Development of a highly sensitive method based on QuEChERS and GC-MS/MS for the determination of polycyclic aromatic hydrocarbons in infant foods.

Polycyclic aromatic hydrocarbons (PAHs) are environmental contaminants that can be found in various food products, including those intended for infants. Due to their potential health risks, it is crucial to develop sensitive analytical methods for the accurate determination of PAHs in infant foods. This study describes the development and validation of a highly sensitive method for the quantification of European PAH markers, namely benzo[a]pyrene, benzo[a]anthracene, chrysene, and benzo[b]fluoranthene, using gas chromatography-tandem mass spectrometry (GC-MS/MS), in baby food samples. The first step was the optimization of the sample preparation procedure, performed using different methods based on the QuEChERS approach, also testing different extraction solvents. Several factors such as extraction efficiency, selectivity, and recovery were evaluated to choose the most effective procedure for sample preparation. Furthermore, the GC-MS/MS method was optimized, evaluating parameters such as linearity, sensitivity, accuracy, and robustness using spiked infant food samples. The method demonstrated excellent linearities with a correlation coefficient higher than 0.999 over a wide concentration range, and limits of detection and limits of quantification in the range 0.019-0.036 µg/kg and 0.06-0.11 µg/kg, respectively. Extraction recoveries were between 73.1 and 110.7%, with relative standard deviations always lower than 8%. These findings are compliant with the indications of the European Commission (Reg. 836/2011). To assess the applicability of the method to official control activities, a survey was conducted on commercially available infant food products. Four markers were determined in commercial samples belonging to different food categories for infants and young children. The outcome of this monitoring showed that PAH contamination, in all samples, was below the quantification limits. In conclusion, the developed GC-MS/MS method provides a highly sensitive and reliable approach for the determination of PAHs in baby foods. The optimized sample preparation, instrumental parameters, and validation results ensure accurate quantification of 4 PAHs even at trace levels. This method could contribute to the assessment of PAH exposure in infants and it could support regulatory efforts to ensure the safety and quality of infant food products with regular monitoring.

Microwave-Assisted Extraction/UHPLC-Q-Orbitrap-MS-Based Lipidomic Workflow for Comprehensive Study of Lipids in Soft Cheese.

In this work, Microwave-Assisted Extraction (MAE) was proposed as an alternative and environmentally friendly technique in lipidomics to study the lipid fingerprint of soft cheeses, such as mozzarella. For method development, a first step concerning an evaluation of extraction solvents was carried out via testing three different mixtures, including methanol/ethyl acetate, isopropanol/ethyl acetate, and ethanol/ethyl acetate, at a 1:2 v/v ratio. The latter was chosen as a solvent mixture for subsequent method optimization. MAE conditions, in terms of solvent volume, time, and temperature, were explored to define their effects on extraction capability through a full factorial experimental design. The best compromise to extract more lipids at the same time was obtained with 24 mL g-1 for solvent-to-solid ratio, 65 °C for temperature, and 18 min for time. Lipid analyses were conducted by UHPLC-Q-Orbitrap-MS associated with multivariate statistics. The developed lipidomic workflow allowed for the extraction of over 400 lipids grouped into 18 different subclasses. The results confirmed that MAE is a suitable technique for lipid extraction in the omics approach with high efficiency, even using low-cost and less toxic solvents. Moreover, a comprehensive structure characterization of extracted lipids, in terms of fatty acid composition and regiochemistry, was carried out.

Red Meat Heating Processes, Toxic Compounds Production and Nutritional Parameters Changes: What about Risk-Benefit?

The heating process is a crucial step that can lead to the formation of several harmful chemical compounds in red meat such as heterocyclic aromatic amines, N-Nitrosamines, polycyclic aromatic hydrocarbons and acrylamide. Meat has high nutritional value, providing essential amino acids, bioactive compounds and several important micronutrients which can also be affected by heating processes. This review aims to provide an updated overview of the effects of different heating processes on both the safety and nutritional parameters of cooked red meat. The most-used heating processes practices were taken into consideration in order to develop a risk-benefit scenario for each type of heating process and red meat.

