Lysergamides LSH Production in Cultured Metarhizium?

[Moldy Plates]

Over the past few years, an ergot alkaloid lab in WV has put out a series of publications about LSH production in Metarhizium species. The Panaccione lab's first genome search paper on this came a couple years ago: https://journals.asm.org/doi/epub/10.1128/AEM.00373-20 Their most recent one from this year (https://bmcresnotes.biomedcentral.com/articles/10.1186/s13104-022-06068-2) includes culture conditions for Metarhizium brunneum that lead to production of LSH, but not a lot of other alkaloids. I thought this could be interesting information for people interested in going further with the theory that LSH is qualitatively different from LSA in effect. (I first became curious when I compared 250 I. tricolor seeds to 50 seedlings, 50 seedlings >>> 250 seeds, much more visual and "up" like other lysergamides I have tried, the seeds were sedating and burpy)

If somebody wanted to obtain a culture of Metarhizium brunneum, Met52 EC is a fungal insecticide where the main ingredient is M. anisopliae, a synonym for brunneum. No, I don't know the history behind this species having two different names. In theory, growing this guy up should result in LSH production and not much else. It's not pretty - involves infecting larve and then extracting. But it is at least a viable path to getting LSH.

ETA: I'm mostly posting this because I want to imagine somebody smashing up bugs to get drugs, but also I wonder if YPD media would be enough to get LSH. Would definitely be easier to spin that shit down and extract. (And to clarify, you'd be extracting from the spent media - see Fig 4 in the first paper for M. brunneum strain ARSEF 9354 - although does that mean you could drink the broth? That just doesn't seem right.)

 

[Jabberwocky]

interesting

@tregar

 

[G_Chem]

I’m highly interested in knowing more on dosing seedlings. This is something that is very easy to do and something that I’d be more than willing to try.

Any and all details on that please! How long did you grow the seedlings? Indoor or outdoor? Nutrients or straight water? What kind of soil?

Thanks

-GC

 

[G_Chem]

On to your OP, what kind of larva need to be infected? How does one do this? (Maybe kill the larva via some mechanism like freezing then infect? Or just infect?)

I want to try both of these lol, great thread!

-GC

 

[Moldy Plates]

I’m highly interested in knowing more on dosing seedlings. This is something that is very easy to do and something that I’d be more than willing to try.

Any and all details on that please! How long did you grow the seedlings? Indoor or outdoor? Nutrients or straight water? What kind of soil?

Thanks

-GC

I threw handfuls of seeds in the yard to see where they would grow. The sprinklers helped them sprout. One morning, I cut down 50 of them that looked like they had just shed their shells, and blended it up with some grapefruit. (the first one didn't taste great) I should have just sprouted between paper towels. IDK how to describe the effect, this is a sample size of one. I've done the morning glory seed thing before, up to 15g chewed or extracted. This was different and seemed like a big improvement. Who knows, set and setting, all that. But it made me curious about the biosynthetic pathway, so it makes sense that actively metabolizing tissue would have more of the compound that is made by the endosymbiote than its breakdown product. If a different compound is responsible for a different trip, what else produces LSH?

And the authors apparently buy their larvae, specifying New York Worms as the supplier for wax worms. The rest, with glass beads to homogenize the dead infected larvae and extract the ergot alkaloids, I leave to your imagination, though I should mention that day 12 post inoculation had the highest LSH for the strain they tested.

 

[G_Chem]

The one thing I’m not seeing in the article after a quick scan is any yield? Or am I blind? I suppose it’s likely a potent alkaloid but still would be nice to know.

“Bead beating in methanol” sounds cool lol.

Also I’ll try the seedlings indoor and report back. I too have tried “LSA extracts” as well as dried seeds all with lackluster effects. One time with CWE I got a light trip similar to mushrooms but too light for my liking.

-GC

 

[ErgotFiend]

this, along with Claviceps Paspali intoxication, needs further investigation. If someone more technically knowledgeable could put together a theoretical procedure for the whole operation, I, and plenty of other dummies, would be willing to be guinea pigs. Sounds like fun, just I am curious to how many bugs would be in a dose, and whether or not they need to be alive during the course of the infection.

