Understanding and improving direct UV detection of monosaccharides and disaccharides in free solution capillary electrophoresis

Abstract

Direct UV detection of carbohydrates in free solution capillary electrophoresis at 270 nm is made possible by a photo-oxidation reaction. Glucose, rhamnose and xylose were shown to have unique UV absorption spectra hypothesizing different UV absorbing intermediates for their respective photo-oxidation. NMR spectroscopy of the photo-oxidation end products proved they consisted of carboxylates and not malondialdehyde as previously theorized and that oxygen thus plays a key role in the photo-oxidation pathway. Adding the photo-initiator Irgacure® 2959 in the background electrolyte increased sensitivity by 40% at an optimum concentration of 1 x 10⁻⁴mM and 1 x 10⁻⁸mM for conventional 50 μm i.d. capillaries and for the corresponding extended light path capillaries, respectively.