© Bioniqs Ltd 2008

Ionic Liquids: towards improved artemisinin extraction

© Bioniqs Ltd 2008

Topics

• Who are Bioniqs and what are ionic liquids?

• The artemisinin extraction challenge

• Ionic liquid - hexane comparison

• Conclusions

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Who are Bioniqs?

• Spin-out from Universities of York and Cambridge, UK

• Founded 2004

• Located on York Science Park

• Supplying solvent technology to the chemical industry

and related sectors

© Bioniqs Ltd 2008

Why use ionic liquids?

To improve performance in 3 areas;

• Technical: more efficient, cleaner extractions

• Economic: low temperature, low cost, simple synthesis

• Environmental: minimalize impact and eliminate hazards

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Protic ionic liquids

• Proprietary range of low melting point ionic liquids based upon secondary and tertiary ammonium salts

• Biodegradable, recyclable, sustainable, low cost

• Excellent “tuneable” solvents for a range of applications

• Designed to combine maximum performance with minimal hazard

NHO

O

OH NH+

HO

O

O-

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Ionic Liquid Design Process

• Computer modelling of required solvent properties

• The new solvent (which may be an IL) is assessed for likely practical, economic and environmental/safety factors

• Efficiency, selectivity, rates of extraction and other required properties are evaluated

• Determine stability fo new solvent

• Develop recovery, purification and recycling systems if required

• Demonstrate process at scale.

The Challenge

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Artemisinin Extraction

Current refluxing hexane extraction suffers from;• co-extraction of many impurities• use of toxic & hazardous solvent• negative environmental impact

O

O

O

CH3

HH

H3C O

O

HCH3

• selectively and rapidly extracts artemisinin• is economically competitive with hexane process• is environmentally benign, non-toxic, non-

viscous, biodegradable and recyclable

Challenge to develop a solvent that:

Bioniqs has designed a new solvent (a protic ionic liquid) specifically for the selective and benign extraction

of artemisinin.

© Bioniqs Ltd 2008

Artemisinin extraction - resultsSolvent Ionic Liquid Hexane

Extraction conditions 30 mins stirring at ambient temp1

45 mins at reflux1

Extraction efficiency 65-75%1 65-75%1

Co-extracted impurities 41 131

Pure artemisinin yield from primary extract

97%1 65%1

Toxicity Non-toxic to algae or Daphnia, non-genotoxic2

High chronic toxicity6, known mutagen6, suspected animal carcinogen6, highly toxic to aquatic life3,4, 5

Environmental profile Fully biodegradable in 24 hours2

Poorly biodegradable6

1 Head-to-head extractions performed in-house2 Testing carried out by Complement Genomics Limited, Sunderland, UK3 Bringmann, G., and Kuhn, R. (1977), Z.Wasser-Abwasser-Forsch. 10(5):161-1664 Bobra, A.M., Shiu, W.Y. and Mackay, D. (1983), Chemosphere 12(9/10):1121-1129 5 Hutchinson, T.C., Hellebust, J.A., Tam, D., Mackay, D., Mascarenhas, R.A., and Shiu, W.Y. (1980) Environ.Sci.Res. 16:577-5866 U.S. Department of Health and Human Services. (1999) Toxicological Profile for n-Hexane.

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High Selectivity of IL for Artemisinin

HPLC-RI trace of ionic liquid primary extract showing artemisinin (centre) and four minor impurities

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Extraction ProcessRoom temperature extraction using a solvent:biomass ratio of 10:1

Up to 5 successive extractions

Extracts 71% of artemisinin per pass

Artemisinin precipitates upon addition of water

Artemisinin can be isolated from this precipitate at greater than 97% yield - overall extraction yield of 69%

Solvent can be recycled by removal of water

© Bioniqs Ltd 2008

Artemisinin ExtractionArtemisinin extraction using an ionic liquid and its precipitation from solution through the addition of water.

Plant Material Precipitation of artemisinin

Primary Ionic Liquid Extract

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Eco-testing of Ionic Liquid

• Toxicity to Daphnia1

• Toxicity to Algae1

• Mutagenicity1

• Flammability2

• Biodegradability1

1 Testing carried out by Complement Genomics Limited, Sunderland, UK 2 Testing carried out by Chilworth Technology Ltd, Southamption, UK

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Toxicity to Daphnia sp.

0 1 2 3 4 5Time 0 hrs 15 15 15 15 15 15 No. of Daphnia

usedTime 48 hrs 1 4 9 10 11 13 No. of Daphnia

immobilized% Immobalized 7% 27% 60% 67% 73% 87%

Calculated EC50 values of >60mg/L.

Testing performed by Complement Genomics Ltd., Sunderland, UK

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Toxicity to Algae.

Growth inhibition data:

Calculated EC50 ~100mg/L

Concentration response curve:

50% growth inhibition rate

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Mutagenicity of IL

Ames Test - mutagenicity index and activity ratios;

REF MI MI (+ S9) MAR MAR (+S9)

Comments

TA97A <1.00 <1.21 0.00 <0.19 Non-mutagenicTA98 <1.24 <1.75 <0.34 <2.47 Non-mutagenicTA100 <0.97 <0.89 <0.34 <0.14 Non-mutagenicTA1535 <1.08 <1.21 <0.1 <0.64 Non-mutagenicWP2 <1.2 <1.12 <0.19 <0.11 Non-mutagenic

Non-genotoxic and is likely to be non-mutagenic and non-carcinogenic.

© Bioniqs Ltd 2008

Biodegradability

Independent assessment of ionic liquid;

Biodegradation with Plant + IL + soil

0.00

20.00

40.00

60.00

80.00

100.00

120.00

140.00

0 3 6 9 24

Time/exposure hours

Con

cent

ratio

n m

M

anion

cation

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Biodegradation ‐ results with autoclaved soil 

0.00

20.00

40.00

60.00

80.00

100.00

120.00

140.00

0 3 21 23 25 27 48

Time/exposure hours

Concen

tration mM

Cation

Anion

Biodegradability

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Flammability Comparison

Ionic LiquidFlashpoint: -7°CLEL: undetectable at 70°CUEL: undetectable at 70°C

HexaneFlashpoint: -26°CLEL: 1.2%UEL: 7.7%

Independent assessment of ionic liquid found that;

“Complete vaporisation of the test substance was not possible at these lower temperatures (50-70°C)”

Strong acid/base relationship means that ionic liquid needs to be operated at well over 100°C to fully vaporise.

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Comparative Risk Information (R Phrases)

HexaneR11 Highly flammable.R38 Irritating to skin.R48/20 Harmful: danger of serious damage to health by prolonged

exposure through inhalation.R62 Possible risk of impaired fertility.R65 Harmful: may cause lung damage if swallowed.R67 Vapours may cause drowsiness and dizziness.R51/53 Toxic to aquatic organisms, may cause long-term adverse

effects in the aquatic environment.

Ionic LiquidR11 Highly flammable.R36/37/38 Irritating to eyes, respiratory system and skin.

© Bioniqs Ltd 2008

• ILs can extracts high yields of artemisinin at

room temperature

• We have designed a particular protic ionic liquid that works

well in extraction of artemisinin

• This IL is low cost and safer to the user & environment

than hexane

• Partners now required for field testing of the IL

Summary

© Bioniqs Ltd 2008

neil.sullivan@bioniqs.com

+44 (0)1904 561538

www.bioniqs.com

Bioniqs Limited, Registered in England and Wales under number 5304761. Registered Office: Heslington Hall, York, YO10 5DD, United Kingdom

“THANK YOU FOR YOUR ATTENTION”

Dr. Neil Sullivan

Commercial Advisor