Emotion regulation depends critically on an individual’s ability to adjust physiological arousal on a momentary basis. Heart rate variability (HRV) is a measure of the continuous interplay between sympathetic and parasympathetic influences on heart rate that yields information about autonomic flexibility and thereby represents the capacity for regulated emotional responding. Although researches still does not provide theoretical and empirical support for the emergence of Heart rate variability as a marker of emotion regulatory ability. The objective of this study was to assess Heart rate variability in response to negatively and positively valenced pictures. One hundred and two International Affective Picture System (IAPS) photographs were grouped into 3 sets of 34 photographs: neutral, positive and negative. Each picture in a set was displayed for 6 seconds followed by 3 second interval before the next picture. The study was conducted in 24 healthy male students [age= 25(24-26)]. Each student was shown three sets of stimuli and their ECG was recorded simultaneously for 5 minutes. Time and frequency domain variables of HRV were calculated and compared among different sets of stimuli. Statistical significance was tested with Friedman test. P value < 0.05 was considered significant. Sympathetic (LF pr, LFnu, LF/HF) and parasympathetic indices (SDNN, RMSSD, NN50cnt, HFpr, HFnu) of HRV analysis in three different sets of stimuli were insignificant when compared among the sets. These data suggest that the affective pictures from International Affective Picture System (IAPS) shown in our study have no effects on cardiac sympathetic and parasympathetic activity.