Quantification of the overall REACTIVE OXYGEN SPECIES scavenging capacity of biological fluids and tissues

• Published in: Free radical biology & medicine 2000-03, Vol.28 (6), p.871-879
• Main Authors: Kohen, Ron, Vellaichamy, Elangovan, Hrbac, Jan, Gati, Irith, Tirosh, Oren
• Format: Article
• Language: English
• Subjects: Animals; Antioxidant profile; Antioxidants - analysis; Ascorbic Acid - analysis; Cyclic voltammetry; Electrochemistry; Free Radical Scavengers - analysis; Free radicals; Humans; Invasive and noninvasive techniques; Liver - chemistry; Low–molecular weight antioxidants; Oxidation-Reduction; Oxidative Stress; Rats; Reactive Oxygen Species - metabolism; Skin - chemistry; Thioctic Acid - analysis; Total antioxidant capacity; Vitamin E - analysis
• ISSN: 0891-5849; 1873-4596
• DOI: 10.1016/S0891-5849(00)00191-X

A method has been developed for measuring and evaluating the overall antioxidant activity derived from the low–molecular weight antioxidants (scavengers). The principle governing this method is based on a common chemical characteristic of the scavengers, their reducing properties. It was hypothesized and then demonstrated that an evaluation of the overall reducing power of a biological sample correlates with the overall scavenging activity of the sample. In order to quantify the total reducing power, the cyclic voltammetry methodology was applied. The resulting measurements correlated with the antioxidant activity of both hydrophilic and lipophilic scavengers. The method is suitable for use in biological fluids and in tissue homogenates, and can supply information concerning the type of antioxidants and their total concentration without having to determine specific compounds. A noninvasive procedure for determining skin overall scavenging activity is also described. This method is based on a well containing an extraction solution that is attached to the skin’s surface. Following incubation time the extraction solution is analyzed using the cyclic voltammeter instrument and other methods. We have found these methods suitable for evaluating the reducing capacity status in various clinical conditions such as diabetes, ionizing and nonionizing irradiation, brain degenerative diseases, head trauma, and inflammatory bowel diseases. This method is also an efficient tool for evaluating the overall antioxidant capacity of mixtures of antioxidant preparations in vitro. The measurements themselves are simple and rapid. Furthermore, they do not require manipulation of the samples.