 

[Moldy Plates]

I, and plenty of other dummies, would be willing to be guinea pigs. Sounds like fun, just I am curious to how many bugs would be in a dose, and whether or not they need to be alive during the course of the infection.

1. I'm not sure if the commercial fungal insecticides produce LSH or other dangerous undesirable compounds, but most of the ones sequenced so far in that species do produce it. I'm sure there will be some isolate that ends up producing a lot, with lots of variation between isolates.
2. No idea on dosage, but the publication in question reported 7 micrograms/larve in figure 5 of the first paper. If a first attempt dose is 100 micrograms, that's what, 14, 15 wax worms?
3. The authors state the wax worms died on day 2 or 3, peak LSH was produced in the 7-11 days post infection range.
4. Again, I imagine growing the fungus up in vitro is going to be a more pleasant experience, especially since fig 4 shows much more LSH secreted into the liquid media than in the hyphae, at a total of 5 micromoles in 5 ml of broth. I'm not sure I am reading the materials and methods correctly, because that is 200 micromoles per liter. A micromole of any lysergamide is something like 0.0003 grams, or 300 micrograms. So, a liter of 200 micromolar secreted LSH would equate to 60 milligrams of LSH? Somebody check that math. If LSH dosage is in the 100 microgram range, I think that a dose would be something absurd, like a third of a mL. That just seems way too good to be true. There's a catch somewhere. Even if the dosage is much higher, it seems too easy and the yield is insane. I guess that's why they included this tidbit "The proportion of alkaloids secreted into the medium was 0.78, 0.85, and 0.84 for ergine, LAH, and ergonovine, respectively. The proportions secreted are more than seven times greater than the proportion of ergot alkaloids secreted from another ergot alkaloid-producing fungus, Neosartorya fumigata (syn. Aspergillus fumigatus)."
5. Don't be a dummy, nobody should be doing anything unless they know the answer to #1. Don't ingest any ergot alkaloid producing fungus directly. This is mainly a shitpost.

 

[G_Chem]

Well according to this article Metarhizium anisopliae produced the ergot alkaloids too and is as you said fairly easy to obtain. What potential dangers do you foresee? Obviously one should be proficient in extracting said product, but I’m wondering the potential of maybe some toxic impurities.

This isn’t a shitpost this a potential goldmine. Especially at the yields you speak of..

-GC

 

[ErgotFiend]

So could one create large sterile dishes of agar substrate and then innoculate it with commercially available Metarhizium? Then scoop it all out after the fact and do a cold water or alchohol extraction. I, and my roomate, would have a lot of fun trying this out.

 

[tregar]

Moldyplates said:

I thought this could be interesting information for people interested in going further with the theory that LSH is qualitatively different from LSA in effect. (I first became curious when I compared 250 I. tricolor seeds to 50 seedlings, 50 seedlings >>> 250 seeds, much more visual and "up" like other lysergamides I have tried, the seeds were sedating and burpy)

Great thread. Thanks snafu in the void for mention. Just wanted to say the combo of LSH + penniclavine is the bomb. I write extensively about claviceps paspali (same exact same alkaloid as the powerful Mesoamerican morning glory) in the link below, long time chemist. I've had 4 very powerful trip experiences over 2 months using LSH + penniclavine extract for only $2 a dose, my latest with 600 morning glory seeds, more powerful than 500ug of acid but has the naturalness of mescaline, link below, highly recommend. I've taken bridgesii cactus tea over 200 times and Ayahuasca over 80 times, and this morning glory extract is at the very top of my favorites list, listed 5 papers showing proof of LSA to LSH conversion. I take this once a week it's so powerful.

With 600 seed (18g) extract, zero nausea, zero sedation, this is stimulating like LSD. 5 hour strong experience where I saw 2 hours of closed eye dancing colorful geometrics that formed completely naked women goddesses at the intersections, mind blowing music enhancement, even tripped on it at the waterpark on my last trip, as it is devoid of anxiety and thought confusion like with LSD. This extract is great for outdoors. The euphoria is extreme as well. This extract is way more powerful than LSD, will never buy LSD again, I prefer this a thousand times over acid. Way more visual, colorful, euphoric, music enhancing with a very deep head space with profound healing and an afterglow the next day that is incredible.

