The overall objective of the proposal is to develop robust, high through-put assays to map the total antioxidant profile for non-enzymatic dietary antioxidants. APPROACH: In Phase I, we will explore the feasibility of extending the most widely used oxygen radical absorbance capacity assay (ORAC) to other ROS/RNS. In particular, we will determine the feasibility of using fluorescein and its derivatives as the probes for superoxide ion, hydrogen peroxide singlet oxygen and peroxynitrite. In addition, a multi-lab validation of the improved ORAC assay developed by Brunswick Laboratories will be conducted using the microplate reader technology in collaboration with Dr. Ronald Prior of the USDA and Dr. Luke Howard of the University of Arkansas. As a result, the ORAC assay should ultimately be accepted as the industrial standard for determining antioxidant capacity. NON-TECHNICAL SUMMARY: Overwhelming scientific evidence suggests that dietary antioxidants scavenge the reactive oxygen/nitrogen species (ROS/RNS) which are known to be involved in the pathogenesis of aging and many common diseases. As considerable scientific and public interest has been directed toward antioxidants and, as more food companies are entering this market, the lack of validated high through-put assays for total antioxidant capacity is negatively affecting the related science and industry. Upon the completion of Phase II, it is anticipated that validated assays will be in place to measure the "total antioxidant capacity" of hydrophilic and lipophilic antioxidants against the common free radicals found in the human body. Brunswick Laboratories will market these assays as part of our analytical chemistry capabilities to our existing customer base of 200+ companies, USA and international-Fortune 100, human food and pet food companies, functional food formulators, fruit and vegetable growers, nutritional supplement companies, and researchers doing clinical trials and bioavailability studies, and eventually on food labels for consumers and regulatory purposes.
Keywords: total antioxidant capacity; fluorescence; assays; high throughput; ros/rns