For gel permeation chromatography, the hot water extract (50 mg) was dissolved in distilled water
(2 ml), and the polysaccharide B (50 mg) was dissolved in 1~ NaOH (2 ml) and acidified to pH
5-6 with glacial acetic acid. The solution was chromatographed on a column (2.9 x 60 cm) packed
with TSK-gel, Toyopearl HW-55s (Toyo Soda Co.) and eluted with distilled water, 30 or 60 ml
h-1. Eluate was detected with a refractive index detector (JAI, model RI-2) and also an aliquot
(5 ml) of the eluate was taken and detected for carbohydrates by the phenol-sulphuric acid m e t h ~ d . ~
The molecular weight was estimated by gel permeation chromatography using dextran T70, T40
and T10 (Pharmacia) as a standard.