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Tulathromycin is a macrolide antibiotic generally used for the treatment of respiratory diseases in cattle and swine. This work proposes an improvement of a previously published LC-MS/MS method for tulathromycin determination in pig serum, here validated in three different bull matrices: plasma, seminal plasma, and urine. The approach is based on a quick protein precipitation with acetonitrile, filtration, and sample dilution before injection, allowing to rapidly process large batches of samples. Analytes separation was obtained using a BEH C18 (50 x 2.1 mm, 1.7 mu m) column, maintained at 40 degrees C with a chromatographic run of 5 min. The method was fully validated over concentration ranges suitable for field levels of tulathromycin found in each matrix (0.01-1 mu g/ml for plasma, 0.05-5 mu g/ml for seminal plasma, and 0.1-10 mu g/m1 for urine), showing good linearity during each day of testing (R-2 always >0.99). Accuracy and precision were within +/- 15% at all QC concentrations in all the three matrices. Furthermore, the use of tulathromycine-d7 as internal standard mitigated the potential impacts of matrix effect. The validated technique was successfully applied to samples collected during a pharmacokinetic study in bulls, allowing to monitor tulathromycin concentrations over time in the three matrices. To our knowledge, this is the first validated approach for LC-MS/MS quantification of tulathromycin in seminal plasma and urine.

Ketamine is a widely used dissociative drug, whose quantification in plasma and urine can be of pharmacological, toxicological, and clinical interest. Although tandem mass spectrometry allows the reliable determination of ketamine and its metabolites in biological matrices, the structural similarity between norketamine (main active metabolite) and dehydronorketamine (a less relevant metabolite) can represent a critical aspect. These compounds differ exclusively in two hydrogen atoms, but the consequent two‐unit difference in their mass/charge ratio is partially nullified by the isotopic abundance of the chlorine atom present in their structure. This, along with their similar fragmentation pattern, can result in the incorrect identification of the enantiomers of these ketamine metabolites even with triple quadrupole instruments, if shared transitions are monitored after chiral chromatography. The key to prevent norketamine overestimation is therefore observing analyte‐specific MS/MS transitions. Here, we describe in detail how we investigated this issue, during the development of an analytical method for ketamine and norketamine enantiomer determination in plasma.

Hive products have numerous beneficial properties; however, the hive’s health is affected by the surrounding environment. The widespread use of herbicides in agriculture, such as glyphosate and glufosinate, has raised alarm among consumers, beekeepers, and environmentalists due to their potential to harm bees and humans through the consumption of bee products. This review aims to provide a comprehensive overview of the presence of glyphosate, glufosinate, and their metabolites in hive products, collecting and comparing available data from peer-reviewed research and surveys conducted across several countries. Moreover, it analyzes and discusses the potential impacts of these substances on human and bee health, analytical aspects, and recent regulatory developments. The data has revealed that these substances can be present in the different matrices tested, but the concentrations found are usually lower than the maximum residue limits set. However, the use of different methodologies with non-uniform analytical performances, together with an incomplete search for regulated analytes, leads to heterogeneity and makes comparisons challenging. In addition to the completion of studies on the toxicology of herbicide active ingredients, further monitoring actions are necessary, harmonizing analytical methodologies and data management procedures.

Oxytetracycline is a broad-spectrum antibiotic, which inhibits protein synthesis and is generally used for the treatment of pneumonia, shipping fever, leptospirosis and wound infections in cattle and swine. The present work proposes a novel liquid chromatography–tandem mass spectrometry (LC–MS/MS) method for oxytetracycline quantification in bull plasma, seminal plasma and urine, requiring limited sample treatment before analysis. Extraction with trichloroacetic acid followed by dilution of the supernatant in mobile phase proved to be effective in all three matrices, allowing to rapidly process large batches of samples. Sharp and symmetrical peak shape was obtained using a BEH C18 reversed-phase column in a chromatographic run of just 3.5 min. The mass spectrometer operated in positive electrospray ionization mode and monitored specific transitions for oxytetracycline (461.1 → 425.8) and the internal standard demeclocycline (465.0 → 447.6). The method was validated over concentration ranges suitable for field concentrations of oxytetracycline found in each matrix, showing good linearity during each day of testing (R2 always >0.99), as also confirmed by analysis of variance (ANOVA) and lack-of-fit tests. Excellent accuracy and precision were demonstrated by calculated bias always within ±15% and CV% below 10% at all quality control (QC) levels in the three matrices. Matrix effect and recovery were investigated for both analytes, which showed consistent and comparable behaviour in each matrix. To our knowledge, this is the first validated approach for mass spectrometric determination of oxytetracycline in seminal plasma and urine. The method was successfully applied to samples collected during a pharmacokinetic study in bulls, allowing to assess the oxytetracycline concentration–time profile in plasma, seminal plasma and urine.

