1.Study on the purification technology of total triterpenoid extracts from Inonotus obliquus and their anti -tumor activity in vitro
Junjing SHAO ; Ying YANG ; Dalong MA ; Zhiqiang LYU ; Jingzhen TIAN ; Xiaoping ZHANG
China Pharmacy 2022;33(18):2198-2203
OBJECTIVE To optimite the purification technology of total triterpenoid extracts from Inonotus obliquus ,and to investigate the anti -tumor activity of its purified products . METHODS Using inotodiol as control ,the method was established for the content determination of total triterpenoid in I. obliquus. The type of macroporous adsorption resin ,sample volume ,sample concentration,sample flow rate ,eluent volume ,eluent dosage and elution flow rate were selected by single factor experiments . The purification technology of the crude extract was determined and verified . The effects of total triterpenoid purified from I. obliquus on the proliferation ,migration and apoptosis of human cervical cancer HeLa cells were detected by cell proliferation test , migration test ,flow cytometry and AO/EB kit . RESULTS The best purification technology of total triterpenoid crude extracts from I. obliquus was as follows :AB-8 macroporous adsorption resin was used ;mass concentration of the sample solution was 2.0 mg/mL;sample volume was 140 mL,and the flow rate was 1.0 mL/min;the impurity was removed with 50% ethanol 40 mL, then eluted with 95% ethanol 160 mL,at the elution flow rate of 3.0 mL/min. After purification ,mass concentration of total triterpenoid from I. obliquus increased from 34.36% to 73.39%. The total triterpenoid of I. obliquus could inhibit the proliferation of HeLa cells ,and the 50% inhibitory concentration was 184.20 μg/mL. Compared with control group ,the purified products could significantly inhibit the migratio n and promote the apoptosis of HeLa cells (P<0.05 or P<0.01). CONCLUSIONS The purification technology of total triterpenoids extracts from I. obliquus is successfully optimited . The purified product could inhibit the proliferation and migration of HeLa cells and induce their apoptosis.