Development and validation of a fluoroimmunoassay for urinary normetanephrine

Ann Clin Biochem. 1991 Sep:28 ( Pt 5):492-6. doi: 10.1177/000456329102800512.

Abstract

The development of a separation fluoroimmunoassay for urinary normetanephrine is described. Antiserum specific to normetanephrine was coupled, using cyanogen bromide, to magnetizable cellulose; and fluorescein labelled normetanephrine was synthesized from fluoresceinthiocarbamylethylene diamine and a previously described normetanephrine derivative. Using these reagents it was possible to construct a reproducible standard curve, covering a wide range of concentrations, and to accurately measure the concentration of this metabolite in acid-hydrolysed urine samples. Cross-reactivities of structurally similar compounds were low and the fluoroimmunoassay showed good correlation with an established gas-chromatographic assay. The procedure is rapid; it is possible to accurately determine the normetanephrine concentration of urine samples approximately 2 h after hydrolysis, resulting in an overall assay time of approximately 4 h. This is the first report of a non-isotopic immunoassay for normetanephrine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cross Reactions
  • Fluorescein
  • Fluoresceins
  • Fluoroimmunoassay / methods*
  • Humans
  • Immunoglobulin G
  • Normetanephrine / urine*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Sheep

Substances

  • Fluoresceins
  • Immunoglobulin G
  • Normetanephrine
  • Fluorescein