Dechlorane Plus is a polychlorinated compound which has exclusively anthropic origin. This compound has been manufactured for close to 60 years for various applications, but mainly as flame retardant. Dechlorane Plus and other Dechlorane-related compounds (DRCs) are currently marketed as a replacement for Dechlorane, also known as Mirex, banned in 1978. These compounds share comparable properties to persistent organic pollutants (POPs), such as persistence in the environment, high lipophilicity, bioaccumulation through the food web and adverse effects on the environment and human health. Despite their long production history, they have been only recently reported in various environmental compartments, such as air, soil, and foodstuff. The aim of this review is to provide a picture of the current state of knowledge on worldwide DRC levels in food, in order to highlight gaps and research needs. The review compares the data on DRC contamination available in literature, considering different food categories and sampling country. In addition, it is specified whether the data were obtained from studies on foodstuff to estimate dietary intake, to evaluate the contamination near the e-waste treatment area or for environmental monitoring purposes.

The analysis of natural cobalamins in dairy products still represents an analytical challenge. The matrix’s complexity, low concentration level, light sensitivity, and binding to proteins are just some of the aspects that make their quantification a difficult goal to achieve. Vitamin B12 plays a fundamental role in human nutrition, and its intake is closely linked to a diet that includes the consumption of food of animal origin. In the current literature, few studies have been carried out on the quantitation of cobalamin in ripened cheeses. A sensitive, selective, and robust ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) method was developed, validated, and applied on ripened cheeses from different species (cow, sheep, and goat) purchased from local Italian markets, highlighting species-dependent differences in vitamin B12 concentrations. The vitamin B12 extraction procedure was performed by converting all cobalamins to the cyanocobalamin form. Furthermore, solid-phase extraction was used for matrix clean-up and analyte preconcentration. The proposed method showed good performance in terms of linearity, sensitivity, reproducibility, and repeatability. The mean vitamin B12 content ranged from

The 12 amino acid peptide derived from the Arabidopsis soluble secretory protein CLAVATA3 (CLV3) acts at the cell surface in a signalling system that regulates the size of apical meristems. The subcellular pathway involved in releasing the peptide from its precursor is unknown. We show that a CLV3-GFP fusion expressed in transfected tobacco protoplasts or transgenic tobacco plants has very short intracellular half-life that cannot be extended by the secretory traffic inhibitors brefeldin A and wortmannin. The fusion is biologically active, since the incubation medium of protoplasts from CLV3-GFP-expressing tobacco contains the CLV3 peptide and inhibits root growth. The rapid disappearance of intact CLV3-GFP requires the signal peptide and is inhibited by the proteasome inhibitor MG132 or coexpression with a mutated CDC48 that inhibits endoplasmic reticulum-associated protein degradation (ERAD). The synthesis of CLV3-GFP is specifically supported by the endoplasmic reticulum cha- perone endoplasmin in an in vivo assay. Our results indicate that processing of CLV3 starts intracellularly in an early compartment of the secretory pathway and that ERAD could play a regulatory or direct role in the active peptide synthesis.

The objective of this investigation was to evaluate the pharmacokinetic parameters of tulathromycin in plasma and semen of beef bulls after administering a single sc dose at two different sites in the neck. Four Simmental bulls with excellent temperament received a comprehensive physical exam that included breeding soundness examination. In addition, blood was collected and analyzed for CBC and chemical panel in order to rule out any subclinical liver or kidney disease. All bulls were diagnosed as healthy and satisfactory potential breeders. The mean plasma levels of tulathromycin for the two neck sites of sc administration were not different between posterior aspect of the ear where it attaches to the head (RP; regio parotidea; 77.9 ± 43.3 ng/mL; X ± SD) and to the middle of the neck (RC; regio collis lateralis; 73.7 ± 39.7 ng/mL; P = 0.84). The mean seminal plasma levels of tulathromycin after administration in the RP was 608 ± 374 ng/mL and for RC was 867 ± 599 ng/mL without differences between both sites (P = 0.29). The mean level of tulathromycin in plasma was 75.8 ± 40.2 ng/mL, which was lower than mean seminal plasma levels of 781 ± 482 ng/mL (P = 0.001). The plasma peak tulathromycin concentration (Cmax) was 160 ± 27 ng/mL at 21 ± 6 h (Tmax) post-administration. The seminal plasma Cmax was 1539 ± 44.4 ng/mL at 33.00 ± 18.00 h (Tmax) post-administration. The Cmax between plasma and seminal plasma were different (P = 0.008) without any differences in Tmax between plasma and seminal plasma (P = 0.35). The terminal half-life for plasma tulathromycin (81.4 ± 27.6 h) showed a tendency to be shorter than in seminal plasma (114.7 ± 21.7; P = 0.10). The plasma area under the curve concentration time from the first to the last sample (AUC0–last) was 15,440 ± 1717 ng/mL/h, which was significatively smaller compared with 171,071 ± 58,556 ng/mL/h for seminal plasma AUC0–last (P = 0.01). The plasma means residence time from the first to the last sample (MRT0-last) was 89.3 ± 5.1 h and it was shorter than for seminal plasma of 96.6 ± 5.0 h (P = 0.05). From the present investigation, it was concluded that tulathromycin is a suitable antibiotic based in its pharmacokinetic properties that could be used for treatment of bull genital infections when its application is indicated.

