柱前衍生-高效液相色谱法测定浒苔多糖的组成

Analysis of monosaccharides in Enteromorpha polysaccharide by pre-column derivatization high performance liquid chromatography

中文关键词: 浒苔多糖  单糖  衍生化  高效液相色谱

英文关键词: Enteromorpha polysaccharide  Monosaccharide  Derivatization  High performance liquid chromatography (HPLC)

中文摘要:

      研究了浒苔多糖样品经三氟乙酸完全水解而无损失的最适条件，并且在优化流动相洗脱程序等色谱条件的基础上，建立了1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生-高效液相色谱法分析浒苔多糖的单糖组成及含量的方法。该方法采用配备紫外检测器的高效液相色谱仪，在250 nm波长下进行测定，外标法定量。结果表明，在此条件下，8种单糖能够良好分离，各单糖在1-100 μg/ml 范围内具有良好的线性关系，相关系数r≥0.999，回收率在76.0%-108%之间，相对标准偏差RSD<10%。采用该方法对浒苔多糖样品进行组成及含量分析，发现无论样品原料为何种浒苔，浒苔多糖均以鼠李糖为主，同时还含有葡萄糖醛酸、葡萄糖、木糖、半乳糖以及少量的甘露糖、半乳糖醛酸和岩藻糖。

英文摘要:

      Nowadays, the function of Enteromorpha polysaccharide has attracted widespread attentions. However, due to the lack of test methods, the product quality and efficacy evaluation of Enteromorpha polysaccharide were compromised. In this study, the optimal conditions under which the samples of Enteromorpha polysaccharide were completely hydrolyzed by trifluoroacetic (TFA) acid without loss. Based on the optimization of the mobile phase elution procedures and other chromatographic conditions, the pre-column derivatization- high performance liquid chromatography (HPLC) using 1-phenyl-3-methyl-5-pyrazolone (PMP) to analyze the composition and content of monosaccharides in Enteromorpha polysaccharide was established. Monosaccharides in Enteromorpha polysaccharide were determined by HPLC under 250 nm wavelength, and were quantified by external standard method. It was found that 8 species of monosaccharides in Enteromorpha polysaccharide can be completely separated under the chromatographic condition. The monosaccharides had good linear relation in the range of 1-100μg/ml, and its correlation coefficient was equal to or greater than 0.999. The average recovery rate was between 76% and 108%, and the relative standard deviation (RSD) was below 10%. The strength of this method is the ability to analyze the composition of Enteromorpha polysaccharide, and it is found in this study that the most important monosaccharides in Enteromorpha polysaccharide were rhamnose; glucuronic acid, glucose, xylose, galactose, and small amounts of mannose, galacturonic acid and fucose were also detected.