Wei Yuyao, Zhu Wen, Tian He, Liu Jinqiu, Chen Lei, Yi Lin, Ouyang Yilan, Zhang Zhenqing
College of Pharmaceutical Sciences and Jiangsu Key Laboratory of Neuropsychiatric Diseases, Soochow University, Suzhou, Jiangsu, 215021, China.
College of Pharmaceutical Sciences and Jiangsu Key Laboratory of Neuropsychiatric Diseases, Soochow University, Suzhou, Jiangsu, 215021, China.
J Chromatogr A. 2024 Sep 27;1733:465242. doi: 10.1016/j.chroma.2024.465242. Epub 2024 Aug 8.
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.
舒洛地希，一种类肝素药物，在临床上广泛用于预防和治疗血栓栓塞性疾病及糖尿病肾病。尽管其应用广泛，但舒洛地希的结构仍未得到充分了解。它由多种多糖组成，这些多糖具有不同的糖组成、连接方式和磺化模式，然而它们具有一些共同特征，如强亲水性、高天然电荷和相当大的多分散性，这给传统的色谱分析和在线质谱（MS）表征带来了重大挑战。在这项工作中，开发了一种结合多中心切割二维液相色谱和源内酸诱导解离（inAID）质谱的新型分析方法。舒洛地希中的三种多糖通过高效强阴离子交换色谱分离，然后在质谱分析前用二维尺寸排阻色谱进行脱盐。采用一种采用inAID技术的新型质谱策略进行在线分析，从而初步鉴定了舒洛地希多糖成分。通过对这三种多糖成分进行离线制备后的二糖组成分析，验证了结果。这种融合了多种技术的先进策略能够对这类复杂药物进行全面的结构解析，并为复杂生物分子的潜在常规分析提供了一种可行的工具。
