Free radical scavenging activity in the nonenzymatic fraction of human saliva: a simple DPPH assay showing the effect of physical exercise.

Free-radicals and other reactive oxygen species (ROS) have been implicated as being major damaging species in pathology and they have been widely investigated. Using 1,1'-diphenyl-2-picrylhydrazyl (DPPH), we estimated total free radical scavenging activity in the low-molecular-weight nonenzymatic fraction (LMNEF) of human whole saliva. The activity of the whole saliva and serum were measured in terms of the rate of decrease in the absorbance at 517 nm in a 40% ethanol DPPH solution (pH 7.4) at room temperature. The DPPH activity of saliva and serum showed a significant linear relationship. The mean DPPH activities of saliva from 257 subjects aged 4-72 was found to be 0.389+/-.190 micromol/ml and bore no relation to age or sex. The activity in saliva of 86 subjects aged 4-11 was significantly different before and after exhaustive aerobic dance exercise for 1 hr. Physical exercise markedly decreased free radical scavenging activity in whole saliva of children. On the basis of the above results, we concluded that DPPH is useful for evaluating the total antioxidant capacity of LMNEF of human saliva.