In this work a sample pretreatment approach assumed liquid-liquid microextraction based on the in situ formation of a hydrophobic natural deep eutectic solvent on a hydrophobic membrane impregnated with natural terpenoid was developed. The procedure included alkaline hydrolysis of a food sample containing fat to form fatty acids, which acted as precursors for the in situ formation of the deep eutectic solvent with natural terpenoid. Two processes were observed on the membrane surface: in situ formation of the hydrophobic deep eutectic solvent and liquid-liquid microextraction of the target analytes. After microextraction, the membrane containing the analytes was easily removed from the sample solution. The developed approach was applied to the separation and preconcentration of hydrophobic organochlorine pesticides (ɑ-hexachlorocyclohexane and γ-hexachlorocyclohexane) from a hydrophobic sample matrix (peanut paste), followed by their determination by gas chromatography with electron capture detection. Under optimal conditions, the limits of detection and quantification for both analytes were 0.3 and 1.0 μg kg−1, respectively. The procedure allowed the separation of fat-soluble analytes from a complex sample matrix with a high content of fat. The extraction recoveries were in the range of 93–95 %. © 2024 Elsevier B.V.