A strategy for screening bioactive components from natural products based on two-dimensional cell membrane...

Area of Science:

• Biochemistry
• Pharmacology
• Analytical Chemistry

Background:

• Cell membrane chromatography (CMC) is a bioaffinity method for screening natural product compounds.
• Limited receptor capacity in CMC columns can lead to column overloading, potentially causing researchers to miss important bioactive components.

Purpose of the Study:

• To develop a novel strategy to comprehensively discover bioactive components from complex matrices.
• To overcome the limitations of traditional CMC, specifically column overloading by high-content or high-affinity compounds.

Main Methods:

• A two-dimensional CMC approach combined with a component-knockout strategy was proposed.
• A CMC model using H9c2 rat cardiac myoblasts (H9c2/CMC) was established for screening cardioprotective compounds from red yeast rice (RYR).
• The method involved two-dimensional H9c2/CMC-HPLC coupled with a QTOF MS system for compound identification.

Main Results:

• The two-dimensional H9c2/CMC-HPLC-QTOF MS method successfully identified three initial compounds.
• After applying the component-knockout approach, an additional four bioactive compounds were discovered.
• The strategy successfully avoided column overloading and enriched trace compounds, identifying one pigment and six monacolins from RYR.

Conclusions:

• The proposed two-dimensional CMC and component-knockout strategy effectively prevents column overloading.
• This innovative approach enables the comprehensive discovery of bioactive compounds, including trace components, from complex natural product samples.
• The strategy holds potential for broader application in identifying active compounds from diverse complex matrices.