Simultaneous measurement of L-dopa, its metabolites and carbidopa in plasma of parkinsonian patients by improved sample pretreatment and high-performance liquid chromatographic determination

J Chromatogr. 1990 Jul 6:511:167-76. doi: 10.1016/s0021-9673(01)93282-0.

Abstract

A procedure is described for the determination of L-3,4-dihydroxyphenylalanine (L-DOPA), its metabolites and carbidopa (CD) in plasma of Parkinsonian patients by high-performance liquid chromatography with dual working-electrode coulometric electrochemical detection. An efficient sample preparation scheme is presented for the isolation of L-DOPA, its metabolites and the catecholamines from the same plasma aliquot. After a simple deproteinization with methanol containing 2% of 0.5 M perchloric acid and evaporation of the solvent, L-DOPA, its metabolites and CD were separated with a 5-micron Nucleosil C18 column. Catecholamines were extracted from the supernatant of the deproteinized plasma by ion exchange on small columns and adsorption on alumina. Recoveries were close to 100% for L-DOPA, its metabolites and CD and 70% for catecholamines. The use of the same mobile phase for the concurrent assay of L-DOPA, its metabolites and catecholamines considerably increased the throughput of samples in the chromatographic system. The dual-electrode coulometric detector afforded peak identification by comparing current ratios. Monitoring of data from patients under L-DOPA therapy is reported.

MeSH terms

  • Carbidopa / blood*
  • Catecholamines / blood
  • Catechols / blood
  • Chromatography, High Pressure Liquid
  • Electrochemistry
  • Humans
  • Hydrogen-Ion Concentration
  • Indicators and Reagents
  • Levodopa / blood*
  • Parkinson Disease / blood*

Substances

  • Catecholamines
  • Catechols
  • Indicators and Reagents
  • Levodopa
  • Carbidopa