LSH Tek:
https://bluelight.org/xf/threads/lsh-extract-tek-500-heavenly-blue-morning-glory-extract-in-1oz-everclear-1-oz-wine-imagine-your-best-3-hit-lsd-experience-x-2.921681/




Adduct formation of d-Lysergic Acid Amide or (LSA) and aldehydes.pdf 1.13MB 0 downloads
2016 Polish morning glory study shows LSH and penniclavine two highest alkaloids in MG.pdf 1.19MB 0 downloads
Tryptophan analogues form adducts with aldehydes alone or with aldehydes in alcohol.pdf 780.97KB 0 downloads
The LSA component of claviceps paspali forms LSH when dunked into fermented liquor (wine).pdf 3.29MB 0 downloads
5 Ergot alkaloids in morning glory just as stimulating as LSD in animal experiments, Yui and Yuji Takeo, 1958.pdf 596.64KB 0 downloads

LSH Tek:
Shroomery link, 4 pages with 7 upvotes and others who have been using it:

How to make LSI or Lysergic Acid Isovaleraldemide at home from LSA in morning glory seeds, ancient LSD - The Psychedelic Experience - Shroomery Message Board

Attachment: 2022 paper on Adduct formation of d-Lysergic Acid Amide or (LSA) and aldehydes.pdf (1,187,857 bytes) --> Update: see page 16 at the end for 8.13.2023 update for 1 page easy tek with

www.shroomery.org

Krystle Cole from the book "Lysergic":

"Isn't Ergot what Socrates used to take at Eleusis?" I thought it was kind of cool to be taking something that the founders of our democracy used to take, but that our current democracy has made illegal. LSD chemist Todd Skinner replied "Yes, the priest who handed down the secret formula from generation to generation did a wine (or water with mint or peppermint leaves added, adds acidity and high in acetaldehyde) infusion of the ergot." Todd had prepared 6 jugs of ergot wine and stored them for many years.

Krystle Cole's "ergot wine" experience (several pages long) in the book "Lysergic", reported that she saw constantly rotating holographic Sanskrit or Arabic & Zodiac symbols, floating in a circle around Todd's head. Even LSD chemist prepared claviceps paspali ergot in wine. He must have known the secret of acetaldehyde adduction to form LSH in the wine from the leftover LSA in the ergot.

LSD chemist often use claviceps paspali ergot to make LSD. They will boil all the extracted alkaloids in a solution of ph=10 potassium hydroxide in the ergot under nitrogen inert atmosphere under low light conditions to convert to Lysergic Acid, and then couple diethylamine to the lysergic acid to form LSD using the modern PyBop peptide coupling agent.

The above report is important, and it shows that LSD chemist such as Todd Skinner and friends have been having very strong psychedelic experiences using wine infusions of claviceps paspali ergot which grows on paspalum distichum L. grass. Many often mow it and don't even realize it. This same ergot grows on affected grass on the famous Rarian plain next to Eleusis, where the kykeon was drank by hundreds of people in the psychedelic ceremony. Wine soaked Ergot could have easily fed hundreds of people. This ceremony went on for 2,000 years. The claviceps paspali ergot contains the exact same alkaloid profile as the powerful Mesoamerican morning glory.
-----------------------------------------------------------------------------------------------------------------------------------------------------
Here is another 18g or 600 seed extract I prepared yesterday, ready to trip on it tonight, this will be my 5th trip with the LSH tek extract.

Visuals = 11 out of 10
mesmerizing euphoria = 11 out of 10
Color enhancement = 11 out of 10
Music enhancement = 11 out of 10
deep head space = 11 out of 10
no thought confusion or anxiety = 11 out of 10
tolerance is much lower than with LSD, you can use this more often, it has a low tolerance like mescaline or cactus, remains super potent even if taken again 7 days later, no loss of strength = 11 out of 10.
500 seeds just as potent as 400ug of acid, but more powerful all around.
600 seeds just as potent as 500ug of acid, but more powerful all around.
Natural = 11 out of 10, no choppiness or frame like nature to the visuals and visions, flowing visions and music enhancement & euphoria extreme. No man made feeling to it at all, extremely potent ancient psychedelic.
Teamwork = 11 out of 10, the combo of penniclavine + LSH hits 6 more receptors than even LSD, and far more aesthetic and beauty enhancing then LSD, identical to mescaline "over the top" beauty enhancement.

