Auditory reaction time (ART) and visual reaction time (VRT ) between males & females during shift duty in hospital employees have been compared. ART and VRT were studied in 286 hospital employees (141 males and 145 females) during day and night shift duty in the age group of 20 to 60 years. Subjects were presented with two auditory stimuli, i.e. high pitch and low pitch sound and two visual stimuli, i.e. red and green light. The significance of difference of ART and VRT during day and night shift duty among males and females were compared with the use of standard error of difference between two means. The statis-tical difference was determined by ‘z’ test. ART during day shift in males (215.15 ± 47.52 ) were less than ART during day shift in females (233.97 ± 44.62), VRT during day shift in males (224.01 ± 30.43) were less than VRT during day shift in females (238.98 ± 29.69). ART during night shift in males (219.96 ± 48.51) were less than ART during night shift in females (237.28 ± 44.01), VRT during night shift in males (229.20 ± 31.92) were less than VRT during night shift in females (240.60 ± 31.71). Our results indicate that in the female, ART and VRT are greater than the male during the day shift and night shift, and the difference was found to be statistically significant. Although the reaction time is found to be more during the night shift as compared to day shift, yet the difference is not signifi-cant.