Per- and poly-fluoroalkyl substances (PFASs) are persistent and bioaccumulative compounds with adverse impacts on the environment and human health. Diet is one of the main sources of exposure to PFASs. Recently, the EFSA established a tolerable weekly intake (TWI) limit (4.4 ng/kg b.w.) for a mixture of the four major PFASs. Eggs and egg products can contribute to this intake, with their contamination possibly dependent on the husbandry system. Monitoring Italian eggs from backyard chickens revealed a relatively uniform PFAS contamination, with perfluoro-1-octanesulfonate being the most abundant. Contamination was detected to be significantly higher in eggs from backyard chickens than in eggs from commercial laying hens, consistent with a previous Italian study. According to the recently set TWI value, the consumption of eggs from backyard chickens could contribute significantly to dietary intake of PFASs (up to 29% of the TWI in children, considering the lower bound approach).

L’invenzione brevettata fa riferimento ad un dispositivo medico impiantabile per il rilascio locale controllato nel tempo di una combinazione di farmaci mai prima associati. L’invenzione ha inoltre aspetti d’uso innovativi: è progettata infatti per il trattamento della fase acuta delle lesioni del sistema nervoso centrale con componente infiammatoria, per la quale non esistono terapie. Si tratta di una nuova soluzione di farmacoterapia costituita dall’inserimento di due farmaci (un antiinfiammatorio ed un promielinizzante) in un materiale polimerico, realizzato mediante elettrofilatura, che consente il rilascio locale controllato nel tempo dei due farmaci, e che va impiegato nella prevenzione della degenerazione secondaria all’evento traumatico.

8 pages, 3 tables, 1 figure, Glycine betaine (GB) is a natural compound with demonstrated metabolic functions in mammals and beneficial effects on the human body: it protects cells under osmotic stress, as an organic osmolyte, and acts as a methylating agent in several vital biochemical pathways. The main dietary sources of GB are vegetables, such as cereals, spinach, chard and beetroot, above all; shellfish is one of the richest animal sources. A simple method for the quantification of GB in the edible portion of Manila clams (Tapes philippinarum) was setup, using ultra-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS). Isotope-labelled internal standards were used during sample extraction, in order to ensure an accurate quantification. The method was successfully validated following the Commission Decision 657/2002/EC as a guideline; since GB is an endogenous compound, present in very high concentrations in the target matrix, appropriate adjustments were taken. The method developed reported very satisfying performances in terms of linearity, trueness and precision; moreover, its applicability was demonstrated on commercial pools of Manila clams.

Perfluoroalkylated substances (PFASs) are a wide cluster of fluorinated molecules largely engaged industrially and commercially for many purposes. Because of the strength of the fluorine-carbon bond, PFASs show a firm tenacity against thermal degradation, hydrolysis, photolysis and biodegradation. On the other hand, such chemical stability gives them persistent environmental pollutant feature. In 2012, EFSA published a scientific report on PFASs in food, mentioning their adverse effects on health. Based on observational studies evidences, EFSA has recommended a tolerable daily intake (TDI) for the two most known PFASs, i.e. PFOS 150 ng/kg b.w./day and PFOA 1500 ng/kg b.w./day. The aim of this study was to monitor, for the first time, the level of contamination of PFASs in chicken eggs laid in Northern Italy. The eggs were collected from different rearing systems, in order to search a correlation between this variable and the contamination of PFASs. In this study four PFASs [perfluoro-nnonanoic acid (PFNA), perfluoro-noctanoic- acid (PFOA), sodium perfluoro-1- hexanesulfonate (PFHxS) and sodium perfluoro- 1-octanesulfonate (PFOS)] were analyzed by liquid chromatography-tandem mass spectrometer (LC-MS/MS). 132 eggs were analyzed, split up in 11 groups according to the geographical origin and rearing system. Results accord with literature data available for chicken eggs: almost all the samples show a PFASs contamination level under the limit of quantification (LOQ) of 0.25 ng/mL. No significant difference results from the rearing system, attesting an equal distribution and a concentration of PFASs detectable under the limit of quantification.

