Background & objective: Yoga is an ancient Indian science practiced since a long time. Breathing exercises in yoga (Pranayama) is believed to have an impact on heart rate of an individual thereby strengthening the cardiac autonomic activity in both physiological & pathological conditions. Reduced heart rate variability (HRV) is considered as a predictor for increased risk of cardiac mortality. Therefore this study was conducted to assess effect of Pranayama on Heart rate variability among Healthy volunteers. Objective: To assess the impact of Pranayama on Heart rate variability among the healthy volunteers. Materials & Methods: This is a cross sectional study donein the department of Physiology Chamarajanagar Institute of medical sciences, Karnataka among 30 healthy volunteers. HRV was measured in a quiet room in supine position after 15mins of rest. The obtained data regarding the effect of Pranayama on the heart rate variability was tabulated. Statistical analysis was done using paired ‘t’ test & Pearson’s correlation was used to find out the significant association between Pranayama & HRV. Results: Heart rate variability was found to be higher among regular Pranayama practitioners. There was a significant correlation between duration of yoga practice and HRV. Conclusion: In this study, it was found that practicing Pranayama on a regular basis will have a beneficial effect on heart rate variability. The balance in the autonomic nervous system can be restored by improving the parasympathetic tone.