Heparan sulfate disaccharide measurement from biological samples using pre-column derivatization, UPLC-MS and single ion monitoring
Article
Antia, I., Yagnik, D., Pantoja Munoz, L., Shah, A. and Hills, F. 2017. Heparan sulfate disaccharide measurement from biological samples using pre-column derivatization, UPLC-MS and single ion monitoring. Analytical Biochemistry. 530, pp. 17-30. https://doi.org/10.1016/j.ab.2017.04.019
Type | Article |
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Title | Heparan sulfate disaccharide measurement from biological samples using pre-column derivatization, UPLC-MS and single ion monitoring |
Authors | Antia, I., Yagnik, D., Pantoja Munoz, L., Shah, A. and Hills, F. |
Abstract | Glycosaminoglycans are a heterogeneous family of linear polysaccharides comprised of repeating disaccharide subunits that mediate many effects at the cellular level. There is increasing evidence that the nature of these effects is determined by differences in disaccharide composition. However, the determination of GAG disaccharide composition in biological samples remains challenging and time-consuming. We have developed a method that uses derivatization and selected ion recording and RP-UPLCMS resulting in rapid separation and quantification of twelve heparin/heparin sulfate disaccharides from 5 μg GAG. Limits of detection and quantitation were 0.02–0.15 and 0.07–0.31 μg/ml respectively. We have applied this method to the novel analysis of disaccharide levels extracted from heparan sulfate and human cancer cell lines. Heparan sulfate disaccharides extracted from biological samples following actinase and heparinase incubation and derivatized using reductive amination with 2-aminoacridone. Derivatized disaccharides were analyzed used UPLC-MS with single ion monitoring. Eight HS disaccharide subunits were separated and quantified from HS and cell lines in eleven minutes per sample. In all samples the most abundant subunits present were the unsulfated ΔUA-GlcNAc, ΔUA-GlcNAc,6S and ΔUA,2S-GlcNS,6S. There was considerable variation in the proportions and concentrations of disaccharides between different cell lines. Further studies are needed to examine the significance of these differences. |
Keywords | Heparan sulfate; Cancer cell lines; Glycosaminoglycan; Proteoglycan; 2-aminoacridone; RP-UPLC-MS |
Research Group | Biomarkers for Cancer group |
Publisher | Elsevier |
Journal | Analytical Biochemistry |
ISSN | 0003-2697 |
Electronic | 1096-0309 |
Publication dates | |
Online | 30 Apr 2017 |
01 Aug 2017 | |
Publication process dates | |
Deposited | 05 May 2017 |
Accepted | 27 Apr 2017 |
Submitted | 24 Jan 2017 |
Output status | Published |
Accepted author manuscript | License |
Copyright Statement | © 2017. This author's accepted manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/ |
Digital Object Identifier (DOI) | https://doi.org/10.1016/j.ab.2017.04.019 |
Web of Science identifier | WOS:000404077200004 |
Language | English |
https://repository.mdx.ac.uk/item/86yz8
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