Preparative separation of EGCG from Korean green tea by high performance liquid chromatography

Abstract

Among catechin compounds abundant mainly in green tea, (−)epigallocatechin gallate (EGCG) has antioxidative and anticarcinogenic properties. Korean green tea (Bosung, Chonnam) was used as a feed sample. The separation was done by a C₁₈ reversed-phase preparative column (22×250 mm) with mobile phases of 0.1% acetic acid in water, acetonitrile, methanol, and ethyl acetate considered. The injection volume and the flow rate of mobile phase were fixed at 400 μL and 16 mL/min, respectively. To purify EGCG preparatively, the effluent was collected from the column outlet and concentrated, and analyzed by analytical column (3.9×300 mm). From the experimental results, the optimum mobile phase condition for separating EGCG from Korean green tea was 0.1% acetic acid in water/acetonitrile/methanol/ethyl acetate (80/16/1/3 vol.%). In the experimental condition, 0.6 mg of EGCG was obtained purely.

Keywords:

• (−)Epigallocatechin gallate
• Green tea
• Preparative high performance liquid chromatography
• Mobile phase composition

Acknowledgments

The authors gratefully acknowledge the financial support of Center for Advanced Bioseparation Technology. This work was performed in the High-Purity Separation Laboratory of Inha University, Inchon, Korea.