Food Safety is an Important Public Health Issue: Chloramphenicol Residues Determination by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) in honey.

BACKGROUND: Honey is used for nutritional, medicinal and industrial purposes and antibiotic residues may harm its quality and constitute a danger to human health. The broad spectrum antibiotic chloramphenicol (CAP) was used for curative purposes in veterinary medicine, but is now forbidden in European Union (EU) because of its many serious side effects (e.g. aplastic anaemia, grey syndrome, severe bone marrow depression and hypersensitivity). The aim of this study was to facilitate analyses of the quality and safety of Croatian honey distributed to whole European Union market; an assessment that has not previously been made. SUBJECTS AND METHODS: CAP in honey was qualifying and quantifying by validated liquid chromatography tandem mass spectrometry with negative electrospray ionisation method (LC-MS/MS). The target antibiotic was separated on chromatographic column Zorbax SB C18 (150 mm x 2.1 mm, 3.5 µm) with a gradient elution using acetonitrile - 0.1% formic acid mobile phase at a flow rate of 0.3 mL/min, with column temperature 35°C for CAP and 5D-CAP as internal standard. Homogenised honey samples were diluted with acetate buffer solution and extracted on Oasis Hydrophilic-Lipophilic-Balanced (HLB) sorbents. The method was used to analyse 280 domestic honey samples collected throughout Croatia between 2005.-2013. RESULTS: Recoveries of the method for real (acacia, chestnut, linden and flower) honey samples were 102% with RSD 8.4%. The value CCα and CCß were 0.09 and 0.12 µg/kg, respectively. Results showed only three subsequent positive detections (1.1%) of CAP in honey. CONCLUSIONS: Analysed honey samples from Croatia showed good quality and safety what is the one of the main objective in consumer health policy in EU.

Chemometric Valorization of Strawberry (Fragaria x ananassa Duch.) cv. 'Albion' for the Production of Functional Juice: The Impact of Physicochemical, Toxicological, Sensory, and Bioactive Value.

Strawberries (Fragaria x ananassa Duch. cv. 'Albion') were harvested at two stages of ripeness (75% vs. 100%) and their physicochemical, sensory, toxicological, and bioactive properties were evaluated before and after processing into juice. The fresh fruits and their by-products were also evaluated. During processing into juice, the color change was higher in the fully ripe fruits, confirming the encouraging prospects for using the less ripe strawberries for processing. The analysis of heavy metals (Cu, Zn, Ni, As, Cd, Pb) was carried out, and in juice and by-product samples of 100% maturity, only Pb was higher than the MDK. Of the 566 pesticides analyzed, only cyprodinil was found in the by-products of the strawberries at 75% maturity, while pyrimethanil was detected in all samples. Fresh strawberries of both ripeness levels were rated similarly to the corresponding juices for all sensory attributes studied, indicating that sensory perception was not affected by processing. However, ripeness was found to be an important factor influencing most sensory attributes. The by-products were the materials with the highest levels of all bioactive compounds. Considering all quality parameters evaluated, the chemometric evaluation confirms the suitability of 75% ripe strawberries for processing into functional juice, which could be important for the juice industry.

Exposure of the Croatian adult population to acrylamide through bread and bakery products.

The aim of the present study was to determine and compare the levels of acrylamide in different types of bread and bakery products using a LC-MS/MS method, before and after the new European regulation on acrylamide reduction (Commission Regulation (EU) 2017/2158) became valid. Also, one of the aim was to estimate the average exposure to acrylamide through this food category. Of the total of 100 analysed samples, acrylamide content ranged from below the limit of quantification (LOQ) to 237 µg/kg in the period before the application of a new European Regulation, and from

Toxicity of nanosilver and fumonisin B1 and their interactions on duckweed (Lemna minor L.).

