124
T. Barclay et al. / Carbohydrate Research 352 (2012) 117–125
ation of the glycosidic linkages of the polymers. The observation of
the difference between the kinetic parameters for the hydrolysis of
short and long chain inulin suggests that these polysaccharides
adopt different conformations in solution.
HHSN272200800039C). Its contents are solely the responsibility
of the authors and do not necessarily represent the official views
of the National Institutes of Health or the National Institute of
Allergy and Infectious Diseases.
This work was also supported by The Australian Research Coun-
cil through
a Linkage Grant (LP0882596) and LIEF Grant
4. Experimental
4.1. General
(LE0668489), the latter used to purchase the NMR spectrometer
used in this study.
The long chain chicory inulin was food grade Orafti InutecÒ ob-
tained from Orafti Group (Belgium) and the short chain inulin was
food grade Fuji FF manufactured by Fuji Nihon Seito Corporation
(Japan). Sucrose (P99.5%), pullulan from Aureobasidium pullulans,
bovine liver glycogen Type IX and trifluoroacetic acid (P99%) were
purchased from Sigma–Aldrich Pty. Ltd (Australia), acetic acid
(analytical reagent) was purchased from Ajax Finechem Pty. Ltd
(Australia) and deuterium oxide (D2O, 99.9%) was obtained from
Novachem Pty. Ltd (Australia). All were used as received.
Supplementary data
Supplementary data associated with this article can be found, in the
References
1. Blecker, C.; Chevalier, J. P.; Van Herck, J. C.; Fougnies, C.; Deroanne, C.; Paquot,
M. Recent Res. Devel. Agric. Food Chem. 2001, 5, 125–131.
2. Stevens, C. V.; Meriggi, A.; Booten, K. Biomacromolecules 2001, 2, 1–16.
3. Franck, A.; De Leenheer, L. Inulin In Biopolymers Polysaccharides II:
Polysaccharides from Eukaryotes; Steinbuchel, A., De Baets, S., Vandamme, E.,
Eds.; Wiley-VCH Verlag GmbH: Berlin, 2002; Vol. 6, pp 439–479.
4. Andre, I.; Mazeau, K.; Tvaroska, I.; Putaux, J. L.; Winter, W. T.; Taravel, F. R.;
Chanzy, H. Macromolecules 1996, 29, 4626–4635.
5. Dan, A.; Ghosh, S.; Moulik, S. P. Biopolymers 2009, 91, 687–699.
6. French, A. D. J. Plant Physiol. 1989, 134, 125–136.
7. Dysseler, P.; Hoffem, D. Determination of Inulin and Oligofructose in Food
Products (Modified Aoac Dietary Fibre Method). In Complex Carbohydrates in
Food; Cho, S., Prosky, L., Dreher, M., Eds.; Marcel Dekker: New York, 1999; pp
213–227.
4.2. NMR
NMR spectra were recorded on a Bruker Avance III 600 operat-
ing at 600 MHz for 1H. 1D and 2D spectra were collected using
standard gradient-based pulse programs. The 1D 1H NMR data
were obtained over 64 scans with a 30° flip angle (90° pul-
se = 8.4 ls), an acquisition time of 2.7 s, a relaxation delay of 2 s
and 65k data points. Temperature was held constant using an in-
built heater that was calibrated using ethylene glycol. All experi-
ments were conducted in D2O using carbohydrate concentrations
of 12.5 or 25 mg mLꢀ1 with chemical shifts reported in parts per
million (ppm) downfield from 3-(trimethylsilyl)propionic acid
sodium salt (TPS) by referencing to the acetic acid internal stan-
dard (acetic acid methyl group at 2.08 ppm70). See Supplementary
data for tables of 1H NMR shifts for each carbohydrate discussed.
8. Koch, K.; Andersson, R.; Rydberg, I.; Åman, P. J. Sci. Food Agric. 1999, 79, 1503–
1506.
9. Barclay, T.; Ginic-Markovic, M.; Cooper, P.; Petrovsky, N. J. Excipients Food Chem.
2010, 1, 27–50.
10. Korbelik, M.; Cooper, P. D. Br. J. Cancer. 2007, 96, 67–72.
11. Cooper, P. D.; Carter, M. Mol. Immunol. 1986, 23, 903–908.
12. Silva, D. G.; Cooper, P. D.; Petrovsky, N. Immunol. Cell Biol. 2004, 82, 611–616.
13. Cooper, P. D.; Steele, E. J. Vaccine 1991, 9, 351–357.
14. Cooper, P. D.; Steele, E. J. Immunol. Cell Biol. 1988, 66, 345–352.
15. Cooper, P. D.; McComb, C.; Steele, E. J. Vaccine 1991, 9, 408–415.
16. Cooper, P. D.; Petrovsky, N. Glycobiology 2011, 21, 595–606.
17. Poulain, N.; Dez, I.; Perrio, C.; Lasne, M.-C.; Prud’homme, M.-P.; Nakache, E. J.
Controlled Release 2003, 92, 27–38.
