Development of a UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon and dosed GLP-1 from human plasma James W. Howard Richard G. Kay Ben Jones Jaimini Cegla Tricia Tan Steve Bloom Colin Creaser 2134/26217 https://repository.lboro.ac.uk/articles/journal_contribution/Development_of_a_UHPLC-MS_MS_SRM_method_for_the_quantitation_of_endogenous_glucagon_and_dosed_GLP-1_from_human_plasma/9390740 © 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. 2017-08-24 13:40:28 Glucagon GLP-1 Endogenous Plasma LC-MS/MS Immunoassay Cross-27 validation Supercharging mobile phase additive m-NBA Chemical Sciences not elsewhere classified