高效阴离子交换色谱-脉冲安培检测法测定多花黄精多糖的单糖组成

Monosaccharide composition analysis on polysaccharides in Polygonatum cyrtonema 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)测定多花黄精多糖的单糖组成。方法多花黄精多糖经提取后,用硫酸水解,用Carbo Pac PA1色谱柱(250 mm×4 mm,10.0μm)进行分离,采用梯度洗脱,HPAEC-PAD测定多花黄精多糖中单糖的组成。结果测得各单糖在1~100 mg/L线性关系良好(r>0.999),最低检出限为0.015~0.025 mg/L,RSD为1.35%~2.80%,加样回收率为88.36%~111.3%。结果表明不同产地的多花黄精多糖均由鼠李糖、阿拉伯糖、半乳糖、葡萄糖、甘露糖和果糖组成。结论本实验所建立的方法分离效果好,灵敏度高,可用于多花黄精多糖的单糖组成和测定的分析研究。

Objective To develop a method for determination of monosaccharide composition of Polygonatum cyrtonema polysaccharides（PCR） by high performance anion-exchange chromatography with pulsed amperometric detection（HPAECPAD）. Methods The water-soluble crude polysaccharides were extracted from the P. cyrtonema Hua, then the polysaccharides were hydrolyzed with sulfuric acid, then separated by Carbo Pac PA1 column（250 mm × 4 mm, 10.0 μm） and with gradient elution, determination of monosaccharide composition in PCP by HPAEC-PAD. Results The results showed that within the range of 1—100 mg/L it has good linearity（r 〉0.999）, The detection limitation was 0.015—0.025 mg/L, 1.35%—2.80% RSD, the recovery rates were 88.36%—111.3%. The results showed that the polysaccharides in P. cyrtonema Hua from different regions were composed of rhamnose, arabinose, galactose, glucose, mannose, and fructose. Conclusion The proposed method has good separation effect and high sensitivity, which can be used for the study on monosaccharide composition and content determination of PCP.