From Environment to Hive: Plasticizer and Bisphenols Contamination in Algerian Honeys

Abstract

Phthalates (PAEs), non-phthalate plasticizers (NPPs) and bisphenols (BPs) were monitored by fully validated GC-MS and HPLC-MS/MS protocols in honeys from diverse Algerian coastal and non-coastal areas. Experimental results showed that no honey was free of these compounds. A higher PAE contamination was evident in coastal honeys, while NPPs were more abundant in non-coastal samples. The revealed PAEs were: dimethyl phthalate (DMP, 28.12–277.14 µg/kg), diethyl phthalate (DEP, 18.20–404.70 µg/kg), dibutyl phthalate (DBP, 29.58–889.71 µg/kg) and bis(2-ethylhexyl) phthalate (DEHP, 20.66–523.16 µg/kg), while bis(2-ethylhexyl) terephthalate (DEHT, 8.95–206.12 µg/kg) and diethyl adipate (DEA, 10.36–97.51 µg/kg) were the NPPs determined. The EU—not Algeria—classifies DBP and DEHP as very high concern substances. Nonetheless, these PAEs were the most abundant and frequently detected contaminants. Even certain honeys showed DEHP outliers compared to the range provided above (1256.53 µg/kg). Coastal and non-coastal honeys were contaminated by bisphenol A (BPA, 2.64–12.73 µg/kg), thus, raising compliance concerns for export in the EU. In fact, the assessment of dietary exposure and toxicological risk derived from the consumption of these honeys highlighted that, while the exposure to plasticizers was within the safety limits, the exposure to BPA raised toxicological concern. Hopefully, these findings will support the constant monitoring of beekeeping activities and products and encourage the adoption of good practices with a view to guide the advancement of the sector and better safeguard consumers.

1. Introduction

Monitoring the environment for pollution, xenobiotic chemicals, and pathogens is crucial for protecting human health, agriculture, and food production, and for assessing the overall ecosystem health status.

Beekeeping has long been a crucial practice for both honey production and pollination services. However, as we become increasingly aware of the global relevance of the environmental pollution phenomenon, beekeeping has become over time also a non-invasive and reliable methodology for detecting and tracking natural or anthropogenic pollutants and providing early-warning signals of environmental stress or emerging pollutants in an area [1,2].

In fact, the honeybee itself, Apis mellifera L., is a sensitive bioindicator of environmental health. Due to their foraging activity over several kilometres, honeybees come into contact with numerous environmental matrices that may contain traces of pollutants. Not only can bees accumulate these substances during the collection of nectar, pollen and water, as well as through the contact with plants and beekeeping materials, but they can also transfer them into the apiary. As a result, hive products such as honey, wax and propolis can be considered excellent indicators of environmental pollution too [2]. Honey is the most renowned natural sugar substitute produced from floral nectar, which the honeybee collects, transforms, and stores in honeycomb for maturation. Its rich composition of sugars, organic acids, vitamins and minerals has given honey significant nutritional and therapeutic properties since the dawn of time. However, these characteristics can be strongly influenced by the botanical and geographical origin of honey, its processing methods, and storage practices [3]. While these factors influence the quality and nutritional value of honey, they can also encourage the accumulation of xenobiotics, such as pesticide residues, antibiotics, toxic or potentially toxic elements, and other anthropogenic chemicals, such as plasticizers and bisphenols (BPs) [4,5,6].

Plasticizers are primarily used in the manufacturing of polymer plastics, such as polyvinyl chloride (PVC), polypropylene, polyethylene, and polyethylene terephthalate, for increasing their flexibility and softness. Based on their chemical structure, they are commonly classified in phthalates (PAEs), such as dibutyl phthalate (DBP), diisobutyl phthalate DIBP), benzyl butyl phthalate (BBP), di-(2-ethylhexyl) phthalate (DEHP), etc., and non-phthalate plasticizers (NPPs), including terephthalates (e.g., di-(2-ethylhexyl) terephthalate [DEHT]), and adipates (e.g., di-(2-ethylhexyl) adipate [DEHA]) [7]. Similarly to plasticizers, BPs, such as bisphenol A (BPA) and its analogues, are a group of additives used in the manufacturing of certain polymers, such as polycarbonate (PC) plastics or epoxy resins, exploitable in a variety of applications, including plastic packaging and the lining of some food and beverage packaging [8].

Both plasticizers and BPs can contaminate the honey via plastic pollution. In this case, the dispersion of plastics, resins and coatings from industrial and urban discharges, agricultural and economic activities (i.e., tourism) and, not least, plastic waste degradation, encourages the leaching of these additives in the soil, water and atmosphere populated by bees during the foraging activity. However, honey can also become contaminated due to improper manufacturing and/or storage practices, i.e., via the direct contact with employed plastic materials, such as synthetic honeycombs, frames, feeders, lids and containers [9,10].

Plasticizers and BPs are known endocrine disruptors, able of interfering with hormone regulation, and have been associated with reproductive toxicity, developmental disorders, metabolic dysfunctions, and increased risk of certain cancers [8,11,11]. Continuous exposure, even at low levels, to these chemicals through honey may have cumulative effects, particularly in vulnerable populations, such as children and pregnant women, making the monitoring of honey safety critical for public health.

In Algeria, beekeeping plays a significant role in agricultural activities and contributes greatly to the socio-economic fabric of the country, being a valuable source of income for farmers and beekeepers [12]. However, the sector is already facing structural challenges, including poor queen rearing practices, climate variability, and declining honey productivity [13]. The emerging problem of plastic pollution and derived contaminants adds a further layer of complexity to the issue, especially given the absence of a regulatory framework, official inspections and monitoring programmes on the honey production process, and the quality and safety of the final product as well. As a result of population growth, rapid urbanization and industrialization, Algeria is experiencing a consistent yearly increase in plastics import, especially in terms of primary polymers rather than finished products. As an example, the plastic imports ramped from 304 Kilotons in 2007 to 931 Kilotons in 2020, and, in 2018, Algeria was the second importer of plastics in Africa. This has implied high rates of industrial processing and production activities, as well as high levels of waste generation in the country. Indeed, Algeria yearly generates around 34 million tons of waste (2018 estimate), and forecasts predict that the overall amount of waste will almost double over the next 17 years, increasing to 73 million tons by 2035. Plastics constitute more than 5 million tons of this waste, and around 0.52 million tons, which are mismanaged yearly, with 0.08–0.21 million tons becoming marine plastic litter. In addition, Algeria has a coastline stretching over more than 1600 km that is constantly threatened by plastic waste, of which 81% is of land-based sources (e.g., tourism, fishing and other economic activities), and it is deliberately or accidentally escaped from collection circuits [14,15]. All these pollution inputs coexist in the Tell region of northern Algeria, a narrow and fertile area featuring marine coasts, plains, and the Tell Atlas Mountains, which is characterized by higher population density, greater anthropogenic activity, and, consequently, higher environmental pollution than the arid Sahara region of southern Algeria [16,17].

In light of the pollution scenario and the legislative and informational gaps, the study of contamination from plasticizers and BPs in the Algerian environment through the monitoring of honey is, therefore, crucial not only for understanding the sources of origin and spread of plastic compounds, but also for developing strategies to protect the public health and the environment as well [18]. Hence, this study aims to monitor plasticizers and BPs in an array of honeys with different botanical origins and collected in diverse areas of the Tell region, which moreover have been already investigated for their inorganic element profiles in previous research [19].

Hopefully, the findings from this research will contribute to provide a detailed overview of the safety and quality of Algerian honey and to guide the advancement of the sector with a view to reducing exposure to hazardous compounds and protecting consumer health. By providing reliable data, the study will also support the future assessment of potential emerging risks associated with honey contamination by these chemicals.

2. Materials and Methods

2.1. Sample Collection

The research involved the same sample set already described in our previous work [19]. Briefly, 36 types of honeys with different botanical origins and produced in diverse coastal and non-coastal regions of northern Algeria during June–December 2024 were considered. For every type of honey, n = 3 honey samples produced from the same apiary but at different times during the semester in question were collected. Hence, a total of n = 108 samples were considered in the study.

The botanical and geographical origin of all honeys was determined based on information provided by the beekeepers and the nutritional labels. In addition, the botanical origin of honeys was experimentally confirmed, as already described by Nava et al. [19].

To facilitate the geographical identification of the samples and the subsequent discussion of the experimental results, the samples were further classified according to the proximity of their production site to the coastal strip. Hence, two sample groups were defined: coastal honeys (n = 42 samples from Annaba, Skikda, Mostaganem and Tizi Ouzou) and non-coastal (or inland) honeys (n = 66 samples from Mascara, Relizane, Tiaret, Chlef, Ain Defla, Tissemsilt, Blida, Naama, El Bayadh, Laghouat, Djelfa, Touggourt and Tebessa). Figure 1 illustrates the map of the coastal and non-coastal areas of north Algeria involved in honey production; meanwhile,

Table 1. Information of honey samples from northern Algeria according to the geographical area of provenance (coastal and non-coastal area), production region, and botanical origin.

