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Understanding and improving direct UV detection of monosaccharides and disaccharides in free solution capillary
James D Oliver1, Adam A Rosser2, Christopher M Fellows2
1University of Western Sydney, Australian Centre for Research On Separation Sciences (ACROSS), School of Science and Health, Parramatta Campus, Locked Bag 1797, Penrith NSW 2751, Australia.
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 1×10(-4) mM and 1×10(-8) mM for conventional 50 μm i.d. capillaries and for the corresponding extended light path capillaries, respectively.
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