Capillary electrophoresis in the evaluation of ischemic injury: Simultaneous determination of purine compounds and glutathione

An understanding of tissue energy metabolism and antioxidant status is of m ajor interest in the field of organ preservation for transplantation. Nucle otide and glutathione are indicators of cell damage occurring during ischem ia and reperfusion. A high performance capillary electrophoresis (HPCE) met hod with UV detection (185 nm) for the simultaneous analysis of intracellul ar free ribonucleotides, nucleosides, bases and glutathione (oxidized and r educed form) in myocardial tissues is described. The method does not involv e thiol derivatization. The separations were carried out in an uncoated fus ed-silica capillary, 60 cm long, 52.5 cm to detector, 75 mu m ID, with 20 m M Na-borate buffer, pH 10.00, at 20 kV voltage and reading at 185 nm. Injec tion was hydrostatic for 12 s and total analysis time was 20 min. The techn ique enables optimum separation of all the compounds examined and has a res olution similar to that of HPLC analysis, with the advantage of fast simult aneous measurement of cell nucleotide metabolism and redox state, not possi ble with HPLC.