An aqueous solution of the partially hydrolyzed
chitosan was chromatographed on a Sephadex G-
75 column (100 mm ¥ 13 mm) and eluted with 1%
(v/v) acetic acid (pH 5.3). The column was calibrated
with 2 mL of Blue Dextran 2000 (4 mg/mL)
and d-glucose (4 mg/mL). Elution was monitored
spectrophotometrically with the phenol-sulfuric
acid reagent for sugars (Chaplin, 1986). The fractions
collected were analyzed under UV light
(480 nm) and the absorbance was graphed as function
of the volume (each 5 mL) (Fig. 1).