Objective: A simple high performance liquid chromatographic method for the determination of ochratoxin A (OTA) in human blood serum is described. Method: The extraction procedure was simple and short, and liquid chromatographic analysis was carried out isocratically on reversed-phase C18 column, with acetonitrile : 0.08M phosporic acid : isopropanol (45:45:10) at a flow rate of 1.5ml/min, and fluorescence detection (excitation at 330 nm and emission at 460 nm). Results: The mean recovery from authentic contaminated samples (n=5) spiked at 0.3, 1, 2, 4 ng/ml OTA was 97.98% (s.d=2.03). Thirty seven Egyptian serum samples were naturally contaminated with OTA in the range 0.1-2.88 ng/ml. Conclusions: The method is technically simple, specific, cost effective, time saving, suitable for large sample and requires small amount of sample and reagents. It fulfills the criteria for routine method and could be a suitable tool for surveying OTA in human blood serum.