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Development of a UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon and dosed GLP-1 from human plasma

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journal contribution
posted on 24.08.2017 by James W. Howard, Richard G. Kay, Ben Jones, Jaimini Cegla, Tricia Tan, Steve Bloom, Colin Creaser
© 2017 Future Science Ltd. Aim: The performance of glucagon and GLP-1 immunoassays is often poor, but few sensitive LC-MS/MS methods exist as alternatives. Experimental: A multiplexed LC-MS/MS method using a 2D extraction technique was developed. Results: The method was established for the quantitation of endogenous glucagon (LLOQ: 15 pg/ml) and dosed GLP-1 (LLOQ: 25 pg/ml) in human plasma, and is the first such method avoiding immunoenrichment. Specificity of endogenous glucagon quantitation was assured using a novel approach with a supercharging mobile phase additive to access a sensitive qualifier SRM. Endogenous glucagon concentrations were within the expected range, and showed good reproducibility after extended sample storage. A cross-validation against established immunoassays using physiological study samples demonstrated some similarities between methods. Conclusion: The LC-MS/MS method offers a viable alternative to immunoassays for quantitation of endogenous glucagon, dosed glucagon and/or dosed GLP-1.

History

School

  • Science

Department

  • Chemistry

Published in

Bioanalysis

Volume

9

Issue

9

Pages

733 - 751

Citation

HOWARD, J.W. ...et al., 2017. Development of a UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon and dosed GLP-1 from human plasma. Bioanalysis, 9(9), pp. 733-751.

Publisher

© 2017 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/

Acceptance date

17/03/2017

Publication date

2017-05-10

Notes

This paper was published in the journal Bioanalysis and the definitive published version is available at http://dx.doi.org/10.4155/bio-2017-0021.

ISSN

1757-6180

eISSN

1757-6199

Language

en

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