LC-HRMS analysis of 13 classes of pharmaceutical substances in food supplements.

Food supplements should not contain substances considered unsafe or pose a health risk to consumers. In recent years illegal adulterants have been found in various functional foods without notification of their presence or amount in the labelling. In this study, a validated method was developed and applied as a screening method to detect 124 forbidden substances belonging to 13 classes of compounds in food supplements. Liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) and a simple and rapid extraction protocol was applied to 110 food supplements collected from the internet market or during official controls in Italy. The percentage of non-compliant samples was 4.5%, relatively high compared with the official control results for these substances usually obtained on other food matrices. The results suggested the need to strengthen controls in this field to detect food supplement adulteration, which represents a potential health risk for the consumer.

Occurrence and migration study of chemicals from baking paper and aluminium foil.

The present work focused on the development of an analytical method suitable to study the presence and the release of organophosphate esters (OPEs) and perfluoroalkylated substances (PFASs) from food contact materials (FCMs), from baking paper and aluminium foil. Although these classes of compounds are attracting increasing attention due to their toxicity the knowledge is still insufficient. The extent of their migration from FCMs to food was estimated using different liquid simulants. Ethanol 95 % was used to simulate the contact with fatty food, whilst acetic acid 3 % to mime contact with acidic aqueous-based food and the contact mode involved the use of the ultrasound-assisted technique. Preliminary results showed the higher migration for baking paper samples in contact with aqueous simulant with contaminations in the ranges 78.30-413.21 ng/dm2 and 1.43-13.87 ng/dm2 for OPEs and PFASs respectively. These findings highlighting the need to monitor particularly OPEs in FCMs.

Detection of Nitrated, Oxygenated and Hydrogenated Polycyclic Aromatic Compounds in Smoked Fish and Meat Products.

Polycyclic aromatic hydrocarbons (PAHs) are present in smoked food products. More toxic nitrated (NPAH) and oxygenated (OPAH) PAHs derivatives are found concomitantly to PAHs and are therefore believed to be found in smoked food products. However, only a few PAH analyses on food include these derivatives. We adjusted and successfully validated a GC-QTOFMS method including 13 NPAHs and 2 OPAHs as well as the 4 regulated PAHs for analysis of 14 smoked (13 fish and one bacon) and one pan fried fish samples.OPAHs were detected in the highest concentrations in 13 of 15 samples. Non-target screening revealed the presence of an additional four OPAHs and two methylated PAHs. Future food analysis should, based on these results, focus on PAH and oxygenated derivatives.

Characterization of organophosphate esters (OPEs) and polyfluoralkyl substances (PFASs) in settled dust in specific workplaces.

An analytical method for detecting flame retardants was slightly modified and optimized for the simultaneous determination of 11 organophosphate esters (OPEs) and 26 polyfluoralkyl substances (PFASs) contained in dust. All the analytes were determined in HPLC/MS-MS, and OPEs were also analyzed in GC/MS, and the results were compared. The study was conducted through the investigation of the Standard Reference Material SRM 2585 of the National Institute of Standard and Technology (NIST). The results were compared with the available reference mass fraction reported in the NIST certificate. The mass fraction obtained for the other OPEs and PFASs was compared to available data in the literature. After verifying the reliability of the results, the method was applied to environmental samples of settled dust, collected in four workplaces, where OPE and PFAS content is expected to be higher than in house dust: a mechanical workshop, an electronic repair center, a disassembly site, and a shredding site of two electronic waste recycling plants. By analyzing both PFASs and OPEs in the same samples, the present work demonstrated that the selected working places were more polluted in OPEs than houses; on the contrary, PFAS content in house dust proved to be more than ten times higher than that in workplaces. Additional research is necessary to confirm these data. Nevertheless, because this preliminary study showed not negligible concentrations of OPEs in some workplaces and of PFASs in houses, their monitoring should be extended to other domestic and selected working sites.

Toxic Organic Contaminants in Airborne Particles: Levels, Potential Sources and Risk Assessment.

