Simultaneous determination of nereistoxin insecticides in foods of animal origins by combining pH-dependent reversible partitioning with hydrophilic interaction chromatography-mass spectrometry

Although nereistoxin insecticides (NIs) are banned for animal husbandry operations, they are still used because of their high insecticidal activities. Therefore, a reliable residue analysis method for the simultaneous detection of cartap, bensultap, thiocyclam, and nereistoxin in foods of animal origins, including beef, pork, chicken, milk, and eggs, was developed using hydrophilic interaction liquid chromatography–mass spectrometry (HILIC–LC–MS/MS). The NIs were extracted with an acidic cysteine and formate buffer solution and hydrolyzed to nereistoxin. The molarity and pH of the buffer were optimized at 20 mM and 3, respectively, to keep the pH of the extracts at 4–5. pH-dependent acid–base partitioning coupled with salting-out-assisted liquid–liquid extraction using acetonitrile was performed for purification and for the direct introduction of the extracts to LC. The optimal pH values were 5 and 9 for the acid–base partitioning. Nereistoxin quantitation was achieved with consistent column retention (RSD < 0.6%) and a high degree of separation (N > 10⁶). The matrix-dependent method limit of quantitation was 2 μg nereistoxin/kg, and the calibration curve showed good linearity (R² > 0.998). The recovery efficiencies were in the range of 89.2–109.9% with relative standard deviations less than 10%, and matrix effects did not exceed ± 10%, which satisfied the criteria outlined in the European SANTE/12682/2019 guidelines.