Oligosaccharide relative quantitation using isotope tagging and normal-phase liquid chromatography/mass spectrometry.
The growing interest in the conversion of plant biomass into biofuels has recently highlighted the lack of analytical techniques that are able to profile the fine structures of plant cell-wall polysaccharides. Here we present a new liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS) platform called Oligosaccharide Quantitation using Isotope Tagging (OliQuIT) developed for profiling the oligosaccharides derived from glycosyl hydrolase digestion of polysaccharides. The method is demonstrated using different arabinoxylan-derived oligosaccharide samples, which are reductively aminated with either the light (12C6) or heavy (13C6) form of aniline. The complex oligosaccharide mixtures are analysed by capillary normal-phase (NP)-LC and ESI-MS. Importantly, arabinoxylan oligosaccharide isomers are separated by NP-LC and their relative abundance in different samples can be determined from the intensities of ions labeled with the different isotopes. OliQuIT will be of use in multiple applications, including screening for plant varieties with improved saccharification properties, characterizing glycosyl hydrolase specificities and analysing plant glycosyl transferase mutants.
Ridlova
Gabriela
G
Chemistry Department, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.
Mortimer
Jenny C
JC
Maslen
Sarah L
SL
Dupree
Paul
P
Stephens
Elaine
E
D000814:Aniline Compounds / Q000737:chemistry; D002247:Carbon Isotopes; D002851:Chromatography, High Pressure Liquid; D007553:Isotope Labeling; D009844:Oligosaccharides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods*
D000814:Aniline Compounds; D002247:Carbon Isotopes; D009844:Oligosaccharides; C023650:aniline