Presenting our research on molecular changes in flower and extract. A visual guide through the production pipeline. With added data sets for better insight.

1. Controlling Terpenes and
Cannabinoids in Flower and
Extract
Cannabis Curing & Extraction Research
by
Dr. Markus Roggen
Complex Biotech Discovery Ventures

2. Agenda
• Introduction
• Data Visualization
• Different Strains = Different Chemovars
• Drying & Storage
• Drying Conditions
• Gasses for Storage
• Extraction
• Particle Size
• DoE
• Post-Processing
• Decarboxylation

3. Cannabis Processing Pipeline
• Drying and curing
• Extraction
• Post processing
CBDA
CBD THCA
THC

4. Cannabis Processing Pipeline
• Drying and curing
• Extraction
• Post processing
Myrcene
a-Pinene
b-Pinene
Limonene
b-Caryophyllene

5. Cannabis Flower and Medicine
• Cannabinoid ratios and concentrations show little variability
• Terpene ratios and concentrations show strong variability
Cannabinoid Chemotaxonomy: DOI:10.1038/s41598-018-31120-2
Terpene Chemovars: DOI: 10.1089/can.2016.0040
Gorilla Glue Sour Diesel
Cookies
Sour Tangie
Lemonade Haze

6. What is in the Plant
6
>140 Cannabinoids
>200 Terpenes
~30 Flavonoids
Alcohols, Phenols
Aldehydes, Ketones
Alkaloids
Carbohydrates, Fatty Acids
Lactones

7. Drying and Curing
• Drying room conditions influence on terpenes
30%
Drying
Conditions
Slower
Faster
Air
Vacuum
N2
Ar
Long Term
Storage
20%
CO2
Storage
Atmosphere

8. 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
• Moisture loss is driven by a difference in vapor pressure
• Balance between inhibiting mold growth and terpene loss

9. Drying and Curing
• Drying room conditions influence on terpenes
• Storage time influences on cannabinoids
• Drying room conditions influence on terpenes
• Storage time influences on cannabinoids
• Curing Gases influence on terpene ratios
30%
Drying
Conditions
Slower
Faster
Air
Vacuum
N2
Ar
Long Term
Storage
20%
CO2
Storage
Atmosphere

10. Drying and Curing
• Drying room conditions influence on terpenes
• Storage time influences on cannabinoids
• Curing Gases influence on terpene ratios
30%
Drying
Conditions
Slower
Faster
Air
Vacuum
N2
Ar
Long Term
Storage
20%
CO2
Storage
Atmosphere

11. Storing under Modified Atmosphere
18.00
20.00
22.00
24.00
26.00
28.00
30.00
0 2 4 6 8 10 12 14 16
w%
Weeks
Cannabinoids
Air Vacuum N2 Ar CO2
0
0.5
1
1.5
2
2.5
0 2 4 6 8 10 12 14 16 18
w%
Weeks
Total Terpens
Air Vacuum N2 Ar CO2

12. Extraction
• Particle size effects yield and precision
2mm
Milling
Method
Whole
Flower
Food
Blender
Extraction
Method
Terpenes
THC
THCA
CBDA

13. Extraction
• Particle size effects yield and precision
2mm
Terpenes
THC
THCA
CBDA
Whole
Flower
Food
Blender
Milling Method

14. Effect of Milling
Does milling destroy THC?
0%
1%
2%
%decarb by Size
0
0.2
0.4
0.6
0.8
1
1.2
1.4
%wt. Terpenes by Size

15. Effect of Milling
But it does give you higher yields!
20
22
24
26
28
30
32
34
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

16. Extraction Parameters
• 3 kg of plant matter
• Terpene Fraction (F1): 1100 psi, 34˚C, 5 h
• Cannabinoid Fraction (F2): 1900 psi, 60˚C, 12 h

17. Extraction Precision Terpenes
30
35
40
45
50
55
Non-Ground Food Blender 2 mm 6 mm 10 mm
%conc.
Terpene Concentration F1
Higher is better, Food Blender wins
0
1
2
3
4
5
Non-Ground Food Blender 2 mm 6 mm 10 mm
%conc.
Terpene Concentration F2
Lower is better, 2mm wins
• 3 kg of plant matter
• Terpene Fraction (F1): 1100 psi, 34˚C, 5 h
• Cannabinoid Fraction (F2): 1900 psi, 60˚C, 12 h

