Although the addition of buffers provides improved stability to flucloxacillin (FLU) solutions, unbuffered solutions are often preferred in clinical practice. The first purpose of this study was to investigate whether a 50 mg/mL solution of FLU in normal saline is stable for 24 h at 33 °C so that it can be applied for outpatient parenteral antimicrobial therapy (OPAT) using portable elastomeric infusion pumps (PEIPs). When the PEIPs were stored in an oven at 33 °C and deflated over 24 h, the volume of the collected solution, pH, and FLU concentration were checked every 4 h. Obtaining better results than expected based on the literature data, other storage conditions, such as refrigeration, room temperature (RT), 37 °C, refrigeration followed by 24 h at 33 °C and 37 °C, and different batches/brands, were also tested. This study confirmed the pronounced effect of temperature on the stability of FLU and also showed the relationship between the stability of FLU and the initial pH of the solution. FLU was quite stable at refrigeration and RT conditions, with more than 99% and 95% remaining. After 24 h at 33 °C, more than 92% of FLU was still present in the solution, while this number decreased to less than 85% when the storage temperature reached 37 °C. The remaining percentage was found to be even lower when the solution was stored at 2-8 °C for 6 days, followed by 24 h storage at 33 °C or 37 °C, with losses of 17% and 30%, respectively. The stability of FLU became worse when the initial pH of the solution was lower than 5.9 since the concentration of FLU dropped to less than 90% after 24 h at 33 °C, and a precipitate started to form when the initial pH of the solution was around 5.3. Therefore, FLU in PEIPs could be employed for 24 h if the temperature was ideally not more than 33 °C, while the pH should be not less than 5.9 upon reconstituting the FLU solution.