In the environment co-contamination of several toxicants commonly occurs. However, toxicological studies usually are focused on only one toxicant. The aim of this study was to investigate toxicity of silver nanoparticles (AgNP) and mycotoxin fumonisin B1 (FB1) and their possible interactions as well as to explore tentative mechanism of their toxic effect. Duckweed (Lemna minor L.) was treated with AgNP or FB1 (at concentrations 0.5 and 1.0 mg L-1) or with their combination at same concentrations for 3 days. Both AgNP and FB1, applied individually significantly affected levels of certain nutrients, reduced growth rate and the levels of photosynthetic pigments though AgNP at a much greater extent compared to FB1. Furthermore, AgNP induced ROS generation, lipid peroxidation and increase of antioxidative enzymes activities, while FB1 induced changes only in the activities of antioxidative enzymes. Those results implicate that phytotoxicity of both AgNP and FB1 can be associated with imbalance of mineral and cell redox status. However, toxic actions of AgNp singly applied were more pronounced. Combined treatment with AgNP and FB1 produced higher degree of changes in all parameters than corresponding concentrations of AgNP or FB1 alone implying their additive effects. Additionally, higher level of FB1 found in medium, and higher level of intracellular Ag following combined treatment indicates interaction of two toxicants at the transport level/uptake in the cell which resulted in higher accumulation of Ag in duckweed cells. The latter in turn exerted higher toxicity to duckweed compared to single treatment of AgNP.

Removing aflatoxin M1 from milk with native lactic acid bacteria, centrifugation, and filtration.

In order to minimise human exposure to aflatoxin M1 (AFM1) the levels of this highly carcinogenic mycotoxin in milk, heat-treated milk, and other dairy products have been limited to <0.05 µg kg-1. However, its removal from dairy products presents a challenge for dairy producers, as commercial additives change organoleptic properties, and filtration alone yields poor results. The aim of this study was to find a strain of lactic acid bacteria (LAB) from milk or dairy products that most effectively binds AFM1 and to see whether heat treatment of the selected LAB affects the binding efficiency. We also wanted to investigate whether centrifugation can improve filtering of the obtained AFM1-LAB complexes from milk. To do that, we isolated and identified 10 native LAB species/strains, incubated their viable or heat-treated cells (108 CFU mL-1) in milk spiked with 0.5 µg L-1of AFM1 at 4 °C for 0, 2, 4, and 24 h, and quantified the amount of unbound AFM1 with HPLC. AFM1 binding efficiency ranged from 21 to 92 % for viable cells and from 26 to 94 % for the heattreated ones. Since both viable and heat-treated Lactobacillus plantarum KM showed the best results, we used it for the next step in AFM1 removal from milk. Heat treatment in combination with filtration and centrifugation yielded removal as high as 96 %.

Monitoring of selected pharmaceuticals in surface waters of Croatia.

Sulfonamides, macrolides, torasemide, fumagillin, and chloramphenicol were simultaneously analyzed in surface water samples by using solid-phase extraction (SPE) and reversed-phase (RP) liquid chromatography-electrospray tandem mass spectrometry (LC-ESI-MS/MS). In the pre-concentration and clean-up process, the pH value of samples and volume of the solvent for extraction of analytes from cartridge were optimized. Extraction recoveries were high with values in the range from 62 to 115%. Limits of quantification (LoQ) were in the range from 0.02 to 0.2 µg L-1. Repeatability of the method was evaluated at LoQ and expressed as relative standard deviation (RSD). Calculated RSDs were low with values in the range from 2.4 to 14.5%. The method was successfully applied for analysis of surface water real samples. Samples were collected along the rivers in Croatia on four sampling sites in 2012 in Danube catchment areas, 19 sampling sites in Danube and Adriatic catchment areas in 2013, and another 19 places in 2014. Altogether, 20 target compounds were analyzed in 148 water samples and detected in 31 samples in range (0.1-5.3) µg L-1 or in 20.1% of samples. The most frequent and highest concentrations were detected for macrolide antibiotics. This is the first attempt of such monitoring in surface waters in Croatia.