North Algeria Area	Apiary Code	N. of Honey Samples	Geographical Origin	Botanical Origin
Coastal area	HS-10	3	Annaba	Eucalyptus globulus
	HS-13	3	Skikda	Eucalyptus globulus
	HS-27	3	Skikda	Citrus
	HS-23	3	Mostaganem	Multifloral
	HS-29	3	Mostaganem	Citrus
	HS-33	3	Mostaganem	Eucalyptus gomphocephala
	HS-34	3	Mostaganem	Pinus halepensis–Rosmarinus officinalis
	HS-35	3	Mostaganem	Citrus
	HS-1	3	Tizi Ouzou	Pinus silvestris
	HS-7	3	Tizi Ouzou	Quercus ilex–Eucalyptus globulus
	HS-11	3	Tizi Ouzou	Quercus ilex
	HS-12	3	Tizi Ouzou	Nasturtium officinalis
	HS-15	3	Tizi Ouzou	Ceratonia siliqua
	HS-32	3	Tizi Ouzou	Erica arborea–Lavandula stoechas
Non-coastal area	HS-5	3	Blida	Citrus
	HS-18	3	Blida	Citrus
	HS-25	3	Relizane	Ziziphus lotus
	HS-24	3	Tiaret	Eruca sativa
	HS-28	3	Tiaret	Multifloral
	HS-17	3	Chlef	Ziziphus lotus–Thymus vulgaris
	HS-36	3	Chlef	Multifloral
	HS-22	3	Ain Defla	Pinus halepensis–Quercus ilex
	HS-26	3	Naâma	Multifloral
	HS-19	3	El Bayadh	Euphorbia officinarum
	HS-2	3	Laghouat	Ziziphus lotus
	HS-6	3	Laghouat	Euphorbia officinarum
	HS-3	3	Djelfa	Ziziphus lotus–Silybum marianum
	HS-8	3	Djelfa	Nasturtium officinalis
	HS-16	3	Djelfa	Multifloral
	HS-20	3	Djelfa	Silybum marianum
	HS-21	3	Djelfa	Eruca sativa
	HS-9	3	Touggourt	Helianthus annuus
	HS-30	3	Tebessa	Ziziphus lotus
	HS-14	3	Tebessa	Rosmarinus officinalis
	HS-31	3	Mascara	Tamarix Africana
	HS-4	3	Tissemsilt	Tamarix africana

reports the apiary codes and number of honey samples obtained per apiary, together with the relative geographical and botanical origins.

All samples were obtained in 150 g glass jars, each labelled with the corresponding botanical source, nutritional label, and provenance, transported under dark conditions and stored at a constant temperature until analysis.

2.2. Materials and Reagents

Solvents (i.e., ultrapure water, n-hexane and acetonitrile) were purchased from Merck (Darmstadt, Germany). The salts used for the extraction step (NaCl and MgSO₄) and the reagents used for purification phase (primary and secondary ammines [PSAs], and octadecyl [C₁₈] sorbent) were purchased from Fluka in Milan, Italy. The analytical standards had a certified purity ≥99%, and were supplied by Sigma-Aldrich (Steinheim, Germany). Specifically, plasticizers were: di-methyl adipate (DMA), di-ethyl adipate (DEA), di-isobutyl adipate (DiBA), di-n-butyl adipate (DBA), bis-(2-ethylhexyl) adipate (DEHA), bis(2-methoxyethyl) adipate (DMEA), di-methyl phthalate (DMP), di-ethyl phthalate (DEP), di-propyl phthalate (DPrP), di-butyl phthalate (DBP), di-isooctyl phthalate (DiHepP), di-cyclohexyl phthalate (DcHexP), bis-(2-ethylhexyl) phthalate (DEHP), di-phenyl phthalate (DPhP), di-isononyl phthalate (DiNP), bis(2-methoxyethyl) phthalate (DMEP), benzyl butyl phthalate (BBP), diisobutyl phthalate (DiBP), benzyl benzoate (BB), bis-(2-ethylhexyl) terephthalate (DEHT), and di(2-ethylhexyl) sebacate (DEHS). BPs included: 4′-sulfonyldiphenol (BPS), 4,4′-methylenediphenol (BPF), 1,1-bis(4-hydroxyphenyl)ethane (BPE), 4,4′-(propan-2,2-diyl)diphenol (BPA), 4-[2-(4-hydroxyphenyl)butan-2-yl]phenol (BPB), 2,2-Bis(4-hydroxyphenyl)hexafluoropropane (BPAF), 1,1-bis(4-hydroxyphenyl)-1-phenyl-ethane (BPAP), 1,1-bis(4-hydroxyphenyl)-cyclohexane (BPZ), and 1,4-bis(2-(4-hydroxyphenyl)-2-propyl)benzene (BPP). Isotopically labelled standards were DBP-d4, DEHP-d4 (both at 100 ng/μL in nonane), ¹³C₁₂-BPA, and ¹³C₁₂-BPS (purity ≥ 99%). They were all provided by Cambridge Isotope Laboratories Inc. (Andover, MA, USA) and used as internal standards (ISs). Stock solutions of plasticizers were prepared at a concentration of 1000 mg/L in n-hexane, while stock solutions of BPs were prepared at 100 mg/L in acetonitrile. Working solutions were subsequently obtained by appropriate dilutions of the stock solutions and stored at 4 °C until use.

2.3. Quality Assurance and Quality Control Procedures

To avoid the background contamination of the analytical workflow, a series of quality control (QC) and quality assurance (QA) processees were considered, as already described in detail by Di Bella et al. [20].

Considering the QC measures, solvents were always of high quality (i.e., SupraSolv^® grade and LC/MS grade) and were daily checked to be free of plasticizers by GC-MS (i.e., target PAEs and NPPs: <LOD) and of BPs by HPLC-MS/MS (i.e., target BPs: <LOD). Plastic materials were avoided throughout the experimental procedures—except for PAE-free nitrile gloves—and laboratory glassware and equipment were always made of polytetrafluoroethylene (PTFE), polyvinylidene fluoride (PVDF), aluminum and stainless steel. Before use, laboratory glassware was pre-washed with a suitable organic solvent and, then, dried in an oven for ~4 h at 120 °C.

Specific QA procedures were also carried out during the validation process and sample analysis. The building up of a precise blank strategy, the use of ISs, and the implementation of method calibration and recovery tests are further discussed in Section 2.5, Section 2.6 and Section 2.7.

2.4. Sample Preparation

The extraction of plasticizer residues from honey samples followed the procedure described by Derrar et al. [21], with minor modifications. Briefly, an aliquot of honey (5.0 g, fresh weight, fw) was weighed into a tube, added with DBP-d₄ and DEHP-d₄ (1 mg/L. each), and extracted with 10 mL of ultrapure water and 10 mL of acetonitrile. The Q-sep QuEChERS salts were then added to promote phase separation. The organic phase was collected and subjected to a dispersive solid-phase extraction (d-SPE) conducted with 0.25 mg of MgSO₄, 0.1 mg of PSA, and 0.1 mg of C₁₈ sorbent. The extract was filtered through a 0.22 µm PTFE syringe filter before analysis.

BP analogues were extracted using a validated procedure [22]. In brief, an aliquot of honey (1.5 g, fw) was placed in a glass tube and mixed with 3 mL of ultrapure water, 3 mL of acetonitrile and 100 µL of the internal standard ¹³C₁₂BPA (100 µg/L). The solution was shaken vigorously and 750 mg of Q-Sep QuEChERS was then added to each sample. The mixture was then shaken and centrifuged at 4000 rpm for 5 min. The upper layer was transferred to a clean-up kit containing 25 mg of C₁₈ sorbent, shaken for one minute and then centrifuged again at 4000 rpm for 5 min. Finally, the 1 mL extract in acetonitrile was filtered through a PVDF syringe filter (0.22 µm) and injected into the HPLC-MS/MS system.

2.5. Analysis by GC-MS and HPLC-MS/MS

The analysis of plasticizers was performed using an Agilent GCMS-5977 single quadrupole mass spectrometer (Agilent Technologies, Santa Clara, CA, USA), equipped with a Supelco SLB-5 MS capillary column (30 m × 0.25 mm i.d., 0.25 μm film thickness). The oven temperature programme was as follows: initial temperature 60 °C; ramp from 60 to 190 °C at 8 °C/min with a 5 min hold; from 190 to 240 °C at 8 °C/min with another 5 min hold; and finally from 240 to 315 °C at 8 °C/min. Helium was used as the carrier gas at a constant linear velocity of 30.0 cm/s. The transfer line and injector temperatures were set at 300 °C and 260 °C, respectively. Injections were performed in splitless mode for 60 s, followed by a split ratio of 1:15; the injection volume was 1 μL. The acquisition was performed in full scan electron ionization (EI) mode, with ionization energy set at 70 eV and an emission current of 250 μA, over a mass range of 40–400 amu. Single Ion Monitoring (SIM) mode was applied for monitoring a subset of fragments with their related mass values in a predefined retention time (RT) range. Specifically, one quantifier and two qualifier ions were monitored for each target plasticizer, as reported in

Table 2. Monitored ions of investigated plasticizers, and main parameters of validation of the GC-MS method employed for their analysis: linearity of calibration curves in neat solvent (R²_solv) and matrix extract (R²_mat), limit of detection (LOD), limit of quantification (LOQ), and matrix effect (t_cal).