18. 0
2
4
6
Non-Ground Food Blender 2 mm 6 mm 10 mm
%conc.
Cannabinoid Concentration F1
Lower is better, 10mm wins
55
60
65
70
75
Non-Ground Food Blender 2 mm 6 mm 10 mm
%conc.
Cannabinoid Concentration F2
Higher is better, 2mm wins
Extraction Precision Cannabinoids
• 3 kg of plant matter
• Terpene Fraction (F1): 1100 psi, 34˚C, 5 h
• Cannabinoid Fraction (F2): 1900 psi, 60˚C, 12 h

19. Extraction
• Particle size effects yield and precision
• SFE can extract terpene fractions
• SFE can extract either THCA or THC
2mm
Whole
Flower
Food
Blender
Terpenes
THC
THCA
Milling Method
Extraction Method

20. Extraction
• Particle size effects yield and precision
• SFE can extract terpene fractions
• SFE can extract either THCA or THC
• SFE can change the THC to CBD ratio
2mm
Milling Method
Extraction Method
Terpenes
THC
THCA
CBDA
Whole
Flower
Food
Blender

21. Analyzing CO2 SFE for THC Recovery (Super)
• a= 14.9 xT + 123 xp + 18.4 xTxp

22. Analyzing CO2 SFE for THC Recovery (Sub)
• a= - 7.75 xT + 126 xp + 59.3 xTxp

23. Analyzing CO2 SFE for %Terpene (Super)
• a= - 14.9 xT - 11.7 xp + 16.8 xTxp

24. Analyzing CO2 SFE for %Terpene (Sub)
• a= 11.8 xT - 16.6 xp - 12.5 xTxp

25. Comparing Sub- and Supercritical
• SFE, 900.0 – 1900 psi, 5.0 – 60 ˚C, 22h max
Response Sub Super
THC Recovery 60% 80%
Terpene Fraction 52% at 34% yield 63% at 82% yield

26. Lessons learned in SFE
• THC extracts faster than THCA
• THC to THCA ratio adjusted by SFE
% Decarboxylation S.M. 7h 22h
Exp. B 8.0% 25% 10%
Exp. J 14% 25% 14%

27. Lessons learned in SFE
• CBD extracts faster than THC
• CBD to THC ratio adjusted by 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

28. Lessons learned in SFE
• Enrichment in minor cannabinoids
• CBG enrichment by 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

29. Lessons learned in SFE
• Changing terpene profiles in SFE
• Sativa: rich in monoterpenes
• Indica: rich in sesquiterpenes
Hazekamp, et al.; Cannabis and Cannabinoid Research 2016, 202

30. Visual Guide to Extracts
30

31. Post-Processing
• Decarboxylation under CO2 vs air
Air
CO2
Decarboxylation
Method
Distillation
Method

32. IR monitoring of decarboxylation
IR as quick and cheap method to monitor decarboxylation

33. IR monitoring of decarboxylation
IR as quick and cheap method to monitor decarboxylation
-10
0
10
20
30
40
50
60
0 10 20 30 40 50 60 70 80 90
Concentration(%)
Elapsed Time (Minutes)
Reference THC Predicted THC Reference THCA Predicted THCA
15
25
35
45
55
65
75
85
15 35 55 75
PredictedTHC(%)
Reference THC (%)
THC
-5
0
5
10
15
20
25
30
35
40
0 10 20 30 40
PredictedTHCA(%)
Reference THCA (%)
THCA

34. Post-Processing
• Decarboxylation under CO2 vs air
• Distillation to purify THC
Air
CO2
Decarboxylation
Method
Distillation
Method

35. Final Product: The Blind Side
• Patient: Male, mid-30, 8 seizures per day
• CBD/THC 3:1 tincture, 40 mg/mL, no terpenes, MCT oil:
• seizure-free for 9 weeks
• Next batch of 3:1 oil:
• seizures re-start immediately
• THC came from two different cultivars: Diamond OG and Lemonade Haze
CBD THC THCA CBDA CBN CBG THCV CBGA CBC
Diamond OG 300 100 15.3 0.00 0.66 1.76 0.73 0.99 0.00
Lemonade Haze 300 100 15.6 0.00 0.45 1.62 1.19 0.00 1.06

36. Research Collaborations
cbdvl.com
slideshare.net/MarkusRoggen email: markus@cbdvl.com
Complex Biotech Discovery Ventures