Etik Wahyuningsih (2020) Pengembangan Metode Kckt-Elsd Dengan Pemisahan Hilic Untuk Penetapan Kadar Glukosamin Hidroklorida Dan Kondroitin Sulfat Secara Simultan Pada Produk Suplemen Kesehatan. Masters thesis, UNIVERSITAS AIRLANGGA.

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Abstract

Glucosamine hydrochloride and chondroitin sulfate are widely used as dietary supplements for osteoarthritis treatment. The analysis of glucosamine hydrochloride and chondroitin sulfate in dietary supplements poses several challenges due to the lack of a sufficient UV chromophore and its high polarity. The purposes of this study were to develop and validate a simple, selective and accurate a HILIC-HPLC-ELSD (high performace liquid chromatography with evaporative light scattering detector-hydrophilic interaction liquid chromatography column) method for determination of glucosamine hydrochloride and chondroitin sulfate in dietary supplements. The chromatographic of glucosamine hydrochloride and chondroitin sulfate was carried on a ZIC-HILIC column 150 mm x 4.6 mm x 5μm in isocratic system mode, the mobile phase consisted of acetonitrile, 30 mM ammonium formate and water (77:20:3, v/v/v) with pH of 4.5, column temperature of 35°C, flow rate of 1 ml/min, and injection volume of 5 μl. Evaporative light scattering was used as detector with the nebulization and evaporation temperature were 50°C and 80°C, respectively. The flow rate of nitrogen was 1.10 standard liter per minutes. A good separation of glucosamine hydrochloride and chondroitin sulfate was obtained, the resolution of the analytes more than 1.5. The retention time of glucosamine hydrochloride and chondroitin sulfate are 18.13 min and 11.28 min, respectively. The linearity of glucosamine hydrochloride and chondroitin sulfate were obtained in 0.4 – 2.5 mg/ml range. The limit of detection and limit of quantification of glucosamine hydrochloride and chondroitine sulfat were 20, 80 μg/mL, 80 and 400 mg/mL. All the validation parameter selectivity, linearity and range, accuracy and precision met the acceptable criteria according to ICH guideline. The method was successfully applied for the determination sample of dietary supplements commercially.

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