Monitoring of met-enkephalin in vivo with 5-min temporal resolution using microdialysis sampling and capillary liquid chromatography with electrochemical detection

J Chromatogr B Biomed Sci Appl. 1997 Dec 19;704(1-2):43-52. doi: 10.1016/s0378-4347(97)00436-2.

Abstract

A method based on microdialysis sampling and capillary liquid chromatography with electrochemical detection that allows in vivo monitoring of met-enkephalin with 5-min temporal resolution is described. Sampling was achieved using a concentric microdialysis probe made from polycarbonate membrane material with a 20 kDa cut-off. This probe had an in vitro relative recovery for met-enkephalin of 63% at a dialysis flow-rate of 0.6 microl/min. Separations were performed using 7 cm x 25 microm I.D. fused-silica capillary columns packed with 5 microm Alltima C18 particles. A carbon fiber microelectrode was used as the detector electrode. The mass detection limit for met-enkephalin with this system was 40 amol. With on-column preconcentration, up to 2 microl of sample could be loaded onto the column resulting in concentration detection limits as low as 20 pM for met-enkephalin. Direct injection of dialysate, collected at 5-min intervals, allowed determination of met-enkephalin concentrations in the rat globus pallidus under basal and K+-induced depolarization conditions.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Brain Chemistry*
  • Chromatography, High Pressure Liquid / methods*
  • Electrochemistry
  • Enkephalin, Methionine / analysis*
  • Enkephalin, Methionine / metabolism
  • Globus Pallidus / chemistry
  • Globus Pallidus / drug effects
  • Globus Pallidus / metabolism
  • Male
  • Microdialysis*
  • Potassium / pharmacology
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Enkephalin, Methionine
  • Potassium