Endophytes from seed on plates & Seed extraction optimization

[lsaresearch]

Been looking into this.

Its known the dynamic in which seed produces its alkaloids is via the endophytic fungus that exists in the seed.

So basically the idea is instead of working with hard to work with wild strains or old under performing production plates of Claviceps ect. ( that need to be mutated) You take a fresh plate from seed to try to isolate the endophyte. Then the idea of taking this to a bio reactor would then be a possibility. This would be possible would it not? and give a an endophyte specimen that we know produces well which can be optimized in a bioreactor .

https://www.researchgate.net/publication/235432750_Endophytic_Fungi_Isolated_from_Oil-Seed_Crop_Jatropha_curcas_Produces_Oil_and_Exhibit_Antifungal_Activity

leads me to believe that isolating the endophyte should be possible.




I have some papers on this ive been looking at, along with more efficient plant matter extraction methods like ultra sound and microwave assisted/microwave synthesis reactors.

Identification and determination of ergot alkaloids in Morning Glory cultivars

Seeds of plants from Ipomoea genera contain numerous ergot alkaloids, including psychoactive ergine and ergometrine, and are often abused as so-called “legal highs.” In this work, an analytical method for determination of ergine and ergometrine, ...

www.ncbi.nlm.nih.gov

which shows a higher yield of ergine potentially due to the conversion of some other alkaloid in the material into ergine caused by the microwave radiation.

additionally, ultrasound or microwave extraction appears to make it easier to do complete extractions in shorter periods.



somethings to talk about, any other interesting ideas surely interested to hear !

Ive been thumbing through director of sound notes and a whole rainbow of related notes for quite some time now, and am very interested in the subject of applying modern applications to the topic at hand.

 

[Aum_Shanti]

and give a an endophyte specimen that we know produces well which can be optimized in a bioreactor .

leads me to believe that isolating the endophyte should be possible.

The isolation of the fungi certainly isn't the problem, but at least the Periglandula fungi on the psychedelic vines have such an extremely interconnected metabolism with the host plant, that any tries to grow these fungi on a medium have completely failed.

But there's also no need for this, as the famous Paspali bioreactor is simple and delivers huge amounts of these ergolines compared to the vessel size and potency of these substances.

which shows a higher yield of ergine potentially due to the conversion of some other alkaloid in the material into ergine caused by the microwave radiation.

Which is likely not what many people want, as LSA itself is mainly a non-visual sedativum, whereas LSH is probably a typical visual psychedelic.

additionally, ultrasound or microwave extraction appears to make it easier to do complete extractions in shorter periods.

I actually use ultrasound cleaners and the microwave for heating a lot for various extractions.

 

[lsaresearch]

Hey alum, I think I saw you write about a defat method using canola oil / house hold oils ? on seed? I was curious if there was any more to that, or any other interesting unique ways to extract that users here have been taking advantage of.

Ergine for my research is more desirable than the rest of the spectrum, I think the LSH infact converts to it. Pretty sure the endophyte consumes trptophan/similar conditions to the claviceps paspali.

The claviceps paspali from my knowledge, Requires you to have access to a performance strange, similar to the steven halls plates ect. Then not only that, you need to maintain your plates and keep them mutated. Which from the people ive talked to who have worked with these plates. Is difficult to get back into a performance setting, if not entirely unsuccessful.

additionally ive been told that working with wild species of claviceps paspali that they yield extremely low alkaloid something like 7mg / liter abysmal .

", that any tries to grow these fungi on a medium have completely failed. "

is this to your own experience / knowledge on the subject? Or have you infact found research that states this and the complications?

I do really appreciate talking to people about these things as its a lonely thing to research. Just from my experience reading forums.. Ive seen alot of " you cant do that" " do this" which the person was totally saying something about not doing , and im not sure if I can just take someones word for it with out having some research that shows someone trying and failing.



the guy who posted about his mixture turning green on the LSA/LSH/ect forum experienced that due to using " spring " water aka water with minerals.

Always use distilled / De ionized.


but I will note I am not here for the seed eating, but the processing, understanding of alkaloid profiles and extracting them in the most efficient manner possible.


I try to state using things that are accessible to all people. Steven halls strains, are not accessible to all people. Which is the idea behind this research. An accessible fungal grown that can produce large volume of alkaloid in bioreactor similar or close to the claviceps paspali that you can only really get from someone else.

Seeds are interesting accessible, renewable. Not the dream boat end game of bio reactors. But as you can see I am attempting to look at all options in that region.

