Development of a UHPLC–MS/MS (SRM) method for the quantitation of endogenous glucagon and dosed GLP-1 from human plasma. (May 2017)

• Record Type: Journal Article
• Authors: Howard, James W; Kay, Richard G; Jones, Ben; Cegla, Jaimini; Tan, Tricia; Bloom, Steve; Creaser, Colin S
• Is Part Of: Bioanalysis. Volume 9:Number 9(2017)
• Page Start: 733
• Page End: 751
• Publication Date: 2017-05
• Subjects: cross-validation -- endogenous -- GLP-1 -- glucagon -- immunoassay -- LC–MS/MS -- m-NBA -- plasma -- supercharging mobile phase additive; Drugs -- Analysis -- Periodicals; 615.19005
• DOI: 10.4155/bio-2017-0021
• Languages: English
• ISSNs: 1757-6180

Abstract

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.