Aims: The present investigation was carried out to study biochemical analysis on human permanent teeth in Dental Fluorosis, a condition caused by excess intake of fluoride. Methodology: In present study 45 human fluorosed posterior teeth were obtained from department of oral surgery and clinically classified according to Dean’s index into mild, moderate and severe forms. Fifteen posterior normal tooth samples were also collected from patients who opted for denture. The samples were than biochemically analyzed for fluoride and calcium contents by Ion Meter and Atomic Absorption Spectrophotometer respectively. Results: Biochemical analysis showed that increase in fluoride content and decrease in calcium content in fluorosed human teeth when compared to normal. Conclusion: The present study showed that pitting, perforation and tooth getting fractured (chipped off) in dental fluorosis were mainly due to accumulation of fluoride, depletion of calcium in the matrix. 

Keywords: Dental fluorosis; Ion meter; Atomic absorption spectrophotometer.