HPLC method development for chitin and chitosan valorisation chemistry

Abstract

Recently, chitin/chitosan chemistry is gaining huge attention to yield chemicals like oligomers, monomers, N-containing chemicals and furan derivatives. However, due to lack of proper analytical technique to separate them due to their almost similar size and polarity hamper the further progress in this domain. In this work, a high performance liquid chromatography method with a variable wavelength and refractive index detector is optimised for simultaneous separation and quantification of amino sugars (D (+)- glucosamine, N-acetyl-D-glucosamine, Glucosamine hydrochloride dimer), other sugars (D (+)-glucose, D-(+)-fructose, 2-Deoxy-D-glucose), N-containing chemicals (D-glucosaminic acid, 2-Aminoethanol, N-acetylethanolamine, Acetamide) and furan derivative (5-Hydroxymethylfurfural). While non-polar C18 column with isocratic elution [acetonitrile/water (1/9v/v)] could show separation of few compounds, but best results were achieved with RPM-Monosaccharide Pb2+ column with water as a mobile phase. The regression equation revealed good linear relationship (R2=0.9904-0.9974) within test ranges. Repeatability of the method and column stability was found to be within acceptable limits.

Keywords

• High Performance Liquid Chromatography
• Method development
• Chitin and Chitosan chemistry
• Separation and quantification