Simultaneous quantification of five marker compounds of Betula utilis stem bark using a validated high-performance...

Area of Science:

• Phytochemistry
• Analytical Chemistry
• Pharmacognosy

Background:

• Betula utilis stem bark contains bioactive compounds with therapeutic potential.
• Accurate quantification of these compounds is crucial for quality control and research.
• Existing analytical methods may lack sensitivity, selectivity, or robustness.

Purpose of the Study:

• To develop and validate a sensitive, selective, and robust densitometric high-performance thin-layer chromatographic (HPTLC) method.
• To enable simultaneous quantitative determination of five key marker compounds: betulin, lupeol, oleanolic acid, 3-acetyloleanolic acid, and β-sitosterol.
• To establish a reliable method for analyzing these compounds isolated from Betula utilis.

Main Methods:

• High-performance thin-layer chromatography (HPTLC) using silica gel 60 plates.
• Mobile phase: n-hexane:ethyl acetate (8:2 v/v).
• Densitometric detection at 500 nm post-derivatization with ceric ammonium sulfate reagent.

Main Results:

• The developed HPTLC method demonstrated good linearity for all investigated analytes (r > 0.9960).
• The method achieved sensitive, selective, and robust separation and quantification of the five marker compounds.
• The marker compounds were successfully isolated from Betula utilis stem bark and characterized.

Conclusions:

• A validated HPTLC method is suitable for the quantitative analysis of betulin, lupeol, oleanolic acid, 3-acetyloleanolic acid, and β-sitosterol.
• This method provides a simple, reproducible, and robust tool for quality assessment of Betula utilis extracts.
• The findings support the potential therapeutic applications of these marker compounds.