Non-planar di-
-substituted PCB 153 (2,2’,4,4’,5,5’-hexachlorobiphenyl), one of the most abundant PCB congeners in the environment and in biological and human tissues, has been identified as ...potential endocrine disruptor affecting the reproductive and endocrine systems in rodents, wildlife, and humans. The aim of this study was to gain a deeper insight into its mode/mechanism of action in Chinese hamster ovary K1 cells (CHO-K1). PCB 153 (10–100 μmol/L) inhibited CHO-K1 cell proliferation, which was confirmed with four bioassays (Trypan Blue, Neutral Red, Kenacid Blue, and MTT), of which the MTT assay proved the most sensitive. PCB 153 also induced ROS formation in a dose-dependent manner. Apoptosis was seen after 6 h of exposure to PCB 153 doses ≥50 μmol/L, while prolonged exposure resulted in the activation of the necrotic pathway. PCB 153-induced disturbances in normal cell cycle progression were time-dependent, with the most significant effects occurring after 72 h.
Concentrations of As, Cd, Cu, Hg and Pb were measured by atomic absorption spectrometry (AAS) in muscle tissues of four fish species: anchovy (
Engraulis encrasicholus), mackerel (
Scomber ...japonicus), red mullet (
Mullus surmuletus) and picarel (
Spicara smaris) from the Croatian waters of the Adriatic Sea during 2008 and 2009. Metal levels measured in anchovy were in the following ranges (mg
kg
−1): As 0.01–54.8, Cd 0.001–0.02, Cu 0.001–6.29, Hg 0.001–0.52 and Pb 0.001–0.34
mg
kg
−1. Metal ranges in red mullet were (mg
kg
−1): As 0.01–70.9, Cd 0.002–0.85, Cu 0.001–57.3, Hg 0.001–2.07 and Pb 0.001–0.27
mg
kg
−1. Metal level ranges measured in mackerel were (mg
kg
−1): As 0.01–36.4, Cd 0.001–0.1, Cu 0.001–15.9, Hg 0.001–0.78 and Pb 0.002–0.24
mg
kg
−1. In picarel, metal level ranges were (mg
kg
−1): As 0.01–54.6, Cd 0.001–0.1, Cu 0.08–32.9, Hg 0.001–0.207 and Pb 0.001–0.46
mg
kg
−1. Significant differences in metal concentrations were found among fish species. The results presented on metal contents in the examined species give an indication of the environmental conditions. Concentrations of Cd, Cu, Hg and Pb obtained were far below the established values by the European Community regulations. However, arsenic levels found in red mullet were higher than the recommended legal limits for human consumption and as such may present a human health issue.
Disruption of any stage of iron homeostasis, including uptake, utilization, efflux, and storage, can cause progressive damage to peripheral organs. The health hazards associated with occupational ...exposure to inhalation anesthetics (IA) in combination with chronic iron overload are not well documented. This study aimed to investigate changes in the concentration of essential metals in the peripheral organs of rats after iron overload in combination with IA. The aim was also to determine how iron overload in combination with IA affects tissue metal homeostasis, hepcidin–ferritin levels, and MMP levels according to physiological, functional, and tissue features. According to the obtained results, iron accumulation was most pronounced in the liver (19×), spleen (6.7×), lungs (3.1×), and kidneys (2.5×) compared to control. Iron accumulation is associated with elevated heavy metal levels and impaired essential metal concentrations due to oxidative stress (OS). Notably, the use of IA increases the iron overload toxicity, especially after Isoflurane exposure. The results show that the regulation of iron homeostasis is based on the interaction of hepcidin, ferritin, and other proteins regulated by inflammation, OS, free iron levels, erythropoiesis, and hypoxia. Long-term exposure to IA and iron leads to the development of numerous adaptation mechanisms in response to toxicity, OS, and inflammation. These adaptive mechanisms of iron regulation lead to the inhibition of MMP activity and reduction of oxidative stress, protecting the organism from possible damage.
