Determination of picomole amounts of dopamine, noradrenaline, 3,4-dihydroxyphenylalanine, 3,4-dihydroxyphenylacetic acid, homovanillic acid, and 5-hydroxyindolacetic acid in nervous tissue after one-step purification on Sephadex G-10, using high-performance liquid chromatography with a novel type of electrochemical detection

J Neurochem. 1981 Apr;36(4):1449-62. doi: 10.1111/j.1471-4159.1981.tb00586.x.

Abstract

A determination of dopamine (DA), noradrenaline (NA), 3,4-dihydroxyphenylalanine (DOPA), 3,4-dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and 5-hydroxyindolacetic acid (5-HIAA) in nervous tissue is described. The method is based on a rapidly performed isolation of DA, NA, DOPA, DOPAC, HVA, and 5-HIAA from one single nervous tissue sample on small columns of Sephadex G-10, followed by reverse-phase high-performance liquid chromatography with electrochemical detection. A new type of electrochemical detector based on a rotating disk electrode (RDE) was used. The rotating disc electrode was found to be a reliable and sensitive amperometric detector with several advantages over the currently used thin-layer cells. The detector appeared very useful for routine analysis. Practical details are given for the routine use of the RDE. Brain samples containing no more than 75-150 pg (DA, DOPA, DOPAC, HVA, and 5-HIAA) or 500 pg (NA) could be reproducibly assayed with high recovery (approx. 85%) and precision (approx. 5%), without the use of internal standards. Endogenous concentrations of DA, NA, DOPA, DOPAC, HVA, and 5-HIAA were determined in eight brain structures.

MeSH terms

  • 3,4-Dihydroxyphenylacetic Acid / analysis
  • Animals
  • Biogenic Amines / analysis*
  • Chromatography, Gel
  • Chromatography, High Pressure Liquid / methods
  • Dihydroxyphenylalanine / analysis
  • Dopamine / analysis
  • Electrochemistry
  • Electrodes
  • Female
  • Homovanillic Acid / analysis
  • Hydroxyindoleacetic Acid / analysis
  • Microchemistry
  • Nerve Tissue / analysis*
  • Norepinephrine / analysis
  • Rats

Substances

  • Biogenic Amines
  • 3,4-Dihydroxyphenylacetic Acid
  • Hydroxyindoleacetic Acid
  • Dihydroxyphenylalanine
  • Dopamine
  • Norepinephrine
  • Homovanillic Acid