Intra‐laboratory validation of microplate methods for total phenolic content and antioxidant activity on polyphenolic extracts, and comparison with conventional spectrophotometric methods

Gloria Bobo‐García; Gabriel Davidov‐Pardo; Cristina Arroqui; Paloma Vírseda; María R Marín‐Arroyo; Montserrat Navarro

doi:10.1002/jsfa.6706

journal article Open Access May 21, 2014

Intra‐laboratory validation of microplate methods for total phenolic content and antioxidant activity on polyphenolic extracts, and comparison with conventional spectrophotometric methods

Gloria Bobo‐García Gabriel Davidov‐Pardo Cristina Arroqui Paloma Vírseda

María R Marín‐Arroyo Montserrat Navarro

Journal of the Science of Food and Agriculture Vol. 95 No. 1 pp. 204-209 · Wiley

View at Publisher Save 10.1002/jsfa.6706

Abstract

AbstractBACKGROUNDTotal phenolic content (TPC) and antioxidant activity (AA) assays in microplates save resources and time, therefore they can be useful to overcome the fact that the conventional methods are time‐consuming, labour intensive and use large amounts of reagents. An intra‐laboratory validation of the Folin–Ciocalteu microplate method to measure TPC and the 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH) microplate method to measure AA was performed and compared with conventional spectrophotometric methods.RESULTSTo compare the TPC methods, the confidence intervals of a linear regression were used. In the range of 10–70 mg L−1 of gallic acid equivalents (GAE), both methods were equivalent. To compare the AA methodologies, the F‐test and t‐test were used in a range from 220 to 320 µmol L−1 of Trolox equivalents. Both methods had homogeneous variances, and the means were not significantively different. The limits of detection and quantification for the TPC microplate method were 0.74 and 2.24 mg L−1 GAE and for the DPPH 12.07 and 36.58 µmol L−1 of Trolox equivalents. The relative standard deviation of the repeatability and reproducibility for both microplate methods were ≤6.1%. The accuracy ranged from 88% to 100%.CONCLUSIONThe microplate and the conventional methods are equals in a 95% confidence level. © 2014 Society of Chemical Industry

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Details

Published: May 21, 2014
Vol/Issue: 95(1)
Pages: 204-209
License: View

Authors

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Gloria Bobo‐García