Analytical method development and validation for monosaccharide profiling in Lignosus rhinocerotis using rP-HPLC.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Pharmacognosy

Background:

• Lignosus rhinocerotis polysaccharides (LRP) exhibit diverse bioactivities.
• Understanding the monosaccharide composition of LRP is crucial for elucidating its structure-function relationship.
• Existing analytical methods may lack the precision or simplicity required for routine LRP analysis.

Purpose of the Study:

• To develop and validate a pre-column derivatization reversed-phase high-performance liquid chromatography (RP-HPLC) method for analyzing monosaccharides in Lignosus rhinocerotis polysaccharides (LRP).
• To quantitatively determine the monosaccharide profile of LRP.
• To establish a reliable analytical tool for LRP characterization.

Main Methods:

• Lignosus rhinocerotis polysaccharides (LRP) were hydrolyzed and derivatized.
• Monosaccharides were separated using reversed-phase high-performance liquid chromatography (RP-HPLC) on a Cosmosil 5C18-MS-II column.
• Detection was performed at 254 nm, with method validation including calibration curves, precision, and accuracy assessments.

Main Results:

• Baseline separation of eight standard monosaccharides was achieved within 45 minutes.
• LRP was identified as a heteropolysaccharide composed of mannose, ribose, rhamnose, glucose, galactose, xylose, and arabinose.
• The method demonstrated high recovery rates, ranging from 100.28% to 111.02%.

Conclusions:

• The optimized RP-HPLC method provides a simple, reproducible, and accurate approach for analyzing monosaccharides in Lignosus rhinocerotis polysaccharides (LRP).
• This validated method facilitates a deeper understanding of LRP's complex structure.
• The findings support further research into the bioactivities and therapeutic potential of LRP based on its detailed monosaccharide composition.