Efficient capillary zone electrophoretic separation of wood-derived neutral and acidic mono- and oligosaccharides.
Neutral and acidic monosaccharides, commonly present as structural units in wood-derived hemicelluloses, were derivatized by reductive amination using 6-aminoquinoline (6-AQ) and subsequently separated as their borate complexes by capillary zone electrophoresis. By using a quite concentrated (420 mmol 1(-1) alkaline borate buffer, a fused-silica capillary column with a small inner diameter (30 microns nominal I.D.) and a constant power of 1200 mW (corresponding to an applied voltage of approximately 21 kV), optimal separation was achieved. Under these conditions, the monosaccharides investigated were separated with a resolution, Rs, of 1.0-1.2 or greater. On-column UV detection at 245 nm was found to provide highly sensitive detection of the 6-AQ-derivatized monosaccharides. The minimum detectable concentrations were on the order of 10(-6) mol 1(-1) (corresponding to an estimated limit of detection of a few fmol). The linear calibration range of the method, including the 6-AQ derivatization step, was found to be about two orders of magnitude. Several neutral beta (1-4)-D-xylooligomers and acidic oligosaccharides containing 4-O-methyl-D-glucuronic acid units, which are common structural elements in hemicelluloses such as birch and spruce xylan, were also efficiently separated as 6-AQ derivatives, using the same buffer system. Finally, the usefulness of this analytical method has been demonstrated using a spruce wood xylan sample subjected to chemical and enzymatic hydrolysis.
Rydlund
A
A
Swedish Pulp and Paper Research Institute, Stockholm, Sweden.
Dahlman
O
O
D000634:Aminoquinolines / Q000737:chemistry; D002240:Carbohydrate Sequence; D019075:Electrophoresis, Capillary / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000737:chemistry / Q000302:isolation & purification*; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification*; D013056:Spectrophotometry, Ultraviolet; D014934:Wood; D014990:Xylans / Q000378:metabolism
D000634:Aminoquinolines; D009005:Monosaccharides; D009844:Oligosaccharides; D014990:Xylans; C032928:6-aminoquinoline