ABSTRACT

Glucosamine hydrochloride and chondroitin sulphate are glycosaminoglycans (GAGs) compound which is a major structural component of bone that form proteoglycans. Both of these compounds can take care of bone health by stimulating the synthesis of synovial fluid and inhibit the degradation of joint cartilage, so it can be used for the treatment of osteoarthritis. The aim of this study was to obtain selective analytical method in determining glucosamine hydrochloride and chondroitin sulphate levels in tablet and cream dosage forms. After derivatization using orthophtalaldehyde and 2-mercaptoethanol (OPA/2-ME), the samples were analyzed using a high performance liquid chromatography (HPLC) with fluorescence detector at excitation wavelength of 335 nm and emission wavelength of 445 nm. Glucosamine has a primary amine group that could be derivatized with OPA/2-ME, while chondroitin has an acetyl group at the amine group, so we required a deacetylation using natrium hydroxide to break the acetyl group. The mobile phase used tetrahydrofuran 0.25% in water-acetonitrile (87:13) with a flow rate 1.5 mL/min. The analysis conditions have been optimized and validated in terms of accuracy, precision, linearity, selectivity, limit of detection, and limit of quantitation. The result showed average levels of glucosamine hydrochloride and chondroitin sulphate in tablet and cream dosage forms were 92.76%; 96.11% and 101.15%; 100.33% and they have fulfilled the acceptance criteria.

Keywords: chondroitin sulphate, derivatization, glucosamine hydrochloride, fluorescence, HPLC, OPA/2-ME, optimization, validation.