Industrial hemp: ‘an old-new’

source of functional ingredients

for nutraceuticals and foods

Prof. Petras Rimantas Venskutonis

Department of Food Science and Technology,

Kaunas University of Technology, Lithuania

UAB Endobiotech: Scientific Consultant1

Department of Food Science and Technology

• Established 1950

• Unique in Lithuania granting BSc, MSc and PhD degrees

• Research group: Functional Food Materials (Plant bioactive

compounds and thermal

reactions in foods)

2

Content

• Introduction: a litle bit of history

• Main uses of hemp

– textile

– seed oil

– proteins

• Main focus – phyto-cannabinoids

• Legal issues

• Extraction/fractionation/purification/analytical

3

Intro: condensed history of hemp

Source: http://www.marketwatch.com/story/hemp-is-getting-hotter-with-help-and-harm-from-marijuana-2016-11-08

When Bayer introduced aspirin in 1899, Cannabis was America’s number one painkiller. Until Cannabis prohibition began in 1937, the US Pharmacopoeia listed Cannabis as the primary medicine for over 100 diseases. 4

Source: https://www.cannarecruiter.com/industrial-hemp-jobs/

Uses: over 25,000 useful products

5

Source: http://www.nkytribune.com/2015/02/bardstown-company-to-sell-industrial-hemp-products-through-vending-machines-in-select-ky-cities/

Modern uses for the hemp plant

6

Source: http://www.coloradohemped.org/

Uses: textile – no legal problems

7

http://www.hemp-technologies.com/page29/page29.html

Uses: seeds for foods and supplements

8

Uses: composition of seeds (in 100 g)Characteristic Value Charact. Value Charact. Value

Energy 553 kcal Maltose 0.07 g Vitamin C 0.5 mg

Proteins 22.72 g Ca 70 mg Thiamine 1.275 mg

Fat 32.12 g Fe 7.95 mg Riboflavin 0.285 mg

Minerals 6.06 g Mg 700 mg Niacin 9.20 mg

Carbohydrates 8.67 g P 1650 mg Vitamin B6 0.60 mg

Dietary fibre 4.0 g K 1200 mg Folates 110 mg

Total sugars 0.85 g Na 5.0 mg Vitamin 1 mg

Glucose 0.2 g Zn 9.90 mg β-Carotene 7 mg

Fructose 0.31 g Cu 1.60 mg Vitamin E 0.80 mg

Lactose 0.07 g Mn 7.60 mg

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Seed oil: fatty acid composition Fatty acid Pressed Soxhlet SFE CO2Palmitic (C16:0) 6.27 7.11 6.55

Margarinic (C17:0) 0.18 0.12 0.11

Stearic (C18:0) 2.35 2.45 2.39

Oleic (C18:1n9c) 9.51 9.99 9.97

Linoleic (C18:2n6c) 53.54 54.85 54.92Arachidic (C20:0) 0.76 0.80 0.82

γγγγ-Linolenic (C18:3n6) 4.11 4.08 4.05cis-11-Eicosanoic (C20:1) 0.42 0.46 0.41

Linolenic (C18:3n3) 18.00 18.07 18.49cis-11,14-Eicosantrienoic (C20:2) 1.45 1.36 1.45

Behenic (C22:0) 0.28 0.29 0.27

Erucic (C22:1n9) 0.14 – –

Lignoceric (C24:0) 0.10 0.07 0.09

Excellent nutritional ratio of PUFA C18:2/C18:3 ≅≅≅≅ 3:1 10

PUFA in seed oil: benefits vs problems

Acknowledgements: Kalsec for donating antioxidants 11

Seed oil: pressing vs extraction

Source:

http://www.mnhempfarms.com/products/w

holesale-cold-pressed-hemp-oil

Press-cake

still contains

lipids

Aprox.

