Deep eutectic solvent extraction of cortisol and cortisone from human saliva followed by LC-UV analysis

A novel extraction method was developed for the determination of cortisol and cortisone. In this study, we prepared a hydrophobic deep eutectic solvent (DES) by mixing trioctylmethylammonium chloride and pentafluorophenol as a hydrogen bond acceptor and a hydrogen bond donor, respectively, for use as the extraction solvent. The extraction of cortisol and cortisone was performed by adding a small volume of the DES to the aqueous sample. After centrifugation, the resulting sedimented DES phase was injected into a reversed-phase liquid chroamtography column, and the analytes were detected with an ultraviolet detector at 254 nm. Under the optimized extraction conditions, the enrichment factors of cortisol and cortisone were 9.3 and 8.5, respectively. Furthermore, the linear dynamic ranges were established over a concentration range of 10-200 pmol mL^-1 (r² > 0.9992), and the limits of detection of cortisol and cortisone were found to be 2.1 and 1.8 pmol mL^-1, respectively. The applicability of this method was evaluated by analyzing the cortisol and cortisone contents of human saliva samples.