In this work, the use of R1234yf as a drop-in replacement for R134a in an air to water refrigeration system used to produce cool water at 7 °C is studied. This equipment was constructed in our laboratory with a minichannel heat exchanger working as condenser and a plate heat exchanger working as evaporator resulting in a 2.5 kW water chiller, which initially was working with R134a as refrigerant. A brief description of the refrigeration system and the installation used to test its behaviour as well as the uncertainty analysis is presented.Then, the test matrix defined in order to evaluate the effect of the different variables of interest on the performance of the refrigeration cycle is also presented. Air inlet temperature in the condenser was varied in order to analyse the influence of a variation in the ambient temperature. Other variables were varied in order to analyse the influence of different operating conditions as a variation in the compressor frequency or in the condenser fan velocity (by varying air inlet velocity). Finally, the subcooling degree was varied in order to compare a system with refrigerant receiver (no subcooling) and a system without refrigerant receiver and different refrigerant charge (different subcooling).According to the results obtained, R1234yf performs worse than R134a, presenting lower cooling capacity and higher compressor consumption in all tests, with a global efficiency around 25% lower. This can be explained by its lower isentropic and volumetric efficiency, which could be related to the type of lubricant used.