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Implementing the Montreal Protocol has limited the use of conventional refrigerants and has led industries to find optimal or economical eco-friendly refrigerants to reduce environmental hazards. The present study analyzes an N2O-based cascade refrigeration system for achieving ultralow cooling temperatures using different eco-friendly refrigerants with low GWP and ODP in a high-temperature cycle. 3E (Energy, Exergy, and Economy) analysis of a cascade refrigeration system using eco-friendly refrigerants such as R290, R1270, RE170, R600, R600a, HFE7100, and HFE7000 in the upper cycle and N2O in the lower cycle was conducted. The R744A/R600 pair performed best among the other refrigerant pairs due to its lower discharge pressure and compression ratio. The refrigerant pair R744A/R600 results in a higher cascade cycle COP of approximately 0.87, with 48 % exergetic efficiency, while the refrigerant pair R41/HFE7100 has the lowest COP (0.68) and lowest exergetic efficiency (42 %). The minimal operational cost varies from 73006 to 76907 USD per annum for refrigerant pair R744A/R600, while for the other pairs, the cost varies from 98625 USD to 114674 USD. The R744A/R600 refrigerant pair provides COP values that are 1.38–5.38 % greater than the others, with a 1.01–3.45 % variation in exergy efficiency.