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ARS Home » Southeast Area » Little Rock, Arkansas » Arkansas Children's Nutrition Center » Research » Publications at this Location » Publication #151453

Title: HIGH-THROUPUT ASSAY OF OXYGEN RADICAL ABSORBANCE CAPACITY (ORAC) USING A MULTICANNEL LIQUID HANDLING SYSTEM COUPLED WITH A MICROPLATE FLUORESCENCE READER IN 96-WELL FORMAT

Author
item HUANG, DEJIAN
item BOXIN, OU
item HAMPSCH-WOODILL, MAUREEN
item FLANAGAN, JUDITH
item Prior, Ronald

Submitted to: Journal of Agricultural and Food Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/23/2002
Publication Date: 7/3/2002
Citation: HUANG, D., BOXIN, O., HAMPSCH-WOODILL, M., FLANAGAN, J.A., PRIOR, R.L. HIGH-THROUPUT ASSAY OF OXYGEN RADICAL ABSORBANCE CAPACITY (ORAC) USING A MULTICANNEL LIQUID HANDLING SYSTEM COUPLED WITH A MICROPLATE FLUORESCENCE READER IN 96-WELL FORMAT. JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY. 2002. v. 50(16). p. 4437-4444.

Interpretive Summary: Oxidation of various chemical components of cells is thought to be the major process leading to tissue damage and diseases of several types. Dietary antioxidants are thought to counteract these actions and provide a degree of protection against this damaging oxidative process. Cells have components that are soluble in lipids (fat) or water. Knowing the levels of antioxidants in the lipid soluble or water soluble portion of cells is important in determining how much antioxidant rich foods are required to provide protection against cellular oxidation. The oxygen radical absorbance capacity (ORAC) assay has been widely accepted as a standard tool to measure the antioxidant capacity in the nutraceutical, pharmaceutical, and food industries. However, the assay has been criticized for a lack of accessibility due to the unavailability of suitable instrumentation for automation. In addition, the manual sample of sample preparation is time-consuming and labor-intensive. The objective of this study was to develop a high-throughput instrument platform that can fully automate the ORAC assay procedure. The new instrument platform consists of a robotic eight-channel liquid handling system and a microplate fluorescence reader. By using the high-throughput platform, the efficiency of the assay is improved with at least a 10-fold increase in sample throughput over the current procedure. This method will provide a tool for the food and neutrceutical industries to standardize antioxidant products and for the nutrition community to better understand relationships between intake of phytochemical antioxidants and health outcomes.

Technical Abstract: The oxygen radical absorbance capacity (ORAC) assay has been widely accepted as a standard tool to measure the antioxidant activity in the nutraceutical, pharmaceutical, and food industries. However, the ORAC assay has been criticized for a lack of accessibility due to the unavailability of the COBAS FARA II analyzer, an instrument discontinued by the manufacturer. In addition, the manual sample preparation is time-consuming and labor-intensive. The objective of this study was to develop a high-throughput instrument platform that can fully automate the ORAC assay procedure. The new instrument platform consists of a robotic eight-channel liquid handling system and a microplate fluorescence reader. By using the high-throughput platform, the efficiency of the assay is improved with at least a 10-fold increase in sample throughput over the current procedure. The mean of intra- and interday CVs was less than15%, and the limit of detection and limit of quantitation were 5 and 6.25 uM, respectively.