A simple, fast and cost-effective liquid chromatographic/tandem mass spectrometry (LC-MS/MS) method for the quantitative determination of flunixin in bovine muscle was developed and validated. The sample preparation procedure involved an extraction with acetonitrile, followed by evaporation and reconstitution. Chromatographic separation was achieved on a reverse phase column under programmed conditions. Flunixin detection was performed with positive electrospray ionization in Selected Reaction Monitoring (SRM) mode, monitoring one precursor and two products ions. For quantification purposes, flunixin-d3 was used as internal standard. The matrix effect on the analysis of flunixin in bovine muscle was evaluated by comparison between calibration curves prepared with standard solution and in blank matrix extracts. The equivalent responses obtained confirmed the absence of signal suppression or/and enhancement. The method was extensively validated according to the parameters requested by European Commission Decision 2002/657/EC in terms of specificity, limit of detection, linearity, trueness, precision, decision limit (CCα) and detection capability (CCβ). Flunixin stability was also investigated in matrix and in sample extracts at different times and storage conditions.

Perfluoroalkyl substances (PFASs) are a family of persistent pollutants of anthropic origin which can reach humans mainly through diet, causing potentially dangerous effects on health. Fish and fishery products are a major source of exposure, but intra- and inter-specific contamination can be extremely variable. In the present study a single species monitoring of the presence of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), the two main compounds of the family, was performed on 140 farmed and wild caught European sea basses (Dicentrarchus labrax) from different places in the Mediterranean area. The results highlight a strong correlation between the level of contamination and the origin of fishes, if wild or farmed: on average, wild caught sea basses (PFOS: 112-12,405 ng/kg, median 1345 ng/kg; PFOA: 9–487 ng/kg, median 28 ng/kg) showed higher levels than intensively farmed sea basses (PFOS: 11–105 ng/kg, median 32 ng/kg; PFOA: 9–51 ng/kg, median 21 ng/kg). Significant differences among the various rearing systems were also observed, with extensively-farmed subjects presenting relatively higher levels of both compounds compared to intensively farmed. Moreover, a certain variability among wild fish caught from different sampling sites was observed, confirming that PFASs contamination, which reached in some cases noticeable concentrations, might be influenced by the geographical origin.

Graphical abstract, Although budesonide is frequently used in veterinary medicine for the treatment of canine respiratory and bowel inflammatory diseases, knowledge is lacking regarding its kinetics in this species. We developed and validated a liquid chromatography–tandem mass spectrometry method for the determination of budesonide and its metabolite 16α-hydroxyprednisolone in dog plasma. The analytes were extracted by solid phase extraction and analysis was performed by high performance liquid chromatography–tandem mass spectrometry, with positive electrospray ionization.•This method allows budesonide and one of its main metabolites to be simultaneously quantified in dog plasma at fairly low concentrations.•The proposed protocol is very easy and fast to execute, without compromising analytical performances.•A small amount (0.5 mL) of plasma is required, making this approach suitable for pharmacokinetic studies also in small sized dogs.

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Rising interest in beneficial health properties of fruit and achievement of the best organoleptic qualities in relation to ripening times, led to focus the attention on peach fruits. Phenolic composition in peach fruit changes during ripening process and influences the final quality. A collection field has been set-up, where different peach genotypes of interest (melting flesh=MF and not melting flesh=NMF) have been grown. After pit hardening, fruits growth and ripening have been monitored, the latter using a non-destructive portable instrument (DA-meter) providing an index linked with the chlorophyll degradation thus related to ripening degree. Classical destructive quality analysis was also carried out during the sampling. The general aim of this research has been to investigate about the possible relation between phenolic compounds and ripening stages in peach fruit. Differences in the amount of phenolic compounds in flesh and peel during the maturation process have been evaluated. Quantification of the most important phenolic compounds was obtained using a LC-MS/MS system. Results reported that the DA index (I(DA)) decreased reaching the ripening stage, in both peach genotypes (MF and NMF) and it also showed a relationship with ethylene production of fruits, demonstrating a good reliability of the instrument. Preliminary results showed that from ‘Oro A’s phenolic analysis in pre-climacteric stage, the amount of quercetin 3-rutinoside and cyanidin 3-glucoside in the peel resulted substantially higher than in the flesh. Among the main flavan-3-ols, catechin resulted in higher content compared to epicatechin, in agreement with previous works. Among hydroxycinnamic acids, chlorogenic acid resulted in higher amount than neo-chlorogenic as reported by other studies.

