Growth parameters:
B+ strain cubensis were colonized on birdseed and transferred to a bulk substrate consisting of coir, manure, straw, vermiculite, and gypsum to which 10mM of melatonin was added giving a ratio of roughly 6.3 grams of melatonin to roughly 4 kg of bulk substrate. Substrate was measured at ph 8.

TLC:
Sample = Cubensis grown on melatonin doped substrate.
PC = Cubensis grown on undoped whole brown rice.
5-meo = 5-MeO-DMT HCL standard.
Mela = Melatonin standard.

Placeholder for analysis information.

Analysis suggests the fungus created a novel, but closely related analogue of the melatonin.

Preparation:
~5 grams of whole fruits were ground to a fine powder and soaked in ~10 ml 96% EtOH for ~2 months to create a tincture.

Bioassay:
With ~1 ml (0.5 grams)
Tincture was diluted with ~2 parts water and held at the back of the throat for ~10 minutes before swallowing. Effects began around the t+20 mark. Pleasant stoning effect on par with standard low dose cubensis ramping up steadily over the course of about 30 minutes to achieve a + on the Shulgin scale. Effects then plateaud into a threshold psychedelic state with some faintly translucent, pulsing, wave-like visual effects behind the eyelids. At ~t+70 the trippy effects began to subside and the characteristic melatonin heaviness set in. The heaviness increased with quite a bit of yawning over the course of ~30 minutes then began to subside as happens when staying awake after taking melatonin. By ~t+120 mark most psychoactive effects had passed apart from a light and easily missed stoney afterglow feeling.

At ~t+150 a standard nightly dose of harmalas + melatonin (~5mg each) was administered and subject went to bed. Sleep came easily and quickly. Dreams were slightly more vivid than usual and followed an extremely cohesive and linear direction, even to the point that getting up in the middle of the night to use the restroom resulted in the dream sequence picking up right where it left off and continuing with the story through the rest of the night. Upon waking subject felt well rested and no residual hangover effects were noted.

With ~3 ml (1.5 grams)
Tincture was diluted moar heavily on the order of ~10 parts water in trying to wash ground mushroom material from filter. Tincture/tea was administered slowly over the course of ~25 minutes taking a sip, holding at the back of the throat for a few minutes, swallowing, and repeating. Effects began at ~t+20 and ramped up slowly over the course of ~30 minutes. It came on as a light stoned feeling and gradually increased over the course of ~30 moar minutes toward a + on the Shulgin scale, at which point the heavy melatonin sedation kicked in. This floaty stoned sleepy state was maintained over the course of the next hour when subject decided to go to bed around the t+120 mark. Dreams were vivid but no moarso than with standard melatonin administration. Subject awoke well rested and no hangover effects were noted.

With ~7ml (3.5 grams)
Tincture was diluted roughly 50/50 with water. Half the dose was held at the back of the mouth for exactly 5 minutes then swallowed. This was repeated with the other half. At t+10 the "dregs" of the ground of mushrooms were soaked in warm water for 5 minutes. This tea was drank over the course of about 10 more minutes. First alerts began around t+20 with unmistakable effects by t+30. The onset ramped up to a nicely stoned feeling that plateaud between t+45 and t+60. This stoned feeling remained more or less for ~60 more minutes when the standard heavy/tired melatonin effects began and became quite strong. At this point ~t+120 the subject went to sleep. Dreams were very linear and coherent as if the entire sleep cycle was spent in one stable dream scenario. Subject slept slightly longer than usual, perhaps an extra 30 minutes and woke up feeling well rested without hangover.

With ~2.5 dry grams eaten:
Pattern was similar to all previous attempts. Effects began around t+30 and built strongly in the manner typical to standard psilocybin mushrooms leveling out a strong stoned melatonin feeling. Upon sleeping dreams were very vivid, but not particularly linear. Refreshed upon waking with no feeling of hangover.

Wow. I love the creative thinking behind this.

Wow, I'm excited to read more. Are you testing out what Shulgin theorized? He talked about making 4-HO-5-MeO-DMT by adding 5-meo-dmt to the growing substrate, although I see you did it with melatonin. What new molecule do you think you've stumbled upon and what kind of doses do you see yourself getting into?

Sasha's hypothesis and the Gartz paper were the inspirations for this avenue of substrate doping.

Melatonin was chosen for it's easy availability, and because the speculated biotransformation was less clear than using other (non-acetylated) tryptamines.

5-MeO-DMT doping is another route we should (and hope to) explore, as well as Mexamine (5-MeO-Tryptamine). Both of which will, hopefully, result in novel 5-MeO-DMT analogues.

I'm hoping someone moar knowledgeable than myself can interpret the mass spectra and give some insight into the possible identity of the novel analogue. Getting an NMR characterization is another project hopefully in the pipeline.

The next sessions will be 1.5 grams and 3.5 grams respectively; assuming favorable results, we'll have to do Terence proud and go for the full 5 dried grams.

I could be very wrong but one thing that could fit is the methylated acetamide (along with the more straight forward 4-OH attachment). It is not clear if the mushroom can pull the amide methylation off (as Mindlusion has said before), but who knows?

The purely hypothetical molecule comes in around 2au below the 264 value (see below), but that may be accounted by the absolute calibration (?). Benzyme, the background peak at 198 - what does it correspond to?

