In current investigation opuntia fruits were analysed for the Physico-chemical properties, Total Phenolic Content and Antioxidant Activity. The findings of thorough investigation show that the fruit contains 11⁰ Bx TSS, 4.6 pH and 0.075 % Titratable Acidity. The fruit founds great source of antioxidants (Betalains, Total Phenolic Content, Vitamin C and β-carotene), and hence it was investigated for the same. The most crucial colouring pigments Betalains were found about 564.645 mg/100g. The Total Phenolic Content was determined using Folin-ciocalteu reagent with varying solvent viz. Methanol (50%) and Distilled Water (100%). Results noted that Methanol as solvent shows 42.454 mg of GAE/100g whereas Distill Water (100%) showed about 38.736 mg of GAE/100g Total Phenolic Content. According to DPPH method the Antioxidant Activity was recorded and results depicted that fruits having 526.238 mg of AAE/100g and 513.764 mg of AAE/100g Antioxidant Activity for Methanol and Distilled Water respectively. Vitamin C was reported 31.95 mg/100g in the fruit pulp. Moreover, β-carotene was analyzed and it founds in negligible amount i.e. 0.85 µg/100g. Finally it is concluded that the Antioxidant Activity of fruit is a combined effect of Betalains, Total Phenolic Content, Vitamin C and β-carotene.