Adriamycin nephropathy (AN) is a widely used nonimmune-mediated rat model of proteinuric chronic glomerular disease and is usually induced by a single intravenous injection of doxorubicin hydrochloride (DX) into the tail vein. However, this route can be associated with local skin necrosis and variability in disease induction and poses an occupational hazard to the investigator. Here we describe a simple technique of administering DX (1.7 mg) to ketamine:xylazine-sedated adult male Wistar rats (mean +/- 1 standard deviation, 238 +/- 8 g; n = 28) by using a single substernal intracardiac injection. The procedure was associated with minimal morbidity and mortality (1 death related to anesthesia). By day 21, severe nephrotic syndrome with effacement of glomerular epithelial cell foot processes and diffuse cortical tubulointerstitial injury was induced in all animals. Therefore, intracardiac injection of DX is a safe and consistent method of inducing AN in the rat and provides an alternative to the tail-vein route.