Abstarct: This dissertation presents the development of a novel, sensitive, and environmentally friendly method for the simultaneous preconcentration and determination of two azole antifungal drugs, clotrimazole and climbazole, in biological and environmental samples. Considering the widespread use of these compounds and the environmental concerns associated with their occurrence, the development of efficient analytical methods in accordance with the principles of green chemistry is of great importance. In this study, a switchable hydrophilicity solvent-baxsed homogeneous liquid–liquid microextraction (SHS-HLLME) method was employed using dipropylamine in the presence of β-cyclodextrin. β-Cyclodextrin enhanced the extraction efficiency and selectivity through the formation of host–guest inclusion complexes. Dipropylamine was used as the extraction solvent, and phase separation was achieved by pH adjustment and the reversible conversion of the extraction solvent between its hydrophilic and hydrophobic states, resulting in the concentration of the analytes in the organic phase. This strategy enabled the efficient extraction and preconcentration of the target compounds within a short period of time and without the need for centrifugation. Finally, the influential parameters, including β-cyclodextrin concentration, type and volume of extraction solvent, ionic strength, and the volumes of acidic and alkaline solutions, were optimized. The results demonstrated that the linear dynamic ranges for clotrimazole and climbazole were 10.0–11000.0 and 40.0–11000.0 μg L⁻¹, respectively. The limits of detection (LODs) were calculated to be 3.0 and 12.0 μg L⁻¹ for clotrimazole and climbazole, respectively. Evaluation of the proposed method in water, milk, plasma, and tablet samples yielded satisfactory recoveries ranging from 98.2% to 101.8%, indicating the excellent accuracy, precision, and reproducibility of the method. Overall, the developed method, characterized by high sensitivity, operational simplicity, low cost, minimal consumption of organic solvents, and compliance with green chemistry principles, represents an efficient approach for the determination of azole antifungal drugs in biological and environmental samples.