title: A rapid, micro-scale preliminary screening method for active components in Galangal with protective effect against hydrogen peroxide induced cell apoptosis through "thin layer chromatography" and "tetrazolium-based colorimetric assay" array correspondence.
abstract: A new method has been established for rapid preliminary screening active ingredients in natural products through thin layer chromatography (TLC) array responding with tetrazolium-based colorimetric assay (MTT assay) along with post LC-MS in micro-scale. The extract of the natural product was first separated by TLC. The separated spots obtained from TLC were visualized in situ with vanillin-ethanolic sulfuric acid agent to define the array correspondence between TLC spots and 384-cell culture plate for MTT cell viability assay. The TLC spots from the replicate TLC plates were then eluted and transferred into the wells of 384-cell culture plates according to the array respondence. The TLC spots with significant antioxidant activities were further screened by MTT assay, and subsequently traced and identified by LC-MS based on the TLC-MTT assay array correspondence. This new method was successfully applied to screen active ingredients in a Chinese medicine known as Galangal. Two major inhibitors for the decline of PC12 cell survival (Galangin, m/z 269.1, and 5-hydroxy-7-(4'-hydroxy-3'-methoxyphenyl)-1-phenyl-3-heptanone, m/z 327.2) were effectively screened and identified by this method.
authors.affiliation: Guangdong Key Laboratory for Research & Development of Natural Drugs, Guangdong Medical College, 524023 Zhanjiang, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Cheng
authors.affiliation: Guangdong Key Laboratory for Research & Development of Natural Drugs, Guangdong Medical College, 524023 Zhanjiang, China.
authors.forename: Yuanting
authors.initials: Y
authors.lastname: Li
authors.affiliation: Guangdong Key Laboratory for Research & Development of Natural Drugs, Guangdong Medical College, 524023 Zhanjiang, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Li
authors.affiliation: Guangdong Key Laboratory for Research & Development of Natural Drugs, Guangdong Medical College, 524023 Zhanjiang, China. Electronic address: dengyfgd@163.com.
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Deng
mesh_terms: D032429:Alpinia / Q000737:chemistry*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002855:Chromatography, Thin Layer; D003124:Colorimetry; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D006861:Hydrogen Peroxide; D013058:Mass Spectrometry; D008516:Medicine, Chinese Traditional; D016716:PC12 Cells; D051381:Rats
chemical_list: D006861:Hydrogen Peroxide
keywords: Array correspondence; Galangal; LC-MS; MTT assay; Screening; TLC