Generally, clinical studies using organoleptic judges, gas chromatography, or a sulfide monitor have been employed in the assessment of treatments for the control of oral malodor. However, these studies can be expensive and time consuming. Also, for agents whose safety has not been proven, these methods are not appropriate. Therefore, in vitro assessment is a critical step toward developing a new technology or implementing changes to an existing formula. The in vitro methods employed in this study combined basic microbiology methods, such as growth inhibition and zone of inhibition experiments. In addition, a newly developed, in vitro, volatile sulfur compounds (VSC) experiment that combined the essential elements of zone of inhibition, head space analysis, and dynamic flow cell techniques was also employed. The in vitro VSC method has been validated using technologies clinically proven to control oral malodor. All in vitro experiments have demonstrated that Colgate Total Advanced Fresh toothpaste has efficacy similar to that of the original Colgate Total toothpaste and that both variants showed superior efficacy to the control toothpaste. These results have been corroborated by clinical study results.

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