Controlling Cannabinoids and Terpenes in Flower and Extract Our latest research results on drying, curing and extraction of cannabis to control the chemical composition of the plant and its extracts.

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Dr. Markus Roggen
VP Extraction
OutCo, San Diego
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Controlling Cannabinoids and
Terpenes in Flower and Extract

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• Background on Cannabis
• Understanding Dry and Cure
• Controls in Dry and Cure
• Optimizing Extraction
• Controls in Extraction
• Thanks / Product Plugs / Veneration
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Abstract

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Background

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Background

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Background

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So many molecules we should keep track of
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What is in Cannabis?
>140 Cannabinoids >200 Terpenes
~30 Flavonoids
Alcohols, Phenols
Aldehydes, Ketones
Alkaloids
Carbohydrates, Fatty Acids
Lactones

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What is Drying and Curing?
Dry Room Conditions: Prevent spoilage
• 60-74˚F
• 40-60% humidity
• Water Activity monitoring
• Aim for <0.65Aw by day 5-7
Curing Conditions: Even out moisture
• Different containers
• bucket, glass, box, bag, …
• Ambient conditions
• 40-50% humidity, 70˚F
• Length of time
• At least 1 week, up to 3 month

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• Moisture loss is driven by a difference in vapor pressure
• Balance between inhibiting mold growth and terpene loss
Water Activity during Dry and Cure
0.45
0.55
0.65
0.75
0.85
0.95
0 5 10 15 20 25 30
WaterActivity
Day
Water Activity
Aw of 0.65 represents the
value in which Botrytis cannot
continue to grow
Terpene Content (% dry weight)
Rapid Dry Slow Dry
Lemonade
Haze 1.42 1.82
Sour Tangie 1.69 2.2

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• Dominated by hydrolytic enzymes, e.g. starch to sugar
• Furans from browning positively influence flavor
• Degrading carotenes negatively influence flavor
Success depends on:
• Leaf maturity
• Temperature
• Humidity
• Water content of leaf 9
Chemistry of Curing
Figure ref.: DOI: 10.1080/00380768.2013.842884

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What gasses are given off in other plants?
How do these correlate with the curing of cannabis
“The Burp”?
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Cannabis vs. post-harvest research

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Plant Material:
• Mendo Breath
• Tangimal
Treatments:
• Control (no burp)
• OutCo Burp: for the first three days lids
will be removed for 15-20 minutes and
flower turned
• Industry Burp: lid opened daily for 1
hour for two weeks and flower turned
The Burp Project

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The Burp Project: CO2
Ambient CO2
Ambient CO2
Burp Burp
• CO2 builds over
time
• A burp releases
CO2 and instantly
encourages the
buildup of CO2
• Industry burp,
although longer,
immediately and
most drastically
increases CO2.
• Allowing in more
O2 for respiration?
Burp

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Burp
End of
work day
Buckets are not air tight
The Burp Project: CO2
Burp Burp

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Conclusions:
• The burp is not a release of moisture
• Humidity in a bucket follows the trend of the humidity in the room
• Humidity stays constant to the starting humidity
• Next step is to prove homogenization…
The Burp Project: RH
Ambient CO2
Burp Burp Burp

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Temperature in the bucket is directly related to the
temperature in the room.
The Burp Project
Burp Burp

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Treatments:
CONTROL: No intentional altering of ethylene levels
PLUS: Liquid ethephon at 200 ppm C2H4
MINUS: 2 Sachets containing 2.5 grams of EthylBloc™
Conclusions and questions:
Ethylene concentrations in post-harvest Cannabis flowers
are extremely high, and were consistent across all
treatment groups.
Is this a stress response due to wounding?
Does ethylene increase respiration?
Gasses during Cure
Ethylene

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Does anything happen to THC, CBD, …?
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Cannabinoids during Cure
15%
20%
25%
0 5 10 15 20 25 30 35
Cannabinoidbywt.
Days of Cure
Potency

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0
2
4
6
0 5 10 15 20 25 30 35
%decarboxylation
Days of Cure
THC/THCA
Does anything happen to THC, THCA, …?
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Cannabinoids during Cure