The aim of this study was to determine the presence and the level of contamination of the most important mycotoxins (deoxynivalenol, fumonisin B1 and B2, aflatoxin B1, B2, G1 and G2, ochratoxin A and zearalenone) in 48 samples of extruded dry dog food found in the Italian market (24 samples from standard economy lines, 24 of premium lines). Analyses were performed using ultra-performance liquid chromatography coupled to tandem mass spectrometry. Although the concentrations of the mycotoxins in all samples proved to respect the European legislation with regards to animal feed, the analyses revealed a substantial presence of deoxynivalenol, fumonisins and ochratoxin A, with values above the limit of quantification (5 µg/kg) in 100%, 88% and 81% of the samples, respectively. In contrast, aflatoxins and zearalenone contamination proved to be very modest, with 88% and 75% of the samples, respectively, showing concentrations below the corresponding limit of quantification (5 µg/kg for aflatoxins and 10 µg/kg for zearalenone). Moreover, despite a very heterogeneous contamination, the concentration of fumonisins and ochratoxin A was significantly higher in standard foods than in premium ones (491 vs. 80.2 µg/kg dry matter for fumonisin B1; 113 vs. 38.5 µg/kg dry matter for fumonisin B2; 599 vs. 103 µg/kg dry matter for total fumonisins; 23.8 vs. 13.0 µg/kg dry matter for ochratoxin A; P < 0.001). Furthermore, a simultaneous presence of different mycotoxins (at concentrations higher than their limit of quantification) was observed in most of the pet foods analyzed; in particular, 19% of the samples were contaminated by no fewer than two different types of mycotoxins, 52% by three, 25% by four and 2% by all the mycotoxins evaluated. These results revealed the need for further investigation into the potential risk deriving from chronic exposure to low doses of the different types of mycotoxins that pet species are subject to today.

Vitamin B12 (cobalamin) is a metal complex composed of a central cobalt ion bonded to six ligands. It is essential for major biological functions such as protein, fat and carbohydrate metabolism, the maintenance of the central nervous system, and the formation of red blood cells. Since mammals cannot synthesize cobalamin, dietary intake represents the only natural source for humans. Dairy products can provide significant levels of cobalamin; moreover, the European Food Safety Authority (EFSA) panel has set the recommended intake at 4 μg/day for adults. Vitamin B12 content was determined in milk and several matrices related to the process of transformation of the residual whey from Parmigiano Reggiano cheese-making to obtain ricotta cheese. In addition, vitamin B12 degradation during ricotta cheese shelf-life was studied. The analyses were performed using an ultra performance liquid chromatography-tandem mass spectrometry method. Results show that vitamin B12 amount in ricotta from dairy and experimental cheese-making brings respectively 1/8 to 1/4 of the adequate intake in adults established by EFSA. In addition, shelf-life experiment shows that cobalamine is fairly rapidly degraded in ricotta: light effect seems to be significant, even if the light exposure is short. The use of photoprotective packaging material increases B12 shelf-life in the early stage of storage.

Due to their widespread diffusion, perfluoroalkyl substances (PFASs) have been frequently found in the environment and in several animal species. It has been demonstrated that they can easily reach also humans, mainly through diet. Being lactation a major route of elimination of these contaminants, their occurrence in human milk is of particular interest, especially considering that it generally represents the unique food source for newborns. Perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), the two most important compounds of this family, have been frequently found in human milk at variable concentrations, but still limited data are available. The present study, the first conducted in Italy capable to detect these pollutants at ultra-trace levels by UPLC-MS/MS, confirmed the role of lactation as a relevant source of exposure for breastfed children. The measured concentrations ranged between 15 and 288 ng/L for PFOS and between 24 and 241 ng/L for PFOA. Moreover, mean concentrations and frequencies of both analytes resulted higher in milk samples provided by primiparous women, suggesting that the risk of intake might be higher for first-borns. Finally, comparing these results with previous data, PFOS gradual decrease over time since year 2000 was confirmed. Keywords: PFASs, PFOS, PFOA, Breast milk, LC–MS/MS

Perfluoroalkyl substances (PFAS) are fully fluorinated compounds widely used during the last 60 yr in the production of multiple industrial and consumer applications, such as food packaging, nonstick cookware, cleaning agents, and many more. These emerging contaminants have recently become of concern for human health because of their potential negative effects. The risk of exposure to PFAS for humans is mainly related to diet, and the increasing interest in food safety has led the European Commission to call Member States to monitor these contaminants in food matrices. The purpose of the present work was to perform the first monitoring on the presence of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA), the 2 main and most widely investigated molecules of this family, in cow milk commercially available in Italy. We used an analytical protocol consisting of liquid-liquid extraction followed by 2 purification steps through solid-phase extraction cartridges and injection on an ultra-performance liquid chromatography-tandem mass spectroscopy system. The analysis of 67 samples of different types of cow milk from Italy demonstrated that contamination by PFOS was often present, although at relatively low concentrations (up to 97 ng/L), whereas PFOA was rarely found. On the basis of these results and data reported in the literature on this matrix, milk does not seem to be a major source of PFAS compared with other food categories such as fish and seafood. However, variability among different types of milk must be taken into account, and surveys of milk-derived products would be helpful to better define the risk for consumers.

