荧光标记糖电泳在新琼寡糖定性和定量分析中的应用

Application of Fluorophore-assisted Carbohydrate Electrophoresis in Qualitative and Quantitative Analysis of Neoagaro-oligosaccharides

目的:建立荧光辅助糖电泳(FACE)对系列新琼寡糖进行定性、定量分析的方法。方法:将系列新琼寡糖用7-氨基-1,3-萘胺二磺酸钾(AGA)氨化还原衍生后,在浓度梯度为18%-25%的聚丙烯酰胺凝胶中电泳分离,于波长264nm直接检测寡糖衍生物,得到聚合度为4-14的新琼寡糖衍生物电泳分析图谱。结果:寡糖衍生物的相对电迁移率与其分子量的负三分之二次方成线性关系;运用图像分析软件对电泳图谱进行数字化转换,发现寡糖衍生物的灰度积分面积与样品浓度呈线形关系。结论:建立了快速、精确的新琼寡糖微量定性、定量分析的方法,为新琼寡糖的质量分析提供了技术支撑。

Objective： To qualitatively and quantitatively analyze neoagaro-oligosacchardes using fluorophore-assisted carbohydrate electrophoresis （FACE）. Methods： The neoagaro-oligosaccharides were ammoniated with 7-amino-1,3-naphthalene-disulfonic-acid monopotassium salt monohydrate （AGA）. The results of the fluorescent derivatives were separated in the polyacrylamide gel. The polyactylamide gel onsists of a linear gradient from 18% to 25% （w/v（ acrylamide and 1.38-1.92% （w/v） N,N＇-methylen bisacrylamide as cross-linker. Neoagaro-oligosaccharide derivatives with polymerization degrees ranging from 4 to 14 were resolved in a single gel. Resuits： The relative electrophoretic mobility of the derivatives was linearly correlated with the negative two thirds power of the molecular weight. Digital analysis software was used to analyse the FACE map, raw volume of fluorescent derivatives were linearly correlated with their contents. Conclusions： This method is simple, rapid and accurate, and it is useful for the exploitation and quality control of neoagaro-oligosaccharides.