Exposure Assessment to Deoxynivalenol of Children over 3 Years Deriving from the Consumption of Processed Wheat-Based Products Produced from a Dedicated Flour.

Wheat-based products are largely consumed by children worldwide. Deoxynivalenol (DON) is known for its acute and chronic toxicity and is the most common contaminant of cereal grains. Since no legal limits are set for DON in wheat-based products and specific foods intended for children over 3 years on the market, a high risk of overexposure to this contaminant may emerge. The main objective of the study, conducted in 2018-2019, was to produce a wheat flour intended for children over three years, characterized by a high level of safety in terms of DON content, to be used to produce wheat-derived products. The dedicated flour was produced by adopting tailored procedures like the selection of wheat suppliers, the predetermination of the safe contamination of DON in the final products, and the evaluation of the transfer rate from the wheat flour to derived products (bread, breadsticks, biscuits, plumcake, and focaccia). The results showed that the daily exposure of children was considered to be safe, in a range between 7% (biscuits) and 67% (bread) of DON tolerable daily intake (TDI) and that only by producing a flour characterized by DON levels much lower than those in force, can "safe" products be marketed.

A validated LC-MS/MS method for quantitative determination of L-dopa in Fagioli di Sarconi beans (Phaseolus vulgaris L.).

An analytical method based on ultrasound assisted extraction (UAE) and liquid chromatography coupled to electrospray tandem mass spectrometry (LC-ESI/MS/MS) was validated and applied for determining L-dopa in four ecotypes of Fagioli di Sarconi beans (Phaseolus vulgaris L.), marked with the European label PGI (Protected Geographical Indication). The selectivity of the proposed method was ensured by the specific fragmentation of the analyte. Simple isocratic chromatographic conditions and mass spectrometric detection in multiple reaction monitoring (MRM) acquisition mode were used for sensitive quantification. The LC-ESI/MS/MS method was validated within a linear range of 0.001-5.000 µg/mL. Values of 0.4 and 1.1 ng/mL were obtained for the limits of detection and quantification, respectively. The repeatability, inter-day precision, and recovery values ranges were 0.6%-4.5%, 5.4%-9.9%, and 83%-93%, respectively. Fresh and dried beans, as well as pods, cultivated exclusively with organic methods avoiding any synthetic fertilizers and pesticides were analyzed showing an L-dopa content ranging from 0.020 ± 0.005 to 2.34 ± 0.05 µg/g dry weight.

Determination of ochratoxin A in pork meat products: single laboratory validation method and preparation of homogeneous batch materials.

Ochratoxin A is one of the most diffused mycotoxin present in a large spectrum of food commodities, mainly produced by Aspergillus ochraceus, Aspergillus carbonarius, Aspergillus niger and Penicillium verrucosum. EU has set maximum limits for a number of matrices such as cereals, wine, spices and liquorice, whilst other commodities such as beer and meat products that are susceptible of OTA contamination and are largely consumed are not included. In 2013, within the framework of the Regulation (EC) 882/2004 on official controls, the European Commission issued the mandate M/520 regarding the standardisation for methods of analysis for mycotoxins in food to the European Committee for Standardisation. Of the 11 priorities of the mandate, the one on "HPLC determination of OTA in meat, meat products and edible offal" was assigned to the Italian National Reference Laboratory for feed and food. The method was single-laboratory validated, and all the performance characteristics of the method were compliant with the corresponding reference values indicated in Regulation (EC) n. 401/2006. The method was applied to characterise a set of 5 pork-based materials (ham, kidney, liver and canned chopped pork) to be used for an inter-laboratory method validation study. Three ham materials (levels of contamination of 0.77, 2.22 and 12.3 µg/kg, respectively), one liver material (contamination level of 2.80 µg/kg) and one chopped pork meat (contamination level of 0.66 µg/kg) were tested for homogeneity and stability.

Ergot Alkaloids in Wheat and Rye Derived Products in Italy.

Genus Claviceps is a plant pathogen able to produce a group of toxins, ergot alkaloids (EAs), whose effects have been known since the Middle Ages (ergotism). Claviceps purpurea is the most important representative specie, known to infect more than 400 monocotyledonous plants including economically important cereal grains (e.g., rye, wheat, triticale). EAs are not regulated as such. Maximum limits are in the pipeline of the EU Commission while at present ergot sclerotia content is set by the Regulation (EC) No. 1881/2006 in unprocessed cereals (0.05% as a maximum). This study aimed to investigate the presence of the six principal EAs (ergometrine, ergosine, ergocornine, α-ergocryptine, ergotamine and ergocristine) and their relative epimers (-inine forms) in rye- and wheat-based products. Of the samples, 85% resulted positive for at least one of the EAs. Wheat bread was the product with the highest number of positivity (56%), followed by wheat flour (26%). Rye and wheat bread samples showed the highest values when the sum of the EAs was considered, and durum wheat bread was the more contaminated sample (1142.6 µg/kg). These results suggest that ongoing monitoring of EAs in food products is critical until maximum limits are set.

Study on the Association among Mycotoxins and other Variables in Children with Autism.

