López-Cervantes J, Sánchez-Machado D I, Delgado-Rosas K E
Departamento de Biotecnología y Ciencias Alimentarias, Instituto Tecnológico de Sonora, Cd. Obregón, Sonora, México.
J Chromatogr Sci. 2007 Apr;45(4):195-9. doi: 10.1093/chromsci/45.4.195.
This work presents a high-performance liquid chromatography (HPLC) method for the quantitation of glucosamine in chitin. The method includes an acid hydrolysis of chitin. The chromatographic separation is achieved using a Hypersil ODS 5-microm column (250 x 4.6 mm) at 38 degrees C, with precolumn derivatization with 9-fluorenylmethyl-chloroformate and UV detection (lambda = 264 nm). The mobile phase is a mixture of mobile phase A [30 mM ammonium phosphate (pH 6.5) in 15:85 methanol-water (v/v)], mobile phase B [15:85 methanol-water (v/v)], and mobile phase C [90:10 acetonitrile-water (v/v)], with a flow rate of 1.2 mL/min. The HPLC method proposed showed adequate repeatability (relative standard deviation, 5.8%), accuracy (92.7% recovery), and sensitivity, with a detection limit of 2 microg/mL. The method is successfully applied to the quantitation of glucosamine for the determination of the purity of chitin from shrimp waste.
本研究提出了一种用于定量测定几丁质中氨基葡萄糖的高效液相色谱（HPLC）方法。该方法包括几丁质的酸水解。色谱分离是在38℃下使用Hypersil ODS 5微米柱（250×4.6毫米）实现的，采用9-芴基甲基氯甲酸酯进行柱前衍生化，并通过紫外检测（λ = 264纳米）。流动相是流动相A [30 mM磷酸铵（pH 6.5），溶于15:85甲醇-水（v/v）]、流动相B [15:85甲醇-水（v/v）]和流动相C [90:10乙腈-水（v/v）]的混合物，流速为1.2毫升/分钟。所提出的HPLC方法显示出足够的重复性（相对标准偏差为5.8%）、准确性（回收率为92.7%）和灵敏度，检测限为2微克/毫升。该方法成功应用于氨基葡萄糖的定量分析，以测定虾壳废弃物中几丁质的纯度。