Vitamin B12 is a water-soluble molecule composed of a tetrapyrrolic complex with a cobalt atom at its centre. It is an essential regulatory element, synthesized only by bacteria; for this reason it is present only in food of animal origin and the daily requirement for humans is about 1 to 2 µg. Since milk and dairy products provide a significant dietary cobalamin intake, an ultra performance liquid chromatographytandem mass spectrometry method was applied to samples collected at different stages along the process of cheese making in order to evaluate the distribution of this molecule. In particular, samples of milk, rennet, whey, ricotta cheese, curd, mozzarella cheese and caciotta cheese were analysed. Results showed a level of vitamin B12 about 10 times higher in whey and ricotta cheese with respect to the milk they are derived from. These data would confirm the tendency of cobalamine to concentrate in the proteic fractions along the cheese production process.

Bisphenol A (BPA) is a man-made compound, mainly used as a monomer to produce polycarbonate (PC), epoxy resins, non-polymer additives to other plastics, which have many food related applications, such as food storage containers, tableware and internal coating of cans, as well as non-food applications such as electronic equipment, construction materials and medical devices. BPA exposure can occur when the residual monomer migrates into packaged food and beverages. Moreover, due to the ubiquitous presence of this compound, the general population can be exposed to environmental sources such as water, air and soil. Many studies have investigated the potential health hazards associated with BPA, which can elicit toxic and cancerogenic effects on humans. According to the European Food Safety Authority opinion, diet is considered to be the main source of exposure, especially canned food; moreover, among non-canned food, meat and fish products have the highest levels of BPA contamination. This review focuses on BPA contamination in seafood, analysing worldwide literature (from January 2010 to October 2015) on BPA contamination of edible parts. The authors try to identify differences between canned and non-canned seafood in literature, and gaps in the state of art. The data evaluated underline that all concentrations for both canned and non-canned seafood were below the specific migration limit set by the European Community Directive for BPA in food. Moreover, the canned seafood is more contaminated than the non-canned one.

Mycotoxins are heterogeneous chemical compounds characterized by a low molecular weight and synthesized by the secondary metabolism of different molds. Fumonisins are water-soluble mycotoxins produced by Fusarium species spoiling corn and derived products. These mycotoxins can enter the food chain also through the consumption of meat of exposed animals. Fumonisins and their metabolites have been associated with several animal and human diseases. They are suspected risk factors for esophageal and liver cancers, neural tube defects and cardiovascular problems. Improved methods are needed to accurately assess fumonisin concentrations in food from vegetable and animal origin to prevent acute and chronic human exposure. The aim of the present work was to develop a sensitive and selective method for quantification and unambiguous identification of fumonisin B1 (FB1), fumonisin B2 (FB2) and their complete hydrolyzed products (HFB1 and HFB2), in order to determine their presence in pig liver. Furthermore, the developed method was applied, in order to test its efficacy, on liver samples of weaned piglets fed a diet in compliance with the FB1 and FB2 contamination limits set by EU. All the samples showed the presence of at least one of the analytes. In particular FB1 was found in 5 out of 7 samples and the average level of contamination found was 28 ppb.

Fumonisins are mycotoxins found primarily in corn and corn products that are produced by Fusarium verticillioides, F. proliferatum, and several other Fusarium species. The toxicity of fumonisin B₁ (FB) from culture material with and without activated carbon was evaluated using weanling piglets. Fifty-six weanling pigs were assigned to one of four treatments diets based on BW. The treatment diets were 1) control = corn-soybean basal diet with

Proximate composition and fatty acid profiles, conjugated linoleic acid (CLA) isomers included, were determined in separable lean of raw and cooked lamb rib-loins with their subcutaneous and intermuscular fat, prepared as roasts or steaks. Two combinations “cooking method × type of cut” were selected: one is a traditional method for this meat (broiling of steaks), while the other (microwaving followed by final grilling of roasts) is far less widely used. The two methods, similar as regards the short preparation time involved, were also evaluated for cooking yields and true nutrient retention values. The cooking yield in microwaving was markedly higher than in broiling. Significant differences between the two methods were also found in the true retention values of moisture, protein and several fatty acids, again to the advantage of microwaving. On the basis of the retention values obtained, with microwaving there was a minimum migration of lipids into the separable lean, consisting almost exclusively of saturated and monounsaturated fatty acids, while there were small losses of lipids in broiling, almost equally divided between saturated, monounsaturated and ω6 polyunsaturated fatty acids. On the whole, the response to cooking of the class of CLA isomers (including the nutritionally most important isomer cis -9, trans -11) was more similar to that of the monounsaturated than the polyunsaturated fatty acids.

