Detection of polyphosphates in seafood and its relevance toward food safety.

Polyphosphates are permitted as food additives (Regulation EC No 1129/2011) but their undeclared utilisation is considered fraudulent. They improve water holding capacity of the seafood, preventing biochemical/physical changes during commercialization. The key objective of this study was the detection of polyphosphate in various seafood categories, by means of high-performance ion-exchange chromatography with suppressed conductometry (HPIEC-SCD) coupled to Q-Exactive Orbitrap high resolution mass spectrometry (HRMS-Orbitrap). Ten frozen cuttlefish samples did not reveal any treatment, while in ten frigate tunas, high concentration of orthophospate was found. Unambiguous hexametaphosphate presence was demonstrated in four prawn samples, while triphosphate was quantified (11.2 ± 4 ug/g) in another four prawn samples that contained orthophosphate (10225 ± 1102 ug/g), as well. Other samples sporadically encompassed polyphosphates profiles that varied according species and processing type. This analytical approach provided sustenance in better understanding regarding utilization of polyphosphates through HRMS fingerprinting of anionic species that would be specific in food safety control.

Exposure to metals and arsenic from yellow and red tuna consumption.

Tuna is one of the most frequently consumed fish and, as a predator, can biomagnify pollutants. Metal and other elements pollution is an important worldwide concern. Based on these considerations, the aim of this work was to investigate the occurrence of As, Cd, Cr, Ni, Hg and Pb in tuna coming from different FAO areas to evaluate human exposure. The analysis was performed on muscle tissues through a quadrupole inductively coupled mass spectrometry. One hundred thirty-one samples were analysed. One red tuna from the Adriatic Sea and 11 yellow tunas exceeded Pb maximum levels (MLs) with a concentration ranging 0.31-0.86 mg kg-1; three red tunas from different Mediterranean sub-areas exceeded Hg MLs, with a concentration range 1.19 to 1.80 mg kg-1. All the Hazard Indexes (HIs) were lower than one, indicating that only a negligible health hazard could derive from the ingestion of tuna, for both average and high consumers. The risk of carcinogenicity from Cr is still under debate at the concentrations detectable in food.

Detection of glyphosate and its metabolites in food of animal origin based on ion-chromatography-high resolution mass spectrometry (IC-HRMS).

Glyphosate and glufosinate are broad spectrum herbicides, widely used in agriculture and in inhabited or industrialised areas, and aminomethylphosphonic acid is a degradation product of glyphosate. In 2015, the International Agency for Research on Cancer reported that glyphosate is a probable carcinogenic. In 2017, however, a scientific opinion of the European Chemicals Agency concluded that glyphosate is not proven to be carcinogenic, mutagenic or to have negative effects on reproduction. Nevertheless, aminomethylphosphonic acid was not considered. Due to their chemical-physical characteristics, these molecules present difficulties that have not yet allowed routine monitoring to be carried out. For these reasons, we developed and validated a simple and versatile liquid extraction, before IC-HRMS analysis, of three different complex matrices: honey, bass fish and bovine muscle. Among the satisfactory validation parameters, the LOQs in the range of 4.30-9.26 ng g-1 demonstrated high method sensitivity, compared to the few works present in literature. Finally, the method was applied to real commercial samples, which showed no traces of the selected pesticides.

Distribution of POPs, pesticides and antibiotic residues in organic honeys from different production areas.

Demand for honey is increasing, especially if it is organic and if its nutritional properties are linked to untreated environments in order to guarantee quality for health. Sources of contamination of honey can be divided into environmental and apicultural. Therefore, the distribution of persistent organic pollutants, pesticides and antibiotic residues from geographical areas with different contamination sources (high anthropic impact, intensive farming, husbandry and low anthropic impact) was investigated in order to confirm the potential transfer of xenobiotics into the supply chain and to give beekeepers tools for the selection of areas dedicated to organic production. The presence of polychlorinated biphenyls, polybrominated diphenyl ether and polycyclic aromatic hydrocarbons was confirmed, not only in proximity to highly urbanised centres, where the concentrations were higher, but in all environment contexts, confirming their ubiquity. No antibiotics or neonicotinoids were detected in 95 organic honeys, demonstrating the absence of apicultural treatments and consequently the good quality of honey of different areas. These results are important due to the undefined regulatory European situation on honey antibiotic limits.

Accelerated solvent extraction by using an 'in-line' clean-up approach for multiresidue analysis of pesticides in organic honey.

The worldwide loss of honeybee colonies may be due to their exposure to several contaminants (i.e., pesticides); such contamination may also have impacts on consumers' health. Therefore, it is essential to develop quick and new methods to detect several pesticide residues in honey samples. In this study, the effectiveness of accelerated solvent extraction (ASE) was compared with QuEChERS methods for the analysis of 53 pesticides in organic honey by gas chromatography-triple quadrupole mass spectrometry. Two simple and rapid ASE methods with 'in-line' clean-up were optimised and then compared with QuEChERS. Hexane-ethyl acetate (Hex:EtAc) and Florisil were chosen as extraction solvent and retainer for the first ASE method respectively; acetonitrile and a primary-secondary amine phase (ACN-PSA) were selected for the second ASE method. The methods were validated according to the European Union SANTE/11945/2015 guidelines. The validation parameters showed that QuEChERS and ASE with PSA as retainer had better repeatability than ASE with Hex:EtAc and Florisil. In particular, QuEChERS and ASE (ACN-PSA) showed good recovery, according to the SANTE criteria, for the majority of investigated pesticides. Conversely, when ASE with Hex:EtAc and Florisil was used as the retainer, several compounds showed recoveries lower than the acceptable value of 70%. The ASE in-line method was finally applied to evaluate pesticide concentration in organic honey samples.

