The aim of this study was to ascertain relationship of physiological parameters with playing ability among softball players. Total 150 male softball players from the various states and universities of India were selected to participate in the study. Vital capacity of the subjects was measured with the help of the computerized spirometer. Cooper’s 12 minutes run/walk test was used to assess the aerobic fitness (VO2max) of the subjects. Speed was measured with the 50m dash. Explosive strength of the subjects was determined by using the standing vertical jump. Sit and reach test was used to measure the flexibility of the subjects. Grip strength was determined with the help of hand dynamometer. Softball playing ability of the players was assessed by AAHPERD softball skill test battery. The results revealed that the physiological parameters i.e. aerobic fitness (VO2max), explosive strength (p<0.05), speed (p<0.05), flexibility (p<0.05) and grip strength of both right (p<0.05) and left (p<0.05) hand demonstrated a significant correlation with the softball playing ability.