Untargeted Lipidomics and Chemometric Tools for the Characterization and Discrimination of Irradiated Camembert Cheese Analyzed by UHPLC-Q-Orbitrap-MS.

In this work, an investigation using UHPLC-Q-Orbitrap-MS and multivariate statistics was conducted to obtain the lipid fingerprint of Camembert cheese and to explore its correlated variation with respect to X-ray irradiation treatment. A total of 479 lipids, categorized into 16 different lipid subclasses, were measured. Furthermore, the identification of oxidized lipids was carried out to better understand the possible phenomena of lipid oxidation related to this technological process. The results confirm that the lipidomic approach adopted is effective in implementing the knowledge of the effects of X-ray irradiation on food and evaluating its safety aspects. Furthermore, Partial Least Squares-Discriminant Analysis (PLS-DA) and Linear Discriminant Analysis (LDA) were applied showing high discriminating ability with excellent values of accuracy, specificity and sensitivity. Through the PLS-DA and LDA models, it was possible to select 40 and 24 lipids, respectively, including 3 ceramides (Cer), 1 hexosyl ceramide (HexCer), 1 lysophosphatidylcholine (LPC), 1 lysophosphatidylethanolamine (LPE), 3 phosphatidic acids (PA), 4 phosphatidylcholines (PC), 10 phosphatidylethanolamines (PE), 5 phosphatidylinositols (PI), 2 phosphatidylserines (PS), 3 diacylglycerols (DG) and 9 oxidized triacylglycerols (OxTG) as potential markers of treatment useful in food safety control plans.

Effect of Different Cooking Treatments on the Residual Level of Nitrite and Nitrate in Processed Meat Products and Margin of Safety (MoS) Assessment.

Nitrite and nitrate are well-known food additives used in cured meats and linked to different food safety concerns. However, no study about the possible effect of cooking treatment on the residual level of these compounds before consumption is available. In this work, 60 samples of meat products were analyzed in order to evaluate the variation in residual nitrite and nitrate level after baking, grilling and boiling. The analyses by ion chromatography demonstrated that meat cooking leads to a decrease in nitrite and an increase in nitrate residual levels in the final products. Meat boiling caused an overall decrease in two additives' concentration, while baking and particularly grilling caused an increase in nitrate and, in some cases, nitrite as well. Some regulatory aspects were also considered, such as the possibility of revising the legal limit of nitrate from the actual 150 mg kg-1 to a more cautious 100 mg kg-1. Indeed, several meat samples (bacon and swine fresh sausage) resulted in a higher nitrate concentration than the legal limit after cooking by grilling (eleven samples) or baking (five samples). Finally, the Margin of Safety evaluation demonstrated a good level of food safety, all values being higher than the protective threshold of 100.

Characterization and Quantification of Arsenic Species in Foodstuffs of Plant Origin by HPLC/ICP-MS.

Arsenic is a well-known carcinogenic, mutagenic and toxic element and occurs in the environment both as inorganic arsenic (iAs) and organoarsenical compounds (oAsCs). Since the toxicity of arsenic compounds depends on their chemical form, the identification and determination of arsenic species are essential. Recently, the European Food Safety Authority, following the European Commission request, published a report on chronic dietary exposure to iAs and recommended the development and validation of analytical methods with adequate sensitivity and refined extraction procedures for this determination. Moreover, the authority called upon new arsenic speciation data for complex food matrices such as seaweeds, grains and grain-based products. Looking at this context, an optimized, sensitive and fast analytical method using high performance liquid chromatography followed by inductively coupled plasma-mass spectrometry (HPLC/ICP-MS) was developed for the determination of iAs (sum of arsenite-AsIII and arsenate-AsV) and the most relevant oAsCs, arsenobetaine, dimethylarsinic acid and monomethylarsonic acid. The method was validated with satisfactory results in terms of linearity, sensitivity, selectivity, precision, recovery, uncertainty, ruggedness and matrix effect, and then successfully applied for the analysis of several matrices, i.e., processed and unprocessed cereal and cereal products, fruits, vegetables, legumes, seaweeds, nuts and seeds. The results obtained indicate that not only seaweed and rice matrices but also many cereals, legumes and plant-based foods for infants and young children contain significant concentrations of iAs and oAsCs. These findings contribute to the data collection necessary to assess the role of these matrices in the total arsenic exposure and if specific maximum limits have to be established.

