Nelesen R A, Dimsdale J E, Ziegler M G
Department of Psychiatry, University of California San Diego, La Jolla 92093-0804.
Circulation. 1992 Aug;86(2):463-6. doi: 10.1161/01.cir.86.2.463.
Atrial natriuretic peptide (ANP) is a hormonal regulator of cardiovascular fluid volume. More than 1,000 scientific articles were written about ANP between 1987 and 1991. Because some articles hinted at problems with storing ANP, this study examined the effect of numerous techniques for storing and processing human ANP samples.
Samples were obtained repeatedly from three patients, treated, and stored under a variety of conditions. Experiment 1 evaluated the effects of different preservatives at 35, 21, 14, 10, and 7 days before assay. Experiment 2 evaluated nonspecific binding of ANP to different storage tubes during 28 days of storage. Experiment 3 evaluated the effect of storage at -20 degrees C, -80 degrees C, and -196 degrees C for 1 month. ANP was very unstable, degrading as much as 30% after 3 days of storage and by more than 50% in 1 month even when stored at -80 degrees C. Only storage at -196 degrees C (in liquid nitrogen) kept ANP stable for 1 month. Extraction and lyophilization of the samples before freezing and assay within 7 days of freezing only partially minimized the amount of degradation. All other processing techniques had little effect on slowing the degradation of ANP.
These findings raise disturbing questions about the interpretation of the substantial literature on ANP.
心房利钠肽(ANP)是心血管液体容量的一种激素调节剂。1987年至1991年间发表了1000多篇关于ANP的科学文章。由于一些文章暗示了ANP储存存在问题,本研究考察了多种储存和处理人ANP样本的技术的效果。
从三名患者身上反复采集样本,进行处理,并在多种条件下储存。实验1评估了在检测前35、21、14、10和7天不同防腐剂的效果。实验2评估了在储存28天期间ANP与不同储存管的非特异性结合。实验3评估了在-20℃、-80℃和-196℃储存1个月的效果。ANP非常不稳定,即使储存在-80℃,储存3天后降解高达30%,1个月内降解超过50%。只有在-196℃(液氮中)储存才能使ANP稳定1个月。在冷冻前对样本进行提取和冻干,并在冷冻后7天内进行检测,只能部分减少降解量。所有其他处理技术对减缓ANP的降解几乎没有效果。
这些发现对大量关于ANP的文献的解读提出了令人不安的问题。