title: Fast counter-electroosmotic capillary electrophoresis-time-of-flight mass spectrometry of hyaluronan oligosaccharides.
abstract: 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·cm(-1). Capillary inner diameters ranging from 5 to 50 μm were investigated, resulting in an optimal value of 15 μm. The influence of capillary dimensions and buffer composition on separation efficiency and sensitivity are discussed. Optimal separations were achieved using a 28 cm × 15 μm 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.
authors.affiliation: Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93040 Regensburg, Germany.
authors.forename: Marco
authors.initials: M
authors.lastname: Grundmann
authors.forename: Martin
authors.initials: M
authors.lastname: Rothenhöfer
authors.forename: Günther
authors.initials: G
authors.lastname: Bernhardt
authors.forename: Armin
authors.initials: A
authors.lastname: Buschauer
authors.forename: Frank-Michael
authors.initials: FM
authors.lastname: Matysik
mesh_terms: D000818:Animals; D002417:Cattle; D053841:Electroosmosis; D019075:Electrophoresis, Capillary; D006820:Hyaluronic Acid / Q000737:chemistry* / Q000378:metabolism; D006821:Hyaluronoglucosaminidase / Q000737:chemistry / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D009844:Oligosaccharides / Q000032:analysis* / Q000378:metabolism; D013737:Testis / Q000201:enzymology; D013997:Time Factors
chemical_list: D009844:Oligosaccharides; D006820:Hyaluronic Acid; D006821:Hyaluronoglucosaminidase