TY - JOUR
T1 - High-throughput synchronous erythrocyte cellular antioxidant activity and protection screening of phenolic-rich extracts
T2 - Protocol validation and applications
AU - Cruz, Thiago Mendanha
AU - Lima, Amanda dos Santos
AU - Silva, Alessandra Oliveira
AU - Mohammadi, Nima
AU - Zhang, Liang
AU - Azevedo, Luciana
AU - Marques, Mariza Boscacci
AU - Granato, Daniel
N1 - Publisher Copyright:
© 2023 Elsevier Ltd
PY - 2024/5/15
Y1 - 2024/5/15
N2 - 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.
AB - 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.
KW - AAPH
KW - Haemolysis
KW - In vitro assays
KW - Phenolic compounds
KW - Reactive oxygen species
KW - Red blood cells
KW - TBARS
UR - http://www.scopus.com/inward/record.url?scp=85182404383&partnerID=8YFLogxK
U2 - 10.1016/j.foodchem.2023.138281
DO - 10.1016/j.foodchem.2023.138281
M3 - Article
C2 - 38160597
AN - SCOPUS:85182404383
SN - 0308-8146
VL - 440
JO - Food Chemistry
JF - Food Chemistry
M1 - 138281
ER -