Separation of capsular polysaccharide K4 and defructosylated K4 by high-performance capillary electrophoresis.
A rapid, highly sensitive and reproducible high-performance capillary electrophoresis (HPCE) method (electrokinetic chromatography with sodium dodecyl sulfate) is described for the determination of the polysaccharide from the uropathogenic Escherichia coli K4 bacteria (05:K4:H4) and its defructosylated product. The two polyanions, K4 and defructosylated K4, are separated and readily determined within 30 min on an uncoated fused-silica capillary using normal polarity at 20 kV and detection at 200 nm. A linear relationship was found for the two polysaccharides over a wide range of concentrations, from approximately 30 ng (0.5 microg/microL) to 210 ng (3.5 microg/microL). The described method was used to evaluate the defructosylation process of K4 under drastic acid conditions.
Volpi
Nicola
N
Department of Biologia Animale, University of Modena and Reggio Emilia, Modena, Italy. volpi@unimo.it
D000143:Acids / Q000737:chemistry; D000838:Anions / Q000737:chemistry; D016667:Bacterial Capsules; D002138:Calibration; D002807:Chondroitin / Q000737:chemistry; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D005632:Fructose / Q000737:chemistry*; D006025:Glycosaminoglycans / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D011135:Polysaccharides, Bacterial / Q000737:chemistry / Q000302:isolation & purification*; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D013995:Time
D000143:Acids; D000838:Anions; D006025:Glycosaminoglycans; D011108:Polymers; D011135:Polysaccharides, Bacterial; C063323:capsular polysaccharide K4; D005632:Fructose; D012967:Sodium Dodecyl Sulfate; D002807:Chondroitin