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Development of a high-throughput UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon from human plasma

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journal contribution
posted on 02.10.2017 by James W. Howard, Richard G. Kay, Tricia Tan, James Minnion, Mohammad Ghatei, Steve Bloom, Colin Creaser
© 2014 Future Science Ltd. Background: Published LC-MS/MS methods are not sensitive enough to quantify endogenous levels of glucagon. Results: An ultra high performance liquid chromatography-MS/MS (SRM) method for the quantitation of endogenous levels glucagon was successfully developed and qualified. A novel 2D extraction procedure was used to reduce matrix suppression, background noise and interferences. Glucagon levels in samples from healthy volunteers were found to agree with radioimmunoassay (RIA) derived literature values. Bland-Altman analysis showed a concentration-dependent positive bias of the LC/MS-MS assay versus an RIA. Both assays produced similar pharmacokinetic profiles, both of which were feasible considering the nature of the study. Conclusion: Our method is the first peer reviewed LC-MS/MS method for the quantitation of endogenous levels of glucagon, and offers a viable alternative to RIA-based approaches.

History

School

  • Science

Department

  • Chemistry

Published in

Bioanalysis

Volume

6

Issue

24

Pages

3295 - 3309

Citation

HOWARD, J.W. ...et al., 2014. Development of a high-throughput UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon from human plasma. Bioanalysis, 6(24), pp. 3295-3309.

Publisher

© Future Science Ltd

Version

AM (Accepted Manuscript)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/

Publication date

2014

Notes

This paper was accepted for publication in the journal Bioanalysis and the definitive published version is available at http://dx.doi.org/10.4155/bio.14.226

ISSN

1757-6180

eISSN

1757-6199

Language

en

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