Honey has been recognized as a reliable indicator of environmental quality because of honeybees’ intense foraging activity, which brings them into contact with many persistent organic pollutants around the hive. In this study, four types of honey (meadow, acacia, chestnut, and honey in comb) collected at three different locations were analyzed for Co, Cr, Cu, Fe, Mn, Pb, and Zn levels. The highest levels of Fe and Cu in chestnut honey, Co and Zn in meadow honey, and Pb in honey in comb were observed in Varaždin County. The lowest levels of Pb in meadow honey and Co in comb honey were found from apiaries in Sisak-Moslavina County. Significant differences in the mean concentrations of Cr, Cu, Mn, and Fe were observed among the four honey types. Conversely, no significant differences in Co, Pb, and Zn levels were found. Most of the significant differences between the elements are related to chestnut honey. While sampling location (Fe) and type of honey (Pb), or both (Cr and Zn), significantly influenced the concentrations of some elements, these factors were found to be irrelevant for Mn, Co, and Cu. The results showed varying degrees of similarities and differences in mineral levels in honey samples, depending on floral and geographical origin.

In this study, the frequency of aflatoxin M1 (AFM1) occurrence in raw milk was investigated across different seasons over a three-year period from 2022 to 2024 in Croatia. Risk assessment was conducted using estimated daily intake (EDI), hazard index (HI), and margin of exposure (MOE) for various age groups and both genders. The frequency of AFM1 detection above the maximum level (ML) ranged from 1.60% to 15.1%. The average incidence of AFM1 exceeding the ML was 5.67%, with the highest incidence recorded in autumn 2024. AFM1 levels within the limit of detection (LOD) and ML were found in 13% of the samples. The average mean value of AFM1 over the three-year period was 19.2 ng/kg. The highest mean AFM1 EDI values were determined for toddlers (0.61–0.67 ng/kg bw/day) and children (0.41–0.43 ng/kg bw/day). The lowest EDI values were observed in elderly females and males (0.058–0.074 ng/kg bw/day). The EDI values for females and males were slightly different. The risk assessment, based on the HI and MOE, indicated that toddlers and children are at the highest risk of exposure to AFM1, which raises significant health concerns. Additionally, consumers of large quantities of milk face a high risk of exposure, particularly during the spring and autumn seasons. For adults and the elderly, milk consumption does not pose a serious health risk.

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.

The rapid translocation of pesticide and metal residues in the environment poses a health risk to honeybees and provides a potential route for consumer exposure to these pollutants through the consumption of honeybee products. Given the high global consumption of honey, especially by immunocompromised individuals, children, and the elderly, quality control highlights the need for ongoing monitoring and risk assessment. In total, 38 comb honey and 22 extracted honey samples were collected in northern Croatia in June 2023 and analysed for 197 pesticides (using LC-MS/MS and GC-MS/MS) and 17 trace metal(loid)s (using ICP-MS) to assess contamination levels. Comb honey generally contained higher concentrations of active substances than extracted honey, with the highest detection frequencies of fipronil-sulfone, trifloxystrobin, and coumaphos in comb honey, and DMF and DMPF in honey. Glyphosate was the only pesticide to exceed the EU MRL of 0.05 mg/kg in three honey samples. Elemental analysis quantified the levels of most target metals, with Al, Cu, Fe, Mn, Ni and Zn being the most abundant, while Ag, As, and Se were the only ones not detected in this study. None of the samples showed Pb content above the regulatory limit for honey established in the EU (0.1 mg/kg).

The rapid translocation of pesticide and metal residues in the environment and their entry into the food chain pose a significant risk to human health. Given the high global consumption of honey, quality control emphasizes the need for continuous monitoring and risk assessment. To evaluate contamination levels in honey from northern Croatia, a region with intensive agricultural land use, 38 comb honey and 22 extracted honey samples were collected by purposive one-time sampling in June 2023. These samples were analyzed for 190 pesticides using liquid chromatography–tandem mass spectrometry (LC-MS/MS) and gas chromatography–tandem mass spectrometry (GC-MS/MS), and for 17 trace metal(loid)s using inductively coupled plasma mass spectrometry (ICP-MS). The highest detection frequencies were observed for fipronil-sulfone, trifloxystrobin, and coumaphos in comb honey, and for N-(2,4-dimethylphenyl)-formamide (DMF) and N-(2,4-dimethylphenyl)-N′-methylformamidine (DPMF) in extracted honey. Glyphosate was the only pesticide to exceed the European Union (EU) maximum residue level (MRL) of 0.05 mg/kg in three honey samples. Elemental analysis quantified most target metals, with aluminum (Al), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni) and zinc (Zn) being the most abundant, while silver (Ag), arsenic (As), and selenium (Se) were not detected in this study. None of the samples contained lead (Pb) above the regulatory limit for honey established in the EU (0.1 mg/kg). To ensure food safety, further efforts are required to assess the health risks associated with exposure to these contaminants through consumption of the evaluated food.

This study evaluated the impact of a fire at a plastic recycling plant in the suburbs of Osijek on the concentrations of per- and polyfluoroalkyl substances (PFAS) in home-produced eggs (HPE) collected from nearby settlements exposed to smoke. The assessment was conducted over three time periods following the fire. Commercial eggs from supermarkets and HPE from northwestern Croatia were also analyzed. Thirteen out of 30 compounds were quantified. In both HPE groups—the one closer to and more exposed to smoke (Zone A) and the one farther from fire (Zone B)—linear perfluorooctane sulfonic acid (L-PFOS) showed the highest detection frequency (91–100%). The highest mean concentrations of L-PFOS and the sum of the four main PFAS (∑4PFAS: PFOS, perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), and perfluorohexane sulfonic acid (PFHxS)), at 1.33 μg/kg, were measured in HPE from Zone A one month after the fire. In Zone B, a lower total ∑4PFAS of 0.93 μg/kg was detected. After eight months, concentrations of all quantified compounds decreased. The sums of ∑4PFAS decreased to 0.41 μg/kg (A) and 0.37 μg/kg (B), respectively. Concentrations are higher than those from northwestern Croatia and the previously determined national average. Weekly intakes of ∑4PFAS exceeded the tolerable weekly intake for toddlers and children even eight months after the fire.