Elsevier

Life Sciences

Volume 41, Issue 7, 17 August 1987, Pages 845-848
Life Sciences

Sample handling techniques when analyzing regulatory peptides

https://doi.org/10.1016/0024-3205(87)90177-9Get rights and content

Abstract

Collection of blood samples in prechilled heparinized tubes, rapid cooling and centrifugation at 4°C were found to be more important than the enzyme inhibitors aprotinin and EDTA in preserving immunoreactive neuropeptide Y. Nine months after storage of plasma in the frozen state at -20°C or -80°C the recovery of NPY was about 50% of the recovery at immediate analysis. Synthetic substance P added to guinea pig plasma at 37°C disappeared almost entirely within 30 seconds as measured by radioimmunoassay while the concentrations of neurokinin A and neuropeptide K decreased only to a minor extent during a 20 min observation period. The total concentration of immunoreactive substance P and neurokinin A in boiled aqueous and acetic acid extracts of rat dorsal spinal cord was on the other hand stable for 72 h at 4°C, 24 h at room temperature and after freezing and thawing three times. However, chromatographic analysis indicated that the immunoreactivity became increasingly more heterogenous in the samples particularly at room temperature. Acid ethanol and Sep Pak extraction of plasma samples resulted in almost 90% recovery of neuropeptide Y, neuropeptide K and calcitonin gene-related peptide while removing crossreacting substances with high molecular weight.

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There are more references available in the full text version of this article.

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