Lecture at the chemistry department of Imperial College, London. A brought overview and specific deep dive into the chemical aspects of cannabis production.
Cannabis Chemistry Industrial Applications of Chemistry & Innovation and Entrepreneurship BONUS lecture
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Dr. Markus Roggen
VP Extraction
OutCo, San Diego
Cannabis Chemistry
Industrial Applications of Chemistry & Innovation and
Entrepreneurship BONUS lecture
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• My gateway drugs
• My descent into drugs
• My daily drug use
• My other habits
• Why you should join in
• How to get your fix
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Abstract
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• Born in Germany
• M/Sci Chemistry, Imperial College London, UK
• PhD Organic Chemistry, ETH Zürich, CH
• PostDoc PhyOrg Chemistry, TSRI, USA
• Laboratory Director, DaVinci Laboratory, CA, USA
• VP Extraction, OutCo, CA, USA
• Climbing Guide, Europe
• Triathlete, USA
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Where do I come from?
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• Germany: Process Optimization
• Imperial College: Design of Experiment
• ETH Zürich: Tolerance to Pain and Suffering
• Scripps Research: Life Perspective
• DaVinci Laboratory: Analytical Chemistry
• OutCo: Research Platform
• Climbing: Teaching, Leadership
• Triathlon: Connections
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Where do I come from?
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• Four options for chemists:
• Academia
• Industry
• Startup
• Traitor
• Three ways to get a job:
• Be amazing
• Be pushy
• Know the right people
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How did I get here?
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• Cannabis Industry is in its infancy
• Great scarcity of scientists
• Potential for large impact and fast rise
• More than just science
• Sin industries pay better
• Prohibition did not work
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Why drugs?
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Outdoor:
•Low facility and operations costs
•Only one harvest per year
Indoor:
•Multiple harvests per year
•High facility and operations costs
Greenhouse:
•Uses sunlight
•High facility costs
Grow Environment
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0.45
0.55
0.65
0.75
0.85
0.95
0 5 10 15 20 25 30
WaterActivity
Day of cure
Water Activity
0
2
4
6
8
10
12
0
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
0 5 10 15 20 25
Day of cure
Terpenes
Limonene Bisabolol Linalool
bCP Myrcene Humulene
• Dry to preserve plant
• ”Cure” plants to develop flavor and make ”smoke-able”
Post Processing
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• Trim to maximize aesthetics and smoke-ability of buds
• 1.5 lbs/day·person (we produce 180 lbs per month)
Post Processing
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So many molecules we should keep track of
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What is in the plant
>140 Cannabinoids >200 Terpenes
~30 Flavonoids
Alcohols, Phenols
Aldehydes, Ketones
Alkaloids
Carbohydrates, Fatty Acids
Lactones
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What is in the plant
THC
• The reason we are all here
• Psychoactive
Decarboxylation
THC Acid (THCA)
• Naturally occurring
• Not Psychoactive
CBN
• Not naturally occurring
• Low Psychoactivity
Oxidation
CBD
• Non-psychoactive
• Assumed beneficial health properties
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What is all the fuss about
Anandamide
From Sanskrit ”ananda” for Joy, Bliss
2-Arachidonoylglycerol
Present in relatively high levels
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• Chronic pain and spasticity: moderate-quality evidence
• Nausea and vomiting due to chemotherapy: low-quality
• Weight gain in HIV infection: low-quality evidence
• Sleep disorders: low-quality evidence
• Tourette syndrome: low-quality evidence
• 79 trials, 6462 participants from >23000 publications
• 4 trials were judged at low risk of bias
Medical Effects of Cannabinoids
JAMA June 23/30, 2015, 313, Number 24 2456
DOI: 10.17226/24625
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• National Institute on Drug Abuse (NIDA) is the only legal
producer of Cannabis for use in US research studies
Medical Studies on Cannabis
DOI: 10.1038/srep46528
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• Over 50,000 death/year in USA due to opioids
• States with legal cannabis have a 25% lower mortality rate
Medical Studies on Cannabis
doi:
10.1001/jamainternmed.2014.4005
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• THC: Partial agonist to CB1 & CB2
• CBD: Antagonist for CB1 & CB2
• 2-AG: Full agonist for CB1
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Terms and Conditions
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• Not synthesized cannabinoid, but synthesized for
endocannabinoid receptor interaction
• John William Hoffman: Full agonist, 5x greater than
THC, death reported
• Alexandros Makriyannis: 100x greater than THC
• Raphael Mechoulam: Initial discovery of
endocannabinoids
• Pfizer: Synthesized in the 1970s and 1980s
• Alan C. Spivey: Solid-Phase Synthesis of
Rimonabant Analogs
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Synthetic Cannabinoids
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Stereodivergent Total Synthesis of THC
Angew. Chem. Int. Ed. 2014, 53, 13898
D9
-Tetrahydrocannabinols**
on Krautwald, David Sarlah, and
D9
-Tetrahydrocannabinols**
on Krautwald, David Sarlah, and
• Allylic Alcohol and Enal
• Two chiral catalysts:
• Ir/(P,olefin) complex
• Secondary amine
• All possible stereoisomers of
γ,δ-unsaturated aldehyde
product
• Ring-closing metathesis
Grubbs 2cd
• Pinnick oxidation/Methylation
• Methylation to 3rd-Alcohol
• Deprotection
• Aryl etherification
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Hemp and CBD Isolate
EIHA
• Fiber: light weight papers, insulation material and bio
composites (Bike parts: Dr. Kunal Masania, IC MechEng)
• Shivs: animal bedding and construction
• Hemp seeds: hemp seed oil, human food and animal feed
• Isolation of CBD
• 33,000 ha cultivated in 2016 in Europe
• Bulk synthesis of CBD please!
