Amberlite Resin Purification of Galactoglucomannan

Two XK 50 columns, (50 mm diameter; 250 mm length) (GE Healthcare Bio-Sciences AB, Uppsala, Sweden) were used of which the first column was loaded with 350 g of Amberlite XAD4 and the second with 400 g of Amberlite IRA958 (Sigma-Aldrich, Saint Louis, MO, USA) [27 (link)]. The flow rate in these columns was 1 mL/min (GP 50 gradient pump, Thermo Fisher Scientific Inc., Waltham, MA, USA) for both the adsorption and wash sequence. After a solution had been pumped through a column, the column was emptied by purging with air using a hose pump. XAD4 was cleaned with 700 mL of 10% ethanol solution and subsequently washed with 1400 mL of deionized water. 5750 g of the AC solution was passed through the XAD4 column after which a sample of 250 g was removed from the adsorption column permeate. The second column (IRA958) was washed with 1400 mL of deionized water and 5500 g of permeate was passed through this column, producing the purest permeate in this series, which was designated adsorption preparation (AD). The yield for each purification step was calculated based on the galactoglucomannan content (g) before and after each purification step and was expressed as % yield. The purification scheme is illustrated in Figure 6.