title: Determination of the composition of the oligosaccharide phosphate fraction of Pichia (Hansenula) holstii NRRL Y-2448 phosphomannan by capillary electrophoresis and HPLC.
abstract: The promising new anticancer agent, PI-88, is prepared by the sulfonation of the oligosaccharide phosphate fraction of the extracellular phosphomannan produced by the yeast Pichia (Hansenula) holstii NRRL Y-2448. The composition of the oligosaccharide phosphate fraction was determined by capillary electrophoresis (CE) with indirect UV detection using 6 mM potassium sorbate at pH 10.3 as the background electrolyte. Further confirmation of the composition was obtained by HPLC analysis of a sample dephosphorylated by treatment with alkaline phosphatase. The structure of the hexasaccharide component has been determined by isolation and NMR spectroscopic analysis of its dephosphorylated derivative. Additionally, the structure of a second, previously undetected tetrasaccharide component (a hexosamine) has been determined by isolation and NMR spectroscopic analysis of the acetate of its dephosphorylated derivative. It is demonstrated that CE is an ideal method for the quality control of the oligosaccharide phosphate fraction for use in the production of PI-88.
authors.affiliation: Department of Research & Development, Progen Industries Ltd, PO Box 28, Richlands BC, Qld 4077, Australia. vito.ferro@progen.com.au
authors.forename: Vito
authors.initials: V
authors.lastname: Ferro
authors.forename: Caiping
authors.initials: C
authors.lastname: Li
authors.forename: Kym
authors.initials: K
authors.lastname: Fewings
authors.forename: Maria C
authors.initials: MC
authors.lastname: Palermo
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Linhardt
authors.forename: Toshihiko
authors.initials: T
authors.lastname: Toida
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D005110:Extracellular Space / Q000737:chemistry; D008351:Mannans / Q000737:chemistry*; D009844:Oligosaccharides / Q000032:analysis* / Q000138:chemical synthesis* / Q000737:chemistry; D010843:Pichia / Q000737:chemistry*; D013451:Sulfonic Acids / Q000737:chemistry
chemical_list: D000970:Antineoplastic Agents; D008351:Mannans; D009844:Oligosaccharides; D013451:Sulfonic Acids; C120158:phosphomannopentaose sulfate; C005437:phosphomannan