Abstract:

A two-dimensional multi-channel chromatographic separator designed by our research group was used to purify
polyphenols extracted from the pinecones of Pinus koraiensis Sieb. et Zucc. from north-eastern China. The chromatographic
conditions were optimized. The stationary and eluent were selected by static adsorption and desorption experiments. Onefactor-
at-a-time design was used to investigate four factors influencing the separation efficiency of polyphenols by dynamic
adsorption, sample loading amount, sample concentration, eluent flow rate, and number of chromatographic columns used.
Results indicated that X-5 macroporous resin was the best stationary phase for polyphenol separation and the best elutent
comprised a mixture of 0.5% formic acid and 60% ethanol in water, providing maximum adsorption quantity of 38.74 mg/g and
maximum desorption rate of 72.08%. The optimal experimental conditions for chromatographic separation of polyphenols
were 256 mg of sample at a concentration of 2.0 mg/mL, eluent flowing at a rate of (200±20) mL/min and using 4
chromatographic columns. Under these conditions, the purity of polyphenols was increased from 2.37% to 23.257%, and the
recovery was 46.81%.

Key words: two-dimensional multi-channel chromatographic separator, pine polyphenols, separation and purification, macroporous resin