Preliminary Study to Develop an Alternative Method for the Non-targeted Determination of Xenobiotics in Food by Means of Ultra High Performance Liquid Chromatography Coupled to High Resolution and Accuracy Mass Spectrometry

This preliminary study describes the use of high resolution and accuracy mass spectrometry techniques combined with new generation chemical software products for detecting and identifying contaminants in food commodities. As a first step, the extracts of routine target analysis samples (obtained in our official laboratory responsible for food residues control) were acquired and processed with this method in order to search unknown and non-targeted contaminants in food. In order to verify the feasibility of the presented method, the research has been firstly addressed to untargeted pesticides and their metabolites in stone fruits commodities and tomatoes. The differential analysis carried with Compound Discoverer 2.0 between the investigated unknown sample and the blank matrix sample allowed to remove all the matrix molecular components; Aggregated Computational Toxicology Resource (ACToR) helped to understand and predict chemical interpretation of substances. The acquisition in FullScan-AIF and FullScanddMS2 allowed the clear detection and identification of isobaric compounds such as quinalphos and phoxim. In order to verify that the proposed method is suitable to the scope of application, the main points of SANTE/11813/2017 Document have been followed. The results demonstrate that no false positives and no false negatives have been detected from the analysis of samples spiked with pesticides at 0.010 and 0.10 mg kg−1. This preliminary study has been also tested with a Proficiency Test (EUPT-FV-SM08) and, according to EUPT-FV-SM08 Final Report, our laboratory has been included in the 67% (56) that clearly detected over 70% pesticides. Finally, this method has been extended to other matrices and contaminants

Ion Chromatography–High-Resolution Mass Spectrometry Method for the Determination of Bromide Ions in Cereals and Legumes: New Scenario for Global Food Security

The new scenario for global food production and supply is decidedly complex given the current forecast of an increase in food fragility due to international tensions. In this period, exports from other parts of the world require different routes and treatments to preserve the food quality and integrity. Fumigation is a procedure used for the killing, removal, or rendering infertile of pests, with serious dangers to human health. The most-used fumigants are methyl bromide and ethylene dibromide. It is important to bear in mind that the soil may contain bromide ions naturally or from anthropogenic source (fertilizers and pesticides that contain bromide or previous fumigations). Different methods (titrimetric, spectrophotometric, and fluorometric approaches) are available to rapidly determine the amount of bromide ion on site in the containers, but these are non-specific and with high limits of quantification. The increasing interest in healthy food, without xenobiotic residues, requires the use of more sensitive, specific, and accurate analytical methods. In order to help give an overview of the bromide ion scenario, a new, fast method was developed and validated according to SANTE 11312/2021. It involves the determination of bromide ion in cereals and legumes through ion chromatography–Q-Orbitrap. The extraction was performed by the QuPPe method, but some modifications were applied based on the matrix. The method described here was validated at four different levels. Recoveries were satisfactory and the mean values ranged between 99 and 106%, with a relative standard deviation lower than 3%. The linearity in the matrix was evaluated to be between 0.010 and 2.5 mg kg−1, with a coefficient of determination (R2) of 0.9962. Finally, the proposed method was applied to different cereals and legumes (rice, wheat, beans, lentils pearled barley, and spelt) and tested with satisfactory results in EUPT-SMR16 organized by EURL

Determination of Pesticides in Wheat Flour Using Microextraction on Packed Sorbent Coupled to Ultra-High Performance Liquid Chromatography and Tandem Mass Spectrometry

An ultra-high performance liquid chromatography coupled to tandem mass spectrometry (UHPLC–MS/MS) method for multiclass analysis of pesticide and fungicide residues in wheat flour based is presented. An efficient and rapid cleanup based on microextraction by packed sorbent (MEPS) has been developed in order to have a good enrichment factor together with a low matrix effect. The target analytes were 25 pesticides widely used in wheat, with different physico-chemical characteristics and different mechanism of action: acetylcholinesterase inhibitors such as organophosphorus, carbamates, neonicotinoids, and inhibitors of ergosterol such as imidazoles and triazoles. MEPS was shown to be successful with reduction of the amount of solvent required and possibility of automation of the cleanup procedure. The whole method was then validated according to the SANCO/12571/2013 guidelines, proving its suitability as confirmation method for the selected analytes

Solid-Phase Extraction of Pesticides by Using Bioinspired Peptide Receptors

A virtual development of hexapeptide receptors bioinspired by the acetylcholinesterase enzyme active site is proposed. A semicombinatorial approach was applied to generate a virtual hexapeptides library with different affinity properties towards organophosphate and carbamate pesticides. The virtual screening process was addressed to obtain peptides able to separate pesticide subclasses in the experimental work. Three hexapeptides, two generated by molecular modeling and one having a scrambled sequence, were used as selective sorbent materials for pesticides in preanalytical solid-phase extraction (SPE) method. Selective adsorption and cross-reactivity were tested directly on a mix of four pesticides (carbaryl, chlorpyrifos-ethyl, malathion, and thiabendazole) having different structures and physico-chemical properties, at a total concentration of 120 ppb (each pesticide at concentration of 30 ppb). The results were compared to traditional sorbent material such as C-18 and strata-X. Data showed that only one of the hexapeptides virtually designed had significant differences in competitive absorption between aliphatic pesticide malathion, fungicide thiabendazole chosen as negative control, and aromatic pesticides. These results partially supported the simulated strategy

Micro-solid-phase extraction (µ-SPE) of organophosphorus pesticides from wheat followed by LC-MS/MS determination

A rapid, selective and effective method of extraction, clean-up and concentration of organophosphorous pesticides from wheat followed by electrospray (ESI) LC-MS/MS analysis was developed. The μ-SPE (micro-solid-phase extraction) procedure resulted in good analytical performance and reduced at the same time matrix effects, analysis time and solvent consumption. Limits of detection (LODs) and quantification (LOQs) were in the range of 0.3–10 and 1–30 μg kg−1, respectively, with good reproducibility (RSD ≤ 13.8) and recoveries between 75% and 109%. Coefficients of determination (r2) were greater than 0.996 for the studied pesticides. Despite the reduced sorbent bed mass of μ-SPE tips (4.2 mg), the analytical data showed that no saturation phenomena occurs in the tested range of concentration both for single compounds and mixtures. Several real samples were analysed and the concentrations of the selected pesticides were found to be below the respective maximum residue limit (MRLs)