title: High-throughput and sensitive screening of compounds with deoxyribonucleic acid-binding activity by a high-performance liquid chromatography-tandem mass spectrometry-fluorescence detection technique using palmatine as a fluorescence probe.
abstract: A high-throughput biochemical detection method based on the combination of high-performance liquid chromatography (HPLC), multiple-stage mass spectrometry (MS(n)) and DNA-binding activity assay was developed and validated for the simultaneous screening and identification of DNA-binding compounds in complex samples. Palmatine was used as a sensitive, nontoxic and environmentally friendly DNA fluorescence probe. HPLC fingerprints, ultraviolet absorption spectra, MS(n) fragments of components, and DNA-binding activity profiles could be simultaneously recorded during real-time analysis. Using the proposed method, 25 compounds were identified from Lophatherum gracile Brongn extracts, of which 18 were novel compounds first identified in these extracts. Nineteen compounds showed DNA-binding activity, most of which were flavone glycosides, with distinct dose-effect and structure-activity relationships. The method was validated and was proven to have a good linearity in the range of concentrations used in the study. The limit of detection was 0.2020nmol. Our study indicated that the proposed method was sensitive, accurate, precise and reliable to be used for simultaneous screening and identification of DNA-binding compounds in complex samples.
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China. Electronic address: echo.huisheng@163.com.
authors.forename: Qingrong
authors.initials: Q
authors.lastname: Fu
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors.forename: Yuexiang
authors.initials: Y
authors.lastname: Lan
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors.forename: Sen
authors.initials: S
authors.lastname: Li
authors.affiliation: Shimadzu (China) Co., Ltd., Shanghai 200052, China. Electronic address: y-hashi@shimadzu.co.jp.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Hashi
authors.affiliation: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China. Electronic address: chenshizhong66@163.com.
authors.forename: Shizhong
authors.initials: S
authors.lastname: Chen
mesh_terms: D001600:Berberine Alkaloids / Q000032:analysis* / Q000737:chemistry*; D002726:Chlorogenic Acid / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D047309:Flavones / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry*; D006027:Glycosides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D009862:Online Systems; D006109:Poaceae / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D001600:Berberine Alkaloids; D047309:Flavones; D005456:Fluorescent Dyes; D006027:Glycosides; D002726:Chlorogenic Acid; D004247:DNA; C005413:palmatine
keywords: DNA binding; Flavone glycosides; Lophatherum gracile Brongn; Online detection; Palmatine