4.3. Hydrolysis studies
18. Castelli, F.; Sarpietro, M. G.; Micieli, D.; Ottimo, S.; Pitarresi, G.; Tripodo, G.;
Carlisi, B.; Giammona, G. Eur. J. Pharm. Sci. 2008, 35, 76–85.
19. Vervoort, L.; Van den Mooter, G.; Augustijns, P.; Busson, R.; Toppet, S.; Kinget,
R. Pharm. Res. 1997, 14, 1730–1737.
20. Hinrichs, W. L. J.; Prinsen, M. G.; Frijlink, H. W. Int. J. Pharm. 2001, 215, 163–
174.
21. Van Drooge, D. J.; Hinrichs, W. L. J.; Frijlink, H. W. J. Pharm. Sci. 2004, 93, 713–
725.
22. Eriksson, J. H. C.; Hinrichs, W. L. J.; de Jong, G. J.; Somsen, G. W.; Frijlink, H. W.
Pharm. Res. 2003, 20, 1437–1443.
4.3.1. Hydrolysis of sucrose
Sucrose was dissolved in D2O (25 mg mLꢀ1) at room temperature
and then the solution was acidified with acetic acid (0.25% v/v,
44 mM) and trifluoroacetic acid (0.025% v/v, 3.3 mM). This solution
was then heated (55, 60, 65, 70, and 75 °C) within the NMR instru-
ment and the hydrolysis was monitored by 1H NMR spectroscopy.
23. Ronkart, S. N.; Deroanne, C.; Paquot, M.; Fougnies, C.; Blecker, C. S. Food Chem.
2010, 119, 317–322.
24. Blecker, C.; Fougnies, C.; Van Herck, J.-C.; Chevalier, J.-P.; Paquot, M. J. Agric.
Food Chem. 2002, 50, 1602–1607.
25. Coussement, P. Inulin and Oligofructose as Dietry Fibre: Analytical, Nutritional
and Legal Aspects. In Complex Carbohydrates in Food; Sungsoo Cho, S., Prosky, L.,
Dreher, M., Eds.; Marcel Dekker: New York, 1999; pp 203–212.
26. Roberfroid, M. B. Br. J. Nutr. 2005, 93, S13–S25.
27. Vanloo, J.; Coussement, P.; Deleenheer, L.; Hoebregs, H.; Smits, G. Crit. Rev. Food
Sci. Nutr. 1995, 35, 525–552.
4.3.2. Hydrolysis of inulin
Inulin was dissolved in D2O (12.5 and 25 mg mLꢀ1) by heating
briefly to 75 °C before cooling to room temperature. The solution
was then acidified with acetic acid (0.25% v/v, 44 mM) and trifluo-
roacetic acid (0.025% v/v, 3.3 mM). Subsequently the acidified solu-
tion was then heated (60, 65, 70, 75 and 80 °C) within the NMR
instrument and the hydrolysis was monitored by 1H NMR
spectroscopy.
28. Koo, H.-N.; Hong, S.-H.; Seo, H.-G.; Yoo, T.-S.; Lee, K.-N.; Kim, N.-S.; Kim, C.-H.;
Kim, H.-M. J. Nutr. Biochem. 2003, 14, 598–605.
4.3.3. Hydrolysis of pullulan and glycogen
29. Petrovsky, N. Vaccine 2006, 24, S26–S29.
30. Fuchs, A. Starch/Stärke 1987, 39, 335–343.
The polysaccharide was dissolved in D2O (25 mg mLꢀ1) by heat-
ing briefly to 85 °C before cooling to room temperature. The solu-
tion was acidified with acetic acid (0.25% v/v, 44 mM) and
trifluoroacetic acid (0.25% v/v, 33 mM). This solution was then
heated to 90 °C within the NMR instrument and the hydrolysis
was monitored by 1H NMR spectroscopy.
31. Carpita, N. C.; Housley, T. L.; Hendrix, J. E. Carbohydr. Res. 1991, 217, 127–136.
32. Matusek, A.; Meresz, P.; Le, T. K. D.; Oersi, F. Eur. Food Res. Technol. 2009, 228,
355–365.
33. Biermann, C. J. Adv. Carbohydr. Chem. Biochem. 1988, 46, 251–271.
34. L’Homme, C.; Arbelot, M.; Puigserver, A.; Biagini, A. J. Agric. Food Chem. 2003,
51, 224–228.
35. Nasab, E. E.; Habibi-Rezaei, M.; Khaki, A.; Balvardi, M. Int. J. Food Eng. 2009, 5,
1–10.
36. BeMiller, J. N.; Steinheimer, T. R.; Allen, E. E. Clin. Chem. 1967, 13, 261–269.
37. BeMiller, J. N. Carbohydr. Res. 1972, 21, 154–155.
Acknowledgements
38. Courtin, C. M.; Van den Broeck, H.; Delcour, J. A. J. Chromatogr. 2000, 866, 97–
104.
39. Buchanan, S.; Kubler, D.; Meigs, C.; Owens, M.; Tallman, A. Int. J. Chem. Kinet.
1983, 15, 1229–1234.
This work was supported by the National Institute of Allergy
and Infectious Diseases, NIH (Contracts U01-AI061142 and