柱前衍生化高效液相色谱法分析淫羊藿多糖中单糖的组成

Analysis of Monosaccharide Compositions in Epimedium Polysaccharide by Precolumn Derivation HPLC

目的测定淫羊藿多糖(EPS)中单糖的组成及其摩尔比。方法将淫羊藿多糖样品用1 mol/L硫酸溶液降解成单糖后,用1-苯基-3-甲基-5-吡唑啉酮(PMP)衍生,通过简化衍生方法,优化分析条件,采用反相高效液相色谱250nm紫外检测和使用梯度洗脱,分离测定各单糖。结果淫羊藿多糖由甘露糖、鼠李糖、半乳糖醛酸、葡萄糖、半乳糖、阿拉伯糖6种单糖组成,其物质的量之比为0.60∶0.74∶1.00∶0.29∶2.29∶1.43。结论该改进后的衍生化方法以及优化的高效液相色谱法具有简单、快速、重现性好等特点,可用于淫羊藿多糖中单糖组成的测定。

Objective To determine the monosaccharides in the Epimedium polysaccharide（EPS） and their molar ratio. Methods The Epimedium polysaccharide were hydrolyzed into monosaccharides with 1 moL/L sulfuric acid. The monosaccharide derivatives obtained with 1 -Phenyl -3 -methyl -5 -pyrazolone （PMP） were separated by reverse -phase HPLC using a developed gradient elution process, and monitored by ultraviolet detector at 250 nm. Results Six monosaccharides derivatives were isolated with high sensitivity, good resolution by HPLC. The Epimedium polysaccharide is composed of mannose, rhamnose, galacturonic acid ,glucose, galactose, and arabonose,with a molar ratio of0.60：0.74 ： 1.00 ： 0.29 ： 2.29 ： 1.43. Conclusion This method has accurate result and good reproducibility and can be used to determine the monosaccharides in Epimedium polysaccharide.