 

[Moldy Plates]

So could one create large sterile dishes of agar substrate and then innoculate it with commercially available Metarhizium? Then scoop it all out after the fact and do a cold water or alchohol extraction. I, and my roomate, would have a lot of fun trying this out.

Liquid Culture, not plates. Sounds like they grew it on yeast extract + sucrose (table sugar). I don't think you would have to extract, just drink the media for LSH. But that seems sketch. When things seem too good to be true they usually are. Could be nothing, could be huge dose, could be dangerous. More info needed.

 

[tregar]

Moldy plates said:

I don't think you would have to extract, just drink the media for LSH.

You can eat the media plain, but it will only contain LSA and the other 4 alkaloids unless it is first dunked into fermented liquor such as wine as shown in 1961 Arcamone study, see paper above. Once the media is dunked into wine, the LSA constituent converts to LSH via adduct/condensation reaction with acetaldehyde, conversion requires acidity as a condition, wine is ph=4 and also contains the acetaldehyde as required.

LSH will decompose to LSA if extracted into plain water, it must be extracted into acidic solutions to remain stable, such as wine which is at ph=4 (see 2016 Polish MG study paper), remains stable indefinitely in wine. See paper above, 1961 Arcamone, LSA converts to LSH in fermented liquor such as wine which contains acetaldehyde, sherry wine is 10mg acetaldehyde per shot or 1oz, there is a attraction + condensation reaction that takes place with LSA and acetaldehyde that converts it to LSH, see 1st paper above from 2022, this is the basis of my TEK. Since acetaldehyde boils off at 70 degree F, it's important to keep the wine and everclear cold in fridge, and keep extract cold in fridge until drank. LSH does indeed reach the brain, toxicology study found it in bloodstream of those who consumed an alcoholic extract of the seeds. The ancient Mayan and Aztec always extracted into liquor, they knew the secret, see photo of the seeds above with how the natives did it using alcohol.

The 2016 Polish morning glory revealed that penniclavine and LSH are two highest alkaloids in morning glory seeds, same in claviceps paspali, LSH Tek above converts any leftover LSA in the seeds back to it's previous fresh off the vine peak LSH levels, bringing the extract to fresh off the vine levels of LSH. Many don't realize it is teamwork between the ultra high levels of LSH + penniclavine in either MG seeds or claviceps paspali that is responsible for the strong trips, both just as stimulating as LSD in animal experiments, wheras LSA is a breakdown product of LSH over time and heat, is sedating and not psychedelic.

 

[Moldy Plates]

LSH will decompose to LSA if extracted into plain water, it must be extracted into acidic solutions to remain stable, such as wine which is at ph=4 (see 2016 Polish MG study paper), remains stable indefinitely in wine. See paper above, 1961 Arcamone, LSA converts to LSH in fermented liquor such as wine which contains acetaldehyde, sherry wine is 10mg acetaldehyde per shot or 1oz, there is a attraction + condensation reaction that takes place with LSA and acetaldehyde that converts it to LSH, see 1st paper above from 2022, this is the basis of my TEK. Since acetaldehyde boils off at 70 degree F, it's important to keep the wine and everclear cold in fridge, and keep extract cold in fridge until drank. LSH does indeed reach the brain, toxicology study found it in bloodstream of those who consumed an alcoholic extract of the seeds. The ancient Mayan and Aztec always extracted into liquor, they knew the secret, see photo of the seeds above with how the natives did it using alcohol.

I assume that Metarhizium is doing some work of its own to adjust the culture pH. If people were able to detect more LSH excreted than LSA, even though LSA is a decomposition product of LSH, then culture conditions must have been sufficient for LSH to survive the trip from the culture tube to the detector. IDK how that would work out at home, but it really doesn't sound like something you would be worrying about extracting or mixing with alcoholic beverages.

 

[G_Chem]

Well didn’t they extract into methanol? Likely 99%? Wouldn’t this negate much of hydrolysis to LSA?

Will the LSH degrade upon evaporation of the solvent/methanol? Methanols low BP would make this a quick process. Then one could dilute in ever clear ethanol to a proper dilution and use.