Analyte	Abbreviation	Monitored Ions (m/z)	R2solv/R2mat	LOD (µg/kg)	LOQ (µg/kg)	Matrix Effect tcal
Dimethyl phthalate	DMP	163, 92,164	0.9991/0.9932	0.046	0.131	0.693
Diethyl phthalate	DEP	149, 177, 176	0.9996/0.9951	0.049	0.174	0.423
Dipropyl phthalate	DPrP	149, 150, 209	0.9999/0.9935	0.072	0.260	0.771
Dibutyl phthalate	DBP	149, 150, 223	0.9990/0.9927	0.068	0.228	1.042
Diisobutyl phthalate	DiBP	149, 150, 223	0.9993/0.9946	0.075	0.236	0.898
Butyl benzyl phthalate	BBP	149, 91, 206	0.9994/0.9979	0.372	1.185	0.662
Diphenyl phthalate	DPhP	225, 226, 104	0.9986/0.9958	0.153	0.480	0.791
Dicyclohexyl phthalate	DcHexP	149, 167, 150	0.9995/0.9975	0.085	0.301	0.411
Di(2-Ethylhexyl) phthalate	DEHP	149, 167, 279	0.9988/0.9881	0.076	0.256	0.745
Diisononyl phthalate	DiNP	293, 149, 167	0.9977/0.9773	0.082	0.270	1.127
Bis(2-Methoxyethyl) phthalate	DMEP	149, 176, 207	0.9991/0.9986	0.077	0.254	0.987
Dimethyl adipate	DMA	114, 101, 111	0.9979/0.9894	0.032	0.101	1.342
Diethyl adipate	DEA	111, 157, 128	0.9993/0.9972	0.146	0.464	0.834
Benzyl benzoate	BB	105, 91, 212	0.9984/0.9898	0.131	0.389	0.351
Dibutyl adipate	DBA	129, 185, 111	0.9981/0.9870	0.177	0.536	0.559
Diisobutyl adipate	DiBA	129, 185, 111	0.9995/0.9923	0.084	0.273	1.732
Di(2-Ethylhexyl) adipate	DEHA	129, 112, 147	0.9988/0.9867	0.140	0.438	0.867
Bis(2-Methoxyethyl) adipate	DMEA	111, 155, 169	0.9991/0.9885	0.120	0.372	0.553
Di(2-Ethylhexyl) terephthalate	DEHT	149, 112, 261	0.9982/0.9874	0.248	0.818	0.498
Di(2-ethylhexyl) sebacate	DEHS	185, 149, 112	0.9995/0.9948	0.275	0.698	0.947

Underlined ions represent quantifier ions.

.

The HPLC system (Shimadzu, Kyoto, Japan) coupled to an LCMS-8040 triple quadrupole mass spectrometer with an electrospray ionization (ESI) source was used for the determination of BPS, BPF, BPE, BPA, BPB, BPAF, BPAP, BPZ, and BPP.

Chromatographic separation was performed on an Agilent Zorbax SB-C18 column (5 µm 4.6 × 250 mm). The flow rate was 0.7 mL/min. The mobile phases were ultrapure water (solvent A) and acetonitrile (solvent B). The following linear gradient was used: 0 min, 20%B; 7 min, 40%B; 25 min, 90%B; 35 min, 20%B. For ESI-MS, the analyses were performed in negative ion mode. The MS operated under the following conditions: nebulizing gas flow 3.0 L/min, nebulizing gas pressure 770 KPa, drying gas flow 15.0 L/min, DL temperature 250 °C, and CID gas 230 KPa. Multiple reaction monitoring (MRM) was used for quantitative and qualitative data. To this purpose, parameters, such as dwell time, Q1 pre bias, collision energy, and Q3 pre bias, were automatically optimized by the instrumental software. For each target BP, the two most abundant product ions were used for qualification and quantification, as reported in

Table 3. Precursor and product ions of investigated BPs, and main parameters of validation of the HPLC-MS/MS method used for their analysis: linearity of calibration curves in neat solvent (R²_solv) and matrix extract (R²_mat), limit of detection (LOD), limit of quantification (LOQ), and matrix effect (t_cal).

Analyte	Abbreviation	Precursor Ion (m/z) [M−H]−	Product Ions (m/z)		R2solv/R2mat	LOD (µg/kg)	LOQ (µg/kg)	Matrix Effect tcal
			Quantifier	Qualifier
2,2-bis-(4-hydroxyphenyl)-propane	BPA	227.29	227.3 → 212.1	227.3 → 133.0	0.9997/0.9923	0.35	1.16	0.657
4,4′-Sulfonyldiphenol	BPS	249.27	249.3 → 107.9	249.3 → 156.0	0.9998/0.9981	0.35	1.16	0.989
4,4′-Methylenediphenol	BPF	199.24	199.2 → 93.1	199.2 → 105.1	0.9994/0.9975	0.35	1.16	0.423
4,4′-sec-Butylidenediphenol	BPB	241.31	241.3 → 212.0	241.3 → 211.0	0.9992/0.9986	0.47	1.56	0.443
4,4′-Ethylidenebisphenol	BPE	213.26	213.3 → 198.0	213.3→194.9	0.9988/0.9888	2.55	8.49	0.771
4,4′-Cyclohexylidenebisphenol	BPZ	267.35	267.3 → 145.0	267.3 → 173.1	0.9989/0.9899	0.47	1.56	0.852
4,4′-(1,4-Phenylenediisopropylidene) bisphenol	BPP	345.47	345.5 → 330.1	345.5 → 133.1	0.9993/0.9946	1.45	4.82	0.834
4,4′-(1-Phenylethylidene) bisphenol	BPAP	289.36	289.4 → 274.1	289.4 → 273.1	0.9985/0.9859	1.45	4.82	0.576
4,4′-(Hexafluoroisopropylidene)diphenol	BPAF	335.24	335.3 → 265.0	335.3 → 177.0	0.9996/0.9977	1.10	3.66	0.972

. The identification criteria were the following: two diagnostic ions (precursor ion and qualifier ion), precision of the mass error ± 5 ppm for the analyte, and retention time shift ± 0.1 min. Quantification was performed considering the most abundant product ion (quantifier ion). Data were acquired by LabSolution software (v. 5.53 SP2, Shimadzu, Kyoto, Japan).

For both plasticizers and BPs, each sample was injected in triplicate, and the potential background contamination was continuously monitored by using procedural and solvent blanks. Specifically, procedural blanks (i.e., ultrapure water) were always prepared and analyzed in parallel with every n = 3 honey samples from a given apiary. Throughout the study period, the analysis of procedural blanks revealed the absence of target analytes (<LOD) or negligible contamination levels (always <LOQ), thus, confirming that the use of materials, solvents, and reagents did not affect the reliability of experimental results. The solvent blanks were n-hexane in the case of PAEs and NPPs, and water/acetonitrile (50:50, v/v) for BPs. They were analyzed along with every honey sample and resulted in findings always free of investigated PAEs, NPPs and BPs (<LOD). Consequentely, any interferences of the solvent or potential carry-over effects from one sample to the next could be excluded.

2.6. Method Validation

The analytical methods described for plasticizers and BPs were checked in terms of linearity, matrix effect (ME), limits of detection and quantification (LOD and LOQ), recovery, and precision, with the help of standard solutions and a matrix blank, namely a sample of commercial organic honey free of target plasticizers and BPs.

The evaluation of linearity occurred by preparing 7-point calibration curves in neat solvent and matrix blank extract. Specifically, calibration curves in solvent were obtained by serially diluting standard mixtures in n-hexane for PAEs and NPPs, and acetonitrile for BPs, and spiking the obtained solutions with known amounts of relative ISs. Matrix-matched curves were prepared by spiking the blank matrix extract with serially diluted standard mixtures and a known amount of IS. In both cases, the same calibration ranges were selected. For PAEs and NPPs, calibration levels were in the range of 5000–10 µg/L and 1000–5 µg/L, respectively. ISs such as DBP-d4 and DEHT-d4 were always spiked at 1 mg/L. Calibration curves from standard solutions in neat solvent had a good linearity with correlation coefficients (R²_solv) from 0.9979 to 0.9999, while calibration curves from matrix extract showed a satisfactory linearity, with coefficients (R²_mat) between 0.9793 and 0.9986. For BPs, the calibration range was 10–0.01 µg/L, and the IS ³C₁₂BPA was always spiked at 100 µg/L. Standard response curves were generated by plotting the ratio of the mean target ion area to the mean IS peak area against the ratio of the mean analyte concentration to the mean IS concentration (n = 6 analytical replicates). In pure solvent, these curves showed high correlation coefficients (R²_solv between 0.9985 and 0.9997); on the other hand, curves from matrix extract were characterized by slightly lower, but still acceptable, coefficients (R²_mat between 0.9859 and 0.9986).

