Effect of Benzoic Acid and its Chemical Analogues on Membrane Resistance against Osmotic Pressure in Isolated Cattle Erythrocytes: A Comparative Study with Other Animal Species
Published on: 2019-04-01
We determined the effects of benzoic acid and its derivatives on osmotic fragility (OF) in cattle red blood cells (RBCs) in vitro. Cattle RBCs were exposed to these substances at 0-100 mM in a buffer solution for 1 hour and then the 50% hemolysis was determined by soaking in 0.2-0.8% NaCl solution. OF was determined as the NaCl concentration inducing 50% hemolysis, for which was colorimetrically measured by concentration of released hemoglobin. Benzoic acid, heptanoic acid and cyclohexanecarboxylic acid tended to decrease OF, but this change was not statistically significant (N.S). Replacement of COOH bound to the benzene ring with PO(OH)2 or SO2OH abolished the OF response obtained by benzoic acid. Replacement with OH did not affect the OF response up to 25mM, but induced hemolysis at 50 and 100 mM. Replacement with CONH2 decreased OF and the degree of the OF-lowering effect was larger than that of benzoic acid. Some derivatives substituted with other groups (OH, CH3 or NH3) or halogens (Cl or Br) decreased or tended to decreased OF, with the degree of change in OF dependent on the positon of the substitution on the benzene ring. 2-, 3- and 4-Hydroxybenzoic acids tended to decrease OF, but these changes were N.S. 2- and 4-Aminobenzoic acids, but not 3-aminobenzoic acid, decreased OF dose-dependently. 3- and 4-Methylbenzoic acids, but not 2-metylbenzoic acid, significantly decreased OF dose-dependently. 2-, 3- and 4-Chrolobenzoic acids, and 2-, 3- and 4-bromobenzoic acids significantly decreased OF dose-dependently. The effects of the substitution with other elements on OF were dependent on the elements and their position on the benzene ring. Regression analysis using the value of all compounds tested revealed that there were significant negative correlations between the partition coefficient of compounds and their effects on OF response at 25, 50 and 100 mM. Although OF responses to benzoic acid and its derivatives in cattle RBCs were similar to those in guinea pig and sheep RBCs, they differed markedly from those in rat RBCs.