Aldehyde and nucleotide separations by high-performance liquid chromatography. Application to phenylhydrazine-induced damage of erythrocytes and reticulocytes.

High-performance liquid chromatographic (HPLC) techniques were used for the determination of aldehydes as lipid peroxidation products and of nucleotide concentrations in rabbit red blood cells exposed to phenylhydrazine hydrochloride. Aldehydes were determined by derivatization to dinitrophenylhydrazones, followed by thin-layer chromatographic and HPLC separation with methanol-water on an ODS column. Nucleotides and their derivatives were measured by ion-pair reversed-phase HPLC gradient elution with 10 mM ammonium phosphate buffer containing 2 mM tetrabutylammonium phosphate on ODS columns. The sensitivity of mature erythrocytes was compared with that of reticulocytes against the toxicological effects of phenylhydrazine. For both cell types an increase in aldehyde concentrations, especially of 4-hydroxynonenal, and a decline in both energy-rich nucleoside triphosphate and NAD+ were demonstrated.