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Some quantitative
methods:
• Taste testing
• TLC (Quantitative?)
• IR (Quantitative?)
• GC-FID (Decarboxylation)
• LC-UV/Vis (Overlap)
• LC-MS/MS (Expensive)
• NMR (still working on it)
Problems:
• 10 Ref. Standards
• No official methods
• Little understanding by
regulatory body
• Little interest by industry
• No pesticide guidelines
• High solvent limits
• Edibles are difficult
matrix to prepare
Thoughts on Testing
US herbal pharmacopoeia: cannabis monography
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Inhalation
Fast acting, high peak concentrations of cannabinoids
Danger of lipoid pneumonia
Ingestion
Slower, gradual uptake of cannabinoids
THC is metabolized to 11-OH-THC in the liver
11-OH-THC is more potent and active than THC
Danger of overdosing due to delayed effect
Topical
Targeted application for localized problems
Will not fail drug test
Delivery
DOI: 10.1089/can.2015.0004
DOI: 10.1089/can.2015.0012
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• Production Yield
• Production Costs
• Extract Constitution/Quality
• Legal Aspects
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What is the best Extraction System?
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For what:
• Yield per Run YpR
• Yield per Week YpW
• Cannabinoid Conc. CC
• Terpene Content TC
• Recovery Rec
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Optimizing your Extraction (SFE)
With what:
• Temperature T
• Pressure P
• Time t
• Flow Rate FR
• Particle Size PS
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For what:
• Yield per Run YpR
• Yield per Week YpW
• Cannabinoid Conc. CC
• Terpene Content TC
• Recovery Rec
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Optimizing your Extraction Process
With what:
• Temperature T
• Pressure P
• Time t
• Flow Rate FR
• Particle Size PS
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For what:
• Recovery Rec
Optimizing your Extraction Process
With what:
• Time t
0
100
200
300
400
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YieldTHC(g)
Time (t)
Optimizing Run TimeExperiment t (h) Rec (g)
1 4 53.0
2 8 103
3 18 263
4 34 367
Raw n/a 500
Mass Transfer
Diffusion
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For what:
• Yield per Run YpR
• Yield per Week YpW
• Cannabinoid Conc. CC
• Terpene Content TC
• Recovery Rec
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Optimizing your Extraction Process
With what:
• Temperature T
• Pressure P
• Time t
• Flow Rate FR
• Particle Size PS
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Design of Experiment to the Rescue
• Test multiple factors simultaneously
• Few test for whole space response surface
• Higher order effects of factors
• Fast and efficient
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Designing the first DoE
1. Defining the problem
• Extracting Cannabis with a 20L, 2000psi SFE
2. Determine the response
• Yield THC
• Percentage Terpenes
3. Determine the factors
• Temperature and Pressure
4. Additional things to keep in mind
• Comparison
• Replication
• Blocking
• Orthogonality
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Critical Point: 304.25 K & 72.9 atm
Factor no. Factors lower level upper level Unit
1 Temperature 34 60 °C
2 Pressure 1100 1900 psi
Expt. no. Temp (˚C) Pressure (psi)
A 34 1100
B 34 1900
C1 47 1500
C2 47 1500
D 60 1100
E 60 1900
Temperature (˚C)
Pressure (psi)
Yield THC (g)
34
60 1100
1900
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1500
0
Executing the DoE
Temperature (˚C)
Pressure (psi)
Yield THC (g)
34
60 1100
1900
47
1500
0
400
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y0+ ixi+ iixi
2+ ijxixj
i
i i j
Calculating response surface of DoE
A Taylor expansion describes the response surface.
Matrix algebra solves the Taylor expansion.
