This study compares the performance according to a working fluid, the number of battery cooling block ports, and header width required for cooling according to the application of the direct contact single-phase battery cooling method in a 1S16P battery module and examines the battery cooling performance according to the flow rate under the standard and summer conditions based on an optimized model. The analysis result verified that R134a showed low-pressure drop and high cooling performance as the working fluid of the direct contact single-phase cooling system in the 1S16P battery module, and R134a showed the best cooling and stability when applied with three ports and a 5 mm header. In addition, under 25 °C outdoor conditions, the maximum temperature of the battery and the temperature difference between the batteries at 3 and 5 lpm excluding 1 lpm are 30.5 °C, 4.91 °C, and 28.7 °C, 3.28 °C, indicating that the flow rate of refrigerant was appropriate for battery safety. In contrast, in the summer condition of 35 °C, the maximum temperature of the battery and temperature difference between the batteries were 38.8 °C and 3.27 °C at the R134a flow rate of 5 lpm or more, which was verified as a stable flow condition for battery safety.