Identification of mechanically separated meat using multivariate analysis of 43 trace elements detected by inductively coupled mass spectrometry: A validated approach.

The European Food Safety Authority asked for novel approaches for identifying mechanically separated meat (MSM) in meat products, due to food safety concern. In this study, a novel approach based on multivariate analysis of 43 trace elements in meat products is described. Overall, 27 trace elements and 16 rare earth elements were determined by using ICP-MS after sample mineralization of 100 meat samples, composed of different percentages of MSM, obtained at low and high pressure, and without MSM. After development and optimization, the multivariate approach was validated by analyzing and then classifying 10 "blind" meat samples, obtaining method accuracy equal to 90%. Thus, the applicability of this new analytical approach was demonstrated. The method represents a significant improvement for this type of determination, especially when MSM is obtained at low pressure, since this product is characterized by chemical characteristics very similar to fresh meat.

Optimizing accelerated solvent extraction combined with liquid chromatography-Orbitrap mass spectrometry for efficient lipid profile characterization of mozzarella cheese.

In this study, a novel Accelerated Solvent Extraction (ASE) procedure combined with UHPLC-Q-Orbitrap-MS was developed for detailed untargeted lipid profile of mozzarella cheese. Response Surface Methodology and Pareto front, using a Central Composite Design (CCD), were employed to define the optimised combination of extraction temperature, number of extraction cycles and mix of solvents. LipidSearch™ software was used for a reliable and accurate lipid identification. A total of 13 subclasses, including ceramides, diacylglycerols, triacylglycerols, lysophosphatidylcholines, lysophosphatidylethanolamines, phosphatidylcholines, phosphatidylethanolamines, phosphatidylinositols, sphingomyelins, bismethyl phosphatidic acids, cholesterol ester, zymosterol ester, hexosyl ceramides were measured. The elaboration of the CCD showed that the solvents ratio was the main factor affecting the extraction efficiency. The optimised ASE method, together with the Folch extraction, synergistically contributed to a complete characterization of lipid profile of mozzarella cheese, confirming ASE technique, associated with high resolution mass spectrometry detection, as an efficient tool for Lipidomics in food science.

Development of a screening analytical method for the determination of non-dioxin-like polychlorinated biphenyls in chicken eggs by gaschromatography and electron capture detection.

A sensitive and reproducible screening analytical method is here proposed for the determination of six non dioxin-like polychlorinated biphenyls (NDL-PCBs, congener 28, 52, 101, 138, 153, 180) in chicken eggs based on accelerated solvent extraction (ASE) procedure for the fat extraction and determination, a solid phase extraction (SPE) sample clean-up process, and a gas chromatography - electron capture detection (GC-ECD) analysis. The optimized chromatographic separation, in less than 25 min, returned good responses for the six NDL-PCBs in the range of 2.5-60.0 µg L-1, with correlation coefficients always higher than 0.9995. Instrumental limits of detection were between 0.08-0.35 µg L-1, corresponding to 0.05 and 0.23 ng g-1 fat in the matrix, while method detection limits, calculated on spiked egg samples, ranged from 1.6 to 3.5 ng g-1 fat. The method has been extensively validated in terms of selectivity, sensitivity, recovery, precision, ruggedness and measurement uncertainty, following the European Directives.

Sensitive Screening Method for Determination of Pyrethroids in Chicken Eggs and Various Meat Samples by Gas Chromatography and Electron Capture Detection.

