Chondroitin Sulfate Testing (AOAC)

Chondroitin sulfate is a major component of connective tissue and is widely used in dietary supplements to promote healthy joints and flexibility. Owing to the complex nature of chondroitin sulfate, many different analytical methods have been developed and used for characterization and determination of purity and label claim verification in finished supplement forms such as cetyl pyridinium chloride (CPC), reverse phase HPLC without hydrolysis, size exclusion HPLC, and carbazole reaction. However, these methods suffer from lack of specificity and selectivity, often resulting in positive interference from impurities and adulterants.

Eurofins’ Success

Eurofins Petaluma has performed an in-house validation of the methodology published in the Journal of AOAC for determination of chondroitin sulfate content in raw materials and dietary supplements by HPLC with UV detection after enzymatic hydrolysis (Enzymatic HPLC). Eurofins has applied this method for determination of chondroitin sulfate in raw materials, human supplements in tablet/capsule/drink forms, and animal supplements in tablet/powder/biscuits/chewables/drink forms with high degree of accuracy and precision.

Method Summary

Chondroitin sulfate in raw materials and dietary supplements is hydrolyzed with chondroitinase AC enzyme after aqueous extraction. This treatment selectively hydrolyses chondroitin sulfate A and C to its unsaturated disaccharide units. The resulting disaccharide units are then separated and quantitated using gradient elution ion-pairing HPLC with UV detection at 240nm. The specific unsaturated disaccharides that respond to the enzyme are found only in chondroitin sulfate A and C. The presence of the disaccharides is confirmation for identity. Disaccharides are summed to determine the level of chondroitin sulfate. Due to it high selectivity, Enzymatic HPLC assay is not affected by possible contaminants and adulterants such as carragennan, dextran sulfate, other related GAGs and proteins, thereby resulting in accurate quantitation of chondroitin sulfate content in the test materials.

Due to it high selectivity, Enzymatic HPLC assay is not affected by possible contaminants and adulterants such as carragennan, dextran sulfate, other related GAGs and proteins, thereby resulting in accurate quantitation of chondroitin sulfate content in the test materials.

Why Enzymatic HPLC?

Enzymatic HPLC method is highly selective, rugged, accuarate, repeatable and precise for quantitation and identification of chondroitin sulfate in raw material and dietary supplement matrices.
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