Effect of particle size in preparative reversed-phase high-performance liquid chromatography on the isolation of epigallocatechin gallate from Korean green tea.
To isolate epigallocatechin gallate (EGCG) of catechin compounds from Korean green tea (Bosung, Chonnam), a C18 reversed-phase preparative column (250x22 mm) packed with packings of three different sizes (15, 40-63, and 150 microm) was used. The sample extracted with water was partitioned with chloroform and ethyl acetate to remove the impurities including caffeine. The mobile phases in this experiment were composed of 0.1% acetic acid in water, acetonitrile, methanol and ethyl acetate. The injection volume was fixed at 400 microl and the flow rate was increased as the particle size becomes larger. The isolation of EGCG with particle size was compared at a preparative scale and the feasibility of separation of EGCG at larger particle sizes was confirmed. The optimum mobile phase composition for separating EGCG was experimentally obtained at the particle sizes of 15 and 40-63 microm in the isocratic mode, but EGCG was not purely separated at the particle size of 150 microm.
Kim
Jung Il
JI
Center for Advanced Bioseparation Technology and Department of Chemical Engineering, Inha University, Incheon, South Korea.
Hong
Seung Bum
SB
Row
Kyung Ho
KH
D002392:Catechin / Q000031:analogs & derivatives* / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D010316:Particle Size; D013056:Spectrophotometry, Ultraviolet; D013662:Tea / Q000737:chemistry*
D013662:Tea; D002392:Catechin; C045651:epigallocatechin gallate