Human serum albumin (HSA) microspheres have been produced in the size range of 200 nm to 10 mum by the controlled addition of acetone to an aqueous solution of HSA, followed by stabilization of the formed microspheres at an elevated temperature. Microspheres produced by this acetone-heat denaturation method could be stabilized at relatively low temperatures (75-degrees-C) over a short time period (15-30 min). The acetone-heat denaturation method is different from the traditional oil/water technique for preparation of HSA microspheres, both in terms of production method and the avoidance of high temperatures (> 100-degrees-C) and extended heating times (> 30 min) for stabilization. This paper describes the influence of process conditions, such as volume of acetone and HSA concentration, on the formation of the microspheres and their morphological characteristics. A loading efficiency of 8 per cent was achieved for the HSA microspheres when using rose bengal as a model compound. The release of rose bengal from the microspheres in phosphate buffered saline at 37-degrees-C was dependent on the presence of the surfactant Tween 80, and varied from a 5 per cent release in 14 days in the absence of surfactant to a 50 per cent release in 8 h in the presence of surfactant.