Selenium (Se), an essential trace element that is toxic when humans and animals are exposed to it in excess, is ubiquitous in coal. For centuries, superhigh-organic-sulfur (SHOS) Raša coal, enriched ...in S, Se, U, V, and Mo, was mined and processed across the Mediterranean Raša Bay area, located in the Istrian peninsula (in the northern Adriatic Sea, Croatia). There is concern that Raša coal mine water is contaminating local water, soil, and crops. The aim of this monitoring study was to determine the levels of Se and selected potentially toxic trace (As, Cd, Cu, Cr, Mo, Pb, U, V, and Zn), and minor (Fe and Mn) elements in Raša coal mine water, surface water, and associated vegetables, one fruit, and wild plants. Levels of Se in coal mine water were increased (up to 12 μg/L) compared to the maximum allowed water Se (10 μg/L). Compared to an EU average soil Se (1.15 mg/kg), Raša garden soil showed a 5-fold increase in Se. Compared to Croatian and Greek vegetable Se levels (low to normal), Raša vegetables showed a 20-fold, and a 50-fold increase in Se, respectively. Although approximative only, estimates of daily intake (EDI) of Se for mixed Raša vegetables (n = 21) showed a high level (0.055 mg/day). Namely, recommended dietary allowances (RDA) of Se for females and males are 0.055 mg/day, and 0.070 mg/ day, respectively. The EDI values of the analyzed vegetables contributed to average RDA levels as follows: garlic (183%), turnip (154%), parsley (147%), onion and gourd (76%), lettuce (74%), kale (62%), radicchio (51%), and potato (20%). Although the calculated EDI for the analyzed Raša vegetables was 1/8 the toxic dose (>0.4 mg/day), these results call for further research on the dietary and nutritional status of the residents in terms of Se.
An outdoor pot experiment was designed to study the potential of poplar (Populus nigra 'Italica') in phytoremediation of cadmium (Cd) and lead (Pb). Poplar was treated with a combination of different ...concentrations of Cd (w = 10, 25, 50mgkg−1 soil) and Pb (400, 800, 1200mgkg−1 soil) and several physiological and biochemical parameters were monitored including the accumulation and distribution of metals in different plant parts (leaf, stem, root). Simultaneously, the changes in the antioxidant system in roots and leaves were monitored to be able to follow synergistic effects of both heavy metals. Moreover, a statistical analysis based on the Random Forests Analysis (RFA) was performed in order to determine the most important predictors affecting growth and antioxidative machinery activities of poplar under heavy metal stress. The study demonstrated that tested poplar could be a good candidate for phytoextraction processes of Cd in moderately contaminated soils, while in heavily contaminated soil it could be only considered as a phytostabilisator. For Pb remediation only phytostabilisation process could be considered. By using RFA we pointed out that it is important to conduct the experiments in an outdoor space and include environmental conditions in order to study more realistic changes of growth parameters and accumulation and distribution of heavy metals. Also, to be able to better understand the interactions among previously mentioned parameters, it is important to conduct the experiments during prolonged time exposure., This is especially important for the long life cycle woody species.
•Poplar could be candidate for Cd phytoextraction from moderately contaminated soils.•Poplar could be only considered for phytostabilisation process for Pb.•RFA pointed out that it is important to conduct the experiments in an outdoor.
A screening and confirmatory method for the determination of 13 antibiotic groups including 91 substances was developed for muscle samples and validated using ultra-performance liquid chromatography ...(UPLC) coupled to time-of-flight mass spectrometry. Optimisation of the parameters was performed for every analyte and personal compound database library was created, comprising the mass spectra for following collision energies; 0, 10, 20 and 40 eV. Validation plan scheming and calculation of the parameters was performed using validation software, which included following performance characteristics; selectivity, trueness, precision, decision limit (CCα), detection capability (CCβ) and relative matrix effect.
