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Development of a potential reference material for evaluating antioxidant activity.

Devanand L Luthria¹, Bryan T Vinyard

¹U.S. Department of Agriculture, Food Composition and Methods Development Laboratory, Beltsville Human Nutrition Research Center, Agricultural Research Service, Bldg 161, BARC (E), 10300 Baltimore Ave, Beltsville, MD 20705, USA. D.Luthria@ars.usda.gov

Journal of AOAC International

|June 24, 2008

Summary

A stable 5-compound phenolic mixture was developed for antioxidant assays. This mixture, containing diverse phenolic phytochemicals, proved stable for three months, enabling robust validation of antioxidant capacity measurements.

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Area of Science:

Analytical Chemistry
Food Science
Nutritional Biochemistry

Background:

Phenolic phytochemicals possess diverse biological functions, including antioxidant activity.
Accurate quantification of antioxidant capacity requires standardized reference mixtures.
Existing methods often use single compounds, limiting comprehensive validation.

Purpose of the Study:

To prepare and evaluate the stability of a multi-component phenolic phytochemical mixture.
To establish a reliable standard for validating antioxidant assays.
To support accurate measurement of antioxidant activity in various matrices.

Main Methods:

A 5-compound mixture including caffeic acid, morin hydrate, hesperetin, catechin hydrate, and epigallocatechin gallate was prepared.
Stability was assessed over 3 months using high-performance liquid chromatography (HPLC) with diode array detection.
Interactions between components and variations in measurements were analyzed.

Main Results:

HPLC analysis showed no significant interactions among the 5 phenolic compounds.
Among-sample relative standard deviation (RSD) for individual compounds and total area was < 1%.
Instrument and among-days RSD were < 2% and < 5%, respectively, indicating high stability.

Conclusions:

The 5-compound phenolic mixture is stable for at least 3 months when stored refrigerated.
This mixture serves as a reliable standard for the Folin-Ciocalteu and oxygen radical absorbing capacity assays.
The developed mixture facilitates robust validation of antioxidant activity quantification.