高效阴离子交换色谱-脉冲安培检测法测定枸杞多糖的单糖及糖醛酸组成

Monosaccharide and uronic acid composition analysis on Lycium barbarum polysaccharides in Fructus Lycii by high performance anion-exchange chromatography with pulsed amperometric detection

目的:建立高效阴离子交换色谱-脉冲安培检测法(HPAEC-PAD)测定枸杞粗多糖及其不同组分单糖和糖醛酸的种类及含量。方法:枸杞子经水提醇沉,Hiprep DEAEFF16/10柱分离,再由HPAEC-PAD法测定枸杞粗多糖及其不同组分中单糖及糖醛酸的组成。结果:测得各单糖及糖醛酸在2.5~50mg/L线性关系良好(r≥0.9990),最低检出限为0.010~0.028mg/L,日内精密度RSD为0.98%~2.20%,日间精密度RSD为1.38%~2.46%,加标回收率为94.60%~112.62%。枸杞粗多糖及其不同组分均由阿拉伯糖、氨基葡萄糖、半乳糖、葡萄糖、木糖、甘露糖、果糖、核糖、半乳糖醛酸和葡萄糖醛酸组成。结论:本研究所建立的方法简便、准确、分离效果好、灵敏度高,可用于枸杞粗多糖及其不同组分单糖及糖醛酸组成和测定的分析研究。

Objective:To develop a method for determination of monosaccharide composition of crude Lycium barbarum polysaccharides and different fractions by high performance anion-exchange chromatography with pulsed amperometric detection(HPAEC-PAD).Methods:The water-soluble crude polysaccharides were extracted from the Fructus Lycii and separated through Hiprep DEAE FF16/10,then determined monosaccharide composition in crude Lycium barbarum polysaccharides and different fractions by HPAEC-PAD.Results:The results showed that within the range of 2.5~50 mg/L had good linearity(r≥0.9990).The detection limitation was 0.010~0.028 mg/L,the intre-day precision RSD was 0.98%~2.20%,the inter-day precision RSD was 1.38%~2.46%,the recovery rates were 94.60%~112.62%.The crude polysaccharides and different fractions in Fructus Lycii were composed of fucose,arabinose,glucosamine,galactose,glucose,xylose,mannose,fructose,ribose,galacturonic acid and glucuronic acid.Conclusion:The proposed method is simple,precise and has good separation effect and high sensitivity,which can be used for the study on monosaccharide composition and content determination of crude Lycium barbarum polysaccharides and different fractions.