Uronic acid determination by high performance liquid chromatography with postcolumn fluorescence derivatization

Area of Science:

• Analytical Chemistry
• Biochemistry
• Chromatography

Background:

• Uronic acids are crucial components in various biological polysaccharides.
• Accurate microanalysis of uronic acids is essential for biochemical and food science research.
• Existing analytical methods may lack sensitivity or require complex sample preparation.

Purpose of the Study:

• To develop a sensitive and reliable fluorimetric high performance liquid chromatography (HPLC) method for uronic acid microanalysis.
• To establish detection limits for key uronic acids using the developed method.
• To validate the method's applicability to real-world samples.

Main Methods:

• Development of a postcolumn fluorimetric detection system.
• Optimization of a saline mobile phase for HPLC separation.
• Analysis of standard uronic acids (D-glucuronic, D-galacturonic, D-mannuronic acids).
• Application of the method to polysaccharide hydrolysates and a beverage sample.

Main Results:

• Achieved low detection limits for D-glucuronic acid (7.19 pmol), D-galacturonic acid (23.88 pmol), and D-mannuronic acid (7.08 pmol).
• Demonstrated successful quantification of uronic acids in a complex polysaccharide hydrolysate.
• Validated the method for uronic acid determination in a beverage sample.

Conclusions:

• The developed fluorimetric HPLC method offers high sensitivity for uronic acid microanalysis.
• The technique is robust and suitable for analyzing uronic acids in diverse biological and food matrices.
• This advancement provides a valuable tool for researchers in carbohydrate chemistry and related fields.