An organic solvent-extractable product was obtained from incubations of primaquine (70 mM) with H2O2 (70mM) and horseradish peroxidase (0.5 mg/ml) in acetate buffer, pH4.2. 2. The metabolite was characterized as 5,5-di-(8-[(4-amino-1-methylbutyl)amino]-6-methoxyquinoline) by 1H-n.m.r., mass, FT-i.r. and u.v.-visible spectroscopy. 3. Incubations of rat erythrocytes with 5,5-di-(8-[(4-amino-1-methylbutyl)amino]-6-methoxyquinoline) led to the formation of methaemoglobin in a time- and metabolite concentration-dependent manner.