10 % lipids

Protein

enriched

powder,

approx. 50 %

Milling, packaging

12

Uses: seed proteinsS

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Source:

https://montereybayholistic.wordpress.com/2014/08/23/

hemp-seed-oil/

13

Fractionation and purification of proteins

ST – standard

1 – ‘albumins’

2 – ‘globulins’

3 – ‘prolamins’

4, 5 – ‘gluteins’

Electophoretic fractionation of hemp protein fractions obtained

by Osborn procedure (originally developed for wheat proteins)

14

Amino acid composition in hydrolysates, µmol/L

15

Protein properties

16

http://hempedification.blogspot.lt/2015/02/

Uses: herb phyto-cannabinnoidsM

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17

No. of publications in Clavirate Analytics WOS

with keywords ‘Cannabis sativa+cannabidiol’

18

Processing of leaves/flowering parts

Extraction

Conventional:

organic solvents

(ethanol)

High pressure:

supercritical CO2,

pressurized liquids

Drying

Microwave,

ultrasound assisted,

ionic liquids, etc.

Dewaxing/fractionation/preconcentration/purification

ChromatographyDistillationWinterization

Optimization of parameters

Recrystallization

Water/solvent removal Decarboxylation

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Extraction with SC-CO2

Response surface plots depicting the influence of SC-CO2 pressure

and temperature (time – 120 min), and pressure and time

(temperature – 70 °C) on hemp SFE-CO2 extract yield

20

The yield and conc. of CBD and CBDA in hemp SC-

CO2 extract at different extraction conditionsSFE-CO2 parameters Yield Cannabidiol (CBD) Cannabidiolic acid (CBDA)

P, bar T, °C t, min g/100 g DW mg/g extract g/100 g DW mg/g extract g/100 g DW

100 35 60 1.76 ± 0.03c 64.18 ± 2.98e 0.11 ± 0.01a 157.56 ± 6.00a 0.28 ± 0.01a

300 70 90 7.93 ± 0.08fg 25.75 ± 1.31d 0.20 ± 0.00ef 185.60 ± 8.26ab 1.47 ± 0.07cd

300 52.5 90 7.62 ± 0.09ef 19.13 ± 0.73ab 0.15 ± 0.01abcd 232.55 ± 0.49bcd 1.77 ± 0.00e

100 52.5 90 0.63 ± 0.00b -n -n -n -n

300 52.5 60 7.65 ± 0.08ef 18.31 ± 0.72a 0.14 ± 0.01abc 236.30 ± 14.01bcd 1.81 ± 0.11e

100 70 120 0.27 ± 0.01ab -n -n -n -n

300 52.5 90 7.70 ± 0.13ef 18.46 ± 3.61ab 0.14 ± 0.03abc 231.55 ± 9.91bcd 1.78 ± 0.08e

300 52.5 90 7.61 ± 0.05ef 21.00 ± 0.73abcd 0.16 ± 0.01cd 233.27 ± 1.51bcd 1.78 ± 0.01e

100 70 60 < 0.00a -n -n -n -n

300 52.5 90 7.56 ± 0.00ef 20.89 ± 0.17abcd 0.16 ± 0.00cd 237.80 ± 3.11cd 1.80 ± 0.02e

500 52.5 90 8.25 ± 0.19g 21.41 ± 0.21 abcd 0.18 ± 0.00de 231.46 ± 2.44bcd 1.91 ± 0.02ef

100 35 120 2.81 ± 0.02d 44.22 ± 1.94e 0.12 ± 0.01ab 203.82 ± 7.88b 0.57 ± 0.02b

500 70 60 9.63 ± 0.26h 24.15 ± 0.89bcd 0.23 ± 0.01f 204.05 ± 7.30b 1.97 ± 0.07ef

300 52.5 120 7.70 ± 0.17ef 20.16 ± 0.96abcd 0.16 ± 0.01bcd 221.12 ± 12.48bc 1.70 ± 0.10de

300 35 90 7.30 ± 0.13e 23.80 ± 0.54bcd 0.17 ± 0.00de 236.23 ± 12.15bcd 1.72 ± 0.09de

