Background: Chewing smokeless tobacco has been a common addiction in Southeast Asia for many years. The present study assessed gutkha's effect on serum levels of vitamin B12 and folic acid. Materials & Methods: 60 subjects of both genders were divided into 3 groups of 20 each. Group I was smokers, group II was tobacco chewers and group III was control. In group I, the cases who smoked daily for >/+ 6 months; in group II, the cases who chewed tobacco or consumed gutkha daily for >/= 6 months. VB12 and FA levels were estimated through the UV-spectrophotometry method. Results: Out of 60 subjects, males were 38 and females were 22. the mean vitamin B- 12 level in group I was 1855.3 pg/ml, in group II was 208.5 pg/ml, and in group III was 235.2 pg/ml The difference was significant (P< 0.05). The mean folic acid in group I was 2.8 ng/ml, in group II was 2.2 ng/ml, and in group III was 16.4 ng/ml. The difference was significant (P< 0.05). Conclusion: Compared to non-smokers, smokers had lower blood levels of folic acid, which may have a role in the onset of vascular and cardiovascular disorders. There is a statistically significant correlation between vitamin B12 deficiency and tobacco usage.