-GC

 

[red22]

There's reason to believe that "LSH" simply decomposes to ergine in the body:

easily decomposes in a weak acid solution to form ergine and acetaldehyde.

Stoll, A. and Hofmann, A. (1965) The ergot alkaloids. In R.H.F. Manske (ed.), The Alkaloids, Vol. VIII, Academic Press, New York, Chap. 21, p. 747


ready cleavage under hot aqueous conditions, yielding D-lysergamide and acetaldehyde.

N. Castagnoli, jun., K. Corbett, E.B. Chain, and R. Thomas, (1970) Biosynthesis of N-(a-Hydroxyethyl)lysergamide, a Metabolite of Claviceps paspali, Biochem. J., 117, 451


The α-hydroxyethylamide spontaneously decomposes to lysergic acid amide, and the naturally-occurring amide may arise by this process.

Heinz G. Floss & John A. Anderson. 'Biosynthesis of Ergot Toxins' (1980) in The Biosynthesis of Mycotoxins (Pieter S. Steyn, ed.), Elsevier, p. 21 (2012)


this material is quite unstable and is easily converted in the process of isolation.

TiHKaL (part 1), Alexander Shulgin, p. 313


These compounds [lysergic acid hydroxyethylamide & isolysergic acid hydroxyethylamide], although well documented as components in the Convolvulaceae, are possibly lost in several of the analyses of alkaloid composition. They are extremely unstable, and are very readily degraded into acetaldehyde and the corresponding amide, ergine or isoergine. In these instances their presence will be measured only by the elevated levels of the derived amides.

Alexander Shulgin, Psychopharmacological Agents, Vol. 4 (ed. Maxwell Gordon), Elsevier, Dec 2, 2012

 

[Moldy Plates]

There's reason to believe that "LSH" simply decomposes to ergine in the body

Well, that's all more evidence of decomposition during extraction. IDK how they managed to get LSH to stay stable in culture media, but from the materials and methods, it didn't sound like anything special. I am unsure if stomach conditions would lead to conversion from LSH to LSA, but I would guess that yes some goes from LSH to LSA when ingested. Lke any other reaction, kinetics and le chatelier are our friend - it takes time, we reach an equilibrium point, and so maybe we experience both molecules. Something that many different labs have observed and deduced from biosynthetic pathways is that LSA is not directly produced by the fungal endosymbionts in morning glory. It's just a breakdown of LSH, which takes place over days, weeks, and months. Not to be an adaptationist, but why would this be the case unless that alkaloid was important for deterring whatever herbivore is a problem for morning glory?

From animal tests, we know that LSH is more stimulating than LSA. (https://www.unodc.org/unodc/en/data-and-analysis/bulletin/bulletin_1971-01-01_1_page003.html)

D-Lysergic acid N-(1-hydroxyethyl) amide induces contractions in the isolated uterus of the guinea pig and in the rabbit uterus in situ, showing about 30-50% of the activity of ergometrine. In mice and rabbits it produced the syndrome of central sympathetic stimulation, such as piloerection, mydriasis, and hyperthermia, which suggests that it could have an LSD-like activity, but this hypothesis has not yet been verified by experiments on humans ( https://www.nature.com/articles/189313a0 ).

The next step of giving LSH to humans via oral and parenteral routes just hasn't been done in medical settings. If I had to hazard a guess as to why this question hasn't been answered yet, it would be that LSD is still available and is much more reliable, so why bother looking at plants? (or parasitic fungi in this case)

Related to the oral route of ingestion, tregar has mentioned Vitamin C, but I don't know what empiric backing that has.

 

[Moldy Plates]

Dunno how I missed it before, but aspergillus sounds much easier to handle in culture. https://pubmed.ncbi.nlm.nih.gov/34586904/

Would be pretty interesting to grow a few ml of the strain used to produce this figure, it seems like an easier route

 

[red22]

The Theoretical Synthesis and in silico Modelling of Lysergic Acid Biscinnamylidene Amide from the Adduct Formation of d-Lysergic Acid Amide and Cinnamaldehyde. Victor Chiruta. The Journal of Chemistry Studies. 2022.

 

[pupnik]

oooh, cinnamon!