1. Don’t Be Neutral About
Psilocybin Testing
Problems with Current Analytical Methods and New
High-Throughput Method
Dr. Markus Roggen
Complex Biotech Discovery Ventures LTD
2. Introduction
Complex Biotech Discovery Ventures
CBDV is a research venture that seeks to add fundamental
scientific insight to the field of cannabis and mushroom
production.
We seek to support the cannabis and mushroom industries by
establishing a centralized hub in Vancouver, BC, for collaborative
research focused on:
• Process Design
• Process Optimization
• Process Analytics
• Formulation Research
3. Collaborative Research
CBDV collaborates with academic, industry and private
groups around the globe. Some highlights of those
collaborations are:
• University of British Columbia, Vancouver
• Loyalist College, Belleville
• Via Innovations by Dr. Monica Vialpando
• Veridient Science by Dr. Linda Klumpers
Fundamental Collaboration
4. Research Topics
• Chemometrics and data analytics for process control and
optimization
• Kinetic studies to understand mechanisms
• In-process analytics for process control
• Computational studies to understand mechanisms
• Process development, like crystallization
Fundamental Cannabis and Mushroom Chemistry
5. Research Topics
• Chemometrics and data analytics for process control and
optimization
• Kinetic studies to understand mechanisms
• In-process analytics for process control
• Computational studies to understand mechanisms
• Process development, like crystallization
Fundamental Cannabis and Mushroom Chemistry
7. Introduction
• Both religious and
recreational use
• Earliest evidence from
Sone Age rock art in
Africa and Europe and
particularly in pre-
Columbian Central
and South America.
• The active compound
are psilocybin, psilocin
and baeocystin, …
Guzmán G. (2008). "Hallucinogenic mushrooms in Mexico: An overview". Economic Botany. 62 (3): 404–412.
8. Introduction
• Psilocybin was first described in the 1950s by Albert
Hofmann
• Potential medical properties have been described
• Most research has been hindered or prohibited by its illegal
status and schedule 1 drug classification across the globe.
a)A. Hofmann, R. Heim, A. Brack, H. Kobel, Experientia 1958, 14, 107– 109; b) A. Hofmann, R. Heim, A. Brack, H. Kobel, A. Frey, H. Ott, T. Petrzilka,
F. Troxler, Helv. Chim. Acta 1959, 42, 1557–1572.
Geiger, H. A., Wurst, M. G., Daniels, R. N. (2018). DARK Classics in chemical neuroscience: Psilocybin. ACS Chemical Neuroscience, 9, 2438-2447.
9. Introduction
• Psilocybin is broken down by the liver to psilocin
• Psilocin is responsible for the psychedelic effects.
• Psilocybin mushrooms are not been known to cause physical or
psychological dependence/addiction.
• Mushrooms are predominantly ingested.
• Around 20 minutes onset time and effect can last up to 6 hours.
• Side effects including nausea, vomiting, euphoria, muscle weakness or
relaxation, drowsiness, and lack of coordination may occur.
10. Compounds of Interest
• Psilocybin is the most
abundant compound in
psychedelic mushrooms,
with concentrations of 0.5-
2%.
• Concentrations of
psilocybin and other
tryptamine derivates vary
widely between mushroom
species and grow
conditions.
11. Psilocybin Mushroom Regulations in
Canada
Controlled Drugs and Substances Act (S.C. 1996, c.
19)
Exemption by Minister
56 (1) The Minister may, on any terms and conditions that
the Minister considers necessary, exempt from the
application of all or any of the provisions of this Act or the
regulations any person or class of persons or any
controlled substance or precursor or any class of either of
them if, in the opinion of the Minister, the exemption is
necessary for a medical or scientific purpose or is
otherwise in the public interest.
12. Psilocybin Mushroom Regulations in
Canada
Email from Health Canada:
As you are aware, activities that are considered production
cannot be considered for a subsection 56(1) exemption or an
authorization under Part J of the Food and Drugs Regulations
(FDR).
