title: Multiple on-line HPLC coupled with biochemical detection methods to evaluate bioactive compounds in Danshen injection.
abstract: On-line high performance liquid chromatography (HPLC) coupled with three biochemical detection (BCD) methods was applied to evaluate bioactive components in Danshen injection. On-line HPLC-photo-diode array-fluorescence detection based on the fluorogenic substrate 7-acetoxy-1-methyl quinolinium iodide, was built to search acetylcholinesterase (AChE) inhibitors in Danshen injection. On-line HPLC coupled with the scavenging assay of 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radicals was developed to screen antioxidants. The three active profiles were obviously different. Radical scavenging profiles revealed seven strong peaks in the chromatographic fingerprint possessing obvious free radical inhibition effects, while some minor peaks exhibited stronger AChE inhibition activities. The main radical scavengers and AChE inhibitors were identified by HPLC-MS. Several unknown ingredients showing strong AChE inhibition activities needed further identification except protocatechuic aldehydrate, salvianolic acid H or I and lithospermic acid. The on-line multiple on-line HPLC-BCD methods will provide powerful tools in the field of pharmacognosy for fast-track identification of interesting and/or novel bioactive compounds.
authors.affiliation: Hubei Institute of Traditional Chinese Medicine Control, Hubei Institute for Food and Drug Control, Wuhan, Hubei, 430075, China.
authors.forename: Wen-Bing
authors.initials: WB
authors.lastname: Peng
authors.affiliation: Hubei Institute of Traditional Chinese Medicine Control, Hubei Institute for Food and Drug Control, Wuhan, Hubei, 430075, China.
authors.forename: Qiu-Hua
authors.initials: QH
authors.lastname: Zeng
authors.affiliation: Hubei Institute of Traditional Chinese Medicine Control, Hubei Institute for Food and Drug Control, Wuhan, Hubei, 430075, China.
authors.forename: Dan-Ping
authors.initials: DP
authors.lastname: Li
authors.affiliation: Hubei Institute of Traditional Chinese Medicine Control, Hubei Institute for Food and Drug Control, Wuhan, Hubei, 430075, China.
authors.forename: Tian-Ming
authors.initials: TM
authors.lastname: Ding
authors.affiliation: Hubei Institute of Traditional Chinese Medicine Control, Hubei Institute for Food and Drug Control, Wuhan, Hubei, 430075, China.
authors.forename: Jing-Ling
authors.initials: JL
authors.lastname: Tan
authors.affiliation: Hubei Institute of Traditional Chinese Medicine Control, Hubei Institute for Food and Drug Control, Wuhan, Hubei, 430075, China. dxp2888@126.com.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Ding
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000737:chemistry* / Q000494:pharmacology; D052160:Benzothiazoles / Q000737:chemistry; D001713:Biphenyl Compounds / Q000737:chemistry; D002800:Cholinesterase Inhibitors / Q000737:chemistry* / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation* / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology; D004867:Equipment Design; D010851:Picrates / Q000737:chemistry; D027561:Salvia miltiorrhiza; D013451:Sulfonic Acids / Q000737:chemistry
chemical_list: D000975:Antioxidants; D052160:Benzothiazoles; D001713:Biphenyl Compounds; D002800:Cholinesterase Inhibitors; D004365:Drugs, Chinese Herbal; D010851:Picrates; D013451:Sulfonic Acids; C028972:dan-shen root extract; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; C004931:1,1-diphenyl-2-picrylhydrazyl
keywords: AchE; DPPH/ABTS free radicals; Danshen injection; HPLC-MS; On-line HPLC-BCD