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

Area of Science:

• Pharmacognosy and Analytical Chemistry
• Natural Product Analysis
• Biochemical Profiling

Background:

• Danshen injection is a traditional medicine with potential bioactive components.
• Identifying these components and their activities is crucial for understanding efficacy.
• Existing methods may not efficiently profile multiple bioactivities simultaneously.

Purpose of the Study:

• To develop and apply on-line high performance liquid chromatography (HPLC) coupled with three biochemical detection (BCD) methods for evaluating bioactive components in Danshen injection.
• To screen for acetylcholinesterase (AChE) inhibitors and antioxidants within Danshen injection.
• To identify and characterize the main bioactive compounds responsible for observed activities.

Main Methods:

• On-line HPLC coupled with photo-diode array-fluorescence detection for AChE inhibitors using 7-acetoxy-1-methyl quinolinium iodide.
• On-line HPLC coupled with 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) free radical scavenging assays for antioxidants.
• HPLC-Mass Spectrometry (HPLC-MS) for the identification of main radical scavengers and AChE inhibitors.

Main Results:

• Three distinct active profiles were observed for AChE inhibition and radical scavenging.
• Seven major peaks with significant free radical inhibition and several minor peaks with strong AChE inhibition were detected.
• Protocatechuic aldehyde, salvianolic acid H or I, and lithospermic acid were identified among the active compounds; several unknown potent AChE inhibitors require further identification.

Conclusions:

• The developed on-line multiple HPLC-BCD methods are effective for rapid screening and identification of bioactive compounds in herbal medicines like Danshen injection.
• This approach facilitates the discovery of novel compounds with significant pharmacological activities.
• The methods provide a powerful tool for pharmacognosy, accelerating the identification of interesting and/or novel bioactive compounds.