Metabolism is essential for the survival of cells with the production of free radicals. There is increasing evidence for the involvement of such species in a variety of normal in vivo regulatory systems. When an excess of free radicals is formed, they cause destructive and lethal cellular effects by oxidizing membrane lipids, cellular proteins, DNA and enzymes. In addition, it has been recognized that oxidative stress plays a significant role in a number of diseases. For example, many studies have shown increased oxidative damage to all the major classes of biomolecules in the brains of Alzheimer’s patients. Furthermore, cancer is probably a consequence of oxidative DNA damage. Therefore, the determination methods of free radical are very important and necessary. The development of determination techniques of hydroxyl and superoxide radicals was reviewed with 62 references in the present paper, the techniques of determination mainly include spectrophotometry, fluorophotometry, chemiluminescence, and electron spin resonance, and their advantages and disadvantages were evaluated.