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Title: Development of a UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon and dosed GLP-1 from human plasma
Authors: Howard, James W.
Kay, Richard G.
Jones, Ben
Cegla, Jaimini
Tan, Tricia
Bloom, Steve
Creaser, Colin S.
Keywords: Glucagon
GLP-1
Endogenous
Plasma
LC-MS/MS
Immunoassay
Cross-27 validation
Supercharging mobile phase additive
m-NBA
Issue Date: 2017
Publisher: © 2017 Future Science Ltd
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.
Abstract: © 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.
Description: This paper is in closed access until 10th May 2018.
Version: Accepted for publication
DOI: 10.4155/bio-2017-0021
URI: https://dspace.lboro.ac.uk/2134/26217
Publisher Link: http://dx.doi.org/10.4155/bio-2017-0021
ISSN: 1757-6180
Appears in Collections:Closed Access (Chemistry)

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