Coupling Liquid Chromatography and Tandem Mass Spectrometry to Electrophoresis for In-Depth Analysis of Glycosaminoglycan Drugs: Heparin and the Multicomponent Sulodexide,Analytical Chemistry

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Coupling Liquid Chromatography and Tandem Mass Spectrometry to Electrophoresis for In-Depth Analysis of Glycosaminoglycan Drugs: Heparin and the Multicomponent Sulodexide
Analytical Chemistry ( IF 6.7 ) Pub Date : 2020-12-28 , DOI: 10.1021/acs.analchem.0c03330
Anran Sheng ₁ , Qingqing Chen ₁ , Mengqi Yu ₁ , Ruiqi Xiao ₁ , Tianji Zhang _{2,

3} , Zhiyu Wang ₄ , Robert J. Linhardt ₅ , Xiaojun Sun ₆ , Lan Jin ₁ , Lianli Chi ₁

Affiliation

Glycosaminoglycans (GAGs) contribute to the treatment of many human diseases, especially in the field of thrombosis, because of their anticoagulant activity. GAGs interrupt the coagulation process by interacting with multiple coagulation factors through defined sequences within their linear and negatively charged chains, which are not fully elucidated. Numerous methods have been developed to characterize the structure of pharmaceutical GAGs, including intravenously or subcutaneously administered heparin and orally administered sulodexide. However, most currently available methods only focus on the oligosaccharide portion or analyze the whole mixture because longer-chain polysaccharides are extremely difficult to resolve by chromatographic separation. We have established two novel electrophoresis–mass spectrometry methods to provide a panoramic view of the structures of pharmaceutical GAGs. In the first method, an in-gel digestion procedure was developed to recover GAGs from the polyacrylamide gels, while in the second method, a strong anion exchange ultrafiltration procedure was developed to extract multiple GAG species from the agarose gels. Both procedures are compatible with liquid chromatography–tandem mass spectrometry, and structural information, such as disaccharide composition and chain length, can be revealed for each GAG fraction. The applications of these two methods on analysis of two different GAG drugs, heparin and sulodexide, were demonstrated. The current study offers the first robust tool to directly elucidate the structure of larger GAG chains with more biological importance rather than obtaining a vague picture of all chains as a mixture, which is fundamental for better understanding the structure–activity relationship and quality control of the GAG drugs.

中文翻译：

液相色谱和串联质谱联用电泳对糖胺聚糖药物：肝素和多组分舒洛糖胺进行深入分析

糖胺聚糖（GAG）由于具有抗凝活性，因此有助于治疗许多人类疾病，尤其是在血栓形成领域。GAG通过其线性和带负电荷的链中尚未明确阐明的序列与多个凝血因子相互作用，从而中断了凝血过程。已经开发出许多方法来表征药物GAG的结构，包括静脉内或皮下给予肝素和口服舒洛地特。但是，大多数当前可用的方法仅关注寡糖部分或分析整个混合物，因为长链多糖很难通过色谱分离法分离。我们建立了两种新颖的电泳-质谱法，以提供药物GAG的结构全景图。在第一种方法中，开发了凝胶内消化程序以从聚丙烯酰胺凝胶中回收GAG，而在第二种方法中，开发了强阴离子交换超滤程序以从琼脂糖凝胶中提取多种GAG。两种方法均与液相色谱-串联质谱法和结构信息（例如二糖组成和链长）兼容，可以显示每个GAG分数。证明了这两种方法在分析两种不同的GAG药物肝素和舒洛地特中的应用。当前的研究提供了第一个强大的工具，可以直接阐明具有更大生物学重要性的较大GAG链的结构，而不是获得所有链作为混合物的模糊图片，这对于更好地了解其结构-活性关系和质量控制至关重要GAG药物。

更新日期：2021-01-26

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