A high-throughput method for the simultaneous determination of multiple mycotoxins in human and laboratory animal biological fluids and tissues by PLE and HPLC–MS/MS

• This is the first method for large-scale testing of mycotoxins in various matrices.
• High-throughput was achieved by using PLE pre-treatment.
• The limits of detection defined as CCα varied from 0.01 μg/kg to 0.69 μg/kg.
• It can be used for seeking a link between mycotoxin exposure and diseases.
• It could be a useful tool in toxicokinetic and toxicological studies.

A high-throughput method for the determination of 28 mycotoxins involving pressurised liquid extraction (PLE) coupled with liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) has been optimised and validated for determination in various biological fluids and tissues of human and laboratory animals. High-throughput analysis was achieved using PLE pre-treatment and without the need for any cleanup. The extraction solvent was acetonitrile/water/acetic acid (80/19/1, v/v/v). The static extraction time was 5 min. The extraction pressure and temperature were 1500 psi and 140 °C, respectively. The flush volume was 60%. The limits of detection, which were defined as CCα, varied from 0.01 μg/kg (μg/L) to 0.69 μg/kg (μg/L). The recoveries of spiked samples from 0.20 μg/kg (μg/L) to 2 μg/kg (μg/L) ranged from 71% to 100.5% with relative standard deviations of less than 17.5%, except FB1 and FB2 recoveries, which were lower than 60%. The method was successfully applied in real samples, and the data indicate that this technique is a useful analytical method for the determination of mycotoxins from humans and animals. To the best of our knowledge, this method is the first for the large-scale testing of multi-class mycotoxins in all types of biological fluids and tissues that uses PLE and HPLC–MS/MS.