A prophylaxis paste is used for removal of exogenous stains, pellicle, materia alba, and oral debris to make tooth surface smoother and cleaner. In addition, raw materials for prophylaxis pastes are available in Thailand. Contrariwise, all the prophylaxis pastes are imported from aboard. Because of this, it will be very worthwhile to start a research on the preparation of prophylaxis paste using local materials. Diatomite, diatomite waste as used as filter aid from brewery industry, local perlite, local silica and pumice were characterized for chemical compositions, mineral phases, particle size and morphology, and used as polishing agents. The ranges of particle size employed were 106-300 microns, 45-63 microns and smaller than 16 microns. The prepared prophylaxis pastes were composed of polishing agent, emulsifier and surfactant. The polishing test of the prepared prophylaxis pastes was performed on enamel and dentin of human teeth to measure the average surface roughness. The assessment of the results confirms that the prepared prophylaxis pastes containing fine to medium diatomite particles and fine perlite are highly competitive with a leading brand.