Integrated Approach for Separation of 2-O-β-D-glucopyranosyl-L-ascorbic Acid From Lycium barbarum L. Fruit Using...

Area of Science:

• Natural Product Chemistry
• Chromatographic Separation Techniques
• Biochemistry

Background:

• 2-O-β-D-glucopyranosyl-L-ascorbic acid (AA-2βG), a derivative of ascorbic acid found in Lycium barbarum, possesses significant biological activities.
• Efficient isolation of AA-2βG is crucial for its further study and application, but complex natural matrices pose challenges.
• Existing separation methods may lack the efficiency and scalability required for preparative isolation of bioactive compounds.

Purpose of the Study:

• To develop and optimize an integrated approach for the preparative isolation of AA-2βG from Lycium barbarum fruit.
• To enhance separation efficiency and reduce processing time and solvent consumption using high-speed counter-current chromatography (HSCCC) and aqueous two-phase systems (ATPS).
• To evaluate the antioxidant activity of the isolated AA-2βG.

Main Methods:

• An integrated method combining HSCCC and ATPS was developed, with an optimized solvent system of ammonium sulfate/ethanol/water/acetonitrile.
• The entropy weight method and response surface methodology were employed to optimize HSCCC parameters for efficient separation.
• An overlapping injection mode was implemented in HSCCC to improve throughput compared to single-injection mode.

Main Results:

• Under optimized conditions, 2.66 mg of AA-2βG with 91.24% purity was isolated from 140 mg of extract using single injection.
• The overlapping injection mode yielded 10.65 mg of AA-2βG with 84.53% purity from 700 mg of extract, reducing separation time by 55.68% and solvent consumption by 36.22%.
• In vitro experiments confirmed the antioxidant activity of the purified AA-2βG.

Conclusions:

• The integrated HSCCC-ATPS approach provides an efficient and robust strategy for the preparative isolation of AA-2βG from Lycium barbarum.
• The optimized overlapping injection mode significantly enhances the efficiency and sustainability of the separation process.
• This hybrid technique demonstrates broad applicability for isolating bioactive compounds from complex natural sources.