Determination of Total Antioxidant Capacity by Oxygen Radical Absorbance Capacity (ORAC) Using Fluorescein as the Fluorescence Probe: First Action 2012.23
An improved method for the measurement of oxygen radical absorbance capacity (ORAC) was developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H], 9'[9H]-xanthen]-3-one) as a new fluorescence probe (ORAC(FL)). Randomly methylated beta-cyclodextrin (RMCD) was...
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| creator | BOXIN OU CHANG, Tony DEJIAN HUANG PRIOR, Ronald L |
| description | An improved method for the measurement of oxygen radical absorbance capacity (ORAC) was developed and validated using fluorescein (3',6'-dihydroxyspiro[isobenzofuran-1[3H], 9'[9H]-xanthen]-3-one) as a new fluorescence probe (ORAC(FL)). Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water-solubility enhancer for lipophilic antioxidants. 7% RMCD (w/v) in 50% acetone-H2O mixture sufficiently solubilized vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). Results indicated that fluorescein shows excellent photostability under the plate reader conditions. This ORAC(FL) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrated that the ORACFL assay is reliable and robust. The mean of intraday and interday CVs were |
| doi_str_mv | 10.5740/jaoacint.13-175 |
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Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water-solubility enhancer for lipophilic antioxidants. 7% RMCD (w/v) in 50% acetone-H2O mixture sufficiently solubilized vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). Results indicated that fluorescein shows excellent photostability under the plate reader conditions. This ORAC(FL) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrated that the ORACFL assay is reliable and robust. The mean of intraday and interday CVs were <15%; for hydrophilic ORAC, LOD and LOQ are 5 and 6.25 microM, respectively; for lipophilic ORAC, LOD and LOQ are 6.25 and 12.5 microM, respectively. It is concluded that unlike other popular methods, the ORAC(FL) assay provides a direct measure of total antioxidant capacity against the peroxyl radicals.</description><identifier>ISSN: 1060-3271</identifier><identifier>EISSN: 1944-7922</identifier><identifier>DOI: 10.5740/jaoacint.13-175</identifier><identifier>PMID: 24645517</identifier><language>eng</language><publisher>Gaithersburg, MD: AOAC International</publisher><subject>Acetone - chemistry ; Antioxidants ; Antioxidants - chemistry ; Area Under Curve ; beta-Cyclodextrins - chemistry ; Biological and medical sciences ; Buffers ; Chromans - chemistry ; Cyclodextrins ; Fluorescein ; Fluorescein - chemistry ; Fluoresceins - chemistry ; Fluorescence ; Fluorescent Dyes - chemistry ; Free Radicals - chemistry ; Fundamental and applied biological sciences. 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Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water-solubility enhancer for lipophilic antioxidants. 7% RMCD (w/v) in 50% acetone-H2O mixture sufficiently solubilized vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). Results indicated that fluorescein shows excellent photostability under the plate reader conditions. This ORAC(FL) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrated that the ORACFL assay is reliable and robust. The mean of intraday and interday CVs were <15%; for hydrophilic ORAC, LOD and LOQ are 5 and 6.25 microM, respectively; for lipophilic ORAC, LOD and LOQ are 6.25 and 12.5 microM, respectively. It is concluded that unlike other popular methods, the ORAC(FL) assay provides a direct measure of total antioxidant capacity against the peroxyl radicals.