamine has a synthesis starting from indole, to gramine, to indole-3-acetonitrile, to tryptamine. It involves formaldehyde, cyanide and LAH - so not relevant for furter discussion here.

I found a somewhat recent paper on gramine that presents an angle that I have not seen mentioned before. I won't comment much on the content of the paper, as I did not study it intensively and it is a little above my head. But it does include a reference for the use of gramine as a starting material for the synthesis of tryptophan and, perhaps interesting to some readers, it involves the alkylation of indolealkylamines. I will leave it for others to comment on the relevance of the paper.

Synthesis, spectroscopy, theoretical and biological studies of new gramine-steroids salts and conjugates
Weronika Kozanecka, Lucyna Mrówczyńska, Tomasz Pospieszny, Beata Jasiewicz, Mateusz Gierszewski
Steroids, Volume 98, June 2015, Pages 92–99
Redirecting

 

[pitubo]

Oh, and I also found this tidbit. It's not exactly peer reviewed science, but the page does list some references. Perhaps it could be useful as a pointer for further research.

Oats

Therapeutics and Pharmacology: Avena is a nourishing herb applicable to any state of debility and exhaustion and during convalescence. It is particularly suited as a long-term treatment in nervous debility, making gradual but sustained progress in fighting off shingles and other forms of herpes, neuralgia, neuritis and even chronic depression. It is also of benefit as part of a regime for people attempting to withdraw from an addiction to alcohol, smoking, tranquillizers or other drugs. The mild sedative and hypnotic properties are due to the indole alkaloid gramine. The alkaloid avenine stimulates the central nervous system and is the component which causes horses fed on large quantities of oats to become highly excitable.

I do remember reading in TiHKAL that there was no proof these toxins effected humans.

But what about the combination with mao inhibition? This needs to be considered also.

 

[nen888]

..just to put a bit in, late in the thread...and reiterate a bit what sauroman1, pitubo and entheogenic-gnosis were saying re Gramine and toxicity :-


things can get a bit buried here in the archives of the halls of the Nexus, so i'll just grab a few things that were posted in the Phalaris = The Way...etc thread..to save people having to go there
(and without wanting to repeat anything alreday said) :-

p3 i wrote:

gramine (also found in Phalaris spp) is speculated as having ephdedrine-like effects.. in sheep it was 'psychotropic' at 10-30mg/kg i.v.; it showed oral toxicity at very large doses (400-600mg/kg) leading to death in some animals [Bourke et al 1988]..
..in the amounts present (if at all) in Desmanthus or Phalaris (less than 5% of total alkaloids) it should present no serious toxicity, and be barely (if at all) noticeable..

p.4

[Gramine] appears in a lot of plants, including phalaris spp which have some history of ingestion. so i would think if gramine were 'acutely toxic' in humans we would have some data on this by now..
the death of animals due to Phalaris aquatica 'staggers syndrome' may be due to MAO inhibition interaction (ß-carbolines), or unknown compounds, and the grazing of massive amounts..from Bourke et al 1988 (Aust Vet J. 1988, Jul; 65(7):218-20.)

The acute toxicity for sheep of 3 alkaloids that occur in Phalaris acquatica was examined by intravenous and oral administration. The lowest tested dose rates that produced clinically observed signs were, for 5-methoxy dimethyltryptamine, 0.1 mg/kg body weight intravenously and 40 mg/kg orally; for gramine, 10 mg/kg intravenously and 500 mg/kg orally; and for hordenine, 20 mg/kg intravenously and 800 mg/kg orally. All induced the clinical signs observed in the nervous form of phalaris toxicity, but none induced the cardiac, sudden death

gramine (3-(dimethylaminomethyl)-indole) was also apparently "behaivourally-active in rats" [Gessner et al. 1961]

Ginkgo wrote: (p4)

On the subject of gramine, I've found some more literature, and it points to what nen888 said, that gramine isn't toxic in small doses. Still I think we should be cautious. It is a medium potent 5HT-2A antagonist, which means it will counteract effects of DMT and similar psychedelics, as they do most of their magic on that very receptor type, so gramine should nevertheless be removed from an extract. We also need a way to separate DMT and 5-MeO-DMT. Maybe one of their salt forms have different solubility properties, enabling one to separate them.