For the assessment of the ME, a comparison between the slope of standard curves in neat solvent and matrix extract was performed for every analyte by a Student’s t-test, as reported elsewhere [23]. For each target compound, the calculated t-value was always lower than the critical tabulated t-value of 2.447, based on a confidence level of 95% and six degree of freedom (Table 2 and Table 3). Therefore, it is possible to assume that the matrix effect was negligible for the determination of PAEs, NPPs and BPs as well.

Due to the negligible ME, instrumental LODs and LOQs were calculated by calibration curves constructed in neat solvent, using the formulae:

LOD = 3.29 σ/b

LOQ = 10 σ/b

where σ is the standard deviation of the solvent blank (n = 6 analytical replicates) and b is the slope of the calibration curve of the analyte. As reported in Table 2 and Table 3, LODs and LOQs were always in the order of µg/kg. Considering plasticizers, the lowest values were obtained for DMP (LOD: 0.046 µg/kg and LOQ: 0.131 µg/kg) and DEP (LOD: 0.049 µg/kg and LOQ: 0.174 µg/kg), while BBP displayed the highest limits (LOD: 0.372 µg/kg and LOQ: 1.185 µg/kg) (Table 2). Among BPs, BPA and BPS had the lowest LOD (0.35 µg/kg) and, hence, LOQ (1.16 µg/kg). Meanwhile, BPP and BPAP revealed the highest values (LOD: 1.45 µg/kg and LOQ: 4.82 µg/kg) (Table 3).

The recovery of target analytes was determined by fortifying the matrix blank with commercial standards at two concentration levels (see

Table 4. Recovery and precision of the analytical methods assessed for plasticizers (DMP, DEP, DBP and DEHP) and BPA revealed in Algerian honeys. Recovery data are expressed as mean recovery percentage, while precision data as the relative standard deviation (RSD%) of the % recovery calculated at the lowest fortification level in the same day (intra-day precision) and in five consecutive days (inter-day precision).

Analyte	Recovery (%)		Precision (RSD%)
			Intra-Day	Inter-Day
Fortification Level	100 μg/L	1000 μg/L	100 μg/L
DMP	96.98	93.66	7.17	11.03
DEP	93.18	90.11	5.29	10.14
DBP	102.59	108.17	7.53	12.27
DEHP	98.60	105.18	9.14	11.44
DEHT	91.99	87.06	7.22	9.85
DEA	93.83	87.66	8.29	10.99
Fortification level	10 μg/L	100 μg/mL	10 μg/L
BPA	96.23	103.55	5.14	9.23

) and processing them in the same way as test samples. At each concentration level, the difference between the mean experimental value (n = 6 analytical replicates) and the spiked concentration value was reported as mean recovery percentage. The precision of the method was determined as the relative standard deviation (RSD%) of the recovery rate calculated at the lowest concentration level, in the same day (intra-day precision) and in five consecutive days (inter-day precision). Table 4 shows the data obtained by the evaluation of recovery and precision of the method only for the plasticizers and BPs determined in Algerian honeys. Overall, the recovery values fell within the range 70–120% and both intra-day and inter-day RSDs% were below 20%, thus meeting the criterion of acceptability [24]. Hence, no significant loss and interference could be ensured during the analytical workflow.

2.7. Statistical Analysis

Statistical processing was performed using SPSS 13.0 software for Windows (SPSS Inc., Chicago, IL, USA). Experimental data were expressed as mean ± standard deviation of n = 3 honeys from the same apiary (HS-1–HS-36), where every honey was analyzed three times. The calculated %RSD for samples from the same apiary was constantly ≤12%, thus confirming a good within-apiary replicability.

The data matrix consisted of n = 108 cases (n = 42 honey samples from the coastal areas and n = 66 honey samples from the non-coastal areas of north Algeria) and n = 8 variables, namely the target analytes determined in all the honey samples (i.e., DMP, DEP, DBP, DEHP, ∑PAEs, DEHT, DEA, ∑NPPs, and BPA).

Once the assumptions of normality of residuals and homogeneity of variances were checked, a one-way ANOVA followed by Tukey’s Honestly Significant Difference (HSD) post hoc test was performed to assess the significance of differences among samples from different provinces (n = 17) in both coastal and non-coastal areas. Statistical significance was accepted at p < 0.01 for each examined variance. In addition, a Spearman rank correlation analysis was employed to investigate potential relationships between the different compounds revealed in honeys samples. For values below the limit of quantification (LOQ), half the value of the limit of detection (LOD/2) was used as a surrogate.

2.8. Assessment of Human Exposure to Plasticizers and BPs via Honey Intake

According to the latest assessment by the European Food Safety Authority (EFSA) in 2019, the tolerable daily intake (TDI) for a group of phthalates including DEHP and DBP was set at 0.05 mg per kg of body weight per day [25]. Additionally, the World Health Organization (WHO) set a higher TDI of 5.00 mg/kg bw per day for DEP in 2003 [26]. The EFSA’s 2023 review significantly lowered the TDI for BPA, established in 2015, to 0.2 ng/kg body weight per day due to growing concerns about the substance’s toxicity even at low concentrations [27]. Based on the results of our study, we calculated the intake of plasticisers and BPA assuming an average daily per capita consumption of honey in Europe and Algeria for adults and children. In Europe, the average per capita consumption is estimated at about 1.6 g of honey per day. Within this average, adults tend to consume slightly more honey, with an estimated intake of around 1.5 to 2.0 g per day, while children generally consume less, with values estimated between 0.5 and 1.2 g per day, as they tend to use honey less frequently compared to other sugars. In Algeria, average consumption is lower and highly variable. Based on available estimates, adults consume approximately 0.6 to 1.0 g of honey per day, while children and adolescents are estimated to consume between 0.2 and 0.6 g per day. To contextualize exposure in terms of potential health impact, average body weights were set at 60 kg for adults and 20 kg for children, aligning with WHO regional reference values for African populations [28].

3. Results and Discussion

3.1. Plasticizers and BPs in the Algerian Honeys

The level of contamination from plasticizers and BPs in the Algerian honeys can be found in

Table 5. Contamination of single plasticizers, total PAEs (ΣPAEs), total NPPs (ΣNPPs), and BPA revealed in the Algerian honeys. Results are expressed in terms of μg/Kg (fw) and as mean ± standard deviation of n = 3 honeys from the same apiary (HS-1–HS-36), where every honey was analyzed three times. Detection frequencies (%) and statistic outputs from one-way ANOVA and Tukey’ HSD tests are also reported.

