FORT and FORD: two simple and rapid assays in the evaluation of oxidative stress in patients with type 2 diabetes mellitus

Abstract

The aim of the study was to evaluate the levels of free oxygen radicals and free oxygen radicals defense in patients with newly diagnosed type 2 diabetes mellitus (T2DM). The disease seems to be involved strongly in the production of reactive oxygen species. Forty-five patients with newly diagnosed T2DM and 20 apparently healthy individuals (control group) were included in the study. Reactive oxygen species were determined using the free oxygen radicals (FORT) test, which is based on the Fenton reaction. In this method, the hydroperoxides reacted with the transition metal ions liberated from the proteins and were converted to alkoxy and peroxy radicals. The radical species produced by the reaction, which are directly proportional to the quantity of lipid peroxides, interact with an additive that forms a radical molecule. Similarly, the free oxygen radicals defense (FORD) test uses preformed stable and colored radicals and determines the decrease in absorbance that is proportional to the blood antioxidant concentration. We found that ( a) FORT levels were increased in diabetic patients (2.86 ± 0.56 mmol/L H 2O 2) compared with controls (1.87 ± 0.26 mmol/L H 2O 2) ( P < .0001) and ( b) FORD levels were lower in diabetic patients (1.23 ± 0.18 mmol/L Trolox) compared with controls (1.34 ± 0.14 mmol/L Trolox) ( P < .01). The intraassay and interassay coefficients of variation were 3.7% and 6.2%, respectively, for FORT and 4.2% and 6.6%, respectively, for FORD. Determination of free oxygen radicals and free oxygen radicals defense seems to play an important role in the generation and evaluation of oxidative stress, an imbalance between oxidants and antioxidants that can lead to oxidative damage and is involved in the pathogenesis of several diseases, such as T2DM.