Stability of glucagon-like peptide 1 and glucagon in human plasma.
Stability of glucagon-like peptide 1 and glucagon in human plasma.
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DOI：
10.1530/ec-14-0126
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发表时间：
2015-03
影响因子：
2.9
通讯作者：
Holst JJ
中科院分区：
文献类型：
--
作者：
Wewer Albrechtsen NJ;Bak MJ;Hartmann B;Christensen LW;Kuhre RE;Deacon CF;Holst JJ
研究血浆中胰高血糖素样肽1（GLP-1）和胰高血糖素在短期和长期储存条件下的稳定性。向其中加入二肽基肽酶4（DPP 4）抑制剂和抑肽酶的合并人血浆（n=20）中掺入合成GLP-1（完整，7- 36 NH 2以及初级代谢产物GLP-1 9- 36 NH 2）或胰高血糖素。在室温或冰上保存1小时和3小时、用各种酶抑制剂处理、经过最多三次解冻-再冷冻循环以及在-20 ° C和-80 °C下储存长达1年后，测量了样品中的肽回收率。 回收率不受冷冻循环或血浆在冰上储存长达3 h的影响，但当样品在RT下放置超过1 h时，回收率受损。 通过添加DPP 4抑制剂（无冰）增加了完整GLP-1的回收率，但中性内肽酶24.11抑制剂或抑制剂混合物未进一步改善。GLP-1（而非胰高血糖素）至少稳定1年。令人惊讶的是，无论储存时间如何，与立即分析相比，冷冻使胰高血糖素的回收率降低了近50%。血浆处理程序可显著影响后续激素分析的结果。我们的数据支持添加DPP 4抑制剂用于GLP-1测量以及GLP-1和胰高血糖素在冰上冷却。冻融循环未显著影响GLP-1或胰高血糖素的稳定性。无论储存温度如何，长期储存可能会影响胰高血糖素水平，应谨慎解释结果。
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.