Y=bX
b=(X’X)-1X’Y
a=bX
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SFE, 900.0 – 1900 psi, 5.0 – 60 ˚C, 22h max
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Compare Sub- and Supercritical
Response Sub Super
THC Recovery 60% 80%
Terpene Fraction 52% at 34% yield 63% at 82% yield
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• THC extracts faster than THCA
• THC to THCA ratio adjusted by SFE
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Lessons learned in SFE
% Decarboxylation S.M. 7h 22h
Exp. B 8.0% 25% 10%
Exp. J 14% 25% 14%
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• CBD extracts faster than THC
• CBD to THC ratio adjusted by SFE
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Lessons learned in SFE
CBD/THC S.M. 7h 22h
Exp. A 4.1E-03 1.3E-02 5.0E-03
Exp. J 1.7E-04 1.3E-03 3.1E-03
Bonus: Decarboxylation of
CBDA slower than for THCA
J. of Supercritical Fluids, 2010, 603
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• Enrichment in minor cannabinoids
• CBG enrichment by SFE
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Lessons learned in SFE
CBG/total Canna. S.M. 7h 22h
Exp. B 2.9% 6.% 2.9%
Exp. J 3.0% 13% 4.9%
J. of Supercritical Fluids, 2010, 603
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Changing terpene profiles in SFE
• Sativa: rich in monoterpenes
• Indica: rich in sesquiterpenes
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Lessons learned in SFE
Hazekamp, et al.; Cannabis and Cannabinoid Research 2016, 202
7h 22h
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Rosin Press
• Optimized for yield and quality by DoE
• ~72% recovery achieved
• Little effect on molecular makeup
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Other Separation Methods
Rosin Yield Rosin Quality
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Further Reading
• Roggen, M.; Clear vs. Pure: How Fallacies and Ignorance
of Extraction Misrepresent the Cannabis Flower. Cannabis
Industry Journal August 3rd, Online Editorial
• Cortina, C.; Roggen, M.; Comply! Terpene and Testing
Aug./Sept. 2017, 36-41
• Grauerholz, B.; Roggen, M.; Journey Down the Production
Pipeline. Terpene and Testing June/July 2017, 35-39
• Roggen, M.; Best Before Dates. Terpene and Testing
Online Editorial
• Roggen, M.; How to optimize your extraction operation. A
case study in supercritical CO2 extraction. Terpene and
Testing May/June 2017, 35-40
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• Cannabis sativa: The Plant of the Thousand and
One Molecules (doi: 10.3389/fpls.2016.00019)
• Changes in Cannabis Potency Over the Last 2
Decades (1995–2014): Analysis of Current Data in
the United States
(doi:10.1016/j.biopsych.2016.01.004)
• American Herbal Pharmacopoeia, Cannabis
Inflorescence, 2013
Further Reading
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• Mature industries have set career paths and
hierarchies
• All your coworkers are PhDs from the same
universities
• There is literature about what you have to do
• People have no idea what you do, but they
admire it
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Why it’s different than biotech?
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• Conversion between THC and CBD is not an
oxidation
• No testing regulation for CA, yet
• Solvent limits are ridiculously high in CO (e.g.
butane: 5000ppm, xylenes: 2170ppm)
• Indica vs. Sativa
• Organic, Artists
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Thoughts on Cannabis vs. Science
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• Better not to know the “right” people
• Join an “established” company
• Start your own thing
• Come from a position of strength
• Don’t beg
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How to get into the cannabis field?
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• Most companies are expanding
• Bring your outside expertise
• Location, location, location!
• Company culture is even more important
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Established cannabis companies
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• What is your idea?
• For ancillary consider Canopy San Diego
• For hands-on research: Talk to me!
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Start your own
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• Licenses
• Legality
• Background
• What is your / their risk limit?
• Company culture
• Look past the window-dressing
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What to look out for?
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• Startups are stressful
• I had to get fingerprinted for my job
• I change clothes and backpack when I travel
• No job security in startups
• Cannabis is schedule 1, there is Jeff Sessions!
• Startups are glorious
• I work with illegal drugs!
• Startups have great perks
• We work in warehouses outside of the city!
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Don’t!
Editor's Notes
NIDA pot is has less variety and lower concentrations than state-legal offerings
Low supply of NIDA material
Limited insights into the chemical, biological and pharmacological properties, and medical potential of cannabis
2kg plant matter, 6 hours run time.
2kg plant matter, 6 hours run time.
Let’s start designing this DoE
Defining the problem
Extracting Cannabis with a 20L, 2000psi SFE
Determine the response
Yield THC
Percentage Terpenes
Determine the factors
Temperature and Pressure
Constant: Time, flow rate,
Additional things to keep in mind
Comparison: Ensure that every experiment is done on the same conditions, e.g. particle size, extract weight, extract type, operator
Replication: Rerun experiments to evaluate variance and standard deviation
Blocking: Do the experiments in one sweep to mitigate unexpected trouble
Orthogonality: Chose factors that complement each other, not overlay, like time and flow rate
Considering all these points, we arrive at a design with the following limits: Temperature 34 to 60˚C and pressure 1100 to 1900 psi. Please note the choice of units, both metric and imperial. Take it as a stand in for the current state of cannabis: Still torn between worlds.
To cover the whole space of possible pressure and temperature combinations we chose a star design. We replicated the central point conditions.
The Taylor expansion includes second order behavior, namely the correlation between temperature and pressure and their influence on the system. All factors are paired with a coefficient ß to scale their influence on the system.
Y is the result (response) matrix, X is the parameter (factor) matrix, and b is the coefficient matrix.
