Five different pulses were taken and in all five pulses the protein content, carbohydrate content and phenolic content were estimated, all the pulses showed different result. All five pulses were treated with the Trypsin enzyme to check the activity of Trypsin inhibitor. The highest activity was shown in kidney bean 5.4 units/ml and then by the chickpea 7.5 units/ml, so the kidney bean was further purified though Dialysis and then SDS PAGE to analyze and to determine the Trypsin inhibitor activity and the molecular weight of Trypsin inhibitor by SDS PAGE .The Trypsin inhibitor is the enzyme which inhibits the activity of Trypsin in legumes. Further the effect of pH and temperature was done on common pulses to check the activity of inhibitor, and kidney bean showed the highest result. The molecular weight of partially purified Trypsin inhibitor was found to be in the range of 24-28kd.