title: Analysis of alpha-amylase-derived pyridylamino-dextran sulfate oligomers by the combination of size-exclusion and reversed-phase high-performance liquid chromatography.
abstract: In a previous study, we reported a novel method for the separation and quantification of a strong negatively charged material, dextran sulfate sodium (DSS), using fluorometric labeling with 2-aminopyridine and size-exclusion high-performance liquid chromatography. In the present study, we developed a method for the separation of pyridylamino-DSS (PA-DSS) using reversed-phase high-performance liquid chromatography (RPLC). In vitro enzymatic degradation of the PA-DSS was carried out using alpha-amylase. In RPLC, depolymerized PA-DSS was eluted on the basis of molecular mass (in the order pentamer, trimer, dimer, and monomer of PA-DSS) and separations were more sharply than in size-exclusion chromatography. The combination of RPLC and size-exclusion chromatography also separated depolymerized PA-DSS as effectively as RPLC alone.
authors.affiliation: Ace Bio Product Co, Tokyo, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Arak
authors.forename: Akira
authors.initials: A
authors.lastname: Andoh
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Fujiyama
authors.forename: Kazunori
authors.initials: K
authors.lastname: Hata
authors.forename: Jin
authors.initials: J
authors.lastname: Makino
authors.forename: Mitsue
authors.initials: M
authors.lastname: Shimada
authors.forename: Hiromichi
authors.initials: H
authors.lastname: Bamba
authors.forename: Takafumi
authors.initials: T
authors.lastname: Okuno
authors.forename: Noboru
authors.initials: N
authors.lastname: Urushiyama
authors.forename: Tadao
authors.initials: T
authors.lastname: Bamba
mesh_terms: D002850:Chromatography, Gel / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D016264:Dextran Sulfate / Q000737:chemistry*; D013058:Mass Spectrometry; D008970:Molecular Weight; D013050:Spectrometry, Fluorescence; D000516:alpha-Amylases / Q000737:chemistry*
chemical_list: D016264:Dextran Sulfate; D000516:alpha-Amylases