Purpose

Glucosamine (C6H13NO5) is an amino sugar and is part of the structure of polysaccharides, chitosan, and chitin. Glucosamine is one of the most abundant monosaccharides in biological systems. It is produced commercially by the hydrolysis of shellfish exoskeletons or, less commonly, by fermentation of grain such as corn or wheat. Glucosamine in nutrient products and other matrices can be separated, identified, and quantified via Hitachi HPLC amino acid analyzers. This method is for accurate determination and quantification of glucosamine in matrices low in free amino acids.

Summary

Samples are prepared for analysis by diluting a portion of the sample with a deproteinizing solution (5-sulfosalicylic acid hydrate), an internal standard (D-Glucosaminic Acid), and (if necessary) a lithium buffer to achieve an appropriate dilution. The mixture is then vortexed, micro-centrifuged, and filtered into an HPLC vial.

Glucosamine analysis is performed on Hitachi HPLC amino acid analyzers. Glucosamine is separated by cation exchange chromatography using various lithium eluents. Absorbance is measured at 440 and 570 nm following post-column color development by Ninhydrin reagent. Appropriate standards are run to ensure each sequence is running properly and used in the quantification of samples using peak areas and applying an external standard technique.

Macromolecules Quantified

Glucosamine