A mitochondria-based method for the determination of antioxidant activities using 2′,7′‐dichlorofluorescin diacetate oxidation

This paper reports a physiologically relevant and rapid mitochondria-based assay for assessing antioxidant properties of pure compounds and herb extracts. The principle of this method is antioxidants can scavenge peroxyl radicals generated in mitochondria thus to inhibit 2′,7′-dichlorofluorescin diacetate oxidation. Ascorbic acid, chlorogenic acid and berberine showed a dose-dependent antioxidative capacity; interestingly, lutein, Rehmannia glutinosa and Scrophularia ningpoensi exhibited pro-antioxidant properties. The antioxidant properties of herb extracts obtained by this method are in good agreement with the results measured by cell-based assay (r = 0.931, p < 0.001). This novel assay has acceptable accuracy, precision, reproducibility and the mitochondria-based assay may be more useful than conventional assay for detecting antioxidant capacity in a variety of foods.

► A new mitochondria-based assay was proposed. ► This method can be used to assess the pro- and antioxidant properties of samples. ► The assay is more physiologically relevant than conventional chemical method. ► The assay has acceptable accuracy, precision and reproducibility.