title: Development of propidium iodide as a fluorescence probe for the on-line screening of non-specific DNA-intercalators in Fufang Banbianlian Injection.
abstract: Fufang Banbianlian Injection (FBI) has been widely used as an anti-inflammatory and anti-tumor prescription. To understand the relationships between its bioactive ingredients and pharmacological efficacies, our previous study has been successfully identified some DNA-binding compounds in FBI using an established on-line screening system, in which 4',6-diamidino-2-phenylindole (DAPI) was developed as a probe. However, DAPI can be only used to screen ATT-specific DNA minor groove binders, leaving the potential active intercalators unknown in FBI. As a continuation of our studies on FBI, here we present a sensitive analytical method for rapid identification and evaluation of DNA-intercalators using propidium iodide (PI) as a fluorescent probe. We have firstly established the technique of high-performance liquid chromatography-diode-array detector-multistage mass spectrometry-deoxyribonucleic acid-propidium iodide-fluorescence detector (HPLC-DAD-MS(n)-DNA-PI-FLD) system. As a result, 38 of 58 previously identified compounds in FBI were DNA-intercalation active. Interestingly, all previously reported DNA-binders also showed intercalative activities, suggesting they are dual-mode DNA-binders. Quantitative study showed that flavonoid glycosides and chlorogenic acids were the main active compounds in FBI, and displayed similar DNA-binding ability using either DAPI or PI. In addition, 13 active compounds were used to establish the structure-activity relationships. In this study, PI was developed into an on-line method for identifying DNA-intercalators for the first time, and thus it will be a useful high-throughput screening technique for other related samples.
authors: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors: Yanyan
authors: Y
authors: Niu
authors: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors: Sensen
authors: S
authors: Li
authors: Department of Pharmaceutical Sciences, University of Tennessee Health Science Center, Memphis, TN 38163, United States.
authors: Zongtao
authors: Z
authors: Lin
authors: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors: Meixian
authors: M
authors: Liu
authors: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors: Daidong
authors: D
authors: Wang
authors: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China.
authors: Hong
authors: H
authors: Wang
authors: School of Pharmaceutical Sciences, Peking University, Beijing 100191, China. Electronic address: chenshizhong66@163.com.
authors: Shizhong
authors: S
authors: Chen
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000032:analysis / Q000737:chemistry; D002726:Chlorogenic Acid / Q000032:analysis / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000737:chemistry*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D005399:Fishes; D005419:Flavonoids / Q000032:analysis / Q000737:chemistry; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis / Q000737:chemistry*; D006027:Glycosides / Q000032:analysis / Q000737:chemistry; D057166:High-Throughput Screening Assays; D007211:Indoles / Q000032:analysis / Q000737:chemistry; D007267:Injections; D007364:Intercalating Agents / Q000032:analysis* / Q000737:chemistry; D008297:Male; D013058:Mass Spectrometry; D011419:Propidium / Q000032:analysis / Q000737:chemistry*; D013094:Spermatozoa; D013329:Structure-Activity Relationship
chemical_list: D000893:Anti-Inflammatory Agents; D004365:Drugs, Chinese Herbal; D005419:Flavonoids; D005456:Fluorescent Dyes; D006027:Glycosides; D007211:Indoles; D007364:Intercalating Agents; D002726:Chlorogenic Acid; D011419:Propidium; C007293:DAPI; D004247:DNA
keywords: DNA binding activity; Fluorescence detector; Fufang banbianlian injection; HPLC−DAD−MS; On-line method; Propidium iodide