BACKGROUND: 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. METHODS AND RESULTS: 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. CONCLUSIONS: These findings raise disturbing questions about the interpretation of the substantial literature on ANP.
BACKGROUND: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 humanANP samples. METHODS AND RESULTS: 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. CONCLUSIONS: These findings raise disturbing questions about the interpretation of the substantial literature on ANP.
Authors: Yan Jiang; Joel Castro; Linda V Blomster; Akello J Agwa; Jessica Maddern; Gudrun Schober; Volker Herzig; Chun Yuen Chow; Fernanda C Cardoso; Paula Demétrio De Souza França; Junior Gonzales; Christina I Schroeder; Steffen Esche; Thomas Reiner; Stuart M Brierley; Glenn F King Journal: ACS Pharmacol Transl Sci Date: 2021-06-07
Authors: Nora E Zois; Dijana Terzic; Kristine Færch; Peter Plomgaard; Jakob S Hansen; Peter Rossing; Jens P Goetze Journal: EBioMedicine Date: 2017-03-01 Impact factor: 8.143
Authors: Estefanía Tarazón; Esther Roselló-Lletí; Miguel Rivera; Ana Ortega; Maria Micaela Molina-Navarro; Juan Carlos Triviño; Francisca Lago; José Ramón González-Juanatey; Placido Orosa; José Anastasio Montero; Antonio Salvador; Manuel Portolés Journal: PLoS One Date: 2014-03-05 Impact factor: 3.240