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

► High-speed countercurrent chromatography is scaled up with three columns in series. ► (−)-Epicatechin gallate is separated from a tea extract using this apparatus. ► (−)-Epicatechin gallate is hydrolyzed by tannase to (−)-epicatechin. ► Flash chromatography on AB-8 macroporous resin purifies (−)-Epicatechin. ► The scaled up apparatus could be used for industrial separation of natural products. 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 10mm and 12mm I.D. tubing to form 12L and 15L 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 1600mL of mobile phase was loaded onto the column. The 12L column gave 7.5L (35g of ECG) and the 15L column gave 9L (40g 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 52g of EC (purity 99.1%). This scaled up apparatus could be used for the industrial separation of natural products.