{"title":"Utilizing <sup>13</sup>C-Labeled internal standards to advance the analysis of heparan sulfate.","authors":"Zhangjie Wang, Jian Liu","doi":"10.1152/ajpcell.00944.2024","DOIUrl":null,"url":null,"abstract":"<p><p>Heparan sulfate (HS) is a highly sulfated and structurally heterogeneous polysaccharide that plays key roles in numerous biological processes. Due to its complex structure and variable sulfation patterns, accurately characterizing and quantifying HS in biological samples poses significant analytical challenges. This review presents an advanced high-performance liquid chromatography-tandem mass spectroscopy (LC-MS/MS) methodology that utilizes isotope-labeled internal standards for the precise quantification of HS disaccharides and rare 3-<i>O</i>-sulfated tetrasaccharides, alongside monitoring 6-<i>O</i>-endosulfatase enzyme activity and the metabolism of synthetic HS oligosaccharides in biological systems. The combination of isotope-labeled standards with LC-MS/MS technology provides a powerful and sensitive approach for comprehensive analysis of HS modifications, offering valuable insights into HS metabolism and its alterations across various biological contexts.</p>","PeriodicalId":7585,"journal":{"name":"American journal of physiology. Cell physiology","volume":" ","pages":"C1091-C1100"},"PeriodicalIF":5.0000,"publicationDate":"2025-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"American journal of physiology. Cell physiology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1152/ajpcell.00944.2024","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/2/19 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"CELL BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Heparan sulfate (HS) is a highly sulfated and structurally heterogeneous polysaccharide that plays key roles in numerous biological processes. Due to its complex structure and variable sulfation patterns, accurately characterizing and quantifying HS in biological samples poses significant analytical challenges. This review presents an advanced high-performance liquid chromatography-tandem mass spectroscopy (LC-MS/MS) methodology that utilizes isotope-labeled internal standards for the precise quantification of HS disaccharides and rare 3-O-sulfated tetrasaccharides, alongside monitoring 6-O-endosulfatase enzyme activity and the metabolism of synthetic HS oligosaccharides in biological systems. The combination of isotope-labeled standards with LC-MS/MS technology provides a powerful and sensitive approach for comprehensive analysis of HS modifications, offering valuable insights into HS metabolism and its alterations across various biological contexts.
硫酸乙酰肝素是一种高度硫酸化和结构异质性的多糖，在许多生物过程中起着关键作用。由于其复杂的结构和多变的硫酸化模式，准确表征和定量生物样品中的HS提出了重大的分析挑战。本文介绍了一种先进的LC-MS/MS方法，该方法利用同位素标记的内标来精确定量HS双糖和稀有的3- o -硫酸化四糖，同时监测6- o -硫酸化酶活性和合成HS低聚糖在生物系统中的代谢。同位素标记标准与LC-MS/MS技术的结合为HS修饰的综合分析提供了一种强大而灵敏的方法，为HS代谢及其在各种生物环境中的变化提供了有价值的见解。
期刊介绍：
The American Journal of Physiology-Cell Physiology is dedicated to innovative approaches to the study of cell and molecular physiology. Contributions that use cellular and molecular approaches to shed light on mechanisms of physiological control at higher levels of organization also appear regularly. Manuscripts dealing with the structure and function of cell membranes, contractile systems, cellular organelles, and membrane channels, transporters, and pumps are encouraged. Studies dealing with integrated regulation of cellular function, including mechanisms of signal transduction, development, gene expression, cell-to-cell interactions, and the cell physiology of pathophysiological states, are also eagerly sought. Interdisciplinary studies that apply the approaches of biochemistry, biophysics, molecular biology, morphology, and immunology to the determination of new principles in cell physiology are especially welcome.
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