GC和HPLC分析金银花多糖的单糖组成方法比较

Comparison of GC and HPLC methods for analysis of monosaccharide composition of polysaccharides from Lonicera japonica

目的:GC法和HPLC法分析金银花多糖的单糖组成,并对两种方法进行比较。方法:金银花多糖经2.0 mol·L-1的三氟乙酸水解后,用糖腈乙酸酯衍生化-GC法和1-苯基-3-甲基-5-吡唑啉酮(PMP)柱前衍生化-HPLC法分别测定其单糖组成及其比例。结果:GC法检测出金银花多糖由鼠李糖(L-Rha)、阿拉伯糖(D-Ara)、甘露糖(D-Man)、木糖(D-Xyl)、葡萄糖(D-Glc)、半乳糖(D-Gal)组成;HPLC法检测到金银花多糖中含有甘露糖(D-Man)、鼠李糖(L-Rha)、葡萄糖醛酸(D-GlcA)、葡萄糖(D-Glc)、半乳糖(D-Gal)、阿拉伯糖(D-Ara)和木糖(D-Xyl),其中半乳糖(D-Gal)与阿拉伯糖(D-Ara)的色谱峰重叠。结论:两种方法均能检测出中性糖,HPLC法不仅能测中性糖,还能检测出葡萄糖醛酸,对金银花多糖的单糖组成分析GC与HPLC联合使用较为合适。

Objective:The monosaccharide composition of Lonicera japonica polysaccharides was analyzed and compared by GC and HPLC.Methods:Lonicera japonica polysaccharide was hydrolyzed by 2 mol·L-1 trifluoracetic acid.The monosaccharide composition was determined by reduction acetylation by GC method and 1–pheny-3-methyl-5-pyrazolone(PMP)pre-column derivatization by HPLC method.Results:The result of GC analysis showed that the monosaccharide composition of Lonicera japonica polysaccharide were L-Rha,D-Ara,D-Man,D-Xyl,D-Glc,D-Gal;The monosaccharide composition of Lonicera japonica polysaccharide which identified by HPLC method was L-Rha,D-Ara,D-Man,D-Xyl,D-Glc,D-Gal,and D-GlcA,where the peaks of D-Gal and D-Ara are overlapped.Conclusion:For the main neutral monosaccharide composition of the Lonicera japonica polysaccharides,the results from the two methods were basically the same,the HPLC method can not only measure neutral sugar,but also detect glucuronic acid.It is more suitable for the analysis of monosaccharide composition by the combination of GC and HPLC.