In the last years, many studies have focused on risk assessment of exposure of workers to airborne particulate matter (PM). Several studies indicate a strong correlation between PM and adverse health outcomes, as a function of particle size. In the last years, the study of atmospheric particulate matter has focused more on particles less than 10 µm or 2.5 µm in diameter; however, recent studies identify in particles less than 0.1 µm the main responsibility for negative cardiovascular effects. The present paper deals with the determination of 66 organic compounds belonging to six different classes of persistent organic pollutants (POPs) in the ultrafine, fine and coarse fractions of PM (PM < 0.1 µm; 0.1 < PM < 2.5 µm and 2.5 < PM < 10 µm) collected in three outdoor workplaces and in an urban outdoor area. Data obtained were analyzed with principal component analysis (PCA), in order to underline possible correlation between sites and classes of pollutants and characteristic emission sources. Emission source studies are, in fact, a valuable tool for both identifying the type of emission source and estimating the strength of each contamination source, as useful indicator of environment healthiness. Moreover, both carcinogenic and non-carcinogenic risks were determined in order to estimate human health risk associated to study sites. Risk analysis was carried out evaluating the contribution of pollutant distribution in PM size fractions for all the sites. The results highlighted significant differences between the sites and specific sources of pollutants related to work activities were identified. In all the sites and for all the size fractions of PM both carcinogenic and non-carcinogenic risk values were below acceptable and safe levels of risks recommended by the regulatory agencies.

Characterization of seven sterols in five different types of cattle feedstuffs.

This paper provides a method for the quantification of sterols in different types of calf feedstuffs based on soy, sunflower, hay, calf feed and a mixture of all of them. The free fraction and the total sterolic fraction, after saponification and acidic hydrolysis of the samples, are extracted by solvent and the sterols are identified/quantified by reversed phase HPLC coupled to tandem mass spectrometry by atmospheric pressure chemical ionization. After the recovery evaluation, the method is validated in terms of linearity (coefficient of determination R2), repeatability (coefficient of variation RSD), limit of detection and quantification. In most of the cases, the most representative phytosterol is ß-sitosterol, followed by campesterol or stigmasterol and by other minor sterols such as fucosterol, and Δ-5-avenasterol. In addition, also cholesterol and ergosterol, if present, are evaluated in all the samples. As far as we know, very little information is available on the investigated feeds, which are commonly used on farms. The results of this survey were compared to other studies, if present in literature, showing good agreement. The proposed method resulted to be simple, fast and suitable for application to other sterols, feedstuffs and derived foods. The knowledge of the sterolic content and composition is getting more and more important, both in terms of comprehension of the vegetal biochemistry and as basis for sterolomic studies.

Occurrence of Halogenated Pollutants in Domestic and Occupational Indoor Dust.

The occurrence of halogenated organic pollutants in indoor dust can be high due to the presence of textile, electronic devices, furniture, and building materials treated with these chemicals. In this explorative study, we focused on emerging organic pollutants, such as novel brominated flame retardants (nBFRs) and some perfluoroalkyl substances, together with legacy polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (BDEs) in settled dust collected in houses and workplaces such as one office and two electrotechnical and mechanical workshops. The total contribution of the investigated pollutants was lower in house and in office dusts except for few nBFRs (such as bis (2-ethylhexyl)-3,4,5,6-tetrabromo-phthalate at a concentration of 464.5 ng/g in a house and hexachlorocyclopentadienyldibromocyclooctane at 40.4 ng/g in the office), whereas in electrotechnical and mechanical workshops a high incidence of PCBs, BDEs, and nBFRs occurred (for example, BDE 209 at a concentration of 2368.0 ng/g and tetrabromobisphenol A at 32,320.1 ng/g in electrotechnical and mechanical workshops). Estimated daily intakes were also calculated, showing that domestic and occupational environments can lead to a similar contribution in terms of human exposure. The higher exposure contribution was associated to nBFRs, whose EDIs were in the range of 3968.2-555,694.2 pg/kg bw/day. To provide a complete view about the indoor contamination, in this investigation, we also included polycyclic aromatic hydrocarbons (PAHs) and their oxygenated and nitrated derivatives. Definitely, dust collection represents a simple, fast, and cost-effective sampling and dust contamination level can be a useful indicator of environment healthiness. Besides, the presented method can be a smart tool to provide a time and money saving technique to characterize 99 pollutants thanks to a single sample treatment.