I need to read more on microwave synthesis, I always brush past notes on it when im looking into things.

 

[lsaresearch]

https://www.researchgate.net/publication/276357002_Phylogenetic_and_chemotypic_diversity_of_Periglandula_species_in_eight_new_morning_glory_hosts_Convolvulaceae#pf5

thought this was interesting stuff. Still related to morning glories not woodrose. But still very interesting.

 

[Aum_Shanti]

Ergine for my research is more desirable than the rest of the spectrum,

How do you know, I assume you never had pure ergine?

I think the LSH infact converts to it.

Yep it does over time, which is likely why fresh seeds have different effects than old seeds.

Pretty sure the endophyte consumes trptophan/similar conditions to the claviceps paspali.

The ergoline biosynthesis starts from there, but the Periglandula fungi don't live on it. Currently it is not known what these fungi really consume for a living. It is assumed from surface lipids on the upper leaf surface.

is this to your own experience / knowledge on the subject? Or have you infact found research that states this and the complications?

Yes results from several scientific researches (e.g. the work from Hellwig is very recommended in this relation). They tried a plethora of known media, and none worked at all. As said, as it seems they are extremely strongly interconnected with the specific metabolism of a certain host. E.g. as it seems so strongly that a fungi living on a specific Ipomoea species is not capable to survive on another Ipomoea species. That is at least how they explain, that every species has their own fungi.

Still related to morning glories not woodrose.

Be aware that HBWR (Argyreia nervosa) is also a morning glory.

I just made a summary on the shroomery. At the end I listed a small sample of research work on this topic, which you could find interesting. But be aware that some very interesting works (e.g. like the one from Hellwig) are only available in german).

Psychedelic Vine Seeds - A small Compendium - The Psychedelic Experience - Shroomery Message Board

 

[lsaresearch]

Pure Ergine Also is not my intention. So Im not really worried about the instability or any issues with it. The most concerning thing is the iso-ergine profile. Though it seems ive answered this myself with a little searching.

As would never have pure ergine isolated for any duration by it self.

I appreciate the notes, am reading the thread you linked. I appreciate your detailed knowledge on the subject.

 

[lsaresearch]

Ive mainly been following Director of sound and other notes relatable.

Extraction and efficiency of procedures are my main focus in my research. Though I like to dream of the possibilities with culturing. I do see focus on more accessible resources as accessibility is part of the point of it all.




simple search yields an answer from one of my favorite contributors benz,

the epimerization of lsa to iso-lsa is irrelevant, because alkaline hydrolysis converts it all to lysergic acid.
doesn't matter if you start with paspalic acid or ET.

@alum,


Do you have experience with the available extraction teks described on your post? I,E cold water extraction, solvent extraction, acid base extraction ?

What's your experience with these separate teks ? What would you note as the purest most efficient extraction to your experience and knowledge ?

 

[Aum_Shanti]

The most concerning thing is the iso-ergine profile.

Why? What concerns you in this relation?

Ive mainly been following Director of sound and other notes relatable.

???

What is "Director of sound"?

the epimerization of lsa to iso-lsa is irrelevant, because alkaline hydrolysis converts it all to lysergic acid.
doesn't matter if you start with paspalic acid or ET.

Well this is the case, if you wanna then synth LSD from it. Then it plays no role, as you anyways convert them to lysergic acid. But if you wanna trip directly from LSA,LSH, Ergmetrine, ... it certainly is a relevant factor, as the isomers are usually almost non-active.

But generally epimerization happens until the natural balance, which for these ergolines is roughly at about 50:50. If you wanna separate the epimers, you need a column. Then you can separate off the active epimer, then again push epimerization to the natural balance (e.g. by making the solution quite acidic, which pushes epimerization), and again separate by column, etc. etc.

Do you have experience with the available extraction teks described on your post? I,E cold water extraction, solvent extraction, acid base extraction ?

Yes and no. I never did an a/b extraction, as IMHO it's really not needed.

What would you note as the purest most efficient extraction to your experience and knowledge ?

Every tek has its advantages and disadvantages. The ones getting a very pure result, need a lot of time and work, wheres a CWE is done superfast. It's a tradeoff of material/time vs. endresult purity.

I personally would recommend a non-polar/polar tek as best practical solution. It gets basically almost all bad stuff out, and is still doable for most people.

I don't see any real benefit in doing an a/b-tek, except you need pure alkaloids for some reason (e.g. further synth, special tests, etc.).