Environmental factors and genetic susceptibility are implicated in the increased risk of autism spectrum disorder (ASD). Mycotoxins are agricultural contaminants of fungal origin that represent real risk factors for human health and especially for children. Thus, the main hypothesis of this work is that the deterioration of the clinical manifestation of autism in children may result from the exposure to mycotoxins through the consumption of contaminated food. Within a cross-sectional study, a group of autistic children (n = 172) and a group of controls (n = 61) (siblings and non-parental) were recruited in North and South Italy. All children had blood and urine samples taken, for testing some mycotoxins by a LC-MS/MS validated method. Blood samples were also tested for assessing specific IgG against food and fungal antigens and cytokines. The analyses outputs highlighted statistically significant differences comparing mycotoxins levels between (i) children groups both in urine (deoxynivalenol and de-epoxydeoxynivalenol, p = 0.0141 and p = 0.0259, respectively) and serum (aflatoxin M1, ochratoxin A and fumonisin B1, p = 0.0072, p = 0.0141 and p = 0.0061, respectively); (ii) a group of selected fungal IgGs, and IgGs against wheat and gluten and (iii) cytokines. These results suggest the need for a deeper examination of the role that mycotoxins may have on the etiology of ASD.

Development of a LC-MS/MS Method for the Multi-Mycotoxin Determination in Composite Cereal-Based Samples.

The analytical scenario for determining contaminants in the food and feed sector is constantly prompted by the progress and improvement of knowledge and expertise of researchers and by the technical innovation of the instrumentation available. Mycotoxins are agricultural contaminants of fungal origin occurring at all latitudes worldwide and being characterized by acute and chronic effects on human health and animal wellness, depending on the species sensitivity. The major mycotoxins of food concern are aflatoxin B1 and ochratoxin A, the first for its toxicity, and the second for its recurrent occurrence. However, the European legislation sets maximum limits for mycotoxins, such as aflatoxin B1, ochratoxin A, deoxynivalenol, fumonisins, and zearalenone, and indicative limits for T-2 and HT-2 toxins. Due to the actual probability that co-occurring mycotoxins are present in a food or feed product, nowadays, the availability of reliable, sensitive, and versatile multi-mycotoxin methods is assuming a relevant importance. Due to the wide range of matrices susceptible to mycotoxin contamination and the possible co-occurrence, a multi-mycotoxin and multi-matrix method was validated in liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the purpose to overcome specific matrix effects and analyze complex cereal-based samples within the Italian Total Diet Study project.

New approach to the differentiation of marble samples using thermal analysis and chemometrics in order to identify provenance.

BACKGROUND: The possibility of applying a novel chemometric approach which could allow the differentiation of marble samples, all from different quarries located in the Mediterranean basin and frequently used in ancient times for artistic purposes, was investigated. By suggesting tentative or allowing to rule out unlikely attributions, this kind of differentiation could, indeed, be of valuable support to restorers and other professionals in the field of cultural heritage. EXPERIMENTAL DATA WERE OBTAINED ONLY USING THERMAL ANALYTICAL TECHNIQUES: Thermogravimetry (TG), Derivative Thermogravimetry (DTG) and Differential Thermal Analysis (DTA). RESULTS: The extraction of kinetic parameters from the curves obtained using these thermal analytical techniques allowed Activation Energy values to be evaluated together with the logarithm of the Arrhenius pre-exponential factor of the main TG-DTG process. The main data thus obtained after subsequent chemometric evaluation (using Principal Components Analysis) have already proved useful in the identification the original quarry of a small number of archaeological marble finds. CONCLUSION: One of the most evident advantages of the thermoanalytical - chemometric approach adopted seems to be that it allows the certain identification of an unknown find composed of a marble known to be present among the reference samples considered, that is, contained in the reference file. On the other hand with equal certainty it prevents the occurrence of erroneous or highly uncertain identification if the find being tested does not belong to the reference file considered.

Exposure assessment for Italian population groups to deoxynivalenol deriving from pasta consumption.

Four hundred and seventy-two pasta samples were collected from long retail distribution chain sales points located in North, Central and South Italy. Representative criteria in the sample collection were followed in terms of number of samples collected, market share, and types of pasta. Samples were analysed by an accredited HPLC-UV method of analysis. The mean contamination level (64.8 µg/kg) of deoxynivalenol (DON) was  in the 95th percentile (239 µg/kg) and 99th percentile (337 µg/kg), far below the legal limit (750 µg/kg) set by Regulation EC/1126/2007, accounting for about one tenth, one third and half the legal limit, respectively. Ninety-nine percent of samples fell below half the legal limit. On the basis of the obtained occurrence levels and considering the consumption rates reported by the Italian official database, no health concern was assessed for all consumer groups, being that exposure was far below the Tolerable Daily Intake (TDI) of 1000 ng/kg b.w/day. Nevertheless, despite this, particular attention should be devoted to the exposure to DON by high consumers, such as children aged 3-5 years, who could reach the TDI even with very low levels of DON contamination.

Characterization of wax manufactures of historical and artistic interest.

Purpose of this scientific research is the physic and chemical characterization of two historical wax manufactures, made at the end of XIX century by Francesco Bianchi, a papal engraver. The chemical and analytical investigation was necessary to complete and to confirm the restorer's work. The IR Spectroscopy, X-Ray and GC-MS, the best technique to characterise wax, allowed us to obtain the following results. The two manufactures were made with commercial beeswax: in fact, the relative chromatograms showed unchanged peaks about esters of palmitic acid with C24 to C32 alcohol molecules; using standard beeswax we determined the same amount of hydrocarbons present in the wax manufactures. We found several hydrocarbons in these beeswax materials so that it is reasonable to think about successive modifications. ZnO (white zinc), a pigment, was added, probably due to its colour and covering capacity. Sb2S3, Anthimoniun vermilion, a red-orange pigment, was added to these manufactures to give them a soft pink-orange colour. By all used techniques we determined some modifications in the original beeswax; surely they were made to get a more malleable, mouldable, and then more able to be modelled wax.