title: Tannase-mediated biotransformation assisted separation and purification of theaflavin and epigallocatechin by high speed counter current chromatography and preparative high performance liquid chromatography: A comparative study.
abstract: A large scale isolation and purification of theaflavin (TF) and epigallocatechin (EGC) has been successfully developed by tannase-mediated biotransformation combining high-speed countercurrent chromatography. After tannase hydrolysis of a commercially available theaflavins extract (TE), the content of TF and EGC in tannase-mediated biotransformation product (TBP) achieved approximately 3 times enrichment. SEM studies revealed smooth tannase biotransformation and the possibility of recovery of the tannase. A single 1.5 hours' HSCCC separation for TF and EGC employing a two-phase solvent system could simultaneously produce 180.8 mg of 97.3% purity TF and 87.5 mg of 97.3% purity EGC. However, a preparative HPLC separation of maximum injection volume containing 120 mg TBP prepared 11.2 mg TF of 94.9% purity and 7.7 mg EGC of 89.9% purity. HSCCC separation demonstrated significant advantages over Prep HPLC in terms of sample loading size, separation time, environmental friendly solvent systems, and the production.
authors.affiliation: Department of Food Science and Nutrition, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang R & D Center for Food Technology and Equipment, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, China.
authors.forename: Guobin
authors.initials: G
authors.lastname: Xia
authors.affiliation: Department of Food Science and Nutrition, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang R & D Center for Food Technology and Equipment, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, China.
authors.forename: Chunfang
authors.initials: C
authors.lastname: Lin
authors.affiliation: Department of Food Science and Nutrition, Fuli Institute of Food Science, Zhejiang Key Laboratory for Agro-Food Processing, Zhejiang R & D Center for Food Technology and Equipment, Zhejiang University, 866 Yuhangtang Road, Hangzhou, 310058, China.
authors.forename: Songbai
authors.initials: S
authors.lastname: Liu
mesh_terms: D044946:Biflavonoids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D001711:Biotransformation; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D002392:Catechin / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003377:Countercurrent Distribution / Q000379:methods*; D008855:Microscopy, Electron, Scanning
chemical_list: D044946:Biflavonoids; C056068:theaflavin; D002392:Catechin; D002265:Carboxylic Ester Hydrolases; C022711:tannase; C057580:gallocatechol
keywords: high-speed countercurrent chromatography; preparative high performance liquid chromatography; scanning electron microscopy; tannase-mediated biotransformation; theaflavins