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A within-subject cross-over study was carried out to examine whether brushing with fluoride toothpastes would alter the composition of approximal plaque. Approximal plaque was collected from 22 subjects following brushing with toothpastes containing sodium fluoride and sodium monofluorophosphate with calcium glycerophosphate and a fluoride-free control toothpaste. Concentrations of plaque calcium, inorganic phosphorus, and fluoride were estimated. Toothbrushing with the fluoride toothpastes resulted in significantly greater concentrations of fluoride in approximal plaque when compared with the control toothpaste. However, there was no significant difference between the two fluoride toothpastes. The use of the sodium monofluorophosphate containing toothpaste resulted in significantly elevated concentrations of calcium and inorganic phosphorus in approximal plaque when compared to both sodium fluoride and control toothpastes. It was concluded that the use of a fluoride toothpaste can alter the chemical composition of approximal plaque.