Fluopyram is used in agriculture to control fungi and its intensive use can lead to a significant risk to living organisms.1 Therefore, monitoring this compound is important to ensure human health and preserve the environment. This work aimed to optimize and validate liquid-liquid extraction with low temperature purification (LLE-LTP) to determine fluopyram in water samples. The optimization was performed through a complete 23 factorial design, and the variables studied were: the composition of the extraction phase (acetonitrile and acetonitrile/ethyl acetate), ionic strength (0.0 and 0.2 g of NaCl) and freezing time (60 and 120 min). The method consists of adding 4.0 mL of water sample and 8 mL of extraction phase. Then, the system is homogenized by vortexing for 1 min and kept at -20°C until complete freezing of the aqueous phase. After this step, 5 mL of the liquid organic phase containing fluopyram were completely evaporated under air flow and immediately resuspended in 400 μL of acetonitrile. The optimal extraction conditions for this work were 8.0 mL of acetonitrile, freezing time of 120 min and without the addition of NaCl. All extracts were analyzed in a liquid chromatograph coupled to a diode array detector (HPLC-DAD). The recovery rate achieved in this condition was 106.29% with a relative standard deviation of 7.54%. The extraction method was selective, precise, and exact (as can be seen in Figure 1), with a linearity range between 6.0 to 200.0 μg L-1 and did not show a matrix effect. The detection and quantification limits reached were 4 and 6 μg L-1, respectively.