The aim of the present work was to develop a sensitive and selective method for identification and quantification of fumonisin B1 (FB1) in bovine milk. FB1 was isolated by immunoaffinity column and was detected using liquid chromatography coupled with tandem mass spectrometry in positive electrospray ionization (ESI+). The LOQ of the method was 0.1 μg/kg that was lower than the others reported in the literature. The high coefficient of determination (R2 > 0.99) obtained in the range of 0.1–10.0 μg/kg, the good recovery (84%) and relative standard deviation (7%) of the proposed method ensure correct fumonisin detection in milk even at relatively low concentrations. The developed method was applied on different commercial samples in order to test its efficacy. FB1 was found above the LOQ in eight out of 10 samples analysed and the average level of contamination found was 0.26 μg/kg.

Mycotoxins are heterogeneous chemical compounds characterized by a low molecular weight and synthesized by the secondary metabolism of different molds. Fumonisins are water-soluble mycotoxins produced by Fusarium species spoiling corn and derived products. These mycotoxins can reach the human also indirectly through the consumption of food products derived from animals fed with contaminated feed. Fumonisins have been associated with several animal and human diseases. They are suspected risk factors for esophageal and liver cancers, neural tube defects and cardiovascular problems. Improved methods are needed to accurately assess fumonisin concentrations in food from vegetable and animal origin to prevent acute and chronic human exposure. The aim of the present work was to develop a sensitive and selective method for identification and quantification of fumonisin B1 (FB1) in milk. FB1 was isolated from milk, by a single step immunoaffinity column and was detected using liquid chromatography coupled with tandem mass spectrometry in positive electrospray ionization (ESI+). The analysis were carried out in multiple reaction monitoring (MRM) mode using the two main product ions. The good performances of the proposed method can assure a correct fumonisin detection in milk even at relatively low concentrations.

The marine dinoflagellates Prorocentrum micans, Gonyaulax polyedra, Gymnodinium sp., and Alexandrium tamarense, collected from the Adriatic Sea during red-tide blooms, were cultured to investigate the 4-methyl sterol constituents. To ascertain a possible influence of cell age on the 4-methyl sterol content, for one strain (Gymnodinium sp.)we investigated the composition of these constituents at exponential and stationary growing phases. The lipid material extracted with acetone from the lyophilized algal samples was fractionated by thin-layer chromatography. The 4-methyl sterols recovered from the layer were converted into the corresponding OTMS derivatives. Nine of 11 constituents were identified by gas chromatography and gas chromatography-mass spectrometry; only two minor constituents were characterized by their gas chromatographic parameters. All free methyl sterols identified in the algal samples had been detected previously in various dinoflagellates. The 4-methyl sterol fractions generally contained very few constituents. Except for the Gymnodinium sp. sample, collected at the exponential growing phase (GyD2 exp), which contains 4,24-dimethylcholestan-3-ol as a unique constituent, dinosterol was the major component. Moreover, 4,24-ethylcholestan-3-ol was also an important constituent of both Prorocentrum and Gonyaulax strains, whereas considerable amounts of dinostanol characterized all the Gymnodinium sp. strains. In addition, the latter contained several minor constituents such as 4-methylcholestan-3-ol, 4,24-dimethylcholesta-22-en-3-ol, and 4-methyl-24-ethylcholestan-3-ol. 4-Methyl-24-methylene-cholestan-3-ol was a constituent of the Gymnodinium sp. sample, collected at the stationary growing phase (GyD2 stat)only, whereas 4-methylgorgostanol was identified only in the Alexandrium tamarense Gt4 strain. Except for 4-methyl-24-ethylcholesta-8(14)-en-3-ol, all the methyl sterol constituents from our algae show a saturated polynuclear system. The pathways by which side-chain modifications occur in dinoflagellate 4-methyl sterols are considered, and a map of the fragmentation pattern of the trimethylsilyl-4-methyl sterols under electronic impact is also reported.

A study has been made on the effects of the dietary administration of a selection of volatile oils from medicinal plants on the polyunsaturated fatty acid composition in the retina of aged (28 month old) rats. Polyunsaturated fatty acids comprised by far the major proportion of the total unsaturated content within the retinal phospholipids with docosahexaenoic acid predominating. The administration daily of 3-9 mg of the essential oils from clove, nutmeg, pepper and thyme over a period of 17 months resulted in the maintenance of very much higher levels of polyunsaturated fatty acids, in particular docosahexaenoic acid, within the retinal phospholipids. This increase occurred mainly at the expense of a reduction in the level of oleic acid. The possible efficacy for the application of the oils from such medicinal plants through their antioxidant capacities in the prevention of age-related macular degeneration is discussed.

