Development of a Fluorescent Probe for Measurement of Singlet Oxygen Scavenging Activity of Flavonoids.

A turn-on fluorescent probe, HOCD-RB, for monitoring singlet oxygen (1O2) was developed by linking rhodamine B as fluorophore with dimethylhomoocoerdianthrone (HOCD) as 1O2 reaction site and fluorescence quencher due to the intramolecular energy transfer (ET) between rhodamine B and HOCD moieties. Upon exposure to 1O2 it rapidly forms endoperoxide with HOCD and turns on the fluorescence of rhodamine B by 18-fold. Taking advantage of the HOCD-RB probe that shows fast response, high sensitivity, and selectivity for 1O2, it is applied for imaging of endogenous 1O2 in living cells and the fluorometric assay for evaluating 1O2 quenching activity of selected common flavonoids found in our daily diets. The results show that the 1O2 scavenging activity of flavonoids depends on not only the structure of individual flavonoid but also the competitive interactions between mixed flavonoids. The best antioxidant capacity for individual and mixed flavonoids is epigallocatechin gallate and the mixture of catechin gallate with kaempferol, respectively. Overall, this work provided a new tool for detection and imaging of singlet oxygen activity in a biological system as well as an efficient fluorometric assay of 1O2 scavenging activity.