Structural elucidation of Sulodexide with multidimensional chromatography and online in-source acid-induced dissociation mass spectrometry

IF 3.8 2区化学 Q1 BIOCHEMICAL RESEARCH METHODS

Journal of Chromatography A Pub Date : 2024-08-08 DOI:10.1016/j.chroma.2024.465242

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引用次数: 0

Abstract

Sulodexide, a heparinoid medicine, is wildly used in clinic for prophylaxis and treatment of thromboembolic diseases and diabetic nephropathy. Despite its widespread use, the structure of Sulodexide remains poorly understood. It consists of various polysaccharides characterized by differing sugar compositions, linkages, and sulfonation patterns, yet they share common features such as strong hydrophilicity, high native charges, and considerable polydispersity, posing significant challenges for conventional chromatographic and online mass spectrometry (MS) characterization. In this work, a novel analytical method combining multiple-heart cut 2D-LC and in-source acid-induced dissociation (inAID) MS was developed. Three polysaccharides in Sulodexide were separated by high efficient strong-anion-exchange chromatography, followed by desalting with the second dimensional size-exclusion chromatography before MS. A novel MS strategy employing inAID technique was utilized for online analysis, leading to the initial identification of Sulodexide polysaccharide components. The results were validated through disaccharide composition analysis of those three polysaccharide components after offline preparation. This advanced strategy, merging various techniques, enable a comprehensive structural elucidation of such complex drugs and provides a viable tool for potential routine analysis of complex biomolecules.

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利用多维色谱法和在线源内酸诱导解离质谱法阐明舒洛地特的结构

舒洛地特是一种肝素类药物，在临床上被广泛用于预防和治疗血栓栓塞性疾病和糖尿病肾病。尽管舒洛地特被广泛使用，但人们对其结构仍然知之甚少。舒洛地特由多种多糖组成，其糖组成、连接方式和磺化模式各不相同，但它们都具有强亲水性、高原生电荷和相当大的多分散性等共同特征，这给传统色谱和在线质谱（MS）表征带来了巨大挑战。本研究开发了一种新型分析方法，将多心切割二维液相色谱和源内酸诱导解离质谱（inAID）相结合。通过高效强阴离子交换色谱分离舒洛地特中的三种多糖，然后用二维尺寸排阻色谱进行脱盐，最后进行质谱分析。在线分析中使用了一种采用 inAID 技术的新型质谱策略，从而初步鉴定了舒洛地特多糖成分。离线制备后，通过对这三种多糖成分进行双糖成分分析，验证了这一结果。这种先进的策略融合了多种技术，能够对此类复杂药物进行全面的结构阐释，并为复杂生物大分子的潜在常规分析提供了一种可行的工具。

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来源期刊

Journal of Chromatography A 化学-分析化学

CiteScore

7.90

自引率

14.60%

发文量

742

审稿时长

45 days

期刊介绍： The Journal of Chromatography A provides a forum for the publication of original research and critical reviews on all aspects of fundamental and applied separation science. The scope of the journal includes chromatography and related techniques, electromigration techniques (e.g. electrophoresis, electrochromatography), hyphenated and other multi-dimensional techniques, sample preparation, and detection methods such as mass spectrometry. Contributions consist mainly of research papers dealing with the theory of separation methods, instrumental developments and analytical and preparative applications of general interest.