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Objective: To evaluate retention of intraoral fluoride in biofilm and saliva, an experimental dentifrice containing hydrocolloid (tara gum) was used as a controlled-release system for fluoride (F)., Materials and Methods: In a triple-blind randomized crossover trial with washout, 18 individuals used the following different dentifrices for a week: 100-TGF (sodium fluoride NaF associated with tara gum, 1100 mg/L), 50-TGF (50% NaF associated with tara gum + 50% free NaF, 1100 mg/L), PC (free NaF, 1100 mg/L), TG (with tara gum and without F), and placebo (without F or tara gum). On the seventh day of dentifrice use, biofilm was collected at 1 and 12 h, and saliva was collected up to 60 min and 12 h after the last toothbrushing. F concentrations were determined by physico-chemical analysis of fluoride using the hexamethyldisiloxane-facilitated diffusion technique. Data were subjected to two-way analysis of variance (repeated measures) and Spearman's correlation coefficient (p < 0.05) testing., Results: No significant difference was observed with the same dentifrice regarding F retention in biofilm at 1 and 12 h after toothbrushing for the 100-TGF, placebo, and TG groups (p > 0.05). The highest area under the curve values in saliva were found for the 50-TGF, 100-TGF, and PC groups., Conclusion: The dentifrice containing hydrocolloid as a controlled-release system for F promoted F retention in the oral cavity, even at 12 h after brushing., Clinical Relevance: Hydrocolloid added to dentifrices as a controlled-release system for F might contribute to a higher anti-caries effect., Trial Registration: NCT02809014.