Sample Code	DMP	DEP	DBP	DEHP	ΣPAEs	DEHT	DEA	ΣNPPs	BPA
Coastal honey
HS-1	<LOD	18.20 ± 0.46 a	32.73 ± 1.43 a	20.70 ± 1.28 a	71.62 ± 2.98 a,b	<LOD	<LOD	<LOD	5.50 ± 0.36 a
HS-7	<LOD	22.13 ± 0.16 a	50.15 ± 2.39 a	<LOD	72.28 ± 2.25 a,b	<LOD	<LOD	<LOD	3.64 ± 0.21 a
HS-10	147.44 ± 8.62 a	404.70 ± 13.21 d	170.90 ± 7.70 c	1256.53 ± 60.54 d	1979.57 ± 64.81 e	84.08 ± 1.70 b	<LOD	84.08 ± 1.70 b	9.64 ± 0.54 a,b
HS-11	<LOD	<LOD	30.57 ± 1.34 a	<LOD	30.57 ± 1.34 a	<LOD	<LOD	<LOD	5.34 ± 0.26 a
HS-12	<LOD	21.89 ± 0.81 a	96.93 ± 2.22 b	<LOD	118.82 ± 2.83 b	<LOD	<LOD	<LOD	7.24 ± 0.52 a,b
HS-13	277.14 ± 10.77 a	50.46 ± 1.04 b	48.84 ± 3.02 a	<LOD	376.44 ± 10.07 c	9.75 ± 0.45 a	<LOD	9.75 ± 0.45 a	12.73 ± 2.51 b
HS-15	<LOD	<LOD	889.71 ± 96.09 d	<LOD	889.71 ± 96.09 d	<LOD	<LOD	<LOD	3.28 ± 0.12 a
HS-23	<LOD	20.13 ± 0.80 a	36.32 ± 1.26 a	60.35 ± 1.52 a	116.80 ± 2.13 b	69.13 ± 1.21 b	<LOD	69.13 ± 1.21 b	8.16 ± 0.67 a,b
HS-27	180.79 ± 4.94 a	43.21 ± 1.75 b	197.55 ± 3.99 c	<LOD	421.55 ± 0.87 c	9.25 ± 0.96 a	<LOD	9.25 ± 0.96 a	8.03 ± 0.70 a,b
HS-29	<LOD	<LOD	32.80 ± 1.05 a	26.29 ± 1.38 a	59.09 ± 1.59 a	22.75 ± 0.88 b	<LOD	22.75 ± 0.88 a	2.69 ± 0.29 a
HS-32	<LOD	<LOD	41.27 ± 2.13 a	<LOD	41.27 ± 2.13 a	<LOD	<LOD	<LOD	6.25 ± 0.34 a
HS-33	<LOD	<LOD	36.05 ± 0.39 a	46.24 ± 1.01 a	82.29 ± 0.99 a,b	52.29 ± 0.51 b	<LOD	52.29 ± 0.51 b	<LOD
HS-34	<LOD	<LOD	239.77 ± 5.84 c	523.16 ± 35.83 c	762.93 ± 41.06 d	<LOD	<LOD	<LOD	<LOD
HS-35	<LOD	<LOD	<LOD	44.06 ± 0.59 a	44.06 ± 0.59 a	77.62 ± 2.78 b	<LOD	77.62 ± 2.78 b	<LOD
Non-coastal honeys
HS-2	<LOD	<LOD	29.58 ± 1.15 a	20.66 ± 0.81 a	50.24 ± 1.00 a	10.55 ± 1.05 a	<LOD	10.55 ± 1.05 a	5.45 ± 0.19 a
HS-3	<LOD	<LOD	37.11 ± 1.93 a	<LOD	37.11 ± 1.93 a	<LOD	<LOD	<LOD	2.79 ± 0.96 a
HS-4	<LOD	<LOD	49.27 ± 0.95 a	<LOD	49.27 ± 0.95 a	<LOD	<LOD	<LOD	<LOD
HS-5	<LOD	<LOD	<LOD	<LOD	<LOD	16.81 ± 0.94 a	<LOD	16.81 ± 0.94 a	<LOD
HS-6	<LOD	<LOD	128.96 ± 4.06 b,c	51.13 ± 1.38 a	180.09 ± 3.21 b	<LOD	<LOD	<LOD	<LOD
HS-8	<LOD	37.18 ± 1.15 a,b	36.58 ± 1.23 a	<LOD	73.76 ± 1.00 a,b	8.95 ± 0.62 a	32.03 ± 1.82 a	40.98 ± 2.42 a	3.01 ± 0.87 a
HS-9	<LOD	34.43 ± 1.63 a,b	258.33 ± 13.50 c	<LOD	292.76 ± 11.93 b	<LOD	<LOD	<LOD	2.64 ± 0.30 a
HS-14	<LOD	101.81 ± 5.07 c	37.34 ± 0.65 a	<LOD	139.15 ± 5.67 b	<LOD	<LOD	<LOD	<LOD
HS-16	44.54 ± 1.41 b	204.22 ± 9.36 c	65.57 ± 0.89 b	123.35 ± 10.73 b,c	438.67 ± 1.91 c	132.74 ± 3.35 c	10.36 ± 1.29 a	143.10 ± 4.10 c	<LOD
HS-17	<LOD	<LOD	<LOD	27.67 ± 1.31 a	27.67 ± 1.31 a	<LOD	<LOD	<LOD	8.65 ± 0.49 b
HS-18	<LOD	<LOD	150.64 ± 5.85 c	<LOD	150.64 ± 5.85 b	<LOD	<LOD	<LOD	<LOD
HS-19	44.90 ± 3.11 b	<LOD	291.82 ± 13.52 c	<LOD	336.72 ± 13.02 c	<LOD	<LOD	<LOD	5.11 ± 0.07 a
HS-20	<LOD	<LOD	278.80 ± 13.22 c	52.89 ± 2.01 a	331.69 ± 13.22 c	<LOD	<LOD	<LOD	<LOD
HS-21	<LOD	<LOD	61.51 ± 1.22 b	134.66 ± 11.12 b	196.17 ± 11.34 b	193.33 ± 8.46 c	97.51 ±2.23 b	290.84 ± 10.64 c	<LOD
HS-22	33.54 ± 1.44 b	<LOD	<LOD	<LOD	33.54 ± 1.44 a	<LOD	<LOD	<LOD	5.28 ± 0.07 a
HS-24	<LOD	<LOD	71.92 ± 5.03 b	102.14 ± 5.47 b	174.06 ± 10.11 b	206.12 ± 7.81 c	<LOD	206.12 ± 7.81 c	<LOD
HS-25	<LOD	28.25 ± 2.07 a	37.35 ± 1.54 a	<LOD	65.60 ± 2.01 a	35.33 ± 1.25 a	<LOD	35.33 ± 1.25 a	4.93 ± 0.19 a
HS-26	252.04 ± 40.24 a	178.76 ± 7.00 c	141.34 ± 3.07 b,c	207.72 ± 7.29 b,c	779.86 ± 52.80 d	<LOD	<LOD	<LOD	6.39 ± 0.02 a
HS-28	<LOD	56.44 ± 4.36 b	45.68 ± 3.08 a	65.24 ± 1.72 a	167.36 ± 5.46 b	38.48 ± 1.77 a	<LOD	38.48 ± 1.77 a	5.97 ± 0.09 a
HS-30	<LOD	<LOD	291.82 ± 13.52 c	<LOD	291.82 ± 13.52 b	<LOD	<LOD	<LOD	<LOD
HS-31	<LOD	20.55 ± 1.31 a	<LOD	<LOD	20.55 ± 1.31 a	<LOD	<LOD	<LOD	5.30 ± 0.19 a
HS-36	28.12 ± 2.08 b	18.69 ± 1.30 a	203.08 ± 16.04 c	300.53 ± 19.28 b,c	550.42 ± 28.32 c	<LOD	<LOD	<LOD	6.81 ± 0.22 a
Df (%)	22	44	86	47	–	44	8	–	64
p-value	0.000	0.000	0.000	0.000	0.000	0.000	0.000	0.000	0.000
Test statistics	89.288	52.280	42.205	66.532	48.355	69.510	60.159	55.937	73.952

a–e: different superscript lowercase letters in the same column indicate significantly different values (p < 0.01 by post hoc Tukey’s HSD test); same superscript letters in the same column indicate not significantly different values (p > 0.01 by post hoc Tukey’s HSD test). Values marked with “–” were below LOD.

. A representative chromatogram of the GC-MS analysis of plasticizers conducted in Algerian honeys can be found in Figure S1.

Of the 21 PAEs and NPPs investigated in the honey samples, the following congeners were found: DMP, DEP, DBP, DEHP, DEHT, and DEA. In terms of detection frequency, DBP was detected in 86% of honeys, DEHP in 47% of products, DEP and DEHT in 44% of samples, and DMP in 22% of cases. DEA had the lowest detection frequency, since it was present in only 8% of samples. Regarding the concentration, DBP and DEHP were the most abundant contaminants, since they were revealed in honey up to 889.71 µg/kg and 1256.53 µg/kg, respectively; meanwhile, DEHT and DEA were the least abundant compounds, being present in samples up to 206.12 µg/kg and 97.51 µg/kg, respectively (Table 5). In addition, as will be discussed shortly, a higher degree of PAE contamination was generally evident in coastal samples than non-coastal counterpart, while NPP congeners were more abundant in non-coastal honeys.

The presence of both PAEs and NPPs in honey may be related to industrial, economic, and urban inputs present in Algeria. The Algerian industry is mainly dominated by the petrochemical, chemical, and agri-food sectors, and concentrated in the coastal strip, where more than 50% of the national industrial units were already registered in 2010 [29]. In the petrochemical sector, specifically, these plasticizers play a crucial role for making flexible PVC polymer, as well as other non-PVC plastics and rubber products used in a variety of applications, while in the agri-food sector, plasticizers are exploited in the production not only of all those plastic films, nets, coatings and irrigation equipment useful for crop management, but also food packaging and containers that protect foodstuff and extend its shelf life. Also, the recent intensification of all those economic activities related to seaside tourism and fishing has been identified as a relevant cause of plastic pollution and, hence, plasticizer contamination [16,17,29,30].

While the anthropogenic pollution is a certain indirect cause of contamination in Algerian honey, an equally important direct cause may be the use of honey frames, uncapping tools, extractors, and filters, made up by materials that easily release these chemicals during the production and processing of honey. In addition, although the possibility of contamination during storage can be ruled out because honeys from this study were stored in glass jars, the prolonged storage of commercial honeys in plastic containers may gradually degrade the polymer, and, consequently, encourage the migration of additives from the packaging into the honey itself [18].

With respect to the single PAEs, DMP was detected in 21% and 22% of coastal and non-coastal samples, respectively. However, significantly higher concentrations (p < 0.01) were generally found in coastal honeys (concentration range: 147.44–277.14 µg/kg, p > 0.01) than inland honeys (concentration range: 28.12–44.90 µg/kg, p > 0.01) (Table 5).

DEP was revealed in 50% of coastal honeys with a concentration range of 18.20–50.46 µg/kg (p < 0.01), and 41% of samples from non-coastal areas with a higher abundance (concentration range: 18.69–204.22 µg/kg, p < 0.01). However, honeys from the coastal area of Annaba (HS-10) were characterized by the highest concentration of this PAE (404.70 µg/kg, p < 0.01) (Table 5).

Considering the Algerian coast, it is interesting to note that the highest levels of both DMP and DEP contamination regarded the samples from Annaba (HS-10) and Skikda (HS-13 and HS-27); conversely, most of the honey productions from Mostaganem were free of both compounds (HS-23, HS-29, HS-33, HS-34, HS-35). According to previous literature, the urban discharges and the industrial activity of the metropolitan area of Annaba, along with the petrochemical poles, and fishing and commercial ports in Skikda make these coastal areas the most exposed to the consequences of organic pollution [17,29,31,32].

