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.

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.

Poaching of endangered species in Mediterranean Sea: Alopias vulpinus.

A thresher shark poached by fishermen in the stretch of sea between the island of Ponza and the coast of Terracina (Lazio, Central Italy, Mediterranean Sea) and confiscated by the Italian health authorities was evaluated for the presence of chemical contaminants. Hg mean concentration of 2.1 mg kg-1 was found in muscle; speciation analysis confirmed it was almost in the toxic organic form of MeHg (2.0 mg kg-1). The Se:Hg molar ratio was <1 not protective against the toxic effect of Hg. 137Cs was detected at a mean concentration of 0.53 Bq kg-1, and 40K, radioisotope of natural origin was found at 91 Bq kg-1. Dioxins and PCBs were found in the common thresher shark at mean concentrations of 0.012 and 0.088 pg TEQ-WHO g-1 w.w. while NDL PCBs at 4.5 ng g-1 w.w., lower than limits set by European Regulation. The major concern is constituted by the presence of Hg twice the maximum limit set by EU Regulation, then the consumption of thresher shark flesh should be strongly discouraged.

Evaluation and Dietary Exposure Assessment of Selected Toxic Trace Elements in Durum Wheat (Triticum durum) Imported into the Italian Market: Six Years of Official Controls.

Durum wheat grains, which are mostly used for the production of pasta and several baked goods, represent a main source of vegetable proteins and calories. Concurrently, many contaminants, including toxic trace elements, may accumulate in them, posing a potential severe hazard to human health. In this context, for official control and food safety purposes, 346 samples of whole durum wheat imported into the Italian market from six countries (Australia, Canada, Kazakhstan, Russia, Turkey, and the United States) during the period 2015-2020 were analysed for cadmium (Cd), lead (Pb), and mercury (Hg) content using inductively coupled plasma mass spectrometry (ICP-MS). All the analysed samples were compliant with Food Agriculture Organization-World Health Organization and European Union regulations. The mean values were 0.0322 and 0.0162 mg kg-1, respectively, for Cd and Pb, while all samples showed levels below the limit of detection (0.004 mg kg-1) for Hg. The results were construed in terms of seasonality, year, and country of production, and compared with reference tolerance values. Confirming previous exposure studies, the obtained data and the dietary intake assessment showed that durum wheat-based products may have a significant impact on exposure to Pb and Cd (20-50%) in the overall population, particularly in more sensitive and/or exposed subgroups (infants, toddlers, and females).

Innovative approaches for identifying a mechanically separated meat: evaluation of radiostrontium levels and development of a new tool of investigation.

Due to food safety concerns, the European Food Safety Authority (EFSA) asked to identify useful parameters for identifying the mechanically separated meat (MSM). In this work, the 90Sr levels of MSM products were assessed for evaluating the suitability of this parameter for MSM identification. Indeed, this contaminant may accumulate in animal bones and bone fragments may be present in MSM. One hundred samples of fresh meats and meat products composed of different MSM percentages and without MSM were analyzed by ultra-low-level liquid scintillation counting. The 90Sr activity concentrations detected in meat products with high percentage of MSM were significantly higher (p ≥ 0.05) (0.099 Bq kg-1) than those of fresh meats (0.024 Bq kg-1) and meat products containing low percentage of MSM (0.011 Bq kg-1). The percentage of correct MSM identification was 61%. In order to increase this percentage and to develop a new tool of investigation, the 90Sr was elaborated in combination with other three parameters: 88Sr, Ca and ash percentage, through a multivariate approach. The precision of this tool of investigation was significantly increased (87%), higher than the reference method (Ca level) (76%), representing a valid novel approach in the identification of MSM products.

Trace elements in raw milk of buffaloes (Bubalus bubalis) from Campania, Italy.

The profile of 18 trace elements was traced in 68 milk samples collected from buffalo farms in the territory known as the "Land of Fires" in the Campania region (Italy). This area has been polluted by the illegal dumping in fields of industrial or domestic waste, wich is sometimes then burned spreading toxic contaminants. Milk from buffaloes raised on rural farms might be a good indicator of environmental contamination risk in the human food chain. Trace element analysis in milk was performed using mass spectrometry. One milk sample was found to be non-compliant due to high Pb concentration. In the absence of threshold values for the elements, established through legislation, the results were compared with similar studies from other countries, and in most cases the content determined in this study was in agreement with values reported elsewhere and do not represent a risk to human health.

