title: A high-resolution peak fractionation approach for streamlined screening of nuclear-factor-E2-related factor-2 activators in Salvia miltiorrhiza.
abstract: Generation of a high-purity fraction library for efficiently screening active compounds from natural products is challenging because of their chemical diversity and complex matrices. In this work, a strategy combining high-resolution peak fractionation (HRPF) with a cell-based assay was proposed for target screening of bioactive constituents from natural products. In this approach, peak fractionation was conducted under chromatographic conditions optimized for high-resolution separation of the natural product extract. The HRPF approach was automatically performed according to the predefinition of certain peaks based on their retention times from a reference chromatographic profile. The corresponding HRPF database was collected with a parallel mass spectrometer to ensure purity and characterize the structures of compounds in the various fractions. Using this approach, a set of 75 peak fractions on the microgram scale was generated from 4mg of the extract of Salvia miltiorrhiza. After screening by an ARE-luciferase reporter gene assay, 20 diterpene quinones were selected and identified, and 16 of these compounds were reported to possess novel Nrf2 activation activity. Compared with conventional fixed-time interval fractionation, the HRPF approach could significantly improve the efficiency of bioactive compound discovery and facilitate the uncovering of minor active components.
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
authors.forename: Li-Ping
authors.initials: LP
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
authors.forename: Hui-Peng
authors.initials: HP
authors.lastname: Song
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
authors.forename: Hai-Ping
authors.initials: HP
authors.lastname: Hao
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China.
authors.forename: Lian-Wen
authors.initials: LW
authors.lastname: Qi
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China. Electronic address: liping2004@126.com.
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.affiliation: State Key Laboratory of Natural Medicines, China Pharmaceutical University, No. 24 Tongjia Lane, Nanjing 210009, China. Electronic address: chenj2002@gmail.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Chen
mesh_terms: D001681:Biological Assay; D002470:Cell Survival / Q000187:drug effects; D005591:Chemical Fractionation; D004224:Diterpenes / Q000737:chemistry* / Q000494:pharmacology; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D057809:HEK293 Cells; D061307:Human Umbilical Vein Endothelial Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D013058:Mass Spectrometry; D051267:NF-E2-Related Factor 2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000737:chemistry; D011809:Quinones / Q000737:chemistry* / Q000494:pharmacology; D027561:Salvia miltiorrhiza / Q000737:chemistry*
chemical_list: D004224:Diterpenes; D004365:Drugs, Chinese Herbal; D051267:NF-E2-Related Factor 2; D011809:Quinones
keywords: Cell-based screening; High-resolution peak fractionation; Natural products; Nrf2 activators; Salvia miltiorrhiza