Aims and Objectives:The aim of the study was to compare and to analyze the antimicrobial efficacy of 0.12% chlorhexidine and new formulated herbal mouthwash after using for 14 days. The objective was to signify whether the noval herbal combination could be a better alternative mouthwash to Chlorhexidine (CHX).Materials and Methods:This is a double-blinded, random controlled research study conducted in the Department of Oral Pathology and Microbiology. A total of 200 dental students were selected randomly, comprising of two groups, 100 in each, aged between 18 and 22 years with gingival index of score II. The first group was advised to oral rinse with 0.12% chlorhexidine mouthwash and the second group with new formulated herbal mouthwash for 14 days. Saliva samples were collected on the day 0 (baseline), followed by day 7 and 14 and microbial colony count was performed. The data obtained was statistically analyzed using SPSS version 16. Student's t-test was applied for comparison of the mean microbial count between the two groups. Repeated measures analysis of variance followed by Tukey's post hoc test was applied to assess the changes from day 0 to day 7 to day 14. The statistical significance level was set at P < 0.05.Results:Microbial colonies were reduced better in chlorhexidine group on the day 7 whereas, on day 14, greater reduction was observed in the herbal group in both gender groups with high statistical significance (P < 0.001).Conclusions:Herbal mouthwash formulation performed effectively well on long-term usage, could be used as an alternative mouthwash to overcome the disadvantages of chlorhexidine.