500 70 120 10.56 ± 0.31i 21.52 ± 0.84abcd 0.23 ± 0,01f 223.65 ± 14.12bc 2.36 ± 0.15g

300 52.5 90 7.50 ± 0.16ef 20.89 ± 0.17abcd 0.16 ± 0.00bcd 239.28 ± 1.02cd 1.80 ± 0.01e

500 35 60 7.39 ± 0.21e 19.42 ± 0.89abc 0.14 ± 0.01abcd 179.90 ± 9.22ab 1.33 ± 0.07c

300 52.5 90 7.63 ± 0.05ef 20.07 ± 0.60abc 0.15 ± 0.01bcd 230.42 ± 9.91bc 1.76 ± 0.08e

500 35 120 7.51 ± 0.04ef 18.28 ± 0.68a 0.14 ± 0,01abc 209.14 ± 7.56b 1.57 ± 0.06cd

Optimal SFE-CO2 extraction conditions:

465 70 120 8.30 ± 0,01g 24.72 ± 0.31cd 0.21 ± 0.00ef 261.40 ± 2.24d 2.17 ± 0.02fg

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Processing: summary of biorefining of hemp threshing (cleaning seeds) residues

1st STEP: SFE-CO2 8.3%

2nd STEP: PLE-Ac 4.3%

3rd STEP: PLE-EtOH/H2O 18.9%

4th STEP: EAE 20.2%

TOTAL 52.4 %

Residue 47.6 %

1

2

3

4

5

CBD = 2.5 %; CBDA = 26.1 %

Phenolics = 14.8 % (5.5 mg/g)

Phenolics = 12.5 % (26.9 mg/g)

PUFA OIL

PROTEINS

5 products; THC conc. ?22

Pharmaceuticals: authorized in Europe

Brand name Form Indications Description

Sativex

(Nabiximols)

Extract of cannabis

(oil): THC and CBD

MS (multiple

sclerosis

Sublingual

spray

Marinol

(Dronabinol)Synthetic delta-9-THC

Cancer

treatment,

AIDS, MS

Gelatin

capsule

Cesamet

(Nabilone)

Synthetic cannabinoid

similar to THC

Cancer

treatmentCapsule

Bedrocan

Dried flower tips

(sometimes powdered);

five strains available

Various Plant material

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Industrial hemp vs medicinal cannabis vs

recreational cannabis (Marijuana)

The same species (Cannabis sativa L. ) -

different composition and applications

Non-psychotropic effects vs Psychotropic effects

Cannabis sativa vs Cannabis sativa vs Cannabis sativa

PHYTOCANNABINOIDS

THC (mainly)CBD and many other structures24

Legal issues: ‘to allow or not to allow’

Leaves, flowering parts: for non-

medicinal products with non-

psycotropic phytocannabinoids:

- to allow or not to allow

- if allow, how much

Recreational purposes

Seeds ant their products:

legal (no question ‘to allow or

not to allow’)

Contains traces of THC

The problem : How much of THC

may be tolerated?

25

High resin hemp: looks

a lot more like

marijuana flowers than

true industrial hemp

Low resin hemp:

Can grow up to 5 m tall

Source: https://www.cbdschool.com/blog/cbd-101/do-cannabis-advocates-hate-cbd-hemp-oil

Industrial hemp vs marijuana

26

The main legal issue: THC vs CBD

Tetrahydrocannabinol Cannabidiol

Medical marijuana can have up to 20% THC and no CBD, where

hemp has less than 0.2-0.3% THC and high levels of CBD 27

Source: http://www.endocafoundation.org/is-cbd-legal/

Legal issues: medicinal and recreational purposes (includes high THC)

28

Medical Cannabis Regulations For CBD Products In Europe

Source: http://www.cannatech.news/2017/07/04/medical-cannabis-regulations-for-cbd-products-in-europe/

EU: in most countries only medicinal and industrial applications

29

European hemp cultivation (ha)

30

Internet business of CBD products

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Should non-psychotropic cannabinoids be

food/food supplement ingredients?

