Research Article
Assay for Glycosaminoglycans by Tandem Mass Spectrometry and its Applications
Shunji Tomatsu1*, Tsutomu Shimada1, Robert W Mason1, Joan Kelly2, William A LaMarr2, Eriko Yasuda1, Yuniko Shibata3, Hideyuki Futatsumori3, Adriana M Montaño4, Seiji Yamaguchi5, Yasuyuki Suzuki6 and TadaoOrii7
1 Nemours/Alfred I. duPont Hospital for Children, Wilmington, DE, USA
2 Agilent Technologies, Inc., Wakefield, MA, USA
3 Central Research Lab., R&D Div. Seikagaku Co. Tokyo, Japan
4 Department of Pediatrics, Saint Louis University, St. Louis, Missouri, USA
5 Department of Pediatrics, Shimane University, Izumo, Japan
6 Medical Education Development Center, Gifu University, Japan
7 Department of Pediatrics, Gifu University, Gifu, Japan
- *Corresponding Author:
- Prof. Shunji Tomatsu, M.D., Ph.D.
Director, Skeletal Dysplasia Center
Nemours Biomedical Research
Nemours/Alfred I. duPont Hospital for Children
1600 Rockland Rd., Wilmington, DE. 19899-0269, USA
Tel: 302-298-7336
Fax: 302-651-6888
E-mail: stomatsu@nemours.org
Received date: December 16, 2013; Accepted date: January 30, 2014; Published date: February 01, 2014
Citation: Tomatsu S, Shimada T, Mason RW, Kelly J, LaMarr WA, et al. (2014) Assay for Glycosaminoglycans by Tandem Mass Spectrometry and its Applications. J Anal Bioanal Tech S2:006. doi: 10.4172/2155-9872.S2-006
Copyright: © 2014 Tomatsu S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Glycosaminoglycans (GAGs) are distributed in the whole body and play a variety of important physiological roles associated with inflammation, growth, coagulation, fibrinolysis, lipolysis, and cell-matrix biology. Accumulation of undegraded GAGs in lysosomes gives rise to a distinct clinical syndrome, mucopolysaccharidoses. Measurement of each specific GAG in a variety of specimens is urgently required to understand GAG interaction with other molecules, physiological status of patients, and prognosis and pathogenesis of the disease.
We established a highly sensitive and accurate tandem mass spectrometry (LC-MS/MS) method for measurements of disaccharides derived from four specific GAGs [dermatan sulfate (DS), heparan sulfate (HS), keratan sulfate (KS), and chondroitin sulfate (CS)]. Disaccharides were produced by specific enzyme digestion of each GAG, and quantified by negative ion mode of multiple reaction monitoring. Subclasses of HS and GAGs with identical molecular weights can be separated using a Hypercarbcolumn (2.0 mm×50 mm, 5 μm) with an aectonitrile gradient in ammonium acetate (pH 11.0).
We also developed a GAG assay by RapidFire with tandem mass spectrometry (RF-MS/MS). The RF system
consists of an integrated solid phase extraction robot that binds and de-salts samples from assay plates and directly injects them into a MS/MS detector, reducing sample processing time to ten seconds. RF-MS/MS consequently yields much faster throughput than conventional LC-MS/MS-based methods.
However, the RF system does not have a chromatographic step, and therefore, cannot distinguish GAGs that
have identical molecular weights. Both methods can be applied to analysis of dried blood spots, blood, and urine specimens.
In this article, we compare the assay methods for GAGs and describe their potential applications.