Optimization and validation of a LC-HRMS method for aflatoxins determination in urine samples.

Mycotoxins' exposure by inhalation and/or dermal contact can occur in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the possible contribution of the occupational exposure to aflatoxins by analysing urine samples for the presence of aflatoxins B1 and M1 and aflatoxin B1-N7-guanine adduct. The study was conducted in 2017 on two groups of volunteers, the workers group, composed by personnel employed in an Italian feed plant (n = 32), and a control group (n = 29), composed by the administrative employees of the same feed plant; a total of 120 urine samples were collected and analysed. A screening method and a quantitative method with high-resolution mass spectrometry determination were developed and fully validated. Limits of detections were 0.8 and 1.5 pg/mLurine for aflatoxin B1 and M1, respectively. No quantitative determination was possible for the adduct aflatoxin B1-N7-guanine. Aflatoxin B1 and its adduct were not detected in the analysed samples, and aflatoxin M1, instead, was found in 14 samples (12%) within the range 1.9-10.5 pg/mLurine. Only one sample showed a value above the limit of quantification (10.5 pg/mLurine). The absence of a statistical difference between the mean values for workers and the control group which were compared suggests that in this specific setting, no professional exposure occurs. Furthermore, considering the very low level of aflatoxin M1 in the collected urine samples, the contribution from the diet to the overall exposure is to be considered negligible.

Determination of Deoxynivalenol Biomarkers in Italian Urine Samples.

Deoxynivalenol (DON) is a mycotoxin mainly produced by Fusariumgraminearum that can contaminate cereals and cereal-based foodstuff. Urinary DON levels can be used as biomarker for exposure assessment purposes. This study assessed urinary DON concentrations in Italian volunteers recruited by age group, namely children, adolescents, adults, and the elderly. In addition, vulnerable groups, namely vegetarians and pregnant women, were included in the study. To determine the urinary DON, its glucuronide and de-epoxydated (DOM-1) forms, an indirect analytical approach was used, measuring free DON and total DON (as sum of free and glucuronides forms), before and after enzymatic treatment, respectively. Morning urine samples were collected on two consecutive days, from six different population groups, namely children, adolescent, adults, elderly, vegetarians and pregnant women. Total DON was measured in the 76% of the collected samples with the maximum incidences in children and adolescent age group. Urine samples from children and adolescent also showed the highest total DON levels, up to 17.0 ng/mgcreat. Pregnant women had the lowest positive samples per category (40% for day 1 and 43% for day 2, respectively), low mean levels of total DON (down to 2.84 ng/mgcreat) and median equal to 0 ng/mgcreat. Estimation of DON dietary intake reveals that 7.5% of the total population exceeds the TDI of 1 µg/kg bw/day set for DON, with children showing 40% of individuals surpassing this value (male, day 2).

Biomonitoring Data for Assessing Aflatoxins and Ochratoxin A Exposure by Italian Feedstuffs Workers.

Mycotoxins exposure by inhalation and/or dermal contact is possible in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the validity of the biomonitoring as a tool to investigate the intake of mycotoxins in a population of workers operating in an Italian feed plant. Serum samples were collected for the determination of aflatoxins B1 (AFB1), AFB1-Lysine adduct and ochratoxin A (OTA). A method based on liquid-liquid extraction coupled with high resolution mass spectrometry determination was developed and fully validated. For AFB1, a high number of non-detected samples (90%) was found and no statistical difference was observed comparing workers and control group. None of the analyzed samples showed the presence of AFB1-Lysine adduct. For OTA, the 100% of the analyzed samples was positive with a 33% of the samples showing a concentration higher than the limit of quantification (LOQ), but no statistical difference was highlighted between the average levels of exposed and control groups. In conclusion, the presence of AFB1 and OTA in serum cannot be attributable to occupational exposure.

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.