American Journal of Clinical Nutrition, Vol 24, 7-13, Copyright © 1971 by The American Society for Clinical Nutrition, Inc.

Mechanism of the hydrogen peroxide hemolysis test and its reversal with phenols

¹ From the Department of Pediatrics, Hospital of the University of Pennsylvania and the University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania

Study of the events surrounding the hemolysis of vitamin E-deficient infant red blood cells after the addition of hydrogen peroxide indicates that hydrogen peroxide is destroyed within 2-5 min of its addition but that hemolysis does not begin for approximately 30 min.

Following the addition and destruction of hydrogen peroxide, lipid peroxidation occurs and precedes hemolysis. Alpha-tocopherol and other phenolic antioxidants such as thymol, alpha-naphthol, and estradiol were found to inhibit both lipid peroxidation and hemolysis when they were added at a time when hydrogen peroxide had already been destroyed, demonstrating that lipid peroxidation is responsible for the hemolysis seen in the hydrogen peroxide hemolysis test.

Morphological studies using interference-phase-contrast microscopy and demonstration of inhibition of hemolysis by high molecular weight dextrans suggest that lipid peroxidation causes hemolysis through membrane damage that results in colloid osmotic lysis.

The studies presented suggest that vitamin E prevents hemolysis in the hydrogen peroxide test by acting as a nonspecific phenolic antioxidant that is capable of neutralizing free radicals and thereby terminates chain reaction-type lipid peroxidation.