摘要
该文建立了激光诱导荧光毛细管电泳(CE-LIF)检测中药多糖和人血清糖蛋白中N-糖链的分析方法。利用8-氨基芘-1,3,6-三磺酸钠(APTS)为柱前荧光衍生试剂,结合CE分析,可在15 min内同时测定7种中性单糖。该方法应用于中药制剂和药物植物中单糖组成和含量的测定,加标回收率为88.0%~102%,相对标准偏差(RSD)小于3.0%。该方法灵敏度高、准确性好、实用可靠,适合于监控中药制剂中糖类物质的种类和含量。同时,基于毛细管凝胶电泳方法(CGE)建立了人血清中糖蛋白的N-糖链指纹图谱。利用中性涂层聚乙烯醇(PVA)有效地抑制电渗流,增加分离时间,实现了糖链异构体的有效分离。对5μL人血清进行毛细管电泳分析,共分离得到17个组分。这为进一步研究复杂生物体系血清蛋白的糖基化变化提供了可能性。
A capillary electrophoresis with laser induced fluorescence detection（CE-LIF） method was developed for the determinations of polysaccharides in traditional Chinese medicines（TCMs） and N-glycome in human serum glycoprotein in this paper.After pre column derivatized with 9-aminopyrene-1,4,6-trisulfonate（APTS） by reductive amination,the seven monosaccharides were sufficiently separated within 15 min.In the analysis of real samples,the average recoveries for the analytes were all in the range of 88.0%-102% with relative standard deviations（RSDs） lower than 3.0%.The method is sensitive,accurate and reliable,and is suitable for the determination of monosaccharide composition in Chinese medicinal preparation.Meanwhile,fingerprints of N-glycome in human serum were established based on capillary gel electrophoresis（CGE）.A poly（vinly alcohol） neutrally coated column was used to suppress the electroosmotic flow which would increase the analysis time,and the excellent separation of glycan isomers was realized.5 μL human sera was analyzed and a total of 17 fractions were isolated（RFU〉0.2）.The analysis of N-glycan profile has made it possible to research the glycosylation alterations of complex biological fluid.
出处
《分析测试学报》
CAS
CSCD
北大核心
2017年第12期1516-1521,共6页
Journal of Instrumental Analysis
基金
西安市科技计划项目(2016CXWL18)
陕西省教育厅专项科研计划项目(17JK1127)
关键词
激光诱导荧光毛细管电泳
中药多糖
N-糖基化修饰
糖蛋白
capillary electrophoresis with laser-induced fluorescence
polysaccharides from tradi-tional Chinese medicines
N-glycosylation
glycoprotein
