title: Stability of glucagon-like peptide 1 and glucagon in human plasma.
abstract: To investigate the stability of glucagon-like peptide 1 (GLP-1) and glucagon in plasma under short- and long-term storage conditions. Pooled human plasma (n=20), to which a dipeptidyl peptidase 4 (DPP4) inhibitor and aprotinin were added, was spiked with synthetic GLP-1 (intact, 7-36NH2 as well as the primary metabolite, GLP-1 9-36NH2) or glucagon. Peptide recoveries were measured in samples kept for 1 and 3 h at room temperature or on ice, treated with various enzyme inhibitors, after up to three thawing-refreezing cycles, and after storage at -20 and -80 °C for up to 1 year. Recoveries were unaffected by freezing cycles or if plasma was stored on ice for up to 3 h, but were impaired when samples stood at RT for more than 1 h. Recovery of intact GLP-1 increased by addition of a DPP4 inhibitor (no ice), but was not further improved by neutral endopeptidase 24.11 inhibitor or an inhibitor cocktail. GLP-1, but not glucagon, was stable for at least 1 year. Surprisingly, the recovery of glucagon was reduced by almost 50% by freezing compared with immediate analysis, regardless of storage time. Plasma handling procedures can significantly influence results of subsequent hormone analysis. Our data support addition of DPP4 inhibitor for GLP-1 measurement as well as cooling on ice of both GLP-1 and glucagon. Freeze-thaw cycles did not significantly affect stability of GLP-1 or glucagon. Long-term storage may affect glucagon levels regardless of storage temperature and results should be interpreted with caution.
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark.
authors.forename: Nicolai J
authors.initials: NJ
authors.lastname: Wewer Albrechtsen
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark.
authors.forename: Monika J
authors.initials: MJ
authors.lastname: Bak
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark.
authors.forename: Bolette
authors.initials: B
authors.lastname: Hartmann
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark.
authors.forename: Louise Wulff
authors.initials: LW
authors.lastname: Christensen
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark.
authors.forename: Rune E
authors.initials: RE
authors.lastname: Kuhre
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark.
authors.forename: Carolyn F
authors.initials: CF
authors.lastname: Deacon
authors.affiliation: NNF Center for Basic Metabolic Research and Department of Biomedical SciencesDepartment of ScienceFaculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK-2200 Copenhagen N, Denmark jjholst@sund.ku.dk.
authors.forename: Jens J
authors.initials: JJ
authors.lastname: Holst
keywords: degradation; glucagon; glucagon-like peptide-1; samples handling; storage