</description><subject>Acetone - chemistry</subject><subject>Antioxidants</subject><subject>Antioxidants - chemistry</subject><subject>Area Under Curve</subject><subject>beta-Cyclodextrins - chemistry</subject><subject>Biological and medical sciences</subject><subject>Buffers</subject><subject>Chromans - chemistry</subject><subject>Cyclodextrins</subject><subject>Fluorescein</subject><subject>Fluorescein - chemistry</subject><subject>Fluoresceins - chemistry</subject><subject>Fluorescence</subject><subject>Fluorescent Dyes - chemistry</subject><subject>Free Radicals - chemistry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hydrogen Peroxide - chemistry</subject><subject>Hydrogen-Ion Concentration</subject><subject>Investigations</subject><subject>Iron - chemistry</subject><subject>Oxygen - chemistry</subject><subject>Peroxides - chemistry</subject><subject>Phosphates</subject><subject>Reactive Oxygen Species - chemistry</subject><subject>Reproducibility of Results</subject><subject>Solubility</subject><subject>Water - chemistry</subject><issn>1060-3271</issn><issn>1944-7922</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkU1rFDEchwdRbK2evUlAhHqYbf55mUx6W1ZXhcJKac8hk8msKTPJmmSh-1X8tGbdrYsgOeSF55f8wlNVbwHPuGD46kEHbZzPM6A1CP6sOgfJWC0kIc_LGje4pkTAWfUqpQeMGTSYvKzOCGsY5yDOq1-fbLZxcl5nFzwKA7oLWY9o7sv-0fXaZ7TQm_JI3qFuh1aPu7X16Fb3zuyxLoXYaW_sibpc3c4XH9F9cn6NluM2RJuMdR7phPIPezrap77H0NlrtHQxZTQ3f0oQDGRG6OvqxaDHZN8c54vqfvn5bvG1vll9-baY39SGY8jlrw2WAtPBkrbDgzVMgmg63lBMBJc9w1RIYWVrhCUNMAOMd0Ba1racdbqnF9Xl4d5NDD-3NmU1udJuHLW3YZsUcCwZYAqsoO8P6FqPVjk_hBy12eNqTksHyWUDhZr9hyqjt5MzwdvBlfN_AleHgIkhpWgHtYlu0nGnAKu9Z_XkWQFVxXNJvDt23naT7f_yT2IL8OEI6FREDbE4cunEtVhIhoH-BuTSryw</recordid><startdate>20131101</startdate><enddate>20131101</enddate><creator>BOXIN OU</creator><creator>CHANG, Tony</creator><creator>DEJIAN HUANG</creator><creator>PRIOR, Ronald L</creator><general>AOAC International</general><general>Oxford University Press</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20131101</creationdate><title>Determination of Total Antioxidant Capacity by Oxygen Radical Absorbance Capacity (ORAC) Using Fluorescein as the Fluorescence Probe: First Action 2012.23</title><author>BOXIN OU ; CHANG, Tony ; DEJIAN HUANG ; PRIOR, Ronald L</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c501t-79609703fe28b0fec49176b56302759d403797e98c7e2614c145b12848854bad3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Acetone - chemistry</topic><topic>Antioxidants</topic><topic>Antioxidants - chemistry</topic><topic>Area Under Curve</topic><topic>beta-Cyclodextrins - chemistry</topic><topic>Biological and medical sciences</topic><topic>Buffers</topic><topic>Chromans - chemistry</topic><topic>Cyclodextrins</topic><topic>Fluorescein</topic><topic>Fluorescein - chemistry</topic><topic>Fluoresceins - chemistry</topic><topic>Fluorescence</topic><topic>Fluorescent Dyes - chemistry</topic><topic>Free Radicals - chemistry</topic><topic>Fundamental and applied biological sciences. 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Randomly methylated beta-cyclodextrin (RMCD) was introduced as the water-solubility enhancer for lipophilic antioxidants. 7% RMCD (w/v) in 50% acetone-H2O mixture sufficiently solubilized vitamin E compounds and other lipophilic phenolic antioxidants in 75 mM phosphate buffer (pH 7.4). Results indicated that fluorescein shows excellent photostability under the plate reader conditions. This ORAC(FL) was validated through linearity, precision, accuracy, and ruggedness. The validation results demonstrated that the ORACFL assay is reliable and robust. The mean of intraday and interday CVs were <15%; for hydrophilic ORAC, LOD and LOQ are 5 and 6.25 microM, respectively; for lipophilic ORAC, LOD and LOQ are 6.25 and 12.5 microM, respectively. 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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE |
| subjects | Acetone - chemistry Antioxidants Antioxidants - chemistry Area Under Curve beta-Cyclodextrins - chemistry Biological and medical sciences Buffers Chromans - chemistry Cyclodextrins Fluorescein Fluorescein - chemistry Fluoresceins - chemistry Fluorescence Fluorescent Dyes - chemistry Free Radicals - chemistry Fundamental and applied biological sciences. Psychology Hydrogen Peroxide - chemistry Hydrogen-Ion Concentration Investigations Iron - chemistry Oxygen - chemistry Peroxides - chemistry Phosphates Reactive Oxygen Species - chemistry Reproducibility of Results Solubility Water - chemistry |
| title | Determination of Total Antioxidant Capacity by Oxygen Radical Absorbance Capacity (ORAC) Using Fluorescein as the Fluorescence Probe: First Action 2012.23 |
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