Multiple on-line HPLC coupled with biochemical detection methods to evaluate bioactive compounds in Danshen injection

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 iodi...

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Bibliographic Details
Published in:Biomedical chromatography 2016-11, Vol.30 (11), p.1854-1860
Main Authors: Peng, Wen-Bing, Zeng, Qiu-Hua, Li, Dan-Ping, Ding, Tian-Ming, Tan, Jing-Ling, Ding, Xiao-Ping
Format: Article
Language:English
Subjects:
AchE
Animals
Antioxidants - chemistry
Antioxidants - pharmacology
Benzothiazoles - chemistry
Biphenyl Compounds - chemistry
Cholinesterase Inhibitors - chemistry
Cholinesterase Inhibitors - pharmacology
Chromatography, High Pressure Liquid - instrumentation
Chromatography, High Pressure Liquid - methods
Danshen injection
DPPH/ABTS free radicals
Drugs, Chinese Herbal - chemistry
Drugs, Chinese Herbal - pharmacology
Equipment Design
HPLC-MS
On-line HPLC-BCD
Picrates - chemistry
Sulfonic Acids - chemistry
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Summary: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.
ISSN:0269-3879
1099-0801
DOI:10.1002/bmc.3772