Application of high-speed countercurrent chromatography to the separation of black tea theaflavins

High-speed countercurrent chromatography (HSCCC) has been applied for the separation of four theaflavins (TFs). The results indicated that pure TF and theaflavin-3,3'- O-digallate (TFDG) can be obtained by HSCCC using a solvent system composed of hexane-ethyl acetate-methanol-water (1.25: 5: 1.25: 5, v/v/v/v). Although the separation of two TF monogallates is incomplete under all the solvent systems examined, peak cutting yielded a small amount of pure theaflavin-3-O-gallate (TF3MG) and theaflavin-3'-O-monogallate (TF3'MG). Thus, preparative HPLC has been employed as a complementary method for the separation of two TF monogallates. The HSCCC isolation of TFs from the solvent extract of black tea infusion has proved to be successful by gradient elution, based on the solvent system mentioned above.