Contamination by Aflatoxins B/G in Food and Commodities Imported in Southern Italy from 2017 to 2020: A Risk-Based Evaluation.

This study reports the results of aflatoxins B/G monitoring in food of vegetal origin, imported in Southern Italy from extra-European Union countries. From 2017 to 2020, we analyzed 1675 samples using an accredited HPLC method with fluorescence detection. We found out 295 samples (17.6%) were contaminated by aflatoxin B1, 204 by aflatoxins B/G (12.2%), while 75 (4.5%) resulted non-compliant to maximum limits set by the European Union law. Most of the batches tested were unprocessed food; the distribution of contamination levels, incidence of non-compliant samples, inference for different kinds of food are reported. The study focuses on the food more susceptible to contamination by aflatoxins; nuts are the food more controlled, showing the higher number of non-compliant samples. Our study confirms that pistachio nuts, hazelnuts and almonds are the major sources of exposure for consumers. Still, other products, such as chili pepper and Brazil nuts, need to get more information about their contamination levels. The study's findings are discussed in the perspective of the last opinion by EFSA about chronic exposure to aflatoxins. A case study to evaluate not compliance of a composed food to the European Union law is reported.

A survey of Δ9-THC and relevant cannabinoids in products from the Italian market: A study by LC-MS/MS of food, beverages and feed.

In 2016, the European Commission recommended the Member States to monitor the content of Δ9-tetrahydrocannabinol and other cannabinoids in food and feed derived from hemp and in food of animal origin for possible transfer from feed. Thus, the Italian Ministry of Health implemented a monitoring plan. To this aim, nine cannabinoids in beverages and food for human consumption and in feed were determined. The method applied, based on rapid clean-up and LC-MS/MS determination, was previously developed and in-house validated, evaluating the analytical performance in the concentration ranges 2-50 µg/L for beverages, 0.020-0.500 mg/kg for food and 0.100-10.0 mg/kg for feed. Then, it was applied to determine the cannabinoids in 78 food, 16 beverage and 6 feed samples, collected from the Italian market since 2017. The results are herein reported, for evaluation of both product characteristics and compliance to national maximum limits. Some study cases are also described.

T-2 and HT-2 toxins in feed and food from Southern Italy, determined by LC-MS/MS after immunoaffinity clean-up.

A method to determine T-2 and HT-2 toxins, both in feed and food, is described. The method is based on sample clean-up by immunoaffinity columns, followed by triple quadrupole LC-MS/MS determination. It was in-house validated in compliance with the European Union legislation, in the concentration range 5.0-750 µg kg-1 and accredited according to the ISO/IEC 17025 standard, for the scope of official control of feed and food. The method was applied to monitor the contamination levels of T-2 and HT-2 toxins in 105 feed samples, both raw materials and compound feed and 17 cereals and derived products for human consumption, collected in Southern Italy between 2015 and 2019. T-2 toxin was quantified in 32 feed (30.5%) and in 2 food samples (11.8%). HT-2 toxin was quantified in 65 feed (61.9%) and in 3 food (17.6%) samples.

A rapid method to determine nine natural cannabinoids in beverages and food derived from Cannabis sativa by liquid chromatography coupled to tandem mass spectrometry on a QTRAP 4000.