Hordenine is a stimulant, by the way, it is well documented as a stimulator of the release of norepinephrine, and is used by body-builders.

..Gramine has also been used as a human health supplement (not sure dosage range..a link is buried somewhere else i think in the corridors of the nexus)

.

 

[entheogenic-gnosis]

So kind of like legos, due to electrons and gaps in electrons, you can add to molecules.

Sorry to keep straying off topic, but no, it's not lego. With lego, you get to pick up the individual parts and choose where to plug on new parts. In chemistry, you cannot expect molecules to react magically where you want them to react.

Say you want to chlorinate the 3 carbon in your above example? How would you stop the action on the 4 carbon happening instead? What about the amine nitrogen? Isn't that going to want to react with halogens? What if side reactions cleave one of the methoxy groups?

Then there are endless subtleties about the conditions favorable to the intended reaction, such as catalysts, solvent effects, etc.

It is like a Lego and it isn't, molecules can be manipulated, but you must understand the rules regarding what you can and can not do...

With the 2,5-phenethylamine/amphetamine example, while there are things you may not be able to do, there are ways to confirm that you did what you set out to, Proton NMR spectroscopy of the hydrochloride salt in D2O should ensure that your bromine (in the case of DOB) atom had entered the four position.

Just like if you have a Lego with out proper "bumps" (electrons) or "indentations" (gaps in atomic shell) and try to mash it together with another Lego without proper bumps and dents, it won't work...

Chemistry is the same way, know the rules of the building blocks and you will be able to construct structures...

I feel there is value in asking "what if", what if we can cleave the 1,2,4-Triazole group from Rizatriptan?...by obtaining the answers you also learn the rules of what your doing...

I guess like shulgin I still mentally visualize the molecules as manipulatable structures, which they are...

When you speak of DMT biosynthesis, I still visualize an amino acid decarboxylase removing the COOH grouping from the tryptophan molecule in the same way that you would picture physically removing the COOH group from a Lego model of the molecule...

In that biosynthesis there is an indole-amine methyl transferase enzyme called S-adenosyl-L-methionine, which will add a methyl group to the amine nitrogen of tryptamine compounds (at which point the SAM becomes S-adenosyl-L-homocysteine)..

So, when I picture an enzyme "adding" a CH3 group to the amine nitrogen of the tryptamine, (simultaneously becoming SAH while the tryptamine becomes methylated) the Lego metaphor really does help...

Or say you understand that tryptamine and methyl-iodide produce N,N,N-trimethyltryptammonium iodide, and that removing a methyl group from N,N,N-trimethyltryptammonium iodide will give n,n-dimethyltryptamine, so, like a Lego, you want to pull a CH3 off of the N,N,N-trimethyltryptammonium iodide, so what do you do? You hit the notebooks, and by knowing the rules and limitations of what you are working with, you eventually find a method for demetyhlating N,N,N-trimethyltryptammonium iodide (reflux the N,N,N-trimethyltryptammonium iodide in thf treated with LiEt3BH in THF for 9 hours, after cooling acidify with HCL, then dry under vacuum...and like a Lego you managed to remove the CH3 group that you set out to remove...giving DMT

Regardless, I see the value of asking "what if", I also see molecules as manipulatable, similar to legos...now, that's not to say that you can just go placing what ever you want where you want it, there are rules, but when you know the rules you can manipulate the matter...

I'll admit I'm still learning, but some of the best lessons I've learned came about by myself suggesting a possibility, and then having someone more knowledgeable explaining to me why that would or would not work , which is why I don't discourage the type of thinking that seems to aggravate those settled in the field...

...it took a long time before I actually read all this, I've been on an academic brake for the last two months or so, and have put very little thought into such concerns. I considered not responding as so much time had passed, but also felt obligated to explain the position I proposed in an earlier post...and I'm not going to argue whether molecules can be viewed as "legos" or not, it's subjective to your understandings... though I would like to remind those who are more knowledgeable to try to be more supportive of those who learning and cut them some slack.

-eg