Bryophyllum pinnatum markers: CPC isolation, simultaneous quantification by a validated UPLC-DAD method and...

Area of Science:

• Phytochemistry
• Pharmacognosy
• Analytical Chemistry

Background:

• Bryophyllum pinnatum (B. pinnatum) is a medicinal plant used in traditional medicine for inflammation.
• Flavonol glycosides are major metabolites, but quality control and marker identification are lacking.
• Standardized B. pinnatum extracts require data on safety, efficacy, and quality.

Purpose of the Study:

• Isolate major flavonoids from B. pinnatum hydroethanolic extract.
• Validate a quantification method for chemical markers.
• Evaluate xanthine oxidase inhibition and antioxidant activity of isolated flavonoids.

Main Methods:

• Centrifugal partition chromatography (CPC) for flavonoid isolation.
• Nuclear Magnetic Resonance (NMR) for structure confirmation.
• Ultra-Performance Liquid Chromatography with Diode Array Detection (UPLC-DAD) for method validation and quantification.
• In vitro assays for xanthine oxidase (XO) inhibition and antioxidant activity (DPPH, ABTS).

Main Results:

• Four major flavonoids were isolated: quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (1), kaempferol 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside (2), quercetin 3-O-α-L-rhamnopyranoside (3), and kaempferol 3-O-α-L-rhamnopyranoside (4).
• A validated UPLC-DAD method demonstrated high linearity, accuracy, and precision for quantification.
• Compound 1 was quantified at 2.43%, compound 2 at 0.25%, and compound 3 at 0.33%.
• Compounds 2 and 3 showed moderate XO inhibition, while compounds 1 and 3 exhibited antioxidant activity.

Conclusions:

• Flavonoid 1 (quercetin 3-O-α-L-arabinopyranosyl-(1→2)-O-α-L-rhamnopyranoside) is proposed as a specific marker for quality control of B. pinnatum hydroethanolic leaf extract.
• The validated UPLC-DAD method provides a reliable tool for standardizing B. pinnatum extracts.
• The study contributes essential data for ensuring the quality, safety, and efficacy of B. pinnatum in medicinal applications.