[Levels of trace elements in the fruits and vegetables from the so-called Land of fires in Campania Region (Southern Italy)]. / Livelli di elementi in traccia tossici e potenzialmente tossici nei prodotti ortofrutticoli della Terra dei fuochi in Campania.

INTRODUZIONE: una vasta area compresa tra le province di Napoli e Caserta nella regione Campania ha negli ultimi anni attirato l'attenzione dei mass media a causa degli interramenti illeciti di rifiuti di varia origine nei campi o del loro abbandono lungo le strade. A tali pratiche illecite si aggiunge il fenomeno dei roghi incontrollati di questo materiale, motivo per il quale la zona è stata soprannominata "Terra dei fuochi". OBIETTIVI: ricerca di elementi in tracce presenti nei prodotti alimentari di origine vegetale prodotti nella Terra dei fuochi. DISEGNO E SETTING: piano di monitoraggio per il rilevamento dei livelli di alcuni metalli tossici ed essenziali nei vegetali coltivati in Campania nella Terra dei fuochi. La determinazione dei microelementi è stata effettuata dopo mineralizzazione della sostanza organica mediante microonde, utilizzando la spettrometria di massa a plasma induttivamente accoppiato (ICP-MS). RISULTATI: durante l'attività di monitoraggio sono stati raccolti 65 campioni di ortaggi e 49 campioni di frutta in 43 comuni rientranti nella Terra dei fuochi. Il contenuto medio di metalli tossici, cadmio e piombo nei prodotti ortofrutticoli è risultato ben lontano dai limiti massimi fissati dalla normativa europea. I valori delle concentrazioni sono significativamente inferiori rispetto a quanto riscontrato in zone ad alto impatto ambientale a causa della presenza di impianti industriali. CONCLUSIONI: la presenza di elementi tossici o potenzialmente tali nei campioni vegetali prodotti nella Terra dei fuochi è attribuibile a una varietà di fattori ambientali rilevanti (caratteristiche geologiche dell'area, inquinamento del suolo, dell'atmosfera e delle acque di irrigazione, pratiche illecite di smaltimento dei rifiuti industriali); non sembra, tuttavia, sussistere un rischio per la salute dei consumatori. Resta comunque l'allerta per la loro presenza nell'ambiente e la necessità che le attività di monitoraggio continuino.

Characterization, chemometric evaluation, and human health-related aspects of essential and toxic elements in Italian honey samples by inductively coupled plasma mass spectrometry.

Concentration values of 24 elements (Al, As, Ba, Be, Ca, Cd, Co, Cr, Cu, Fe, Ge, Hg, Mn, Mo, Pb, Sb, Se, Sn, Sr, Ti, Tl, U, V, and Zn) were determined in 72 honey samples produced in Italy by inductively coupled plasma mass spectrometry (ICP-MS). Considering the recommended established heavy metal daily intakes for humans, in this perspective, an equilibrated and ordinary honey consumption should not be considered matter of concerns for human health, even if particular attention should be addressed if honey is consumed by children, due to different maximum daily heavy metal intakes. Chemometric analysis of the results obtained highlights heavy metal content differences in honey samples obtained from notoriously polluted zones, confirming then that honey can be considered a bio-indicator of environmental pollution. Finally, Pearson coefficients highlighted correlations among element contents in honey samples.

Environmental monitoring of the area surrounding oil wells in Val d'Agri (Italy): element accumulation in bovine and ovine organs.

