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Using high-performance liquid chromatography (HPLC), small amounts of liquid samples in which 25 premolar human teeth were immersed were evaluated. Each tooth was immersed separately in 800-ml flasks with distilled ultra-pure deionized water and remained there for 1678 h after the filling of their canals with Ca(OH)2 associated with different vehicles: group 1: polyethylene glycol and colophon (Calen); group 2: glycerin and camphorated paramonochlorophenol; group 3: camphorated paramonochlorophenol; group 4: glycerin and tricresol formol; and group 5: anesthetic solution (Citanest). Five polyethylene tubes were filled with each of these pastes and placed unsealed in similar flasks. At the end of this period, HPLC analyses of the aqueous medium related to each group were performed to detect other substances that had diffused from the pastes used in the canals of the teeth other than calcium and hydroxyl ions. Although the groups presented different maximum peaks when there was no barrier, they all showed higher values than when the tooth was present. At least 15 substances other than Ca2+ and OH− were detected in the aqueous medium of group 4. Analyzing the HPLC graphs, we concluded that not only Ca2+ and OH−, but also a considerable quantity of other components of the pastes diffused through the dentine and reached the external root surface.