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  • Author: Monika J Bak x
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Nicolai J Wewer Albrechtsen NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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Monika J Bak NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty 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 Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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Bolette Hartmann NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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Louise Wulff Christensen NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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Rune E Kuhre NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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Carolyn F Deacon NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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Jens J Holst NNF Center for Basic Metabolic Research and Department of Biomedical Sciences, Department of Science, Faculty of Health Science, University of Copenhagen, Blegdamsvej 3B, 12.2, DK‐2200 Copenhagen N, Denmark

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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.

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