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FACE Analysis as a Fast and Reliable Methodology to Monitor the Sulfation and Total Amount of Chondroitin Sulfate in Biological Samples of Clinical Importance

机译:FACE分析是一种快速可靠的方法可监测具有重要临床意义的生物样品中的硫酸盐和硫酸软骨素的总量

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摘要

Glycosaminoglycans (GAGs) due to their hydrophilic character and high anionic charge densities play important roles in various (patho)physiological processes. The identification and quantification of GAGs in biological samples and tissues could be useful prognostic and diagnostic tools in pathological conditions. Despite the noteworthy progress in the development of sensitive and accurate methodologies for the determination of GAGs, there is a significant lack in methodologies regarding sample preparation and reliable fast analysis methods enabling the simultaneous analysis of several biological samples. In this report, developed protocols for the isolation of GAGs in biological samples were applied to analyze various sulfated chondroitin sulfate- and hyaluronan-derived disaccharides using fluorophore-assisted carbohydrate electrophoresis (FACE). Applications to biologic samples of clinical importance include blood serum, lens capsule tissue and urine. The sample preparation protocol followed by FACE analysis allows quantification with an optimal linearity over the concentration range 1.0–220.0 µg/mL, affording a limit of quantitation of 50 ng of disaccharides. Validation of FACE results was performed by capillary electrophoresis and high performance liquid chromatography techniques.
机译:糖胺聚糖(GAG)由于其亲水特性和高阴离子电荷密度在各种(病理)生理过程中起着重要作用。生物样品和组织中GAG的鉴定和定量在病理条件下可能是有用的预后和诊断工具。尽管在开发灵敏而准确的GAG测定方法方面取得了显着进展,但是在样品制备和可靠的快速分析方法(可同时分析多个生物样品)方面仍存在明显的方法不足。在本报告中,已开发出用于分离生物样品中GAG的方案,以使用荧光团辅助碳水化合物电泳(FACE)分析各种硫酸软骨素硫酸盐和透明质酸衍生的二糖。具有临床重要性的生物样品的应用包括血清,晶状体囊组织和尿液。样品制备方案随后进行FACE分析,可在1.0–220.0 µg / mL的浓度范围内以最佳线性进行定量,从而提供了50 ng二糖的定量限。 FACE结果的验证通过毛细管电泳和高效液相色谱技术进行。

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