Quality Testing For Psilocybin
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Psilocybin is having a revival. Modern clinical trials have lifted the magic mushroom drug above its ‘60s-psychedelia trappings and once more demonstrated its medical potential for treating depression and other mental health disorders. Corporate interest has followed; there are more psychedelic start-up companies than you can shake an incense stick at these days.
Right now, many of these companies are investing in regions of North America that have some form of legal access to psilocybin therapy, such as Canada and the US state of Oregon.
With this legality comes rigor. Restrictions. Regulations. These words may be taboo in the more anarchic corners of the psychedelics community, but they help form a protective net around the vulnerable patients receiving psilocybin. Because before it can become a medicine, the fungus must be tested to ensure its safety and potency.
But how exactly does one test psilocybin? Decades of prohibition have prevented scientists from understanding the deeper chemical nature of psychedelic fungi, so where do the scientists of today start when tasked with analyzing its unique chemicals? Analytical Cannabis asked some of them to find out.
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Magic mushroom machinery
“We have been testing psilocybin for a little while. So when Oregon decided to announce that they were going to bring this program on, it was like, ‘Hey, why not us?’,” says Dan Huson, CEO of Rose City Laboratories, a commercial lab based in Portland, Oregon.
Back in April, Rose City became the first lab in Oregon to receive a psilocybin testing license from the state health department. But to get that license, the team had to get a testing method together. And to do that, they first had to figure out what machinery they needed.
“The state were primarily looking [for labs to test] for the psilocybin and the psilocin content,” Bjorn Fritzsche, senior chemist at Rose City, tells Analytical Cannabis.
“[We] did some research into the methodologies that were available, and what amounts of active ingredients vs. active compounds we were expecting in the samples. Based on that, we went with high performance-liquid chromatography [HPLC] with a UV detector. Nothing super fancy.”
“I found a published method that wasn’t necessarily great – it didn't have the column or the mobile phase – but it was a starting point,” Fritzsche adds. “And we developed from there.”
The crux of this method development proved to be extracting the core hallucinogenic chemicals – the tryptamines psilocybin and psilocin – from the fungi, so they can be tested in isolation.
“[We] developed extraction methodologies to make sure that we get the best recoveries,” Fritzsche says, “which actually turned out to be the most difficult part of the development: extracting the psilocybin and psilocin into a solvent without doing any conversion of the compounds and also [to] get close to 100% recoveries.”
While they may have been endeavoring into this slim avenue of science, Fritzsche and his colleagues weren’t the first to discover the challenges of magic mushroom extraction; over the national border in Vancouver, the team at Delic Labs have been developing their own psilocybin sampling protocols since 2019.
“For the tryptamines, we actually have to start with extraction or sample preparation,” Dr. Markus Roggen, president and chief science officer of Delic Labs, told Analytical Cannabis during a recent webinar.
“We have the [following] problem,” he added. “How do we get everything off from the mushroom into our liquids, and then actually inject it into the test instrument? It seems all over the place. When we did a literature review on what is available—all solvents, mostly polar solvents, because psilocybin is water soluble.”
By experimenting with soaking the fungi in different solvents, Roggen and his team first found out what not to do when it came to extraction.
“We [did] everything – soaking it, sonication, checking vortex - everything,” he explained. “You have temperatures, from low temperature to high temperature, you have soaking times from an hour to two days. And there’s a paper that shows that the longer you soak it, the more you get out.”
“But then we looked at it,” he added. “And when you do sonication or use heat – when sonication normally heats up your sample – you dephosphorylate and you don’t test for psilocybin that’s in the mushroom, you actually test psilocin that you just extracted.”
For Roggen and his team – who also happen to test and research cannabis – the extraction–degradation hurdle was a familiar one.
“If you recall back, the cannabis world had the problem that the first cannabinoid was CBN [cannabinol], which was a degradation product,” he said. “And we have the same problem now in the mushroom world: that our sample preparation methods are an issue.”
And they still are. After having conducted their own experiments and read through the limited literature, the Delic team concluded that, while several psilocybin sample preparation methods seem viable, there ultimately is no standard to swear by.
“There’s a lack of standard method, which is the main pitfall of this whole industry,” Callum Teevens, a neuroscience student at the University of Victoria – and co-presenter of Markus Roggen’s webinar – told the Analytical Cannabis audience in May.
“It’s hard to continue studies without knowing what exactly to use and how to do it and what the outcome will be,” he added. “So that’s the worst challenge.”
But it’s a challenge Fritzsche and his Rose City team had to overcome to get their Oregonian license.
“We actually ended up developing isocratic methods to go out and find exactly the right column,” Fritzsche says.
“It was a little bit of combination of just research and trial and error. But in the end, we found something that worked really well for us – a fairly short run time, eight minutes per sample or so. And we have shown that we have no coelutions with any other compounds and the numbers we’re getting are in the range that we expected from literature.”
Moving forward in Oregon
Confident with their sampling technique and their HPLC-UV method, the Rose City team didn’t waste time getting their validation.
“We presented all that data to our state recognition body here, known as ORELAP, Oregon Environmental Laboratory Accreditation Program,” Fritzsche says. “They were very happy with our data and accepted it and we got our accreditation from them.”
“The only next step after that was to actually get the licensure from the Oregon Psilocybin Program,” he continues, “which costs a stack of money. But we ended up being the first to the finish line and, on the speciation side, still the only lab [licensed to test for the species of psilocybin].”
“We have the marked market pretty much cornered right now.”
But capturing the Oregonian psilocybin testing market in 2023 isn’t as rewarding as one might expect, it turns out.
“The program currently is going on pretty slow here,” Fritzsche says. “It is somewhat cost-prohibitive to get treatment, and there’s not really enough treatment facilities.”
And even if more treatment centers open, there won’t be that much more psilocybin to test.
“There is sufficient supply of mushrooms available,” Fritzsche continues, “due to the fact that a single grower can produce quite large amounts and the actual amounts consumed in sessions are a few grams.”
So, right now, there isn’t a great demand for psilocybin testing in Oregon. But the Rose City team are still hoping the sector will grow soon enough, and not just in Oregon; efforts to set up a legal psilocybin licensing system are currently underway in Colorado and the lab’s CEO is keeping a close eye on developments.
“I’ve actually been asked to be on the advisory board’s committee for Colorado,” says Dan Huson. “And I feel like Colorado is a little bit ahead of the game, even though they’re behind right now.”
“We have a system that is the first and so it’s a little bit rough,” he adds. “I think everybody else in the future will be able to roll out a much better, more comprehensive system for patients to be able to access medicine.”
This wider access will also invite more scrutiny of Rose City’s current psilocybin testing method. But, for now, the lab seems confident its technique can weather the appraisal.