top of page
Image by Luke Chesser

Potency Analysis on Major and Minor Hydroxy-Tryptamines of Psilocybe Mushrooms

Updated: Dec 30, 2021


Mycelium Growth on a Petri Dish

The recent cultural shift on psychedelics, especially “magic mushrooms”, have created a rather unique space that opens the discussion on the overall properties and characteristics of psychoactive compounds. It has been long known that psilocybin and psilocyn are major compounds that produce the psychoactive effects of “magic mushrooms”, but with recent research, it has been discovered that there are several other minor compounds that also likely contribute to this effect. This document describes the analytical approach to the quantification of known major and minor compounds of psilocybe mushrooms, also known as hydroxy-tryptamines, using in depth analytical testing methods.



This research utilizes Liquid Chromatography-Mass Spectrometry/ Mass Spectrometry (LCMS/MS) technology rather than High-Pressure Liquid Chromatography (HPLC) for potency analysis of the hydroxy-tryptamines. As a testing platform, LCMS/MS has a much higher sensitivity than HPLC, and also provides additional modes of distinguishing peaks of active ingredients from one another. This includes multiple mass transitions per compound as well as normalization by deuterated internal standards for accurate quantitation. In essence, LCMS/MS are able to detect compounds present in trace amounts.

Through the use of rigidly defined testing methods, two major and two minor hydroxy-tryptamines commonly present in psilocybe cultivars have been quantified . Refer to Table 1. For these testing compounds.

Table 1. Potency Analysis Testing Standards

Testing Method

The rapid oxidation of psilocin and dephosphorylation of psilocybin during sample preparation are common issues encountered when testing potency. This degradation makes it paramount to prepare samples within a short, analytical window for accurate quantitation. The sample preparation procedure developed allows for at least 48 hours confirmed stability. Total psilocin was chosen as a mode of reporting to account for transformation of psilocybin to psilocin.



Table 2. Hydroxy-Tryptamines Content vs strain Chart

As shown in Table 2, all minor compounds are present in comparatively low concentrations. In some cases, such as with the Melmac Penis Envy, the “Total Psilocin” content is 180 times larger than the minor compound concentration.

Figure 1. Hydroxy-Tryptamine Content vs Strain Graph

Fig. 1 represents all tested hydroxy-tryptamines as a percent of dried weight.

Figure 2. Major Hydroxy-Tryptamine Content Vs Strain Graph

Fig. 2 represents a closer view on all tested major hydroxy-tryptamines as a percent of dried weight.

Figure 3. Minor Hydroxy-Tryptamine Content vs Strain Graph

Fig. 3 represents a closer view on all tested minor hydroxy-tryptamines as a percent of dried weight. Note that all these compounds are present in concentrations less than a tenth of a percent of total biomass.



The data clearly defines that different strains of psilocybe cultivars can have different chemical compositions of their major and minor active compounds. Furthermore, a sample derived from the same harvest at the same time can have large variances in hydroxy-tryptamine content, as shown with the set of “Avary Albinos”.


Future Developments

The world is becoming ever more open to the idea psychedelics. With continued research and development, a greater understanding of mushroom chemistry will not be too far off into the future.

Further research is still needed to provide an encompassing view on the chemical composition and attributes of psychedelics, including "magic mushrooms". This additional research should include a psycoactive panel on norpsilocin, a safety panel on mycotoxins, fungi/bacterial contamination panel, and many more.

Disclaimer: This article is for informational purposes only and is not intended to be a solicitation, offering or recommendation of any service, commodity, or derivative. We do not claim or own any of the information provided.

Connect with us and learn more!

bottom of page