Herein, we have demonstrated the extraction, analysis, and medicinal use of the essential oil available in the Cestrum nocturnum plant. Gas Chromatography-Mass Spectrometry (GC-MS) report of the extracted oil reveals 18 different chemical constituents such as Diethylphthalate (31.88%), α-terpineol (31.52%), benzyl acetate (11.61%) and γ-terpineol (9.78%) as major products along with several other compounds. 2,2’-diphenyl-1-picrylhydrazyl (DPPH) assay is used to measure the antioxidant yields (IC50 value of 15.8±0.51μg/ml) of the extracted oil. Furthermore, the oil (6.25-100μg/ml) was assessed for its antibacterial activity against human pathogens such as Proteus mirabilis, Klebsiella pneumonia, Shigella flexneri, and Clostridium botulinum The oil revealed the highest inhibition in C. botulinum (ZOI= 38.1±0.65 mm), followed by K. pneumoniae, S. flexneri, and P. mirabilis. Further the oil was used to formulate herbal toothpaste. Various aspects of toothpaste, such as pH, stability, moisture content, homogeneity, foaming power and spread-ability, among others, were assessed and found to be quite promising. The toothpaste was also checked for its activity against Porphyromonasgingivalis, a bacterium associated with gingivitis. The formulation displayed substantial effects (ZOI=40.5±1.2 mm) with a MIC of 27 μg/mL, surpassing both the Dabur toothpaste (36±0.7 mm; MIC= 24μg/mL) and amoxicillin (38±1.2 mm). In conclusion, this study suggests that the herbal toothpaste derived from C. nocturnum seems very competitive for commercialization to prevent gingivitis.