• ‘nutraceuticals’,

• ‘designer foods’,

• ‘f(ph)armafoods’,

• ‘medifoods’,

• ‘vitafoods’, etc.,

• but also the more traditional ‘dietary

supplements’ and

• ‘fortified foods’.32

Legal issues: will phytocannabinoids

appear in the list of EU regulation?

33

Health claim – where food meets medicine

“Reduces cholesterol"

phytocannabinoids

34

Legal issues: THC guide in Europe• Seeds were reintroduced for human consumption in the mid-

1990s, it was not uncommon to find THC levels > 100 ppm in oil

from Chinese and European seeds. Until the mandatory farming

of low-THC var. combined with the seed cleaning, that THC levels

in oil from European producers were effectively reduced.

• Currently European THC limits only exist for hemp cultivation:

permitted if contains less than 0.2/03% THC in the upper 1/3 of

the mature crop (Regulation 1420/98)

• EU General Food Law: “ based on UN Convention on Narcotic

Drugs (1961) and the UN Convention of psychotic substances

(1971).

• Article 2(g) (EC) No 178/2002): narcotic or psychotropic

substances covered by the conventions should not be considered

as “food” and consequently, they should not be allowed to be

incorporated into the food…….. ”35

High doses (> 200 mg oral/day): regulated as a medicinalproduct.

Physiological doses (20–200 mg oral /day): OTC-product (over

the counter) or a food supplement equal to substances, such as

valerian, ginkgo, some vitamins, iron products, etc.

Low doses (

Legal issued: USA• Dietary Supplement Health and

Education Act of 1994 (DSHEA)

• Ingredients are not “food additives”

• “Grandfathered” ingredients–Old ingredients, on the U.S. market before Oct. 15, 1994

• “New dietary ingredients”–Require a notification to FDA, with a summary of safety information, at least 75 days before marketing

• Hemp ingredients are not “Grandfathered”

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http://www.thecannabist.co/2017/08/15/industrial-hemp-farming-act-

congress-comer-polis/85923/

Farmers and politicians are

trying to convince the

government as well as the

DEA to separate the two

plants and allow the

cultivation and production

of hemp for economic and

natural resource benefits

Legal issues: the debate today (USA)

38

U.S. cannabis retail sales estimates, $ 109

Vending machines selling

industrial hemp products could

be coming to a Kentucky city

near you

http://www.nkytribune.com/2015/02/

bardstown-company-to-sell-industrial-

hemp-products-through-vending-

machines-in-select-ky-cities/

39

Federal Institute for Health Protection of Consumers and Veterinary Medicine (BgVV)

BgVV calculations for THC guidance values in foodstuffs

40

Legal issues: other countries

• Canada: 2nd country to pass THC limits, after Switzerland, in 1998: ‘products containing hemp are exempt from further regulation if they contain semifinished or derivative hemp goods that were found to contain 10 mg or less THC per g

• THC limit for ind. hemp: 0.3% of the dw of leaves and floweringparts

• The US: no regulations on THC in hemp foods. Technically, “zero tolerance” (0% THC) is expected on imported hemp goods. However, products with proper HS codes are allowed to be imported, including those under the Canadian 10 ppm legislation.

• Hemp foods are not permitted for human consumption in Australia and New Zealand under laws regulated by FSANZ (Food Standards Australia New Zealand). From 2014: seeds = 5, oil = 10, beverages = 0,2; other = 5. C. sativa is allowed to contain < 0.5% THC in the leaves and flowering heads

41

Scientifically Sound Guidelines for THC

in Europe, July 2015

Comparison of LOEL and uncertainty factor of THC in hemp foods

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Conclusion

• It is just a plant, just a very useful plant

• Existed and has been used for millennia

• Very useful and deserves 21st century research

• Hemp is not intoxicating

• Growers can be reasonably regulated until a system can be established

• All processes could be located on site reducing need for transportation

‘The judges are not toxicologists’ (Huub Lelieveld)

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