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Playing venue conditions is a prominent contributor to the performance of players in a competition. Hence, we designed the present study to evaluate the effect of playing venue on the hormonal responses, psychological state and perception of effort in inter-university basketball players. Ninety-six male basketball players from eight different public and private sector universities were monitored during eight competitive matches played against each other on a home and away ground. Testosterone and cortisol levels were measured before and after each match. The psychological state test was conducted before each match and the perception of effort score was taken after the match. Elevated testosterone levels, before playing a match, as evidenced at home ground, however, a 4% decline was recorded in testosterone level at away venue, even before playing the match. Both testosterone and cortisol levels depicted the same trend on home as well as away ground. Pre-match testosterone concentration and the percent changes in these hormones were related to self-confidence, especially when playing at home and cortisol pre and post concentration related to cognitive anxiety, somatic anxiety and perception of effort. In conclusion, the playing influenced hormonal responses before the commencement of basketball matches at away ground. These hormonal responses were related to players' psychological state, which might contribute to players’ behavior and outcomes of matches.