This study defines the total sodium-23 magnetic resonance spectroscopy (23Na MRS) signal from in vivo heterotopic rat heart transplants in the early post-transplant period and examines the utility of this noninvasive method for monitoring allograft rejection. Measurements were performed at 4.7 T. Syngeneic (n = 6) and allogeneic (n = 6) donor hearts were transplanted into the neck of recipient rats. There were 27 MRS observations between days 0 and 29 post-transplant. Heart grafts were excised at various intervals post-transplant for histologic examination. Allogeneic heart grafts rejected between days 4 and 5 post-transplant while syngeneic grafts continued to beat. All hearts showed ischemic damage. Allogeneic hearts showed cellular rejection by Day 1. 23Na MRS showed a steady elevation in signal in the 3 days prior to rejection and a sharp rise after rejection. 23Na MRS accurately identified full rejection and was also sensitive to the rejection process.