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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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Where do I come
from?

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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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How did I get into
drugs?

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How did I get into
drugs?
Serendipity!

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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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Cannabis Science

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• Biology: Plant shape, strains, cultivation
• Biochemistry: Active Molecules, biosynthesis
• Organic Chemistry: Synthesis
• Medicinal Chemistry: Receptors, synthetic cannabinoids
• Analytical Chemistry: Testing
• Physical Chemistry: Melting Point, shatter
• Food Chemistry: Consumption methods
• Chemical Engineering: DoE
• Mechanical Engineering: Rosin
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Cannabis Science

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CannaBio

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The Cannabis Plant

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The Cannabis Plant

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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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CannaMolecules

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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
Biosynthesis: Eur. J. Biochem. 268, 1596 (2001)

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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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• Indica vs. Sativa
• Strain and Cultivars
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Cannabis strains

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• Let’s think about Chemovars
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Cannabis strains
DOI: 10.1089/can.2016.0017
DOI: 10.1089/can.2016.0040

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Cannabis strains

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CannMed

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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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What is all the fuss about
Cell 167, 750, October 20, 2016

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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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Synthetic Cannabinoids

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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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CannOrganic

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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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CannAnal

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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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CannaPhys

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What extracts are out there?
Kief Dry Sift
BHO
Glass
ShatterCrumble
Ethanol CO2 Oil
Terp Sauce

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Munchies

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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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Pick your poison:
Buds
Oil, Shatter, Glass, Wax (budder, crumble, honeycomb)
Sublingual, Cream, Tablet, Drink, Edible
Product Variations

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What is an Extract?

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CannExtract

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Extraction Methods
Butane (Hydrocarbons) Alcohol

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Extraction Methods
Butane (Hydrocarbons) Alcohol (Ethanol) Carbon Dioxide (SFE)

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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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For what:
• Recovery Rec
Optimizing your Extraction Process
With what:
• Particle Size PS
Experiment PS (mm) Rec (g)
1 6 215
2 4 230
3 2 235
4 1 253
210
220
230
240
250
260
0 1 2 3 4 5 6 7
YpR(g)
PS (mm)
Optimizing Particle Size

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Optimizing your Extraction Process
0
0.05
0.1
0.15
0.2
0.25
0.3
0 2 4 6 8 10 12 14 16 18
%Weight
Terpene
Effect of Milling on Terpene Cocentrations
Food Blender 0.5mm 1mm 2mm 4mm 6mm 10mm 1mm (700rpm) 10mm (700rpm) Reference

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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
47
1500
0
Executing the DoE
Temperature (˚C)
Pressure (psi)
Yield THC (g)
34
60 1100
1900
47
1500
0
400

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y0+ 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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a= 14.9 xT + 123 xp + 18.4 xTxp
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Analyzing the DoE, THC Recovery (Super)

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a= - 14.9 xT - 11.7 xp + 16.8 xTxp
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Analyzing the DoE, %Terpene (Super)

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Visual Guide to Extracts

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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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CannEngineering

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Extraction Methods (Mechanical Manipulation)
Kief Tumbling Rosin Press Water Extraction

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Other Separation Methods
Ice Water Hash
• ~80% cannabinoid concentration
• ~25% recovery achieved
• Little effect on molecular makeup
150 micron 25 micron

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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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X
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Other Separation Methods

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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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QA: Chris Cortina
Formulation:
Taylor Trah
Extraction:
Blake Grauerholz
Production:
Matt Zbiec
mroggen@outco.com

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Why it’s different
than biotech?

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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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How to get into the
cannabis field?

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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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What to look out for?

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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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Why do it?

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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!