RATIONALE: Phytocannabinoids are natural compounds produced by Cannabis spp. Some of these compounds show psychotropic effects on humans and are therefore used as drugs of abuse. These compounds are present in food and beverages containing ingredients from hemp, and thus can reach consumers. The Italian Ministry of Health planned to evaluate the intake of cannabinoids from food containing hemp ingredients. Thus, we were asked to develop and validate a multi-residue test method to determine nine phytocannabinoids in beverages and food. METHODS: Nine natural phytocannabinoids, hereafter called cannabinoids, were cleaned up from food by solid-liquid extraction, while beverages were simply diluted prior to analysis. The cannabinoids were separated by reversed-phase high-performance liquid chromatography, and on-line determination was carried out by tandem mass spectrometry using a 4000 QTRAP mass spectrometer with a TurboIonSpray source, in multiple-reaction monitoring mode, using both positive and negative ionization. RESULTS: Each compound was determined down to 0.25 ng/mL in solvent. In-house validation was carried out; the mean recoveries ranged from 83.4% to 101.2% in food, and from 84.5% to 104.5% in beverages. The limits of quantification were 20 µg/kg for food and 2 µg/L for beverages. CONCLUSIONS: A reliable and rapid method for the identification and quantification of the psychotropic Δ9 -tetrahydrocannabinol, its non-psychoactive precursor Δ9 -tetrahydrocannabinolic acid A, and seven other cannabinoids was developed and validated, to monitor the content of these substances in food and beverages produced using hemp seeds, flour and oil as ingredients.

Mediterranean Azadinium dexteroporum (Dinophyceae) produces six novel azaspiracids and azaspiracid-35: a structural study by a multi-platform mass spectrometry approach.

Azadinium dexteroporum is the first species of the genus described from the Mediterranean Sea and it produces different azaspiracids (AZA). The aims of this work were to characterize the toxin profile of the species and gain structural information on azaspiracids produced by the A. dexteroporum strain SZN-B848 isolated from the Gulf of Naples. Liquid chromatography-mass spectrometry (LC-MS) analyses were carried out on three MS systems having different ion source geometries (ESI, TurboIonSpray®, ESI ION MAX) and different MS analyzers operating either at unit resolution or at high resolution, namely a hybrid triple quadrupole-linear ion trap (Q-Trap MS), a time of flight (TOF MS), and a hybrid linear ion trap Orbitrap XL Fourier transform mass spectrometer (LTQ Orbitrap XL FTMS). As a combined result of these different analyses, A. dexteroporum showed to produce AZA-35, previously reported from Azadinium spinosum, and six compounds that represent new additions to the AZA-group of toxins, including AZA-54 to AZA-58 and 3-epiAZA-7, a stereoisomer of the shellfish metabolite AZA-7. Based on the interpretation of fragmentation patterns, we propose that all these molecules, except AZA-55, have the same A to I ring system as AZA-1, with structural modifications all located in the carboxylic side chain. Considering that none of the azaspiracids produced by the Mediterranean strain of A. dexteroporum is currently regulated by European food safety authorities, monitoring programs of marine biotoxins in the Mediterranean area should take into account the occurrence of the new analogues to avoid an underestimation of the AZA-related risk for seafood consumers. Graphical Abstract A multi-platform MS approach reveals known and new azaspiracids in a Mediterranean strain of Azadinium dexteroporum.

Monitoring the Presence of Domoic Acid in the Production Areas of Bivalve Molluscs.

Algal biotoxins, chemical compounds produced by some microscopic algae, constitute the phytoplankton. The mussels, feeding on phytoplankton, can accumulate these compounds to become themselves toxic. There have been several cases of food poisoning by consumption of contaminated shellfish. Such food poisoning have pushed our health care system to provide monitoring of shellfish in the framework of the monitoring plans carried out by AASSLL. In this paper we report the results obtained monitoring the presence of amnesic shellfish poisoning biotoxins, like domoic acid and its isomers, produced by Pseudonitzschia algae. The analyses were carried out by using both the high-performance liquid chromatography-ultraviolet official method and an experimental method performed with a time-of-flight mass spectrometer (ESI-TOF). The 100% of samples analysed by the official method have always been below the limits of sensitivity (except one sample), and the 65% of samples analysed by ESI-TOF showed the presence of domoic acid.

[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.

A new potentially toxic Azadinium species (Dinophyceae) from the Mediterranean Sea, A. dexteroporum sp. nov.

