Determination of glucosamine in impure chitin samples by high-performance liquid chromatography.

A simple, rapid, selective, and specific high-performance liquid chromatography (HPLC) method was developed to quantitate glucosamine, and its application for estimating purity of chitin was investigated. The chromatographic separation was achieved using a reversed-phase C8 column, pre-column derivatization with 9-fluorenylmethoxycarbonyl chloride (Fmoc-Cl) and ultraviolet detection (lambda=254 nm). The mobile phase consisted of CH3CN and H2O. The optimum conditions of acid hydrolysis of chitin (concentration of HCl, temperature, and heating time) was obtained by performing the orthogonal array design (OAD) procedure and the released glucosamine was determined by the above HPLC method. The accuracy of the method was checked by the standard addition technique. The method was found to be specific with good linearity, accuracy, precision, and well suited for quantitation of glucosamine and determination of the purity of chitin in biological materials and food products.