Three postharvest icing treatments were implemented on cuttlefish specimens (overall number = 243) after they were hauled onboard: T1-ICO, storage under a 15-mm-thick layer of contact crushed freshwater ice; T2-ICA, storage over a 15-mm-thick layer of contact crushed freshwater ice; T3-NOI, storage without any contact ice, but placed over a 15-mm-thick layer of contact crushed freshwater ice once ashore. All storage trials (n = 6) were conducted at 2 ± 2 °C and were terminated on the sixth day of storage because the aim of the study was to evaluate the effect of realistic icing practices on the freshness quality and shelf -life of cuttlefish, as assessed by sensory (a non-destructive Quality Index Method, QIM), physical (dielectric properties through the Fish Freshness Meter, “295 – Torrymeter”) and chemical (the breakdown products of adenosine 5′–triphosphate, or ATP, as well as relevant ratios among them) methods. The early use of contact ice, either above (T1-ICO) or underneath (T2-ICA) single-layered cuttlefish placed in self-draining polystyrene boxes, was able to slow down ATP degradation. Nevertheless, because three visual traits were significantly worsened in the former treatment, the T2-ICA treatment was suggested as the most proper icing procedure for this cephalopod. The Torrymeter readings had no utility either for predicting the cuttlefish actual/remaining shelf-life for each icing treatment, or for discriminating among them. The percentage ratio between hypoxanthine (Hx) and adenosine 5′–monophosphate (AMP) proved to be the chemical index most highly correlated with ice days. A useful, non-destructive, QIM scheme for cuttlefish was successfully tried and tested in the process.

A method using ultra performance liquid chromatography coupled with tandem mass spectrometry was developed to measure cobalamins in naturally enriched raw milk and to evaluate their fate during thermal treatments and along the process of cheese making. After addition of methotrexate as internal standard, samples were submitted to heat treatment in the presence of cyanide, which converts all the less-stable cobalamins into cyanocobalamin; then, purification was performed by a solid-phase extraction step. Reverse-phase ultra performance liquid chromatography separation coupled with tandem mass spectrometry provided a fast and reliable determination. Mass spectrometric analysis was carried out in multiple reaction monitoring mode. The monitored transitions were m/z 678.36 → 147.10 and 678.36 → 359.30 for vitamin B 12 and m/z 455.22 → 175.13 and 455.22 → 308.22 for methotrexate (internal standard). The limit of quantification was 2ng/g. The method showed good linearity from 2 to 20ng/g (R 2 ≥0.98) and intra- and interday precisions were always less than 19%.

Ribavirin has proved to be effective in vitro against several RNA viruses responsible for encephalitis in humans and animals. However, the in vivo efficacy towards the cerebral viral load seems to be limited by the blood-brain barrier. Since the nose-to-brain pathway has been indicated for delivering drugs to the brain, we investigated here the distribution of ribavirin in the central nervous system (CNS) after intranasal administration. We first tested in vitro ribavirin diffusion from an aqueous solution across a biological membrane, using Franz cells and rabbit nasal mucosa. About 35% of ribavirin permeated in 4. h across the mucosa, after reaching steady-state flux in less than 30. min. In the first in vivo experiment, ribavirin aqueous solution was administered intranasally to Sprague Dawley rats (10. mg/kg). Animals were sacrificed at 10, 20 or 30. min after administration to collect brain areas (cerebellum, olfactory bulb, cerebral cortex, basal ganglia and hippocampus) and biological fluids (cerebrospinal fluid and plasma). Ribavirin, quantified by LC-MS/MS spectrometry, was detected at each time point in all compartments with the highest concentration in olfactory bulb and decreasing in rostro-caudal direction. Two subsequent in vivo experiments compared the nasal route (ribavirin solution) with the intravenous one and the nasal administration of ribavirin solution with ribavirin powder (10. mg/kg). It was found that 20. min after administration, ribavirin concentration in olfactory bulb was similar after intravenous or nasal administration of the ribavirin solution, whereas the powder led to significantly higher levels. Ribavirin was also present in deeper compartments, such as basal ganglia and hippocampus.Even if the mechanisms involved in ribavirin nose-to-brain transport are not clear, these results suggest a rapid extracellular diffusive flux from the nasal epithelium to the olfactory bulb and different CNS areas. © 2011 Elsevier B.V.

A liquid chromatography–tandem mass spectrometry method for the quantitative determination of fumonisin FB1, FB2 and their respective hydrolysed metabolites HFB1 and HFB2 in pig liver has been developed and validated. The method was based on an easy extraction procedure followed by SPE purification. Chromatographic separation of the analytes was performed on a C18 column using 0.3% of formic acid in water and acetonitrile as elution solvent. The mass spectrometer operated in the positive electrospray ionisation mode (ESI+) using multiple reaction monitoring (MRM). An intralaboratory validation was carried out with fortified samples for determining precision, trueness, specificity, limit of detection (LOD) and limit of quantification (LOQ). The method showed good performance characteristics and proved to be sensitive, selective and reliable. The LOD was 0.05 ng g −1 and the LOQ was 10 ng g −1 for all the analytes. The developed method has been applied to seven pig liver samples in order to test its applicability to evaluate exposure of food animals to fumonisins.