In this work, environmental heavy metal contamination in the Val d'Agri area of Southern Italy was monitored, measuring the accumulation of 18 heavy metals (U, Hg, Pb, Cd, As, Sr, Sn, V, Ni, Cr, Mo, Co, Cu, Zn, Ca, Mn, Fe, and Al) in the organs of animals raised in the surrounding area (kidney, lung, and liver of bovine and ovine species). Val d'Agri features various oil processing centers which are potentially a significant source of environmental pollution, making it essential to perform studies that will outline the state of the art on which any recovery plans and interventions may be developed. The analysis was carried out using official and accredited analytical methods based on inductively coupled plasma mass spectrometry, and the measurements were statistically processed in order to give a contribution to risk assessment. Even though five samples showed Pb and Cd concentrations above the limits defined in the European Commission Regulation (EC) No 1881/2006, the mean concentrations of most elements suggest that contamination in this area is low. Consequently, these results also suggest that there is no particular risk for human exposure to toxic trace elements. Nevertheless, the findings of this work confirm that element accumulation in ovine species is correlated with geographical livestock area. Therefore, ovine-specific organs might be used as bioindicators for monitoring contamination by specific toxic elements in exposed areas.

Trace elements in free-range hen eggs in the Campania region (Italy) analyzed by inductively coupled plasma mass spectrometry (ICP-MS).

Eggs from hens raised on rural or domestic farms are a good indicator of environmental contamination, as the hens are in close contact with the ground and the air and can therefore accumulate heavy metals and other toxic contaminants from the environment as well as from the diet. In this paper, we report the results of the determination of 19 trace elements (As, Be, Cd, Co, Cr, Cu, Hg, Mo, Mn, Ni, Pb, Sb, Se, Sn, Sr, Tl, U, V, Zn) in 39 hen egg samples collected from domestic poultry farms in the territory dubbed the "Land of fires" in the Campania region (Italy). This area is characterized by environmental problems caused by the illegal dumping of industrial or domestic waste in fields or by roadsides. In some cases, these wastes have been burned, thereby spreading persistent contaminants into the atmosphere. The content of trace elements in whole egg samples was determined by mass spectrometer after a microwave-assisted digestion procedure. Because European legislation does not indicate maximum values of these elements in this foodstuff, the results were compared with the content of trace elements reported in literature for eggs, in particular home-produced eggs, in various countries. In some cases (Cd, Cu, Ni, Mn), the content determined in this study was in line with those reported elsewhere, in other cases (Pb, Cr), lower values were found.

Distinctive Pattern of Serum Elements During the Progression of Alzheimer's Disease.

Element profiling is an interesting approach for understanding neurodegenerative processes, considering that compelling evidences show that element toxicity might play a crucial role in the onset and progression of Alzheimer's disease (AD). Aim of this study was to profile 22 serum elements in subjects with or at risk of AD. Thirtyfour patients with probable AD, 20 with mild cognitive impairment (MCI), 24 with subjective memory complaint (SMC) and 40 healthy subjects (HS) were included in the study. Manganese, iron, copper, zinc, selenium, thallium, antimony, mercury, vanadium and molybdenum changed significantly among the 4 groups. Several essential elements, such as manganese, selenium, zinc and iron tended to increase in SMC and then progressively to decrease in MCI and AD. Toxic elements show a variable behavior, since some elements tended to increase, while others tended to decrease in AD. A multivariate model, built using a panel of six essential elements (manganese, iron, copper, zinc, selenium and calcium) and their ratios, discriminated AD patients from HS with over 90% accuracy. These findings suggest that essential and toxic elements contribute to generate a distinctive signature during the progression of AD, and their monitoring in elderly might help to detect preclinical stages of AD.

Assessment of lead, cadmium and mercury in seafood marketed in Puglia and Basilicata (Italy) by inductively coupled plasma mass spectrometry.

Lead, cadmium and mercury are non-essential heavy metals that may interfere with biological systems, exhibiting high toxicity to human and marine biota. Due to bioaccumulation of heavy metals in the food chain, seafood may concentrate high levels of these contaminants since they are often at the top of aquatic food chain. In this study, 342 seafood samples, subdivided into four categories (bivalve molluscs, cephalopod molluscs, blue-fish and other sea fish), were analysed by inductively coupled plasma mass spectrometry, in order to assess the levels of lead, cadmium and mercury. Contamination levels higher than allowable limits were verified. In particular, two bivalve mollusc samples were non-compliant for lead and cadmium, four cephalopod mollusc samples non-compliant for cadmium and 14 samples (4 blue-fish and 10 other sea fish) non-compliant for mercury. This survey confirmed the necessity to perform routinely controls related to this type of food inspection.