采用高效液相色谱法测定不纯几丁质样品中的氨基葡萄糖。
Determination of glucosamine in impure chitin samples by high-performance liquid chromatography.
作者信息
Zhu Xiaolan, Cai Jibao, Yang Jun, Su Qingde
机构信息
Department of Chemistry, University of Science and Technology of China, Hefei 230026, China.
出版信息
Carbohydr Res. 2005 Jul 25;340(10):1732-8. doi: 10.1016/j.carres.2005.01.045.
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.
开发了一种简单、快速、选择性和特异性的高效液相色谱（HPLC）方法来定量测定氨基葡萄糖，并研究了其在估算几丁质纯度方面的应用。使用反相C8柱、用9-芴甲氧羰基氯（Fmoc-Cl）进行柱前衍生化以及紫外检测（波长=254nm）实现色谱分离。流动相由乙腈和水组成。通过进行正交试验设计（OAD）程序获得了几丁质酸水解的最佳条件（盐酸浓度、温度和加热时间），并通过上述HPLC方法测定释放的氨基葡萄糖。通过标准加入技术检查了该方法的准确性。发现该方法具有特异性，线性、准确性、精密度良好，非常适合定量生物材料和食品中的氨基葡萄糖以及测定几丁质的纯度。
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