Development of a potential reference material for evaluating antioxidant activity

J AOAC Int. 2008 May-Jun;91(3):506-10.

Abstract

Phenolic phytochemical are known to perform several functions ranging from phytoprotectants, to protecting lipids in food products, to antioxidant activity in animals and humans. The need for a common standard mixture containing multiple phenolic phytochemicals is critical for the development of a robust validation assay for accurately quantifying antioxidant activity in various matrixes. Different research groups have used A wide array of single purified reference phenolic compounds in this regard. A 5 compound mixture (caffeic acid, morin hydrate, hesperetin, catechin hydrate, and epigallocatechin gallate) containing phenolic compounds from 4 subgroups (phenolic acid, flavone, flavanone, and flavan-3-ol) was prepared. The mixture was assayed for stability evaluation by high-performance liquid chromatography (HPLC) using a diode array detection procedure for a 3 month time interval. HPLC analysis confirmed that there was no significant interaction between different components of the mixture. The among-sample relative standard deviation (RSD) of all 5 phenolic compounds, as well as the total HPLC area, was < 1%. The RSD due to instrument variation was < 2% and the total RSD among-days was < 5%. These results unambiguously suggest that the sample was stable for a 3 month time interval in an amber vial stored in a refrigerator below 5 degrees C. This mixture is currently being used for the single-laboratory validation study for the assay of total phenolic content by the Folin-Ciocalteu method and the antioxidant capacity by oxygen radical absorbing capacity procedure.

MeSH terms

  • Antioxidants / analysis*
  • Antioxidants / standards*
  • Caffeic Acids / analysis
  • Caffeic Acids / standards
  • Catechin / analogs & derivatives
  • Catechin / analysis
  • Catechin / standards
  • Chromatography, High Pressure Liquid / methods
  • Chromatography, High Pressure Liquid / standards
  • Flavonoids / analysis
  • Flavonoids / standards
  • Food Analysis / methods
  • Food Analysis / standards
  • Hesperidin / analysis
  • Hesperidin / standards
  • Phenols / analysis*
  • Phenols / standards*
  • Reference Standards

Substances

  • Antioxidants
  • Caffeic Acids
  • Flavonoids
  • Phenols
  • morin
  • Catechin
  • epigallocatechin gallate
  • Hesperidin
  • hesperetin
  • caffeic acid