A previous study mapping the air pollution in the wilaya of Annaba found that the intense industrial activity was responsible for high levels of environmental pollutants in some areas [32]. Additionally, the accumulation of plastic waste on the coast of Annaba and Skikda is reported as a growing environmental problem due to not only the tourism and fishing activities, but also the proximity of both cities to wastewater outlets and river mouths [17,33]. In this respect, the Seybouse river near Annaba has shown sediments and water heavily contaminated with plasticizers and heavy metals due to urban and industrial discharges [17]. Similarly, the Saf Saf river near Skikda has resulted in being severely polluted by spills of oil, tar and other refinery waste [32].

DBP was found in most products from coastal and non-coastal areas, with such a quantitative variability (concentration range: 29.58–889.71 µg/kg, p < 0.01) that it was not possible to define a spatial distribution between investigated areas (Table 5). The strong presence of PAE in Algerian honeys is the result of its common use in a vast array of plastic products, leading to the widespread contamination of the environment and, consequently, human exposure through diverse routes, including food. However, the highest levels of this plastic additive were found in certain honeys from the Tizi Ouzou area (HS-15, 889.71 µg/kg, p < 0.01), an area heavily impacted by a significant waste mismanagement, and related issues of air, soil and water quality. Indeed, according to the daily newspaper El Watan, approximately 350,000 tonnes of organic and inorganic waste—88% of which is plastics—are produced each year in the wilaya of Tizi Ouzou, and over 44% of which ends up in illegal or uncontrolled landfills [34].

DEHP showed similar frequency and amount in samples from coastal and non-coastal zones (df: 50% and 45%, respectively). However, a higher contamination level could be observed in coastal (concentration range: 20.70–523.16 µg/kg, p < 0.01) than non-coastal (concentration range: 20.66–300.53 µg/kg, p < 0.01) products (Table 5).

Interestingly, a spatial distribution of this PAE could be defined among the coastal sites under study. In fact, honeys from Annaba (HS-10) were characterized by a very high level of DEHP, which, to be precise, was outside the range described above (1256.53 µg/kg, p < 0.01). In addition, all samples from Mostaganem (HS-23, HS-29, HS-33, HS-34 and HS-35) revealed the presence of DEHP, which, on the contrary, was not detected in honeys from Skikda (HS-13 and HS-27) and Tizi Ouzou (HS-7, HS-11, HS-12, HS-15 and HS-32) (Table 5). Hence, it could be assumed that both the metropolitan areas of Annaba and Mostaganem represent relevant sources of this PAE.

Overall, the coastal samples most affected by PAE contamination were the Eucalyptus globulus honeys from Annaba (HS-10), characterized by the sum of DMP, DEP, DBP, and DEHP equal to 1979.57 µg/kg (p < 0.01), the Ceratonia siliqua honeys from Tizi Ouzu (HS-15), with the sole DBP contamination amounting to 889.71 µg/kg (p > 0.01), and the Pinus halepensis–Rosmarinus officinalis honeys from Mostaganem (HS-34), with DBP and DEPH totalling 762.93 µg/kg (p > 0.01). Conversely, the bifloral (Erica arborea–Lavandula stoechas) honeys from Tizi Ouzou (HS-32) and the Citrus honeys from Mostaganem (HS-35) were the least contaminated products, since they showed the sole presence of DBP (41.27 µg/kg) and DEHP (44.06 µg/kg) at non significantly different levels (p > 0.01) (Table 5).

When considering inland honeys, multifloral honeys from Naâma (HS-26), Chlef (HS-36) and Djelfa (HS-16) were the most contaminated products, since they showed all PAE congeners amounting to a total of 779.86 µg/kg, 550.42 µg/kg, and 438.67 µg/kg, respectively (p < 0.01) (Table 5). Although the plasticizer contamination of honey is typically determined by its geographical origin and the quality of the surrounding environment, in this case, a relationship between plasticizer contamination and botanical origin of the honey may be highlighted. This could be explained by the fact that plants have a varying ability of taking up, translocating, and metabolizing environmental contaminants [35], including plasticizers [36]. The different abilities of plants to cope with pollution can also be recognized in the different abilities of pollen to accumulate pollutants [37]. Hence, it is reasonable to hypothesize that multifloral honeys deriving from a broader floral source are likely to have higher levels of contamination than monofloral honeys.

On the other hand, many non-coastal products were contaminated by low and comparable levels of only one PAE congener, thus showing very low ΣPAEs. For example, the Tamarix africana honeys from Mascara (HS-31), the bifloral Ziziphus lotus–Thymus vulgaris honeys from Chlef and Pinus halepensis–Quercus ilex honeys from Ain Defla (HS-22) showed only DEP (20.55 µg/kg), DEHP (27.67 µg/kg) and DMP (33.54 µg/kg), respectively (p > 0.01). In the Ziziphus lotus–Silybum marianum honeys from Djelfa (HS-3), and the Tamarix africana honeys from Tissemsilt (HS-4), only DBP was revealed (37.11 µg/kg, 49.27 µg/kg, respectively, p > 0.01). The Citrus honeys from Blida (HS-5) were the least contaminated product, being free of all investigated PAEs (Table 5).

Among detected NPPs, DEHT was present in 57% of products from coastal areas, with concentrations ranging from 9.25 µg/kg (HS-27) to 84.08 µg/kg (HS-10). Interestingly, DEHT showed a spatial distribution similar to that of DEHP, since all honeys from the Annaba (HS-10) and Mostaganem (HS-23) areas were among the most contaminated products (concentration range: 69.13–84.08 µg/kg, p > 0.01); conversely, products from Skikda had similar and low levels of DEHT (HS-13 and HS-27, 9.75 and 9.25 µg/kg, p > 0.01), and none of the samples from Tizi Ouzou (HS-1, HS-7, HS-11, HS-12, HS- 15 and HS-32) contained this plasticizer (Table 5).

Although fewer samples from the inland zones revealed the presence of DEHT (36%), they showed a greater abundance of this congener. In fact, high and non-significantly different amounts (p > 0.01) of DEHT were found in honeys from Djelfa (HS-16 and HS-21, 132.74 µg/kg and 193.33 µg/kg) and Tiaret (HS-24, 206.12 µg/kg), while lower and non-significantly different levels (p > 0.01) of this NPPs were present in samples from Djelfa (HS-8, 8.95 µg/kg) and Laghouat (HS-2, 10.55 µg/kg) (Table 5).

None of the coastal honeys contained DEA. For this reason, these samples showed ΣNPP values equal to the DEHT concentrations. However, DEA was revealed only in honeys from the non-coastal area of Djelfa at varying concentrations (i.e., HS-8, HS-16 and HS-21 concentration range: 10.36–97.51 µg/kg, p < 0.01) (Table 5). As a result, honeys from Djelfa showed the highest ΣNPPs (i.e., HS-8, HS-16 and HS-21 concentration range: 40.98–290.48 µg/kg, p < 0.01) among all the samples analyzed.

A literature review from the last decade pointed out that few studies focused on the monitoring of plasticizers and BPs in honey. Only one study delineated the profile of plasticizers in Algerian honeys. Specifically, Derrar and colleagues investigated an array of Algerian honeys with different botanical and geographical origin collected during 2022 and 2023 (n = 54 samples) and highlighted the presence of PAEs, such as DEP, DBP, DiBP and DEHP, and NPPs, such as DEHT and DEA. Except for DEP, which was at higher concentrations than those reported in Table 5 (concentration range: <LOQ-1656 µg/kg), DBP, DEHP, DEHT and DEA generally contaminated honeys at lower levels than those reported in this study (respective concentration ranges: <LOQ-97 µg/kg; 45–118 µg/kg; 38–144 µg/kg; 13–100 µg/kg) [18]. Recently, Moroccan monofloral honeys displayed a worst contamination scenario, with a greater number of congeners determined at higher concentrations. Revealed compounds included DEP (940–3100 μg/Kg), DBP (450–1050 μg/Kg), DEHP (350–1060 μg/Kg), DEA (1300–5650 μg/Kg) and DEHT (520–1140 μg/Kg) [9].

EU honeys generally showed lower levels of contamination. A study conducted by Lo Turco et al. [38] highlighted that in Sicilian and Calabrian honeys the most abundant plasticizer was DEHP (up to 202.7 µg/Kg) followed by DBP (mean concentration: 40.3 µg/Kg), whereas other PAEs, such as DMP and DEP, ranged from <LOD to 68.2 µg/Kg.

Different honeys collected in 2018 from central Italy were characterized by DMP (up to 6.0 μg/Kg), DEP (range: <2–371 μg/Kg), DBP (range: <10–551 μg/Kg) and DEHP (range: <6–960 μg/Kg) [39].

Of all BPs tested, only BPA was determined in most honey samples, and none of the BP analogues were detected (Table 5). A representative chromatogram of the HPLC-MS/MS analysis of BPs conducted in Algerian honeys can be found in Figure S2.

