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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
Author(s) -
Xia Guobin,
Lin Chunfang,
Liu Songbai
Publication year - 2016
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.22715
Subject(s) - chromatography , tannase , chemistry , biotransformation , countercurrent chromatography , theaflavin , high performance liquid chromatography , hydrolysis , enzyme , biochemistry , polyphenol , gallic acid , antioxidant
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.