A novel method for differentiating dextran sulfate from related sulfated polysaccharides.
A method is described for unequivocal identification of dextran sulfate, based on combined chemical desulfation and dextranase enzymolysis of dextran sulfate moieties to isomaltose, a specific indicator of dextran-type precursors. The method was developed using high-resolution (300 MHz) 1H NMR spectroscopy for assurance of the molecular transformations, identification, and estimation of the hydrolysis products. Overall conversion of approximately 80% of highly sulfated and moderately sulfated dextran sulfates was realized. Both 2-D 1H and 13C NMR spectra of a dextran sulfate (MW 500,000) clarified the extent of sulfation (75%) at C-4 and confirmed that sulfation at positions C-2 and C-3 was virtually complete. Estimation of the hydrolysis products (isomaltose, major; alpha-D-glucose, minor) is not restricted to 1H NMR now that the desulfation-enzymolysis methodology has been established; rather, it can be performed using HPLC or GLC (with derivatization).
Ludwig-Baxter
K G
KG
Department of Chemistry, McGill University, Montreal, Quebec, Canada.
Rej
R N
RN
Perlin
A S
AS
Neville
G A
GA
D002384:Catalysis; D016264:Dextran Sulfate / Q000032:analysis*; D003910:Dextranase / Q000737:chemistry; D004121:Dimethyl Sulfoxide; D005947:Glucose / Q000032:analysis; D006868:Hydrolysis; D007534:Isomaltose / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy; D008970:Molecular Weight; D011134:Polysaccharides / Q000032:analysis*; D013429:Sulfatases
D011134:Polysaccharides; D007534:Isomaltose; D016264:Dextran Sulfate; D013429:Sulfatases; D003910:Dextranase; D005947:Glucose; D004121:Dimethyl Sulfoxide