高效液相色谱法分析猪屎豆种子胶多糖中的单糖组成

Determination of carbohydrates in seed gum of Crotalaria mucronata by HPLC

关键词:  高效液相色谱法  半乳甘露聚糖  猪屎豆  种子胶  多糖分类号

Key words:  HPLC  galactomannans  Crotalaria mucronata  seed gum  polysaccharide

摘要:

建立了一种采用SUGAR SP G 0810（Pb型）糖分析色谱柱，以纯水为流动相，利用示差折光检测器的高效液相色谱外标法直接分离分析半乳甘露聚糖胶中单糖组成的方法。木糖、葡萄糖、半乳糖和甘露糖的分离在20 min内完成，检出限分别达到2.0 μg，20 μg，1.0 μg和20 μg，线性范围为2～10 mg/mL。该方法简便、快速、重现性好，用于猪屎豆种子胶多糖中单糖组分的测定，并进行回收率试验，结果5次测定的半乳糖和甘露糖的回收率分别为95.83%和103.68%，RSD分别为1.53%和1.50%。

Abstract:

A new approach by which the free monosaccharides of galactomannan were separated directly and determined by means of high performance liquid chromatography(HPLC)of external standard method on a SUGAR SP G 0810 colunm with a differentail refract meter detector and a mobile phase of pure water. The separation time for xylose(xyl)，glucose(glc)，galactose(gal)and mannose(man)was within 20 min；and the detection limits were 2.0 μg，20 μg，1.0 μg and 20 μg，respectively，while the linear dynamics varied within the range of 2～10 mg/mL. The method is rapid，simple and reproducible and can be used to the determinations of the monosaccharides in seed gum galactomannans of Crotalaria mucronata. Moreover，the recovery experiment was also conducted. The results showed that the relative standard deviations(RSD)of six time determinations to Gal and Man were 1.53% and 1.50% and the recoveries were 95.83% and 103.68%，respectively.