High-throughput synchronous erythrocyte cellular antioxidant activity and protection screening of phenolic-rich extracts: Protocol validation and applications

Thiago Mendanha Cruz, Amanda dos Santos Lima, Alessandra Oliveira Silva, Nima Mohammadi, Liang Zhang, Luciana Azevedo, Mariza Boscacci Marques, Daniel Granato

Research output: Contribution to journalArticlepeer-review

Abstract

Oxidative/nitrosative damage takes part in chronic disease development, which generates an urgent need for intervention and better therapies to manage them. The scientific community has demanded easy-to-run, cheap, and reliable methods for cellular antioxidant activity assays. This work standardised and validated an erythrocyte cellular antioxidant activity and membrane protection/injury (HERYCA-P) protocol to study food-derive extracts. The method measures intracellular reactive oxygen species (ROS) generation, lipoperoxidation, and haemolysis induced by 2,2′-azobis(2-amidinopropane) dihydrochloride. Quercetin decreased ROS generation by 50.4% and haemolysis by 2.2%, while ascorbic acid inhibited lipid peroxidation by 40.1%. Total phenolic contents of teas were correlated with decreased ROS generation (r = −0.924), lipoperoxidation (r = −0.951), and haemolysis (r = −0.869). The erythrocyte ROS generation and lipoperoxidation were also associated with CUPRAC (r = −0.925; r = −0.951) and hydroxyl radical scavenging activity (r = −0.936; r = −0.949). The precision rates of antioxidant standards and tea samples were below 15%. HERYCA-P is feasible as a complementary antioxidant assay for food matrices.

Original languageEnglish
Article number138281
JournalFood Chemistry
Volume440
DOIs
Publication statusPublished - 15 May 2024

Keywords

  • AAPH
  • Haemolysis
  • In vitro assays
  • Phenolic compounds
  • Reactive oxygen species
  • Red blood cells
  • TBARS

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