A new photosynthetic planktonic marine dinoflagellate, Azadinium dexteroporum sp. nov., is described from the Gulf of Naples (South Tyrrhenian Sea, Mediterranean Sea). The plate formula of the species, Po, cp, X, 4', 3a, 6″, 6C, 5?S, 6‴ and 2″″, is typical for this recently described genus. Azadinium dexteroporum is the smallest rep-resentative of the genus (8.5 µm average length, 6.2 µm average width) and shares the presence of a small antapical spine with the type species A. spinosum and with A. polongum. However, it differs from all other Azadinium species for the markedly asymmetrical Po plate and the position of the ventral pore, which is located at the right posterior end of the Po plate. Another peculiarity of A. dexteroporum is the pronounced concavity of the second intercalary plate (2a), which appears collapsed with respect to the other plates. Phylogenetic analyses based on the large subunit 28S rDNA (D1/D2) and the internal transcribed spacer (ITS rDNA) support the attribution of A. dexteroporum to the genus Azadinium and its separation from the other known species. LC/MS-TOF analysis shows that Azadinium dex-teroporum produces azaspiracids in low amounts. Some of them have the same molecular weight as known compounds such as azaspiracid-3 and -7 and Compound 3 from Amphidoma languida, as well as similar fragmentation patterns in some cases. This is the first finding of a species producing azapiracids in the Mediterranean Sea.

Palytoxin in seafood by liquid chromatography tandem mass spectrometry: investigation of extraction efficiency and matrix effect.

Blooms of Ostreopsis spp. have been recently reported along the Mediterranean coasts of Spain, France, Italy, and Greece posing serious risks to human health. Occurrence of Ostreopsis spp. may result in palytoxin contamination of seafood and, in order to prevent sanitary risks, the need exists to develop efficient extraction procedures to be coupled to rapid and sensitive monitoring methods of palytoxin-like compounds in seafood. In the present study, the best conditions for both extraction of palytoxin from seafood and palytoxin quantification by using liquid chromatography tandem mass spectrometry (LC-MS/MS) were investigated. Three seafood matrices (mussels, sea-urchins, and anchovies) were selected and five different extraction systems were tested, namely: the official protocol for extraction of lipophilic toxins and various aqueous methanol or acetonitrile solutions (MeOH/H(2)O 1:1, MeOH/H(2)O 8:2, MeCN/H(2)O 8:2 and MeOH 100%). Extraction with MeOH/H(2)O 8:2 provided the best results in terms of accuracy and matrix interference on LC-MS/MS detection of palytoxin. Accuracy and intra-day reproducibility (n = 3) were evaluated for all the selected matrices but only for mussels at three spiking concentration levels, including the provisional limit proposed by the Community Reference Laboratory for marine biotoxins (250 µg kg(-1)). Limits of quantitation of palytoxin in mussels, sea-urchins and anchovies tissues were calculated using matrix-matched standards; taking into account extraction efficiency of MeOH/H(2)O 8:2, they resulted to be 228, 343, and 500 µg kg(-1), respectively.

New palytoxin-like molecules in Mediterranean Ostreopsis cf. ovata (dinoflagellates) and in Palythoa tuberculosa detected by liquid chromatography-electrospray ionization time-of-flight mass spectrometry.

A rapid, high resolution liquid chromatography coupled with ElectroSpray Ionization Time-Of-Flight Mass Spectrometry (ESI/TOF/MS) method was developed for the determination of the toxin pattern in cultured cells of Ostreopsis cf. ovata from the Mediterranean Sea. The samples were separated on a Phenomenex Luna 3µ HILIC 200A (150 × 2.00 mm) and analyzed by LC/TOF/MS with electrospray ionization (ESI) interface in positive ion mode. The method developed here provides the capability for a fully automated analysis, which requires relatively easy sample preparation and gives clean and simple chromatograms. The method was successfully applied to the determination of ovatoxin-a, mascarenotoxin-a and four new palytoxins in O. cf. ovata. Another new palytoxin was detected in the standard material from Palythoa tuberculosa provided by Wako Chemicals.