A sensitive and reproducible screening analytical method is described for the determination of six pyrethroids (phenothrin, permethrin, cyfluthrin, cypermethrin, deltamethrin, fenvalerate) in egg and meat samples by gas chromatography and electron capture detection (GC-ECD). A fast cleanup procedure, based on solid-phase extraction has been used, ensuring reduced solvent consumption and time and allowing the simultaneous preparation of multiple sample extracts. Under the optimal chromatographic conditions, an efficient separation was obtained with a total analysis time of less than 60 min, including the extraction-purification steps. Good responses for the six pyrethroids were obtained in a range of 50-500 µg L-1, with linear coefficients higher than 0.9992. Instrumental limits of detection were between 0.22 and 0.63 µg L-1, corresponding to 0.04 and 0.13 µg kg-1 in the matrix. Detection limits in chicken eggs and various meat samples, calculated on spiked samples, were in the range 0.05-0.25 µg kg-1 and 0.07-0.23 µg kg-1, respectively. The validation results confirmed that the proposed GC-ECD method can be used as a reliable screening tool for the determination of pyrethroids in official check analyses. The method was extensively validated following the European directives, demonstrating its conformity in terms of selectivity, sensitivity, recovery, precision, and measurement uncertainty.

Intake estimates of dioxins and dioxin-like polychlorobiphenyls in the Italian general population from the 2013-2016 results of official monitoring plans in food.

The implementation of the European Union strategy for polychlorodibenzo-dioxins and -furans (PCDD/Fs), and dioxin-like polychlorobiphenyls (DL-PCBs) is determining a general reduction of their presence in the environment and in the food chain. The most important route for human exposure to these substances is food consumption and, as a consequence, a progressive decrease of their dietary intake has been observed in the last decades. In this context, it seemed worth updating the PCDD/F and DL-PCB intake estimation for the Italian population. A total of 2659 samples of food of animal and vegetable origin analyzed for PCDD/Fs and DL-PCBs in the period 2013-2016 by accredited official laboratories and the national food consumption database were considered for the dietary intake assessment in different age groups of the Italian general population The median cumulative intake estimates expressed as pg WHO-TEQ/kg body weight per day and computed with a deterministic and a probabilistic approach were 1.40-1.52 for children, 0.82-0.85 for adolescents, and 0.64-0.61 for adults, respectively. Such results confirm the decreasing trend of PCDD/F and DL-PCB dietary intake even though the Tolerable Daily Intake (TDI) value of 2 WHO-TEQ/kg body weight per day is exceeded at the 95th percentile for all age groups, with children as sensitive group. Most contributing food categories to the intake resulted fish, food of vegetable origin, and cheese. A sensitivity analysis was also performed to calculate the target contamination levels able to keep the dietary exposure below the TDI. Computed target levels fall between P50 and P97 of the occurrence distribution of the main food groups, meaning that most of the Italian food production can be considered safe.

Multi-residue method for the determination of organochlorine pesticides in fish feed based on a cleanup approach followed by gas chromatography-triple quadrupole tandem mass spectrometry.

A multi-residue method for the determination of organochlorine pesticides in fish feed samples was developed and optimized. The method is based on a cleanup step of the extracted fat, carried out by liquid-liquid extraction on diatomaceous earth cartridge with n-hexane/acetonitrile (80/20, v/v) followed by solid phase extraction (SPE) with silica gel-SCX cartridge, before the identification and quantification of the residues by gas chromatography-triple quadrupole tandem spectrometry (GC-MS/MS). Performance characteristics, such as accuracy, precision, linear range, limits of detection (LOD) and quantification (LOQ), for each pesticide were determined. Instrumental LODs ranged from 0.01 to 0.11 microg L(-1), LOQs were in the range of 0.02-0.35 microg L(-1), and calibration curves were linear (r2>0.999) in the whole range of explored concentrations (5-100 microg L(-1)). Repeatability values were in the range of 3-15%, evaluated from the relative standard deviation of six samples spiked at 100 microg kg(-1) of fat, and in compliance with that derived by the Horwitz's equation. No matrix effects or interfering substances were observed in fish feed analyses. The proposed method allowed high recoveries (92-116%) of spiked extracted fat samples at 100 microg kg(-1), and very low LODs (between 0.02 and 0.63 microg kg(-1)) and LOQs (between 0.05 and 2.09 microg kg(-1)) determined in fish feed samples.