Grundmann, Marco and Rothenhoefer, Martin and Bernhardt, Guenther and Buschauer, Armin and Matysik, Frank-Michael (2012) Fast counter-electroosmotic capillary electrophoresis-time-of-flight mass spectrometry of hyaluronan oligosaccharides. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 402 (8). pp. 2617-2623. ISSN 1618-2642,
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Fast capillary electrophoresis-mass spectrometry measurements under counter-electroosmotic analyte migration conditions are presented. Efficient separations of a homologous series of six hyaluronan oligosaccharides (comprising 1-6 hyalobiuronic acid moieties) could be completed in 65 s. Separations were achieved in short-length fused silica capillaries under high electric field strengths of up to 1.25 kV center dot cm(-1). Capillary inner diameters ranging from 5 to 50 mu m were investigated, resulting in an optimal value of 15 mu m. The influence of capillary dimensions and buffer composition on separation efficiency and sensitivity are discussed. Optimal separations were achieved using a 28 cm x 15 mu m capillary, a separation high voltage of 35 kV, a background electrolyte of 25 mM ammonium acetate adjusted to pH 8.5, and negative ionization mode. The optimized method was successfully applied to a bovine testicular hyaluronidase digest of hyaluronan. Only minimal sample pretreatment for protein-containing samples is required. The simple manual injection procedure and fast separations allow for a sample throughput of 35 samples per hour.
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