The presence of BPs in honey has long been primarily linked to the migration of this undesirable contaminant from PC packaging and other materials used during honey production and processing (i.e., epoxy resins) [40]. In this study, the possibility of BPA contamination during storage has been excluded by using glass jars. However, BPA may still be released from the processing equipment. In addition, BPA is known as a ubiquitous environmental contaminant, and the honey contamination is possible since honeybees interact with polluted plants, air, soil, and water in proximity to the hive [41].

Although samples from coastal areas showed a df higher than non-coastal honeys (i.e., 78.8% and 54.5%, respectively), the two groups had fairly comparable concentration ranges (i.e., 2.69–12.73 µg/kg [p < 0.01] and 2.64–8.65 µg/kg [p < 0.01], respectively). For coastal honeys, high and comparable levels of BPA were generally found in samples from Annaba (HS-10, 9.64 µg/kg) and Skikda (HS-13 and HS-27, 12.73 µg/kg and 8.03 µg/kg) (p > 0.01). Relevant and non-significantly different amounts of BPA were determined also in the multifloral honeys from Mostaganem (HS-23, 8.16 µg/kg, p > 0.01). However, the other samples from Mostaganem (i.e., HS-29, HS-33, HS-34, HS-35) showed the lowest concentrations of this BP (concentration range: <LOD-2.69 µg/kg, p < 0.01). Concerning inland products, the highest levels of BPA were found in honeys from Chlef (HS-17 and HS-36, 8.65 µg/kg and 6.81 µg/kg, p < 0.01). On the other hand, all honeys from Dijelfa resulted in the lowest levels of contamination (i.e., HS-3 and HS-8, 2.79 µg/kg and 3.01 µg/kg, p > 0.01) or were not even contaminated (i.e., HS-16, HS-20 and HS-21, <LOD). Moreover, based on provided samples, zones such as Tissemsilt, Blida, and Tebessa produced BPA-free honeys (i.e., HS-4, HS-5, HS-14, HS-18, HS-30, <LOD) (Table 5).

Overall, the most contaminated coastal products were the Eucalyptus globulus honeys from Annaba (HS-10 and HS-13, 12.73 µg/kg and 9.64 µg/kg), followed by the multifloral honeys from Mostaganem (HS-23, 8.16 µg/kg) (p > 0.01); meanwhile, the non-coastal honeys with the highest abundance of BPA were the bifloral Ziziphus lotus–Thymus vulgaris honeys from Chlef (HS-17, 8.65 µg/kg, p < 0.01). Interestingly, most multifloral honeys from non-coastal areas, such as Naâma (HS-26), Tiaret (HS-28) and Chlef (HS-36), contained high and non-significantly different amounts of such additive (concentration range: 5.97–6.81 µg/kg, p > 0.01) (Table 5). Hence, according to experimental data, the multifloral origin of the honey would imply a greater susceptibility of honey not only to the accumulation of plasticizers but also BPA.

In the last decade, few works monitored the presence of BPA and its analogues in honey. Specifically, Derrar and colleagues found out that the concentration of BPA and its analogues was always <LOQ in Algerian honeys with different botanical and geographical origin collected during 2022 and 2023 (n = 54 samples) [18]. Massous and coworkers demonstrated that Moroccan monofloral honeys were contaminated with BPA at levels comparable to those in this study (<LOQ-8.07 µg/Kg). However, in the same samples, BP analogues, such as BPB and BPAF, were also determined [9].

Potortì et al. demonstrated that BPA and its analogues were not detected in most Algerian and Tunisian honey samples analyzed (n = 25). However, when detected, BPA and some analogues (i.e., BPAP, BPF, BPS, and BPZ) were at levels comparable to those found in this study [22].

Considering the EU, an array of 39 Calabrian and Sicilian honeys collected in 2016 were free of BPA (<LOD) [38], while a total of 24 honeys from Central Italy sampled in 2018 had BPA at concentrations varying between <18 µg/Kg and 997 µg/Kg [39]. Additionally, 36 commercial honeys from EU and non-EU countries collected between 2015 and 2016 contained BPA in a highly variable range (<LOD– 107 µg/Kg), and other BPs, such as BPAF, BPE, BPF, BPS, and BPZ, were revealed [40].

Overall, the present study highlighted that BPA was detected in a greater number of samples and with greater consistency than previous studies, thus suggesting a more widespread environmental contamination. This difference may be attributed to a more pronounced use of plastic materials in the beekeeping equipment or during honey processing, as well as a greater anthropogenic pressure in the study areas investigated.

Regarding the latter point, the epoxy resin industry appears well integrated into Algeria’s local manufacturing sector, whereas the PC market relies more on imports. Specifically, epoxy resins are widely used in Algeria for coatings, adhesives, and advanced composites. Companies producing epoxy-based products are in various wilayas, including study areas such as Skikda and Tizi Ouzou. However, several local companies serving the region’s needs, but no major primary production facilities, are within the Annaba area [42]. On the other hand, the Algerian PC industry features a growing demand, driven also by the packaging sector, with diverse local companies producing PC sheets, though export/import data and the presence of international suppliers indicate that it relies more on imports for raw resins. However, among the study areas, Blida and Chlef represent relevant hubs for PC sheets manufacturing and distribution [42].

3.2. Considerations on the Contamination of Algerian Honey by Plasticizers and BPs in Light of the Current EU and Algerian Regulatory Contexts

To date, neither the EU nor Algerian legislation has set maximum concentration limits for plasticizers and BPs in honey or food in general.

However, the EU has a more restrictive and evolving regulatory framework for the use and placing on the market of plasticizers and BPs in consumer products and food contact materials (FCMs) than Algeria, which, adhering to a less stringent and detailed approach, has significantly less restrictive national standards.

Considering plasticizers, the Regulation (EC) No. 1907/2006 currently classifies certain PAEs, including DEHP and DBP, which have been found in Algerian honeys (Table 1), as substances of very high concern (SVHCs), due to reproductive toxicity and endocrine disruption for human health and/or the environment. Consequently, under Annex XVII, both chemicals shall not be used nor placed in the market in toys, childcare articles, and any article made with plasticized material, either individually or in any combination with other SVHC PAEs, at a concentration equal to or greater than 0.1% [43].

With the Interministerial Order of 11 July 2016, Algeria has demonstrated to be in the process of strengthening its food safety and consumer health regulations regarding the exposure to endocrine disrupting chemicals, including plasticizers and BPA [44].

However, the Algerian legislation does not recognize SHVC PAEs and applies the same measures described above by targeting only childcare articles rather than all consumer goods, thus, confirming the risk of a still widespread exposure to these substances. This inevitably creates a regulatory gap.

On the other hand, NPPs, such as DEHT—an isomer of DEHP—and DEA, are currently being considered as an alternative to PAEs, and they are expected to be increasingly used in the EU for the manufacturing of a variety of PVC goods, since they are under Regulation (EC) No. 1907/2006, and have already shown a favourable toxicological profile [45]. Hence, in line with the EU transition from SHVC PAEs to safer alternatives, the presence of DEHT and DEA in many honey samples (Table 1) may reflect the use of NPP-based materials also in Algeria to mitigate the risk of exposure to certainly harmful PAEs.

Considering BPA and BP analogues, the recent Regulation (EU) 2024/3190 bans the use of BPA, other hazardous BPs and derivatives in most FCMs, aiming to prevent “regrettable substitutions” with similarly harmful alternatives (e.g., BPS and BPF) [46]. From the latest scientific assessments by the European Food Safety Authority (EFSA), a tolerable daily intake (TDI) for BPA has been set at the extremely precautionary level of only 0.2 ng/kg of body weight per day [27]. Although BPA was determined in Algerian honeys at concentrations that appear low in terms of µg/kg (Table 1), its very presence may raise concerns of non-compliance within the EU regime.

On the other hand, the Algerian Interministerial Order of 11 July 2016, has banned BPA in childcare articles, but not in other FCMs. In addition, Algerian legislation on other BPs and derivatives remains largely absent, thus highlighting, again, a gap compared to the stringent EU framework.

Basically, it is evident that the EU prioritizes consumer health and environmental safety, leading to bans and restrictions, while Algeria might face challenges balancing these with economic development. However, Algeria needs to develop robust and stringent regulations, potentially harmonizing with the current EU standards, not only to protect its consumers but also to ensure product compliance for export.

3.3. Statistical Correlation

Spearman’s rank correlation analysis was performed to evaluate the relationships between the concentrations of individual plasticizers, their sum, and BPA in analyzed honeys to better delineate the distribution trends of these pollutants in the areas under study. Specifically, results referring to samples from coastal areas are presented in

Table 6. Spearman’s rank correlation matrix used to examine the relationships among individual PAEs and NPPs, their sum, and BPA, present in honey samples from Algerian coastal areas. The ‘*’ indicates significance at p < 0.05 level and ‘**’ indicates significance at p < 0.01 level.

