New insights into the search of meat quality biomarkers assisted by Orbitrap Tribrid untargeted metabolite analysis and chemometrics.

Metabolite profiles of normal and defective dry, firm and dark (DFD) meat extracts with known ultimate pH (pHu) values were determined by Orbitrap Tribrid ID-X untargeted analysis coupled to chemometrics. An intelligent MS3 AcquireXTM workflow firstly approached the unambiguous characterization of detected features that were subsequently quantified by a complementary MS1 study of biological replicates. Chemometric research revealed how threonylphenylalanine (overexpressed in normal meats) together to tetradecadienoyl- and hydroxydodecanoyl-carnitines (both overexpressed in DFD meats) appropriately grouped meat groups assayed. Robustness of such biomarkers was confirmed through a time-delayed study of a blind set of samples (unknown pHu) and evidenced limitations of pHu as an isolated parameter for accurate meat quality differentiation. Other acyl-carnitines also characterized DFD samples, suggesting interferences induced by pre-slaughter stress (PSS) on lipid catabolism that would explain accumulation of such intermediate metabolites. Results achieved can ease understanding of biochemical mechanisms underlying meat quality defects.

Review of the state of the art of acrylamide human biomonitoring.

Human biomonitoring (HBM) is a very useful tool for assessing human exposure to acrylamide (AA). In the framework of the Human Biomonitoring Initiative (HBM4EU) AA was included in its second list of priority substances due to the potential threat to human health. HBM data on AA are scarce, but the use of specific and sensitive biomarkers represents a reliable indicator of exposure. In this review an overview of available knowledge on HBM of AA is provided in terms of: i) preferred exposure biomarkers and matrices for the HBM of AA; ii) analytical methods for determining its biomarkers of exposure in the most used specimens; iii) current HBM data available; and iv) tools for interpreting HBM data for AA in relation to risk assessment. Finally, future trends in this field are discussed.

Children's exposure to polycyclic aromatic hydrocarbons in the Valencian Region (Spain): Urinary levels, predictors of exposure and risk assessment.

Polycyclic aromatic hydrocarbons (PAHs) are pollutants that are released into the environment during incomplete combustion of organic matter and which can have a negative effect on human health. PAHs enter the human body mostly through ingestion of food or inhalation of tobacco smoke. The purpose of the present study is to evaluate the internal levels of PAHs that children living in the Valencian Region (Spain) are exposed to. In total, we measured eleven biomarkers of exposure to naphthalene, fluorene, phenanthrene, pyrene, and benzo(a)pyrene in the urine of 566 children aged 5-12. The analytical method was based on a liquid-liquid extraction of the PAH metabolites from the urine samples, followed by their determination by liquid chromatography coupled to tandem mass spectrometry. In addition, we used a questionnaire to collect the socio-demographic characteristics and 72 h dietary recall information of the participants in our study. Overall, we detected PAH metabolites in more than 78% of the children, with the exception of 3-hydroxyfluorene and 3-hydroxybenzo(a)pyrene, which were found in less than 37% of the analyzed samples. The most abundant biomarker found was 2-hydroxynaphthalene, with a geometric mean of 10 ng·ml-1. Reference values (RV95) ranging from 0.11 (4-hydroxyphenanthrene) to 53 ng·ml-1 (2-hydroxynaphthalene) in urine of Spanish children were derived from the present study. According to the statistical analysis, the factors that were significantly associated with the internal exposure to PAHs were province of residence, body mass index (BMI), children's age, consumption of plastic-wrapped food, and dietary habits. The estimated daily intakes in geometric mean terms ranged from 5 (fluorene) to 204 ng·kg-bw-1·day-1 (naphthalene). Risk assessment calculations showed higher hazard quotients and hazard indexes for children aged 5-8 than those aged 9-12, but all were below 1. In conclusion, no potential non-cancer health risk due to PAH exposure was observed in children living in Spain.

Do background levels of the pesticide pirimiphosmethyl in plant-based aquafeeds affect food safety of farmed Atlantic salmon?

The substitution of fish oil and fishmeal with plant-based ingredients in commercial aquafeeds for Atlantic salmon, may introduce novel contaminants that have not previously been associated with farmed fish. The organophosphate pesticide pirimiphos-methyl (PM) is one of the novel contaminants that is most prevalent in commercial salmon feed. In this study, the feed-to-fillet transfer of dietary PM and its main metabolites was investigated in Atlantic salmon fillet. Based on the experimental determined PM and metabolite uptake, metabolisation, and elimination kinetics, a physiologically based toxicokinetic (PBTK) compartmental model was developed. Fish fed PM had a relatively low (~4%) PM retention and two main metabolites (2-DAMP and Desethyl-PM) were identified in liver, muscle, kidney and bile. The absence of more metabolised forms of 2-DAMP and Desethyl-PM in Atlantic salmon indicates different metabolism in cold-water fish compared to previous studies on ruminants. The model was used to simulate the long term (>1.5 years) feed-to-fillet transfer of PM + metabolite in Atlantic salmon under realistic farming conditions including seasonal fluctuations in feed intake, growth, and fat deposition in muscle tissue. The model predictions show that with the constant presence of the highest observed PM concentration in commercial salmon feed, fillet PM+ metabolite levels were approximately 5 nmol kg-1, with highest levels for the metabolite 2-DAMP. No EU maximum residue levels (MRL) for PM and its main metabolites exist in seafood to date, but the predicted levels were lower than the MRL for PM in swine of 32.7 nmol kg-1.

LC-MS/MS method for the determination of organophosphorus pesticides and their metabolites in salmon and zebrafish fed with plant-based feed ingredients.

The composition of Atlantic salmon feed has changed considerably over the last two decades from being marine-based (fishmeal and fish oil) to mainly containing plant ingredients. Consequently, concern related to traditional persistent contaminants typically associated with fish-based feed has been replaced by other potential contaminants not previously associated with salmon farming. This is the case for many pesticides, which are used worldwide to increase food production, and may be present in plant ingredients. Earlier studies have identified two organophosphorus pesticides, chlorpyrifos-methyl and pirimiphos-methyl, in plant ingredients used for aquafeed production. In the present study, we developed a reliable and sensitive analytical method, based on liquid chromatography coupled to tandem mass spectrometry, for the determination of these pesticides and their main metabolites in warm water (zebrafish) and cold water (Atlantic salmon) species, where possible differences in metabolites could be expected. The method was tested in whole zebrafish and in different salmon tissues, such as muscle, bile, kidney, fat, and liver. The final objective of this work was to assess kinetics of chlorpyrifos-methyl and pirimiphos-methyl and their main metabolites in fish tissue, in order to fill the knowledge gaps on these metabolites in fish tissues when fed over prolonged time.