Natural hydrophobic deep eutectic solvent based dispersive liquid-liquid microextraction followed by liquid chromatography with tandem mass spectrometry analysis of multi-class metabolites of pesticides, phthalates, and polycyclic aromatic hydrocarbons in animal-originated foods

Demonstrated a new natural hydrophobic deep eutectic solvent (NADES) based dispersive liquid-liquid microextraction (DLLME) approach/method to extract and enrich 37 multi-class metabolites (MCM) such as pesticides, phthalates, and polycyclic aromatic hydrocarbons metabolites from a complex animal- originated food matrices. The extraction solvent (NADES) synthesized from an equimolar mixture of the terpenes, thymol and menthol. The analysis was performed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) employing electrospray ionization, and an enzymatic deconjugation of metabolites was carried out with beta-glucuronidase. The parameters were optimized using the design of experiments (DoE). Further, the method was validated, and measurement uncertainty was also calculated. The detection and quantitation limits of the method were 0.03 to 0.4 ng/mL and 0.1 to 1.32 ng/mL, respectively. Recovery and expanded uncertainty ranged from 81.3 to 112.3 % and 8.9 to 19.7 %. The pertinence of the method was proven in identifying/finding metabolites in egg and meat samples ranging from 0.31 to 7.83 ng/g and 0.51 to 9.78 ng/mL in milk samples.