Aims and Objectives: The aim of this study is to assess the presence of nicotine in saliva among smokers and non-smokers. Materials and Methods: Unstimulated whole saliva was collected by asking the patients to spit in a cup that is able to measure 1ml of saliva. Testing was done with One (+) step strip as per the manufacturer’s instructions, the presence of nicotine was identified after 10 minutes.s Results: The saliva One (+) step strip seems to be valid, highly sensitive and specific method for validating smoking status and may have clinical applications in selected medical settings. In this study the participants were divided into 3 groups and a review was conducted once in 5 days. The results obtained were statistically significant. Conclusion: Smokers had significantly higher values of nicotine compared to that of nonsmokers. Nicotine presence proved to be a useful biomarker of recent smoking and can be used in epidemiological studies and smoking cessation programs.