Quantitative Analysis of Oligosaccharides Derived from Sulfated Glycosaminoglycans by Nanodiamond-Based Affinity Purification and Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry

详细信息    查看全文

• 作者：Chih-Chien Hsieh ; Jiun You Guo ; Shain-Un Hung ; Rui Chen ; Zongxiu Nie ; Huan-Cheng Chang ; Chih-Che Wu
• 刊名：Analytical Chemistry
• 出版年：2013
• 出版时间：May 7, 2013
• 年：2013
• 卷：85
• 期：9
• 页码：4342-4349
• 全文大小：377K
• 年卷期：v.85,no.9(May 7, 2013)
• ISSN：1520-6882

文摘

Degraded fragments of sulfated glycosaminoglycans (GAGs) are key reporters for profiling the burden of mucopolysaccharidosis (MPS) disease at baseline and during therapy. Here, we present a high-throughput assay, which combines microwave-assisted degradation, solid-phase affinity purification, and matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS), for quantitative analysis of sulfated oligosaccharides in biological samples. First, sulfated oligosaccharides such as chondroitin-4-sulfate (CS) were efficiently isolated from highly diluted solutions or spiked artificial cerebrospinal fluid (aCSF) using polyarginine-coated nanodiamonds (PA-coated NDs) as affinity sorbents. Next, they were degraded to disaccharides through microwave-assisted methanolysis or enzymatic digestion for subsequent MALDI-TOF MS analysis. The reaction times for GAG depolymerization were significantly reduced from a few hours to less than 7 min under the microwave irradiation. Deuterium-labeled internal standards were then mixed with the CS-derived disaccharides for quantitative analysis by MALDI-TOF MS using the N-(1-naphthyl) ethylenediamine dihydrochloride (NEDC) matrix. The new assay is facile, specific (with distinct chlorine-isotope trait markers), sensitive (with a detection limit of 70 pg), and potentially useful for clinical diagnosis of MPS.