Analysis of Total Human Urinary Glycosaminoglycan Disaccharides by Liquid Chromatography鈥揟andem Mass Spectrometry
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- 作者：Xiaojun Sun ; Lingyun Li ; Katherine H. Overdier ; Lee Anne Ammons ; Ivor S. Douglas ; Clay Cothren Burlew ; Fuming Zhang ; Eric P. Schmidt ; Lianli Chi ; Robert J. Linhardt
- 刊名：Analytical Chemistry
- 出版年：2015
- 出版时间：June 16, 2015
- 年：2015
- 卷：87
- 期：12
- 页码：6220-6227
- 全文大小：486K
- ISSN：1520-6882
文摘
The determination of complex analytes, present at low concentrations, in biological fluids poses a difficult challenge. This study relies on an optimized method of recovery, enzymatic treatment, and disaccharide analysis by liquid chromatography鈥搕andem mass spectrometry to rapidly determine low concentrations of glycosaminoglycans in human urine. The approach utilizes multiple reaction monitoring (MRM) of glycosaminoglycan disaccharides obtained from treating urine samples with recombinant heparin lyases and chondroitin lyase. This rapid and sensitive method allows the analysis of glycosaminoglycan content and disaccharide composition in urine samples having concentrations 10- to 100-fold lower than those typically analyzed from patients with metabolic diseases, such as mucopolysaccharidosis. The current method facilitates the analysis low (ng/mL) levels of urinary glycosaminoglycans present in healthy individuals and in patients with pathological conditions, such as inflammation and cancers, that can subtly alter glycosaminoglycan content and composition.