Effect of Dissolved Oxygen on the Reduction of Stable Nitroxide by Ascorbic Acid: In Vitro Model for Acute Hypoxia Ischemia

The reaction between 3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (PCAM), which is a blood–brain-barrier (BBB) permeable stable cyclic nitroxide, and ascorbic acid was examined in a low dissolved oxygen-phosphate buffer (low DO-PB, 0.1 mM DO) under 8% oxygen in a glove bag or in an ordinal dissolved oxygen-phosphate buffer (ordinal DO-PB, 0.3 mM DO) under 21% oxygen in an open atmosphere with and without diethylenetriaminepentaacetic acid (DTPA). In PB with DTPA, no difference was observed between the results in the low DO-PB and those in the ordinal DO-PB. In the ordinal DO-PB without DTPA, the rate constant of the PCAM decrease was half that in the low DO-PB without DTPA. The present results indicate that high DO and coexisting transition metal ions caused the oxidation of ascorbic acid, which degraded the PCAM reduction. The stable nitroxide-ascorbic acid system without DTPA was found to be applicable to an in vitro model for the acute hypoxia ischemia.