Production is defined in the Controlled Drugs and Substances
Act (CDSA) as follows:
“produce” “production” means, in respect of a substance
included in any of Schedules I to IV, to obtain the substance by
any method or process including:
(a) manufacturing, synthesizing or using any means of altering
the chemical or physical properties of the substance, or
(b) cultivating, propagating or harvesting the substance or any
living thing from which the substance may be extracted or
otherwise obtained,
13. The Mission
• Develop an HPLC method to quantify psilocybin
• Suitable for high-throughput testing at cultivation site
• Easy to operate for non-chemists
• To be developed with pure psilocybin, not mushrooms
14. Current State
Column Mobile A Mobile B Flow, Mix (B) Total Time
Poroshel 120 SB-C18,
100mm
0.1% Formic Acid,
H2O
ACN 0.2 mL/min, 5-20% 12 min
Synergi 4 mm Max-
RP C12, 150mm
10 mM ammonium
formate, H2O
MeOH 0.5 mL/min, 95% 5 min
Symmetry C18,
150mm
10 mM ammonium
formate, H2O
ACN 0.2 mL/min, 5% ???
Phenomenex Gemini
C18, 150mm
0.2% Formic Acid,
H2O
0.2% Formic Acid,
ACN
0.2 mL/min, 5-95% 30 min
Mightysil RP-18 GP,
100mm
50mM ammonium
acetate, H2O
ACN 0.15 mL/min, 27% ???
Acquity UHPLC HSS
C18, 150mm
5 mM ammonium
formate, H2O
0.3% Formic Acid,
ACN
?, 5-90% 16.50 min
15. Method Development
• Our tools:
• Agilent 1220 Infinity II / Chemstation
• Agilent Poroshell 120 EC-C18 2.7µm 3.0x50mm (50C)
• Reference Material: Psilocybin (Cerilliant P-097-1mL)
• Test Material: Psilocybin by Psygen Labs Inc.
• Injection Volume: 15µL
22. Method Development
Run # Mobile Phases Flow
(ml/min)
Time
(min)
Pump A Pump B
10H-59 A: 10mM NH4HCO2, H2O
B: ACN
1.0 5 3% B hold for 20s then ramp to
95% B in 1min, hold till 2min, then
bring back to 5% B by 4min and
hold for 1min
10H-66 A: 0.025mM NH4HCO2, H2O
B: ACN
1.0 3.5 3% B hold for 20s then ramp to
95% B in 1min, hold till 2min, then
bring back to 5% B by 3min and
hold for 0.5min
26. Summary
• 3.5 min method for psilocybin quantification
• Choose your solvent carefully: ACN works better than MeOH
• Choose your acid carefully: NH4HCO2 over HCOOH
• Choose your acidity carefully: Do not degrade to psilocin
27. Where Next?
• Sample preparation / extraction of mushrooms
• Wildly different sample prep methods
• Other solvents explored: ethyl acetate, acetonitrile, chloroform, and
diethyl ether
Solvent Time Temperature
acetic acid, pH4 1 hour 70˚C
acetic acid minutes r.t.
methanol 15 min r.t.
methanol overnight r.t.
methanol 48 hours r.t.
Anastos, N., Lewis, S. W., Barnett, N. W., and Sims, D. N. (2006). Journal of Forensic Science, 51, 45-51.
doi:10.1111/j.1556-4029.2005.00033.x
28. Expertise
CEO: Dr. Markus Roggen
Dr. Roggen has been actively involved in the cannabis industry for over 5 years in executive
positions overseeing production, R&D and process optimization for multiple producers. Dr.
Roggen is also a trusted advisor and mentor for multiple startups, startup accelerators and
organizations.
Co-Founder: Prof. Glenn Sammis
Prof. Sammis is an Associate Professor in the Chemistry Department at the University of British
Columbia. He has built an internationally recognized research group working on the
development of novel synthetic methods for the preparation of natural products and
pharmaceuticals.
CBDV Team
Our team covers a wide range of expertise,
including analytical chemistry, process
chemistry, engineering physics, data science
and statistics.
