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Testing psilocybin mushrooms, research demands a full suite of magic

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Basing a psychedelic mushroom’s potency on a single prodrug and its hallucinogen is marred for various reasons. Labs have a lot more work to do beyond psilocybin and psilocin as they explore vast amounts of chemical, therapeutic, and toxicological data after testing mushrooms.

A previous story documented other psychedelics related to psilocin, and don’t forget about terpenes found in fresh mushrooms. Psychedelic research exemptions are, however, a global novelty. Eric Janusson, Ph.D. in analytical chemistry, is the Lead Chemist at Delic Labs. His tone chimes with excitement as he touches on numerous unknowns in the expansive field of psychedelic research,

We are looking at the full suite. I keep saying, Fear and Loathing in Las Vegas, the trunk of their car – that’s what we are analyzing. We don’t know what we are looking for besides the obvious analogues. But there is still lots to figure out in the [mushroom].

Dr. Janusson

The full suite of tryptamine compounds in psychedelic mushrooms, featuring DMT and melatonin.

Delic Labs is currently one-third fixated on the accurate analysis of psilocybin when testing mushrooms. This was according to their CSO and President, Dr. Markus Roggen. He essentially broke down their approach into three stages and precise quantification of the main ingredients is the first. To take the initial step, Delic Labs had to first improve chromatography and extraction to be able to analyze those ingredients.

Everyone, right now, wants to know how much psilocybin and psilocin are in the mushroom. That, [Delic Labs] can do, that is the paper (1) we published, and we are constantly improving our methods. And we are adding [the full suite] of other compounds to our analytical capabilities.

Dr. Roggen.

A different psychedelic research lab, also on Canada’s west coast, is engaging with similar areas of psilocybin mushroom testing as well. Numinus uses highly advanced equipment to analyze the full suite of psilocybin analogues, such as triple-quad mass spectrometers. An email response from Science Officer and General Manager at Numinus, Sharan Sidhu, confirmed their tests include 4-hydroxytryptamine. But that type of targetted analysis is still missing a step.

Stack sixteen high-resolution mass spectrometers in a series and labs still cannot scan the fruiting body of an intact mushroom and expect accurate results. This is because compounds need to be separated and isolated to gain insights from mass spectrometry data. Unfortunately, psilocybin has for decades been accidentally converted to psilocin and then destroyed during the crucial separation process known as chromatography. Thus, labs were previously quite inaccurate when testing mushrooms for psilocybin content before recent upgrades to extraction. (1) With better extraction and chromatography methods, scientists can say, for example, what’s truly more potent: caps or stems.

HPLC (high-pressure liquid chromatography) is displayed as a schematic in the figure. A substance, or mushroom, is dissolved in a solvent and then drawn through a column filled with filter media. Until recently, however, poor buffers and reagents would convert or simply destroy a sample of psilocybin before it entered a detector – such as a mass spectrometer. (2)

Exploring uncharted chemistry – other mushrooms are magic, too

The second step for testing is the untargeted analysis of mushrooms that peaks Dr. Janusson’s enthusiasm.

We want to look at everything, the complete [fungi], and try to characterize it the best we can because there is still a lot of research to be done. And developing analytical workflows to accurately tell us more about the mushroom is a lot of our focus right now. 

Dr. Janusson

We want to find out what else is the mushroom… untargeted analysis. So [Delic Labs] takes functional mushrooms as our test platform for that because these have their own group of interesting compounds. We can then find out if those same compounds are in psychedelic mushrooms, so translate and marry these worlds. Similar to the cannabis world, everyone talks about synergistic effects but no one really looked for much else other than THC and CBD.  

Dr. Roggen

Fruiting psychedelic research beyond fungi

Dr. Markus Roggen described other important therapeutics in the realm as the next saga of Delic Lab’s research.

As a third area, psilocybin is a psychedelic compound. So as we touch the psychedelics, let’s keep touching the psychedelics. And there, we want to look at other compounds that are not in the mushrooms themselves but are psychedelic in their own right.

Dr. Roggen

Despite an application to study DMT, LSD, and other psychedelics, though, the chemists at Delic Labs turned down the idea of synthetic psilocybin. After all, science has far more work left before modifying those magical molecules. Or perhaps, this simply reflects the founder of Delic Labs, Dr. Markus Roggen, who is based in the field of organic chemistry. Yes, lysergic acid and modified psilocybin can each start from organic fungi, but it’s also true that psychedelic science is a monumental task.

Discovering a new molecule – the current role of synthetics

Psychedelic research labs sometimes rely on reference standards synthesized from modified psilocybin. An example is the acetylated, 4-ACO-DMT, which can be used to synthesize standards of a rare relative of psilocybin known as 4-HO-TMT. (2) Rather than consume peculiar molecules, though, synthetic copies are better intended for clinical research or discovering substances in nature for the first time.

Zooming back to the first and second steps, Numinus recently identified 4-HO-TMT in psychedelic mushrooms. According to Sidhu, this was achieved using HRMS (High-Resolution Mass Spectrometry.) Essentially, this molecule has an extra group of atoms compared to psilocin but is more related to aeruginascin – an analogue found in a limited few varietals. Lastly, Sharan Sidhu confirmed that metabolism is next to toxicology in Numinus’s 2022 mushroom testing plans.

From this, does it appear that synthetic and organic chemists need to marry their research for psychedelic science to truly bloom? And let us know in the comments if you are excited for new research in this pyschedelic space.

Sources

  • Wright, S., Malcolm, A., Wright, C., O’Prey, S., Crichton, E., Dash, N., Moseley, R. W., Zaczek, W., Edwards, P., Fussell, R. J., & Syms, R. R. (2015). A microelectromechanical systems-enabled, miniature triple quadrupole mass spectrometer. Analytical chemistry87(6), 3115–3122. https://doi.org/10.1021/acs.analchem.5b00311Hatakeyama, Shiro & Akatsuka, T.. (2014). Measurements of Gaseous Peroxides in the Oceanic Lower Atmosphere.
  1. Samuelsson, Alexzander & Janusson, Eric & Shah, Sajni & Roggen, Markus. (2021). Rapid quantification of Psilocybin with reversed-phase HPLC and single-wavelength detection. 10.33774/chemrxiv-2021-70mm1.
  2. Chadeayne, A. R., Pham, D., Reid, B. G., Golen, J. A., & Manke, D. R. (2020). Active Metabolite of Aeruginascin (4-Hydroxy-N,N,N-trimethyltryptamine): Synthesis, Structure, and Serotonergic Binding Affinity. ACS omega5(27), 16940–16943. https://doi.org/10.1021/acsomega.0c02208





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