EXPANDED STARCH AS A NOVEL STATIONARY PHASE FOR CHROMATOGRAPHY

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EXPANDED STARCH AS A NOVEL STATIONARY PHASE FOR
CHROMATOGRAPHY

• FABIEN DESWARTE
• Supervisors Prof. James Clark, Dr. Jeff Hardy

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PLAN

• Background of the project
• Characterisation of expanded starch comparison
  with silica
• Use in Column Chromatography
• Use in Flash chromatography
• Hydrothermal stability issues solutions

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WHAT IS STARCH ?

• One of the most abundant biopolymer on the planet
• Biopolymer of D-glucose
• 2 types of molecules
• Amylose
• linear, a-1,4 linkages
• Amylopectin
• Branched, a -1,6 linkages
• at branch points

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EXPANSION OF STARCH
granules
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PHYSICAL PROPERTIES
EXPANSION
Expanded starch
Type IV
Relative pressure (P/P0)
Relative pressure (P/P0)
Silica K60
Nitrogen adsorption isotherms conducted at 70K
Type IV
IUPAC recommendations, Pure Appl. Chem., 1994
(66) 1739
Relative pressure (P/P0)
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PHYSICAL PROPERTIES
Microporous volume was calculated through the
Dubinin-Radushkevich model
Surface polarity measurement ( )obtained
using Reichardts dye as a indicator
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PHYSICAL PROPERTIES
Microporous volume was calculated through the
Dubinin-Radushkevich model
Surface polarity measurement ( )obtained
using Reichardts dye as a indicator
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PHYSICAL PROPERTIES
Microporous volume was calculated through the
Dubinin-Radushkevich model
Surface polarity measurement ( )obtained
using Reichardts dye as a indicator
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SEPERATION MODEL

• Model system

20 cm
Ferrocene
Diacetyl ferrocene
Monoacetylferrocene
Elution system
- 30 ml hexane
- 45 ml hexane/acetone
(955, vv) - 30 ml acetone
Polarity
5 ml

• Collection of 5ml fractions and analysis by GC

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EXPANDED STARCH GRAVITY CHROMATOGRAPHY
GC response (Arb. Units)
No separation
Native starch (SA lt 1 m2/g)
EXPANSION
Expanded starch (SA 138 m2/g)
Hexaneacetone (955)
Acetone
Hexane
Fractions
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EXPANDED STARCH GRAVITY CHROMATOGRAPHY
GC response (Arb. Units)
Expanded starch (SA 138 m2/g)
Silica K60 (SA 458 m2/g)
Hexaneacetone (955)
Acetone
Hexane
Fractions
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EXPANDED STARCH GRAVITY CHROMATOGRAPHY
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WHEAT STRAW WAXES
Wheat straw
COSMETICS, COATINGS, ETC..
Liquid extraction
WAXES
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PURIFICATION OF WHEAT STRAW WAXES
SILICA K60 400 mL of a (8515, vv) mixture of
hexane/ether
EXPANDED STARCH 200 mL of a (928, vv) mixture
of hexane/ether
Purification of a crude methyl-tetrahydrofuran
extract of wheat straw
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EXPANDED STARCH FLASH CHROMATOGRAPHY
Conditions 10 mL/min Hexane ? (9010, vv)
hexane/acetone (51 mL) (9010, vv)
hexane/acetone ? acetone (51 mL)
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HYDROTHERMAL STABILITY ISSUES
GC response (Arb. Units)
Expanded starch (SA 138 m2/g)
Saturated water vapour
50C / 24h
Hexaneacetone (955)
Acetone
Hexane
Fractions
Degraded starch (SA lt 1 m2/g)
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HYDROTHERMAL STABILITY SOLUTION (1)
GC response (Arb. Units)
Expanded starch (SA 138 m2/g)
(a) Stored under hexane
(b) Stored in a dessicator
Hexaneacetone (955)
Acetone
Hexane
Fractions
Starch stored under hexane (SA 132 m2/g)
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HYDROTHERMAL STABILITY SOLUTION (2)
Solvent exchange
granules
drying
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HYDROTHERMAL STABILITY SOLUTION (2)
GC response (Arb. Units)
Expanded starch (SA 138 m2/g)
Solvent-exchanged retrograded starch
Fractions
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HYDROTHERMAL STABILITY SOLUTION (3)
THERMOLYSIS

• Heated under vacuum
• Doped with a catalytic amount of
  p-toluenesulfonic acid

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CONCLUSIONS

• Application of expanded starch to chromatography
• Potential separation of very polar compounds such
  as proteins
• More work is needed to fully understand
  chromatographic properties of material
• Physical and chemical modifications can be
  applied to modify/improve properties
• Same methodology could be applied to other
  biomaterials

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Budarin V., Clark, J.H., Deswarte F.E.I., Hardy
J.J.E., Hunt A.J., Kerton F.M., Chem. Comm., 23
(2005) 2903-2905 PATENT PENDING Clark, J.H.,
Deswarte F.E.I., Hardy J.J.E., Hunt A.J., Kerton
F.M., and Milkowski K., PCT Patent Application,
PCT/GB2004/003276. July 27th 2004
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THANKS TO
THANKS FOR YOUR ATTENTION
feid100_at_york.ac.uk