Preparation of high purity monodisperse oligosaccharides derived from mannuronan by size-exclusion chromatography followed by semi-preparative high-performance anion-exchange chromatography with pulsed amperometric detection.
Oligosaccharides of ([4)-beta-d-ManpA-(-->](n)) with a degree of polymerisation (DP) of 5, 10 and 15 were generated by partial acid hydrolysis of alginate mannuronan. These were subsequently purified by a combination of size-exclusion chromatography and semi-preparative high-performance anion-exchange chromatography with pulsed amperometric detection. The purity of the isolated oligosaccharides was greater than 96%. With automated operation of the chromatography system, milligram quantities can be generated over a period of a few days. Thus, our methodology now offers some significant advantages over earlier, including our own, protocols focused on uronic acid oligomers, where the final products are either not as pure or more starting material is needed to generate an equivalent yield of product. Removal of ammonium ions in collected fractions after size-exclusion chromatography and prior to freeze-drying was found to be essential to prevent the formation of imines and subsequent Maillard browning products.
Ballance
Simon
S
NOBIPOL, Department of Biotechnology, Norwegian University of Science and Technology, N-7491 Trondheim, Norway. simon.ballance@nofima.no
Aarstad
Olav Andreas
OA
Aachmann
Finn Lillelund
FL
Skjåk-Braek
Gudmund
G
Christensen
Bjørn E
BE
D002850:Chromatography, Gel / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D009844:Oligosaccharides / Q000737:chemistry*
D009844:Oligosaccharides