Taming the Zwitterion
Taming the Zwitterion
With the advent of psychedelic research finally opening in the U.S., a new industry appears to be forming. Recent medical studies have shown so many positive results that progress in this burgeoning field is unlikely to slow any time soon.
As with any new field, we start out not knowing what we don’t know. The early days of the cannabis industry were a perfect example of that. Fortunately, this is a curable condition and the cure is information/knowledge.
Psilocybe cubensis offers some not-so-obvious problems. Psilocin (4-hydroxy tryptamine) is a nice stable molecule that can be analyzed easily in a variety of ways. Psilocybin (4-phosphoryloxy tryptamine), which is the most abundant (and very unstable) active, is known as a zwitterion. This means that it has a positively charged portion and a negatively charged portion within the same molecule. Amino acids fit this description as well.
Psilocybin is quickly converted to psilocin by the acidic environment and enzymatic dephosphorylation.
In our digestive tracts, psilocybin is quickly converted to psilocin by the acidic environment and enzymatic dephosphorylation. Dephosphorylation is the highly active process in our bodies that helps regulate several processes, all of which are essential to health and growth. So in the end, psilocin is the actual active that can cross the blood-brain barrier.
The passing of Prop122 in Colorado as well as Oregon legislation will have a lot of researchers working out ways of isolating the active compounds of known composition for accurate dosing of patients. The polar extractables (water/alcohol soluble) comprise a large majority of the fungal mass. It’s made up of amino acids, carboxylic acids, mono and disaccharides, and a very small amount of psilocin and psilocybin. So in short, it’s quite a mess. With water around, psilocybin gets very nervous– particularly where varying pH is concerned. I can verify this as I’ve destroyed plenty. The ability to analyze the entire matrix is essential to provide researchers with potential paths toward the isolation of actives.
The early days of analytical research on psilocybe mushrooms were shown to be plagued with errors due to the dephosphorylation of psilocybin to psilocin during sample preparation. HPLC with tightly controlled pH buffering systems has been successfully applied, though sometimes problematic. HPLC/MS improved the situation a great deal allowing greater sensitivity and discrimination. In either case, you still have to work through a sample preparation that’s potentially sensitive to matrix differences.
Psychedelic therapy has opened wonderful new doors that could help minimize the use of daily medications or medications with side effects that are sometimes worse than the actual conditions
My analytical background consists of highly active compounds. Organometallic catalysts, pyrophorics, and other oxygen, water, and pH-sensitive compounds being the majority of them. My first passes at analyzing mushrooms consisted of failures stemming from dephosphorylation or some other decomposition pathway. Luckily this was a familiar red flag given my background so I immediately went back to my roots and started handling these things as catalysts. That was the ticket.
So here at Prop122Labs, I set out on a mission. I wanted to know exactly what samples look like as they are, so I developed a methodology that freezes the psilocybin into a fixed configuration while it’s being extracted from the fungal mass. This gives us a thermally stable moiety (necessary for GC analysis) to analyze. It provides great chromatography, easy sample preparation, and a wealth of analytical information in one preparation. The same sample can be analyzed under different acquisition parameters to yield information on the balance of the matrix. Utilizing GC/MS I’m able to get razor-sharp peaks, fast acquisition, and a high degree of separation with the ability to measure the minors and majors on the same preparation. The method is also universally applicable to all tryptamines as isolates or in a wide array of biological matrices.
The acceptance of psychedelic therapy has opened wonderful new doors that could help with the minimizing of having to use daily medications or medications with side effects that are sometimes worse than the actual conditions. We’ve learned a great deal through mistakes made by other industries. Let’s just make sure we learn from them and not re-learn in a way that could risk a great new opportunity.
So here’s hoping that this industry progresses smoothly and without incident. The key is knowing exactly what you have. Prop122Labs is here to help in any way we can.
Richard Crawford is a B.S. chemist, process developer, and a 30 year veteran of various spectroscopic techniques. The attraction to GC/MS analysis was quite immediate as the ability to separate and characterize compounds was so vast. 20 years of industry experience in the manufacturing of highly sensitive compounds was the final impetus for developing the methodology to test Psilocybin mushrooms.