Determination of extracellular glutathione in livers of anaesthetized rats by microdialysis with on-line high-performance liquid chromatography.
An on-line analytical system for the continuous in vivo monitoring of extracellular glutathione (gamma-glutamyl-cysteinylglycine) (GSH) concentrations in the livers of anaesthetized rats was developed. Microdialysates perfused through implanted microdialysis probes were collected with a loading loop of an on-line injector for direct and automatic injection into a high-performance liquid chromatographic system equipped with an electrochemical detector with Au-Hg electrodes. This method shortened the analysis time and circumvented the sample preparation process which is essential for accurate determination of GSH levels in biological samples. Additionally, this method provided continuous and real-time monitoring of extracellular GSH levels. Basal extracellular GSH concentrations in the livers of anaesthetized rate were found to vary over a wide range (from 4.16 to 76.5 microM). The method was applied to study the effect of global liver ischaemia on extracellular GSH concentrations and it was found that extracellular GSH levels in livers increased immediately with the onset of ischaemia and remained elevated for the 30-min ischaemic period. Ensuing reperfusion did reduce the GSH increase; however, the GSH levels did not return to the basal value.
Yang
C S
CS
Department of Medical Research, Taichung Veterans General Hospital, Taiwan.
Tsai
P J
PJ
Chen
W Y
WY
Liu
L
L
Kuo
J S
JS
D000758:Anesthesia; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004563:Electrochemistry; D005978:Glutathione / Q000378:metabolism*; D007511:Ischemia / Q000378:metabolism; D008099:Liver / Q000098:blood supply / Q000378:metabolism*; D008297:Male; D017551:Microdialysis; D051381:Rats; D017207:Rats, Sprague-Dawley
D005978:Glutathione