Hexafluoroisopropanol-based deep eutectic solvents for high-performance DNA extraction

In this study, hexafluoroisopropanol (HFIP)-based deep eutectic solvents (DESs) were developed and used for DNA extraction from human whole blood samples for the first time. HFIP-based DESs were prepared using HFIP and choline chloride (ChCl)/tetrabutylammonium chloride/cetyltrimethylammonium bromide as the hydrogen bond donor and acceptor, respectively. The two-phase system formation was promoted with different inorganic salts as the phase-forming component. According to the strong phase separation capability and high DNA extraction efficiency, DESs consisting of HFIP/ChCl-(NH₄)₂SO₄, HFIP/ChCl-Na₂SO₄ and HFIP/ChCl-MgSO₄ were then selected for application in DNA extraction. The factors that could have impacted the DNA extraction process, including molar ratio of DES, DES addition, salt species and addition, and sample pH, were systematically investigated via single-factor experimental analysis. Furthermore, we selectively examined bovine serum albumin and RNA to assess the specificity of the HFIP-based DESs for DNA extraction. Conclusively, 93.9%, 96.7% and 99.8% DNA could be extracted using the HFIP/ChCl-(NH₄)₂SO₄, HFIP/ChCl-Na₂SO₄, and HFIP/ChCl-MgSO₄ systems, respectively. Moreover, the developed systems were successfully used to extract DNA from human whole blood with satisfactory results. The DNA secondary structure was stable after DES extraction with the electrostatic interaction between DES and DNA as the main force driving DNA adsorption by DES.