Bovine teeth as a novel matrix for the control of the food chain: liquid chromatography-tandem mass spectrometry detection of treatments with prednisolone, dexamethasone, estradiol, nandrolone and seven ß2-agonists.

Veterinary drugs usually have rapid clearance rates in the liver and kidney, hampering their detection in conventional matrices such as the liver or urine. Pharmacological principles such as esterification may be applied to facilitate the administration of veterinary drugs and increase drug half-life. Prednisolone, whose therapeutic administration is regulated for food producing animals in the EU, is available in its acetate form as well as nandrolone, a banned anabolic steroid, which may be obtained as nandrolone phenylpropionate and estradiol as a benzoyl ester. While the distribution and accumulation of lipophilic and hydrophilic substances in human teeth have been well documented, studies on residues in bovine teeth are lacking. We hypothesised that analysis of bovine teeth could be used to detect both regulated and banned veterinary drugs. Steroids may be illegally used as growth promoters in food producing animals, alone or combined with ß2-agonists; therefore, we developed, and validated, in accordance with the Commission Decision 2002/657/EC, two analytical confirmatory LC-MS/MS methods to detect these classes of compounds following a unique liquid extraction procedure. Finally, we analysed teeth from three male Friesian veal calves treated with intramuscular estradiol benzoate, oral prednisolone acetate or intramuscular nandrolone phenylpropionate in combination with oral ractopamine, respectively, and from seven bovines from the food chain. Teeth from treated animals were positive for their respective drugs, with the exception of nandrolone phenylpropionate. One sample from a food chain bovine was positive for isoxsuprine, one of the seven ß2-agonists studied. Non-esterified forms of the steroids were not found. These results demonstrate that bovine teeth are a suitable matrix for the determination of pseudoendogenous substances or illicit administration of veterinary drugs.

A headspace solid-phase microextraction gas-chromatographic mass-spectrometric method (HS-SPME-GC/MS) to quantify hexanal in butter during storage as marker of lipid oxidation.

A method using headspace solid-phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC/MS) was developed and validated for the extraction and quantification of hexanal content in butter (at ngg(-1) level) during storage at 4°C. The variability of hexanal content among seasons of production and the influence of high extraction temperature on ex-novo formation of hexanal were also evaluated. The HS-SPME conditions were optimised and analytical parameters of the method (linearity, accuracy, and precision) demonstrate its usefulness. The reproducibility and accuracy of the quantitative analysis was assured by the use of D(12)-hexanal as internal standard. For the applications, the headspace was extracted using CAR/PDMS fiber for 180min at 4°C. Hexanal contents in samples during all storage period (shelf-life) and from butters produced in different seasons were analysed. Butter samples at the end of shelf-life and samples produced in August showed highest values of hexanal, confirming that the temperature both in storage and distribution phases represents a critical factor to maintain the quality of butter.

Determination of carbon monoxide in tuna by gas chromatography with micro-thermal conductivity detector.

The suitability of a portable gas chromatograph equipped with a micro-thermal conductivity detector for the head-space determination of carbon monoxide (CO) in tuna samples is evaluated; CO is estimated after its liberation from tissue by acidic treatment at 70 degrees C. Using the tested technique, the CO contents in untreated and suspected treated samples are analyzed. A limit of detection of approximately 13 ng/g is reached. The results demonstrate that this apparatus has performances similar to more expensive and sophisticated instruments.

Synthesis and characterization of 4-hydroxy-2-nonenal derivatives for gas chromatographic analysis with electron capture detection (GC-ECD).

4-Hydroxy-2-nonenal (HNE) has been prepared from the corresponding dimethylacetal (HNE-DMA), in turn synthesized by a conventional approach with a few modifications of the experimental protocol and some improvements in the purification of the final product. In order to exploit the sensitivity of gas-chromatography with electron capture detector (GC-ECD) in the analysis of HNE derivatives, reaction of HNE with 2,4,6-trichlophenylhydrazine (TCPH) and 3,5-dichloro-phenylhydrazine (DCPH) was tested. Reaction with TCPH afforded a mixture of products, whereas with DCPH a single major product was formed that was prepared on a millimolar scale and purified. (1)H-NMR analysis established that the derivative of HNE with DCPH is HNE 3,6-dichloro-phenylhydrazone, that can be used as standard for GC-ECD analysis.

Improved procedure for the determination of malonaldehyde by gas-chromatography with electron-capture detection as 2,4,6-trichlorophenylhydrazine derivative.

A previously described derivatization method using trichlorophenylhydrazine was developed for the estimation of malonaldehyde measured by gas-chromatography (GC) with electron-capture detection. The precision and reliability of the procedure are improved here by the use of methylmalonaldehyde as internal standard and by the introduction of a diverter valve at the end of the capillary column to protect the electron-capture detector, respectively. The method was applied to determine malonaldehyde content in bovine plasma samples.