Studies On Isolation & Purification Of Theaflavins And Its Physiological Activity

Theaflavins (TFs ) is the key influence factor on black tea .A great deal of research including both home and overseas reveals that it has bright development prospect due to its multiple pharmacological function and health-protection. Its further use is blocked because it is very difficult to purify .To solve this problem, NKA-macro-resin is first used to accumulate TFs in this paper .With this method , content of TFs is up to 85%(mainly including four kinds) and the rate of preparation is 14.38%. The monomers isolation of TFs is the key point in this research ,so polyamide is applied for the first time to isolate TFs by HPTLC (High Performance Thin Layer Chromatography) and column chromatography. By HPTLC, the results reveal that the solvent system is the best one when methanol: acetone: acetic acid glacial is 8 : 5 : 3 and saturate time is 20min. The Rf of the former 7 peaks are 0.14,0.22,0.28,0.31,0.35,0.4,0.48 respectively. This result shows polyamide is effective isolation material of TFs. Based on the above results, TFs is further isolated by column chromatography when chloroform: acetone: acetic acid glacial is 3 : 5 : 8 : 0.5.There are seven color strips to be isolated. The latter four strips are detected by HPLC and the results show that the forth and fifth strips are still mixture; the sixth strip is TF-3'-G and its content is 85% ; the seventh strip is TF and its content is 93%.Then TF is further purified ,and its content surpasses 98%.The isolation effect of column chromatography is always limited when the complex compound are isolated. HP-HPLC is used in this research to isolate the forth, the fifth and the sixth strips can't be isolated by column chromatography. The results show that ECG and unknown compound (may be new-TF-3-G) are acquired in the forth strip; the ECG and TF-3-G is isolated in the fifth strip; the TF-3'-G is isolated in sixth strip and the content of TF-3'-G is 95%.For the first time self-preparation TFs and its monomers are used to detect its effect on salivary amylase, lipase and cyclooxygenase (COX: including...