Analysis of alpha-amylase-derived pyridylamino-dextran sulfate oligomers by the combination of size-exclusion and reversed-phase high-performance liquid chromatography.
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.
Arak
Yoshio
Y
Ace Bio Product Co, Tokyo, Japan.
Andoh
Akira
A
Fujiyama
Yoshihide
Y
Hata
Kazunori
K
Makino
Jin
J
Shimada
Mitsue
M
Bamba
Hiromichi
H
Okuno
Takafumi
T
Urushiyama
Noboru
N
Bamba
Tadao
T
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*
D016264:Dextran Sulfate; D000516:alpha-Amylases