Determination of levels of glucosamine hydrochloride and chondroitin sulfate in mixtures in tablet and cream forms using high-performance liquid chromatography with fluorescence

Objective: Glucosamine hydrochloride (HCl) and chondroitin sulfate are glycosaminoglycan compounds and major structural components of bones in the form of proteoglycans. These compounds maintain bone structure by stimulating the synthesis of synovial fluid and inhibiting the degradation of joint cartilage, and they can be used for the treatment of osteoarthritis. The aim of this study was to identify a selective analytical method for determining glucosamine HCl and chondroitin sulfate levels in tablet and cream forms. Methods: After derivatization using ortho-phthalaldehyde and 2-mercaptoethanol, the samples were analyzed using high-performance liquid chromatography (HPLC) with a fluorescence detector at an excitation wavelength of 335 nm and an emission wavelength of 445 nm. Deacetylation using sodium hydroxide was required to break the acetyl group bond. The mobile phase used tetrahydrofuran 0.25% in water-acetonitrile (87:13) with a flow rate of 1.5 ml/minute. Results: The average levels of glucosamine HCl and chondroitin sulfate were 92.76% and 96.11% in tablets and 101.15% and 100.33% in creams, which fulfilled the acceptance criteria. Conclusions: Our validation method for glucosamine HCl and chondroitin sulfate met the acceptance criteria of accuracy, precision, selectivity, and linearity.