This study aimed to evaluate if the change of vehicle for CTZ (chloramphenicol, tetracycline, zinc oxide, and eugenol) paste improves the inhibition of Enterococcus faecalis in vitro. The vehicles evaluated alone and mixed with CTZ were eugenol, propylene glycol (PG), chitosan, su-per-oxidized solution (SOS), grapefruit-seed extract (GSE), and as a negative control, 0.9% saline solution. A clinical isolated of E. faecalis was morphological and biochemical characterized, in-cluding antimicrobial susceptibility was tested using 20 antimicrobial agents. Once character-ized, the clinical isolated was cultivated to performed Kirby–Bauer disc diffusion method with paper discs embedded with the different vehicles mixed or used alone, incubating at 37 ºC for 24 h. Data were analyzed by One-way ANOVA and the means were compared using Tukey test with a significance level of p<0.05. For vehicles used alone, GSE presented the greatest inhibition showing a statistically significant difference with the rest of the vehicles. When vehicles were mixed with the CTZ paste, PG showed a greater inhibition with a statistically significant differ-ence from the rest of the vehicles. In conclusion, the vehicle used to mix the CTZ paste plays an im-portant role in the inhibition of E. faecalis in vitro, therefore we consider that this can be an im-portant factor to achieve success in the use of this technique.