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激光诱导荧光作为肝素/硫酸乙酰肝素二糖毛细管电泳分析的强大检测工具。

Laser-induced fluorescence as a powerful detection tool for capillary electrophoretic analysis of heparin/heparan sulfate disaccharides.

作者信息

Militsopoulou Maria, Lecomte Céline, Bayle Christopher, Couderc Francois, Karamanos Nikos K

机构信息

Department of Chemistry, Section of Organic Chemistry, Biochemistry and Natural Products, Laboratory of Biochemistry, University of Patras, 261 10 Patras, Greece.

出版信息

Biomed Chromatogr. 2003 Jan;17(1):39-41. doi: 10.1002/bmc.207.

Abstract

In quest for high sensitivities necessary for determining the disaccharide composition of heparin/heparan sulfate present in trace amounts in biologic samples, an ultrahighly sensitive capillary electrophoresis (CE) method using laser-induced fluorescence (LIF) detection was developed. Heparin/heparan sulfate-derived Delta-disaccharides were derivatized with the fluorophore 2-aminoacridone and resolved by a reversed-polarity CE method. Estimation of the limit of detection in concentration term and limit of quantitation showed that LIF detection of AMAC-derivatives of Delta-disaccharides resulted in 27-744 times higher sensitivity as compared to those detected by UV at 255 nm. These data suggest that CE-LIF is a powerful tool to quantify minute amounts of heparin/heparan sulfate disaccharides.

摘要

为了获得测定生物样品中痕量存在的肝素/硫酸乙酰肝素二糖组成所需的高灵敏度,开发了一种使用激光诱导荧光(LIF)检测的超高灵敏度毛细管电泳(CE)方法。肝素/硫酸乙酰肝素衍生的Δ-二糖用荧光团2-氨基吖啶进行衍生化,并通过反相CE方法进行分离。浓度检测限和定量限的估计表明,与在255nm处通过紫外检测相比,Δ-二糖的吖啶酰胺衍生物的LIF检测灵敏度提高了27-744倍。这些数据表明,CE-LIF是定量痕量肝素/硫酸乙酰肝素二糖的有力工具。

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