title: An in vitro AChE inhibition assay combined with UF-HPLC-ESI-Q-TOF/MS approach for screening and characterizing of AChE inhibitors from roots of Coptis chinensis Franch.
abstract: In this study, an in vitro acetylcholinesterase (AChE) inhibition assay based on microplate reader combined with ultrafiltration high performance liquid chromatography-electrospray quadrupole time of flight mass (UF-HPLC-ESI-Q-TOF/MS) was developed for the rapid screening and identification of acetylcholinesterase inhibitors (AChEI) from roots of Coptis chinensis Franch. Incubation conditions such as enzyme concentration, incubation time, incubation temperature and co-solvent was optimized so as to get better screening results. Five alkaloids including columbamine, jatrorrhizine, coptisine, palmatine and berberine were found with AChE inhibition activity in the 80% ethanol extract of C. chinensis Franch. The screened compounds were identified by HPLC-DAD-ESI-Q-TOF/MS compared with the reference stands and literatures. The screened results were verified by in vitro AChE inhibition assays, palmatine showed the best AChE inhibitory activities with IC50 values of 36.6μM among the five compounds. Results of the present study indicated that the combinative method using in vitro AChE inhibition assay and UF-HPLC-ESI-Q-TOF/MS could be widely applied for rapid screening and identification of AChEI from complex TCM extract.
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China. Electronic address: hqzhao2007@163.com.
authors.forename: Hengqiang
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China; College of Food Science and Technology, Nanjing Agricultural University, Nanjing 210095, PR China.
authors.forename: Siduo
authors.initials: S
authors.lastname: Zhou
authors.affiliation: College of Life Science, Shandong Normal University, Jinan 250014, PR China.
authors.forename: Minmin
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China.
authors.forename: Jinhong
authors.initials: J
authors.lastname: Feng
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China.
authors.forename: Shanshan
authors.initials: S
authors.lastname: Wang
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China.
authors.forename: Daijie
authors.initials: D
authors.lastname: Wang
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Geng
authors.affiliation: Key Laboratory of TCM Quality Control Technology, Process Control Research Center of Traditional Chinese Medicine of Shandong Academy of Sciences, Shandong Analysis and Test Center, Jinan 250014, PR China. Electronic address: wxjn1998@126.com.
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
mesh_terms: D002800:Cholinesterase Inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D039323:Coptis; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000737:chemistry / Q000302:isolation & purification*; D018517:Plant Roots; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014462:Ultrafiltration / Q000379:methods
chemical_list: D002800:Cholinesterase Inhibitors; D004365:Drugs, Chinese Herbal
keywords: Acetylcholinesterase inhibitor; Coptis chinensis Franch; In vitro AChE inhibition assay; UF-HPLC-ESI-Q-TOF/MS