Scaling up of high-speed countercurrent chromatographic apparatus with three columns connected in series for rapid preparation of (-)-epicatechin

In the present study, compact high-speed countercurrent chromatographic apparatus was constructed with three columns connected in series. Two sets of columns were prepared from 10 mm and 12 mm I.D. tubing to form 12 Land 15 L capacities, respectively. Performance of these columns was compared for the separation of (-)-epicatechin gallate (ECG) from a tea extract by flash countercurrent chromatography (FCCC). In each separation, 200g of the tea extract in 1600 mL of mobile phase was loaded onto the column. The 12 L column gave 7.5 L(35 g of ECG) and the 15 L column gave 9 L(40 g of ECG) of ECG solution without impurities. The ECG solution was directly hydrolyzed by tannase into (-)-epicatechin. The hydrolysate was purified by flash chromatography on AB-8 macroporous resin to give 52 g of EC (purity 99.1%). This scaled up apparatus could be used for the industrial separation of natural products. (C) 2012 Elsevier B.V. All rights reserved.