I guess another option is a double -OH attachment (4 position plus somewhere else).

Have you compared it yet to just taking cubensis and melatonin simultaneously?

N',4-dihydroxymelatonin would have about the right MW. Seems perhaps more likely than the N'-methylated version.

Or of course the other hydroxy group rather than being on the N' atom could instead be somewhere else on the benzene ring, perhaps. Maybe you already said this.

Yes, agreed, double hydroxy may be more likely since amide methylation is not all that clear.

For what it is worth, there are examples of N-Methylamides (called NMA) in the salvinorin family. See below for an example molecule and attached for an example paper. Infundibulum also gave some info here for Salvinorin B methylamide.

So if the unlikely molecule does turn up we could call it 4-OH-5-MeO-NMAT (the N-acetylated version of this I believe).

I have taken that combination at active doses a handful of times in the past (and many many times as harmelacybin low/microdoses before bed).

With higher dose melatonin and moderate dose cubensis the melatonin seems to kind of "grease the wheels" so to speak. The experience seems moar fluid and comfortable moving into the deeper psychedelic spheres. It's similar to harmalas and psilocyin in synergsitic effect, but without the intense potentiation.

The presentation of this one seemed almost the opposite, it came on like psilocybin, but just as it started getting trippy, it shifted gears into a very heavy melatonin state and basically totally overpowered the psychedelic effects just leaving me sleepy and stoned.

I'm really quite intrigued with this one, but I'm hesitant to draw too many conclusions at this point. We'll see how it goes at higher doses.

If it is indeed some kind of mutant melatonin, I'm curious about the downstream metabolism. How does a hydroxy on the 4 position effect metabolism to mexamine, pinoline, tryptoline, etc?

I will venture that I'm already suspecting this one is less a psychedelic and moar of a oneiorogen.

It's probably worth pointing out that that particular 'Salvinoid' N-methylacetamide was obtained through laboratory synthesis; the fact that a molecule exists is not an indication of an existing biosynthetic pathway in an organism.

It would be worth seeing what a mushroom would do to N'-acetyltryptamine (i.e. like melatonin sans methoxy group) to explore metabolic transformation at this end of the molecule. Maybe mushrooms do like methylating secondary amides; we don't really know.

And, getting a bit tangential now, what would the mushroom do with harman and other β-carbolines? Cyclisation of melatonin produces 6-methoxyharmalan, so it's worth thinking about. There's a whole array of metabolic studies to be done... We could potentially be dealing with a cyclised thing and that would account for the 2 "missing" mass units.

DWFZ - Yep, completely agree - lab synthesis does not mean an organism could do it. Mindlusion did find info on natural methylation of amides and posted a link here. Of course, it could be that the mushroom cannot do this anyway.

Good idea on the 2au shift (ring closing). Benzyme may be able to let us know about the absolute calibration of the MS data.

This post has a PhD dissertation and other documents describing several biotransformations from fungi which you may have reviewed already. If may give other ideas for other more candidates here.

This is a really strange coincidence. I had a really vivid dream that i was trying a new compound, i even remember the name : 5-MEO-Lysergique-acid-oxide (the effect was profound like 5 MEO, with the long lasting, gentle, subtle magic of LSA) . The next morning, i open nexus and found this thread. What's the odd?

Thanks for all of this. This year i will try to feed some MHRB to the cubes, and maybe to wood lovers?!

Most of this is over my head, but from what little i understand, this seems like a pretty valid experiment.

I wonder how well ps cyan myc would spawn to some acacia or mimosa wood? Would the math workout with the potential levels of alks, would it be worthwhile?

I gave my neighbor some acacia bark to put into one of his mushroom projects and it did seem to be stronger than some of the same batch grown without additives.

That enzymatic methylation ML linked was on an aromatic amide ('anilide') nitrogen; an aliphatic amide in the same molecule was not methylated. Just an observation.

And, of course, fungal biotransformations are utterly vast in scope!

Re-reading, I'm not entirely sure what has the N-2 molecular mass. Was this something that's showed up with the GC-MS? I'll knuckle down and do the calculations when my brain starts working

Yes there is a peak at 264 in the mass spec image that dreamer posted at the top. Scratching our heads about it.

Brain back in gear (i.e. it's not way past the middle of the f***in´ night!)

m/z 264 corresponds, potentially, to the dihydroxylation of melatonin; this would (in the 5,7 4,7 positions) easily produce a quinone of m/z 262. High res MS would be instructive here, NMR at least as good.

Thanks DWFZ, but is it 5,7 or 4,7?

You're right, I meant 4,7! I had a cognitive flatulence moment.

(So much for the brain being in gear!)

This is really quite fascinating. we've managed to take a psychedelic mushroom and make it no longer psychedelic.

The effects do seem to be consistent throughout the dose range. Effect strength seems to moar or less increase linearly with dose size. It's very very similar to melatonin but more stoning in general.

The dream effects are intriguing. A good dose of melatonin generally increases how vivid my dreams are, but they remain fairly chaotic and unstable, and I usually remember multiple dreams a night. This effect of one continuous stable dream scenario is something unique I've never encountered using any other oneirogen. I'd be curious to explore this more and see if others would experience the same effect.

I'd also be curious to know what effect these would have on a full dose of harmalas or in combination with another psychedelic.