A Novel in vitro Assay for Anti-Inflammatory Agents Based on Stabilization of Erythrocytes

Abstract

Summary

An in vitro technique for screening nonsteroidal anti-inflammatory compounds based on their ability to inhibit heat-induced hemolysis of canine erythrocytes has been demonstrated by incubating red blood cells (3.0 ml) as a 40% (v/v) suspension in M 15 sodium phosphate, pH 7.4. for 20 minutes at 53 °C with nonsteroidal anti-inflammatory agents (3.0 ml) dissolved in 0.9% saline. pH 7.4. Lysergic acid diethylamide, phenylbutazone, flufenamic and mefenamic acids, oxyphenbutazone, indomethacin, and aspirin inhibit hemolysis by 40 to 60% at concentrations of 5 × 10^-4 M or less. An inhibition of hemolysis of 28% was significantly different from zero at the 95% confidence level. To determine the specificity of the test, more than 48 nonanti-inflammatory agents were tested.