用于生物样品以及药物/营养保健品中半乳糖氨基聚糖二糖组成分析的经过验证的毛细管电泳测定法。

Validated capillary electrophoretic assays for disaccharide composition analysis of galactosaminoglycans in biologic samples and drugs/nutraceuticals.

作者信息

Asimakopoulou Athanasia P, Malavaki Christina, Afratis Nikolaos A, Theocharis Achilleas D, Lamari Fotini N, Karamanos Nikos K

机构信息

Laboratory of Biochemistry, Department of Chemistry, University of Patras, 1414, 26504, Patras, Greece.

出版信息

Methods Mol Biol. 2015;1229:129-41. doi: 10.1007/978-1-4939-1714-3_13.

DOI:10.1007/978-1-4939-1714-3_13

PMID:25325950

Abstract

Capillary electrophoresis is a separation technique with high resolving power and sensitivity with applications in glycosaminoglycan analysis. In this chapter, we present validated protocols for determining the variously sulfated chondroitin or dermatan sulfate-derived disaccharides. These approaches involve degradation of the polysaccharides with specific chondro/dermato-lyases and electrophoretic analysis with capillary zone electrophoresis in a low pH operating buffer and reversed polarity. This methodology has been applied to drug/nutraceutical formulations or to biologic samples (blood serum, lens capsule) and has been validated. Analysis of biologic tissue samples is often more demanding in terms of detection sensitivity, and thus concentration pretreatment steps and/or a derivatization step with 2-aminoacridone are often advisable.

摘要

毛细管电泳是一种分离技术，具有高分辨率和灵敏度，可应用于糖胺聚糖分析。在本章中，我们介绍了用于测定不同硫酸化程度的硫酸软骨素或硫酸皮肤素衍生二糖的经过验证的方案。这些方法包括用特定的软骨素/皮肤素裂解酶降解多糖，并在低pH操作缓冲液和反相极性下通过毛细管区带电泳进行电泳分析。该方法已应用于药物/营养制剂或生物样品（血清、晶状体囊），并已得到验证。就检测灵敏度而言，生物组织样品的分析通常要求更高，因此通常建议进行浓缩预处理步骤和/或用2-氨基吖啶进行衍生化步骤。