title: Development of a high-throughput UHPLC-MS/MS (SRM) method for the quantitation of endogenous glucagon from human plasma.
abstract: 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: LGC Limited, Cambridgeshire, CB7 5WW, UK.
authors.forename: James W
authors.initials: JW
authors.lastname: Howard
authors.forename: Richard G
authors.initials: RG
authors.lastname: Kay
authors.forename: Tricia
authors.initials: T
authors.lastname: Tan
authors.forename: James
authors.initials: J
authors.lastname: Minnion
authors.forename: Mohammad
authors.initials: M
authors.lastname: Ghatei
authors.forename: Steve
authors.initials: S
authors.lastname: Bloom
authors.forename: Colin S
authors.initials: CS
authors.lastname: Creaser
mesh_terms: D000595:Amino Acid Sequence; D001774:Blood Chemical Analysis / Q000379:methods*; D001800:Blood Specimen Collection; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D005934:Glucagon / Q000097:blood* / Q000737:chemistry; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D005934:Glucagon