This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.Objectives: The aims of this study were to determine the total fluoride concentration and bioavailable fluoride concentration in different toothpastes, based on a newly suggested method by the International Organization for Standardization (ISO), and to compare the measured concentrations with the concentrations written on the packaging. Methods: The concentrations of total fluoride (TF) and bioavailable fluoride (BF) were measured in six toothpastes. For the TF measurement, 1 g of each toothpaste was mixed with dipotassium hydrogen phosphate (K 2 HPO 4 ), and hydrogen chloride (HCl) was placed. After 24 hours, the samples were centrifuged and total ionic strength adjustment buffer (TISAB) solution was added. For the BF measurement, the toothpaste was mixed with K 2 HPO 4 for only 1 minute. The samples were centrifuged, and then HCl was placed and allowed to stand for 24 hours. The TISAB solution was added subsequently. The concentration of fluoride ions was measured using a fluoride ion-selective electrode and calculated against a standard curve. Results: The six toothpastes were composed of different fluoride compounds and abrasives. The measured TF concentration ranged from 624.99 ppm to 1,353.00 ppm, and the similarity to the declared fluoride concentration ranged from 53.48% to 93.31%. The measured BF concentration ranged from 587.61 ppm to 1,360.05 ppm, and the similarity to the expected fluoride concentration ranged from 41.97% to 93.80%. Two samples were clearly separated when the samples were centrifuged, whereas the remaining four samples had unclear supernatants. The clearly separated toothpastes (i.e., toothpastes 5 and 6) had BF concentrations that were similar to or lower than the declared fluoride concentrations and the measured TF concentrations. However, the unclearly separated toothpastes showed inconsistent relationships between the measured TF and BF concentrations. Conclusions: The measured TF and BF concentrations of the six toothpastes did not reach the expected fluoride concentration. This finding resulted from the different compositions and forms of the toothpastes. Therefore, the properties of toothpastes need to be considered when measuring their fluoride concentrations.