High resolution mass spectrometry operated in a narrow mass window and resolving power of 50,000 FWHM (2 GHz). Accurate mass measurements with mass accuracy below 10 ppm for all analytes were obtained in spiked muscle samples. The validation experiments showed good selectivity and specificity. The method was validated in the measurement range from 0.5 to 1000 μg/kg, where a specific analyte mixed standard solution was prepared depending on the defined maximum residue limit (MRL) or lowest calibration level (LCL). For screening analysis, detection capabilities (CCβ) below the set MRL values or at LCL concentration levels were defined. CCβ, as the concentration above which the sample can be considered suspect, ranged from 0.63 to 207.8 μg/kg. For confirmation of the analyte, CCα was calculated at LCL or at the MRL, where it ranged from 0.38 to 14.3 μg/kg for unauthorised substances and from 13.8 to 1492.6 μg/kg for authorised substances. Relative within-laboratory reproducibility standard deviation ranged from 5.1 to 34.2%. Satisfactory z-score values in the range from −0.87 to 1.88 were obtained as a result of participating in four proficiency tests organized by European Union Reference Laboratories.
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•Screening and confirmatory HRMS method for 91 antibiotics.•Wide validation range around the established residue limits.•CCβ and CCα as parameters for screening and confirmatory analysis.•Satisfactory proficiency results as confirmation of good method performance.
Monitoring beehive health and timely prevention of possible infections by bacteria, mould, viruses or parasites is exceptionally important in beekeeping. Antibiotics originating from the environment ...can be found in honey and due to improper beekeeping practices. Enabling the detection of antibiotic residues in honey and suppressing the development of antibiotic resistance require the development of a sensitive multi-class method for the determination of antibiotics. For the purpose of analysing honey, a screening and confirmatory method for the determination of 36 antibiotics was developed. The QuEChERS procedure and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was selected to achieve high sensitivity and selectivity. Validation according to the new Commission Implementing Regulation (EU) 2021/808 included the following performance characteristics: selectivity, trueness, precision, decision limit (CCα), detection capability (CCβ) and relative matrix effect. The method was validated in the measurement range from 1 to 10 μg/kg, where maximum trueness and acceptable coefficients of variation were achieved. The detection capability (CCβ) equals the lowest concentration level for all analytes, ranging from 0.1 μg/kg to 2.5 μg/kg, as the false compliant rate less than 5% was confirmed at this level of interest. For unauthorised pharmacologically active substances, the CCα was determined and ranged from 0.16 μg/kg for sulfaquinoxaline to 3.67 μg/kg for difloxacin. The high matrix influence of floral and chestnut honey indicated the need for quantitative analysis using the matrix calibration curve.
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•Screening and confirmatory UHPLC-MS/MS method for 36 antibiotics was validated.•QuEChERS extraction procedure allows for fast and specific extraction.•Highly sensitive method for the determination of residues in the range from 0.1 to 10 μg/kg.•Compliance of samples according to the decision limit (CCα).
Glyphosate is the most used herbicide in agriculture. Its major metabolite is AMPA (aminomethylphosphonic acid), but N-acetyl-AMPA and N-acetylglyphosate are also metabolites of interest. For risk ...assessment, a general residue definition was proposed as the sum of glyphosate, AMPA, N-acetyl-glyphosate and N-acetyl-AMPA, expressed as glyphosate. A confirmatory method for glyphosate in fat, liver and kidneys, as well as a confirmatory method for AMPA and N-acetyl-glyphosate in all matrices, are still missing. In this paper, we present a method for the quantitative determination of glyphosate residues and its metabolites AMPA, N-acetyl-AMPA and N-acetyl-glyphosate by liquid chromatography–mass spectrometry (LC-MS/MS) in adipose tissue, liver, eggs, milk and honey without derivatization. Different chromatographic columns were tested, with the Hypercarb column providing the best results. The analytes were eluted with mobile phases of acidified water with 1.2% formic acid and 0.5% formic acid in acetonitrile. Sample purification procedures were also optimized by varying the solvent extraction mixtures (water, methanol and mixture ψ (methanol, water) = 1:1, each with the addition of 1% formic acid (v/v)), using different sorbents in solid phase extraction (SPE) (polymeric cationic (PCX) and anionic (PAX)) and using dispersive solid phase extraction (dSPE) (C18 and PSA) by modifying the extraction procedures. Finally, the analytes were extracted from the samples with 1% formic acid in water (v/v). Milk and adipose tissue were purified by the addition of dichloromethane, while liver and egg samples were purified by SPE with a mixed cation exchange sorbent and ultrafiltration with cut-off filters. The proposed analytical procedures were validated according to SANTE/11312/2021 guidelines: linearity, limits of quantification, precision and accuracy were determined for all matrices. The limits of quantification (LOQs) ranged from 0.025 to 0.2 mg kg−1. Precision, expressed as relative standard deviation, was <20%, while accuracy, expressed as analytical recovery, ranged from 70% to 120%. During method validation, the measurement uncertainty was estimated to be <50% for all analytes. Good validation parameters according to the SANTE document were achieved for all analytes. Therefore, the method can be considered reliable and sensitive enough for routine monitoring of polar pesticides. The application of the accredited method in routine analysis will provide data that are useful for the re-evaluation of risk assessment studies in foods of animal origin.
