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Highlights • 1 Five kinds of hydrophobic DESs were designed and prepared for the first time. • 2 The prepared DESs showed high extraction efficiency for formaldehyde (HCHO). • 3 The DES was applied to extract and determine of HCHO in biological and gas samples. • 4 Hydrophobic DESs based VA-LLME combined with HPLC method was developed. Abstract Hydrophobic deep eutectic solvents (DESs) are a new class of green solvents, and their research and application are a brand new area. So far, only a few hydrophobic DESs have been reported as extraction solvents. In this paper, five kinds of hydrophobic DESs composed of trioctylmethylammonium chloride and hydroquinone, 4-phenylphenol or 4-cyanophenol are designed and prepared for the first time. Melting point / glass transition temperature and density are also determined for these DESs. Coupled with HPLC, these hydrophobic DESs have been used as extractants in vortex-assisted liquid-liquid microextraction for the selective enrichment and indirect determination of formaldehyde. The optimization of reaction and extraction conditions, such as pH value, reaction time and temperature, concentration of 2,4-dinitrophenylhydrazine, type and dosage of the extraction solvent, speed and time of vortex, salt addition and centrifugation time are investigated in detail. It is found that under the optimized experimental conditions, the calibration curve of this method is linear in the range of 1.0–200.0 μg L−1 with the linear correlation coefficient (r2) of 0.9994. The limit of detection (S/N = 3) and limit of quantification (S/N = 10) are 0.2 μg L−1 and 1.0 μg L−1, and the intra-day and inter-day precision (RSD, n = 5) are 1.1% and 3.5%, respectively. The interference of some common coexisting ions and neutral molecules have been also tested. The proposed method has been successfully used for the determination of formaldehyde from biological and indoor air samples. The recovery of formaldehyde is in the range from 83.1% to 104.4%, and the relative standard deviation is lower than 5.9%. Furthermore, the analytical parameters of the proposed method have been compared with other extraction and microextraction methods. [ABSTRACT FROM AUTHOR]