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Does anything happen to terpenes? Please!
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Terpenes during Cure
0%
1%
2%
3%
0 5 10 15 20 25 30 35
Terpenesbywt.
Days of Cure
Terpenes

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Maybe to individual terpenes?
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Terpenes during Cure
0
2
4
6
8
10
12
0
2
4
6
0 5 10 15 20 25
Terpenesbywt.
Days of cure
AutoCure Terpenes
Limonene Bisabolol Linalool Humulene bCP Myrcene

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Now we can start to optimize
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Terpenes during Cure
0%
100%
200%
300%
Change MAP Change VAC Change Control
Changeovertime
Terpenes in Cure
Limonene Myrcene Terpinolene Total

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Extraction: Removal of a substance from a
matrix. Needs a solvent, e.g. butane, CO2,
ethanol.
Separation: Mechanical force, not chemical
properties, dictates success, e.g. Rosin, Ice
Water Hash, Kief
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What is an Extract?

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What is an Extract?

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What is an Extract?

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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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Does milling destroy THC?
Optimizing your Extraction Process
0%
1%
2%
%decarb by Size
0
0.2
0.4
0.6
0.8
1
1.2
1.4
%wt. Terpenes by Size

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But it does give you higher yields!
Optimizing your Extraction Process
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Non-Ground Food Blender 2 mm 6 mm 10 mm
%Recovery
Cannabinoid Recovery by Size
75
80
85
90
95
Non-Ground Food Blender 2 mm 6 mm 10 mm
%Recovery
Terpene Recovery by Size

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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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Expt. No. Temp.
(˚C)
Pres. (psi)
A 34 1100
B 34 1900
C1 47 1500
C2 47 1500
D 60 1100
E 60 1900
Factor No. Factor Super - Super + Unit
1 Temperature 34 60 ˚C
2 Pressure 1100 1900 psi
Temperature (˚C)
Pressure (psi)
Yield THC (g)
34
60 1100
1900
47
1500
0
Executing the DoE
Critical Point: 304.25 K & 72.9 atm
Factor No. Factor Super - Super + Sub - Sub + Unit
1 Temperature 34 60 5.0 29 ˚C
2 Pressure 1100 1900 900.0 1900 psi
Expt. No. Temp.
(˚C)
Pres. (psi)
F 5.0 900.0
G 5.0 1900
H1 17 1400
H2 17 1400
I 29 900.0
J 29 1900

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A Taylor expansion describes the response surface.
Matrix algebra solves the Taylor expansion
Y=bX
b=(X’X)-1X’Y
a=bX 31
Calculating the Response Surface
y0+ ixi+ iixi
2+ ijxixj
i i i j

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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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Analyzing the DoE, THC Recovery (Sub)
a= - 7.75 xT + 126 xp + 59.3 xTxp

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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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Analyzing the DoE, %Terpene (Sub)
a= 11.8 xT - 16.6 xp - 12.5 xTxp

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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% conc. at 34% yield 63% conc. 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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Thanks
Nate Controls
AutoCure
Henkelman Vacuum
Systems

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Apeks Supercritical
• Andy Joseph, President
• Jeremy Sexton, Director of Engineering
Fritsch
• Barry Schubmehl, Application Consultant
• Leos Benes, Global Lab. Serv. Manager
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Thanks

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The Crew
Blake Grauerholz
Taylor Trah
Matt Zbiec
Chris Cortina
and team
Thank You

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Thank You!
See expanded results from our
experiments on IG:
@outcosd & @dr.justice_grows
And our blog:
http://outco.com/blog/

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Oversees all Extraction, Formulation and Production at
OutCo. jointly leads OutCo’s R&D
● PhD in Organic Chemistry, ETH Zürich, Switzerland
● Postdoc in Physical Organic Chemistry, The Scripps
Research Institute, San Diego
● Mentor, Board Member, Scientific Advisor to Cannabis
Startups, Journals and the NCIA
Public Presentations:
● Emerald Conference 2017
● CannaCon Seattle 2017
● ACS Conference 2017
● CannMed (Harvard Medical School) 2017
● Terpenes & Testing Magazine 2017
Dr. Markus Roggen
Vice President, Extraction
www.slideshare.net/MarkusRoggen