title: Bioactivity-based liquid chromatography-coupled electrospray ionization tandem ion trap/time of flight mass spectrometry for β₂AR agonist identification in alkaloidal extract of Alstonia scholaris.
abstract: Although chromatographic fingerprinting combined with chemometrics, is a rational method for the quality control of traditional Chinese medicine (TCM), chemometrics cannot fully explore the relationship between chemical information and the efficacy of the potential activity. In the present work, a cell-based β₂ adrenergic receptor (β₂AR) agonist functional evaluation model coupled with high-performance liquid chromatography was developed to screen the potential β₂AR agonist components in the alkaloidal extract of Alstonia scholaris leaves. Using a liquid chromatography with ion trap time-of-flight mass spectrometry (LCMS-IT-TOF) system, the potential bioactive compounds in the prescription were identified and deduced based on the mass spectrometric fragmentation patterns, tandem mass spectrometry (MS/MS) data, and relevant literature. Several new β₂AR agonists of indole alkaloids were successfully found, and their activities were confirmed through an in vivo relaxant test on guinea pig tracheal muscles. The developed method is rapid and reliable compared with conventional fingerprinting and showed high sensitivity and resolution for the identification of β₂AR agonists in TCM prescriptions. This strategy clearly demonstrates that bioactivity-integrated fingerprinting is a powerful tool not only in screening and identifying potential lead compounds and in determining the therapeutic material basis of Chinese herbal prescriptions, but also in supplying suitable chemical markers for their quality control.
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin 300071, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Hou
authors.forename: Xuelin
authors.initials: X
authors.lastname: Cao
authors.forename: Linyi
authors.initials: L
authors.lastname: Dong
authors.forename: Liqiang
authors.initials: L
authors.lastname: Wang
authors.forename: Binfeng
authors.initials: B
authors.lastname: Cheng
authors.forename: Qian
authors.initials: Q
authors.lastname: Shi
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Luo
authors.forename: Gang
authors.initials: G
authors.lastname: Bai
mesh_terms: D058666:Adrenergic beta-2 Receptor Agonists / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D000470:Alkaloids / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D031051:Alstonia / Q000737:chemistry*; D000704:Analysis of Variance; D000818:Animals; D002853:Chromatography, Liquid / Q000379:methods*; D055808:Drug Discovery / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000737:chemistry; D006168:Guinea Pigs; D057809:HEK293 Cells; D006801:Humans; D009130:Muscle, Smooth / Q000187:drug effects; D018515:Plant Leaves / Q000737:chemistry; D018343:Receptors, Adrenergic, beta-2 / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D058666:Adrenergic beta-2 Receptor Agonists; D000470:Alkaloids; D004365:Drugs, Chinese Herbal; D018343:Receptors, Adrenergic, beta-2