Determination of average degree of polymerisation and distribution of oligosaccharides in a partially acid-hydrolysed homopolysaccharide: a comparison of four experimental methods applied to mannuronan.
The average degree of polymerisation (DP) and distribution of oligosaccharides in partially acid hydrolysed mannuronans were quantitatively evaluated by 1H NMR, electrospray ionisation mass spectrometry (ESI-MS), micellar electrokinetic capillary chromatography with UV detection (MEKC-UV), and high-pressure anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). Our investigation shows that 1H NMR, MEKC-UV and, in particular, HPAEC-PAD can be used as quantitative tools to aid the investigation of polysaccharide structure, function and synthesis. For the latter two techniques, especially, this represents a significant new development as it enables calculation of the quantity of individual oligomers of nominal DP by direct analysis of a defined oligomer mixture. Appropriate statistical averages of number and weight distributions were also calculated and found to fit very well to predicted Kuhn distributions that assume random depolymerisation.
Campa
Cristiana
C
Department of Biochemistry, Biophysics and Macromolecular Chemistry University of Trieste, 1-34127 Trieste, Italy.
Oust
Astrid
A
Skjåk-Braek
Gudmund
G
Paulsen
Berit Smestad
BS
Paoletti
Sergio
S
Christensen
Bjørn E
BE
Ballance
Simon
S
D000143:Acids / Q000737:chemistry; D001704:Biopolymers / Q000737:chemistry*; D002852:Chromatography, Ion Exchange; D020374:Chromatography, Micellar Electrokinetic Capillary; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry*; D012015:Reference Standards; D021241:Spectrometry, Mass, Electrospray Ionization
D000143:Acids; D001704:Biopolymers; D009844:Oligosaccharides