Top marine predators present high mercury concentrations in their tissues due to biomagnification in the marine food chain. This study reports mercury (Hg) and selenium (Se) status, and the Hg:Se ...molar ratio assessment in bottlenose (Tursiops truncatus), striped (Stenella coeruleoalba) and Risso's dolphins (Grampus griseus). Total Hg and Se concentrations were determined in muscle, liver, kidney, lung, spleen, adipose tissue and skin collected from 186 specimens stranded in the Croatian part of Adriatic Sea from 1995 to 2014. Total Hg concentrations in tissue samples ranged from 0.001 in the spleen to 2238 mg/kg wet weight in liver. Se concentrations in dolphin samples ranged from 0.010 to 2916 mg/kg ww. Minimum Se concentration was found in muscle and maximum Se concentration were found in liver of bottlenose dolphin. Hg and Se levels in Risso's dolphins showed higher concentrations in all tissues in comparison to bottlenose and striped dolphins. Significant and positive correlations were observed between age and Hg concentrations (P < 0.05). In 66.6 % of Risso's, 15.3 % of bottlenose dolphins and one stranded striped dolphin in this study, the hepatic concentration of Hg exceeded the higher toxic thresholds (400 mg/kg w.w.) previously defined as evidence of liver damage in marine mammals. The Hg:Se molar ratio in the liver of Risso's dolphin was 0.670. The liver of adult bottlenose dolphins showed expected values (0.870), while the liver of young dolphins had a high ratio (0.750), non-specific for the age group. The Hg:Se molar ratio in the liver of striped dolphins was 0.390, which is lower than the literature values.
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•Hg and Se were determined in 186 specimens stranded in the Adriatic Sea.•64 individuals exceeded the lower critical level (Hg = 100 mg/kg) in liver.•The upper critical level in the liver (Hg = 400 mg/kg) exceeded 29 dolphins.•Bioaccumulation of Hg and Se was found in the liver of dolphins.
Aflatoxin M 1 in raw milk in Croatia Bilandžić, Nina; Varenina, Ivana; Solomun, Božica
Food control,
09/2010, Letnik:
21, Številka:
9
Journal Article
Recenzirano
In this study the levels of aflatoxin M
1 (AFM
1) of 61 milk samples delivered from small milking farms were determined in January, February, March and April (winter–spring season), and June, July ...and September (summer–autumn season) of 2009. The AFM
1 concentration was determined by competitive enzyme-immunoassay method. The maximum mean concentrations of AFM
1 recorded in winter–spring season were in the range of 35.8–58.6
ng/l and in summer–autumn season in the range of 11.6–14.9
ng/l. The AFM
1 levels determined in January, February, March and April were significantly higher in accordance with concentration of AFM
1 in June, July and September (
P
<
0.05 to
P
<
0.0001, respectively). Also, there was significant difference (
P
<
0.0001) between the mean concentrations of AFM
1 in samples taken all together in winter–spring and summer–autumn season. Only in one sample delivered in February the level of AFM
1 was higher than the maximum tolerance limit (50
ng/l). Therefore, it was concluded that in 98.4% of milk samples in Croatia the levels of AFM
1 were below maximum tolerance level accepted by the European Union.