Validation of a single liquid chromatography-tandem mass spectrometry approach for oxytetracycline determination in bull plasma, seminal plasma and urine

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

PRELIMINARY DATA ON FUMONISINS PRESENCE IN PIG LIVER

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

Occurrence of mycotoxins in extruded commercial dog food

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 \ub5g/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 \ub5g/kg for aflatoxins and 10 \ub5g/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 \ub5g/kg dry matter for fumonisin B1; 113 vs. 38.5 \ub5g/kg dry matter for fumonisin B2; 599 vs. 103 \ub5g/kg dry matter for total fumonisins; 23.8 vs. 13.0 \ub5g/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

Perfluoroalkyl substances in human milk: A first survey in Italy

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

BACTERICIDAL ACTIVITY OF ELECTROLYZED OXIDIZING WATER ON MEAT AND POULTRY

Electrolyzed oxidizing water (EOW) has potential application as a residue free sanitizing agent for food of animal origin. Meat and poultry were contaminated with microorganism, pathogens or not, and different types of electrolyzed oxidizing water treatement were investigated to evaluate the activity of each of these method. In detail, this study is aiming at evaluating the effectiveness of EOW in reducing microbial count, including total bacterial count, Salmonella Typhimurium, Staphylococcus aureus, Listeria monocytogenes and Escherichia coli on meat and poultry. EOW has a very strong disinfectant activity which, along with its easy and safe use, makes a good alternative to many other more widely used disinfectants

A New Biocomposite Material Based on Wheat Waste and Suitable for 3D Printing Applications

Biopolymers, such as poly(lactic) acid (PLA), which is obtained through green synthesis pathways from renewable resources, has attracted considerable interest in recent years because of the increasing need to reduce petroleum-based plastic pollution and bringing their prices comparable with conventional thermoplastic commodities’ price (e.g., polyethylene, polypropylene, and polystyrene). The present work investigates the employment of 10% wt of natural materials, deriving from wheat milling process, as biofiller of PLA to develop a biocomposite filament suitable for 3D-printing technique. The inclusion of a cost-free natural material leads to a strong reduction of the whole material cost. Implementing this new class of composite material to additive manufacturing technique allows to dramatically reduce the environmental impact of 3D printed products

Occurrence of Mycotoxins in Extruded Commercial Cat Food

The occurrence of the most important mycotoxins (deoxynivalenol, fumonisin B1 and B2, aflatoxins B1, B2, G1, and G2, ochratoxin A, zearalenone, T-2, and HT-2 toxins) was determined in 64 extruded cat foods purchased in Italy through ultra-performance liquid chromatography coupled with tandem mass spectrometry. Deoxynivalenol and fumonisins were the most common contaminants (quantified in 80 and 95% of the samples, respectively). Conversely, aflatoxins B2, G1, and G2 were not identified in any sample. Some cat foods exceeded the regulatory limit for aflatoxin B1 (n = 3) or the guidance values for zearalenone (n = 3), fumonisins (n = 2), ochratoxin A (n = 1), and T-2 (n = 1) recently established for pets in the European Union. A widespread co-occurrence of mycotoxins was observed (28, 42, and 8% of the samples contained quantifiable amounts of two, three, and four mycotoxins, respectively). This study describes criticisms regarding the mycotoxin issue in pet food and suggests an improvement of the monitoring of the pet food chain

DETERMINATION OF FUMONIS FB1 IN MILK BY LC-MS/MS

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

Design and in vitro study of a dual drug-loaded delivery system produced by electrospinning for the treatment of acute injuries of the central nervous system

Vascular and traumatic injuries of the central nervous system are recognized as global health priorities. A polypharmacology approach that is able to simultaneously target several injury factors by the combination of agents having synergistic effects appears to be promising. Herein, we designed a polymeric delivery system loaded with two drugs, ibuprofen (Ibu) and thyroid hormone triiodothyronine (T3) to in vitro release the suitable amount of the anti-inflammation and the remyelination drug. As a production method, electrospinning technology was used. First, Ibuloaded micro (diameter circa 0.95–1.20 µm) and nano (diameter circa 0.70 µm) fibers were produced using poly(L-lactide) PLLA and PLGA with different lactide/glycolide ratios (50:50, 75:25, and 85:15) to select the most suitable polymer and fiber diameter. Based on the in vitro release results and in-house knowledge, PLLA nanofibers (mean diameter = 580 ± 120 nm) loaded with both Ibu and T3 were then successfully produced by a co-axial electrospinning technique. The in vitro release studies demonstrated that the final Ibu/T3 PLLA system extended the release of both drugs for 14 days, providing the target sustained release. Finally, studies in cell cultures (RAW macrophages and neural stem cell-derived oligodendrocyte precursor cells—OPCs) demonstrated the anti-inflammatory and promyelinating efficacy of the dual drug-loaded delivery platform