	DMP	DEP	DBP	DEHP	ΣPAEs	DEHT	DEA	ΣNPPs	BPA
DMP	1	0.6141 **	−0.0197	0.0131	−0.0801	0.4291 **	-	0.545 **	0.5596 **
DEP		1	0.07587	−0.02888	0.1435	0.1817	-	0.2176 *	0.5623 **
DBP			1	−0.0985	0.6583 **	−0.2937 **	-	−0.3000 **	−0.03706
DEHP				1	0.2497 *	0.5621 **	-	0.4854 **	−0.1746
ΣPAEs					1	−0.05507	-	−0.1699 *	0.3165 *
DEHT						1	-	0.9265 **	0.05492
DEA							1	-	-
ΣNPPs								1	0.1736
BPA									1

, while data in

Table 7. Spearman’s rank correlation matrix used to examine the relationships among individual PAEs and NPPs, their sum, and BPA, present in honey samples from Algerian non-coastal areas. The ‘*’ indicates significance at p < 0.05 level and ‘**’ indicates significance at p < 0.01 level.

	DMP	DEP	DBP	DEHP	ΣPAEs	DEHT	DEA	ΣNPPs	BPA
DMP	1	0.5314 **	0.1762 *	0.4588 **	0.6149 **	−0.06415	−0.02939	−0.05221	−0.005118
DEP		1	0.4023 *	0.3153 **	0.5314 **	0.1238	0.03237	−0.006198	0.1404
DBP			1	0.3018 **	0.6619 **	−0.071	−0.04516	−0.1115	−0.1479
DEHP				1	0.7169 **	0.3513 **	0.1644 *	0.04071	−0.08236
ΣPAEs					1	0.1203	−0.02998	−0.2009 *	−0.1971 *
DEHT						1	0.4429 **	0.2566 **	−0.06965
DEA							1	0.1744 *	−0.09261
ΣNPPs								1	−0.3407 **
BPA									1

refers to honeys from inland areas. However, one should always bear in mind that (i) correlation does not necessarily imply causation and (ii) interpreting these relationships is somewhat troublesome, because the honey contamination can reflect the widespread environmental pollution as well as the incorrect beekeeping practices, or even both phenomena.

For the coastal honey samples, the analysis revealed strong and significant pairwise positive correlations between DMP and DEP (p < 0.01), as well as between ∑PAEs, on the one hand, and DBP and DEHP on the other (p < 0.01 and p < 0.05). This could mean that DMP and DEP may share the same production and distribution patterns in northern Algeria, and that the PAE industry and market still relies heavily on the presence of SVHC compounds, such as DBP and DEHP (Table 6). For inland honeys, significant positive correlations were established between all single PAEs (p < 0.01 and p < 0.05), as well as between these congeners and ∑PAEs (Table 7).

Considering NPP congeners in coastal honeys, DEHT displayed positive and significant correlations (p < 0.01) with DMP and DEHP due to potential similarities in the distribution pattern, and a significant negative correlation with DBP (p < 0.01). In light of the results reported in Table 5, this inverse relationship may stress again the still widespread use of DBP in Algeria. However, the significant correlation (p < 0.01) between DEHT and ∑NPPs may underline the relevance of this congener in shaping the NPPs industry and market in coastal areas of northern Algeria (Table 6).

The situation is slightly different for non-coastal honeys. Beyond a significant positive relationship between DEHT, DEA and DEHP (p < 0.01 and p < 0.05), which still confirms the use of this SVHC compound alongside alternative plasticizers, there are also significant positive correlations between DEHT and DEA (p < 0.01), as well as between these congeners and ∑NPPs (p < 001 and p < 0.05). These findings may highlight the increasing use of DEA, along with DEHT, in the inland cities under study, and the influence of these congeners in outlining the NPPs industry of these areas (Table 7).

Significant negative relationships were present between ∑PAEs and ∑NPPs both in products from coastal and non-coastal areas (Table 6 and Table 7). Based on experimental data, these findings may suggest that NPPs are still facing challenges in establishing themselves as an alternative to conventional PAEs, which remain predominant.

Interpreting the relationships between BPA and plasticizers becomes even more challenging due to the diverse uses and applications of the classes of additives. Overall, significant positive correlations between BPA and DMP, DEP and ∑PAEs (p < 0.01 and p < 0.05) and non-significant positive correlations with NPPs were highlighted in coastal honeys. This could mean that the pollution from plasticizers and BPA go hand in hand in coastal areas due to the presence of multiple and relevant urban, industrial and economic inputs (Table 6).

On the other hand, in inland honeys, BPA mostly established negative correlations with individual PAEs, NPPs and their sum. Based on the experimental data, this may be explained by the lower detection frequency of this BP in the study areas, and, consequently, a lower number of BPA inputs and/or the adoption of better practices in honey production and processing (Table 7).

3.4. Dietary Exposure and Risk Assessment

The interpretation of the results was carried out in light of the consumption scenarios and anthropometric parameters considered in the calculation of the Estimated Daily Intake (EDI). In this study, the average honey consumption was assumed to be 1.6 g/day for European adults and 0.8 g/day for European children, whereas for Algeria the values were halved (0.8 g/day for adults and 0.4 g/day for children). Reference body weight was set at 60 kg for adults and 20 kg for children. The EDI was expressed in mg/kg body weight/day for plasticizers and in ng/kg body weight/day for BPA.

In this context, although children consume lower absolute amounts compared to adults, their body weight is three times lower; consequently, the consumption-to-body-weight ratio is higher in children. This results in systematically higher EDI values in the pediatric population compared to adults, both in Europe and in Algeria. Moreover, since European consumption is double that of Algeria, assuming equal contaminant concentrations in honey, the EDI is approximately doubled in the European scenario, as observed in the data reported in

Table 8. EDI (mg/Kg_bw/day and ng/Kg_bw/day) and HQ calculated for plasticizers (honey concentrations expressed in mg/kg) and for BPA (honey concentrations expressed in ng/kg), calculated for adults and children in Algeria and Europe.

Sample Code	DEP								DBP + DEHP
	Algerian Population				European Population				Algerian Population				European Population
	Adults		Children		Adults		Children		Adults		Children		Adults		Children
	EDI	HQ	EDI	HQ	EDI	HQ	EDI	HQ	EDI	HQ	EDI	HQ	EDI	HQ	EDI	HQ
HS-1	0.00024	<1	0.00036	<1	0.00049	<1	0.00010	<1	0.00071	<1	0.00107	<1	0.00142	<1	0.00214	<1
HS-7	0.00030	<1	0.00044	<1	0.00059	<1	0.00089	<1	0.00067	<1	0.00100	<1	0.00134	<1	0.00201	<1
HS-10	0.00540	<1	0.00809	<1	0.01079	<1	0.01619	<1	0.01903	<1	0.02855	<1	0.03806	<1	0.05710	1.142
HS-11	-	-	-	-	-	-	-	-	0.00061	<1	0.00061	<1	0.00082	<1	0.00122	<1
HS-12	0.00029	<1	0.00044	<1	0.00058	<1	0.00088	<1	0.00129	<1	0.00194	<1	0.00258	<1	0.00388	<1
HS-13	0.00067	<1	0.00101	<1	0.00135	<1	0.00202	<1	0.00098	<1	0.00098	<1	0.00130	<1	0.00195	<1
HS-15	-	-	-	-	-	-	-	-	0.01186	<1	0.01779	<1	0.02373	<1	0.03559	<1
HS-23	0.00027	<1	0.00040	<1	0.00054	<1	0.00081	<1	0.00193	<1	0.00193	<1	0.00258	<1	0.00387	<1
HS-27	0.00058	<1	0.00086	<1	0.00115	<1	0.00173	<1	0.00263	<1	0.00395	<1	0.00527	<1	0.00790	<1
HS-29	-	-	-	-	-	-	-	-	0.00118	<1	0.00118	<1	0.00158	<1	0.00236	<1
HS-32	-	-	-	-	-	-	-	-	0.00083	<1	0.00083	<1	0.00110	<1	0.00165	<1
HS-33	-	-	-	-	-	-	-	-	0.00110	<1	0.00165	<1	0.00219	<1	0.00329	<1
HS-34	-	-	-	-	-	-	-	-	0.01017	<1	0.01526	<1	0.02034	<1	0.03052	<1
HS-35	-	-	-	-	-	-	-	-	0.00059	<1	0.00088	<1	0.00117	<1	0.00176	<1
HS-2	-	-	-	-	-	-	-	-	0.00067	<1	0.00100	<1	0.00134	<1	0.00201	<1
HS-3	-	-	-	-	-	-	-	-	0.00049	<1	0.00074	<1	0.00099	<1	0.00148	<1
HS-4	-	-	-	-	-	-	-	-	0.00066	<1	0.00099	<1	0.00131	<1	0.00197	<1
HS-5	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
HS-6	-	-	-	-	-	-	-	-	0.00240	<1	0.00360	<1	0.00480	<1	0.00720	<1
HS-8	0.00050	<1	0.00074	<1	0.00099	<1	0.00149	<1	0.00049	<1	0.00073	<1	0.00146	<1	0.00146	<1
HS-9	0.00069	<1	0.00069	<1	0.00138	<1	0.00138	<1	0.00517	<1	0.00517	<1	0.00689	<1	0.01033	<1
HS-14	0.00204	<1	0.00204	<1	0.00271	<1	0.00407	<1	0.00050	<1	0.00075	<1	0.00100	<1	0.00149	<1
HS-16	0.00272	<1	0.00408	<1	0.00545	<1	0.00817	<1	0.00252	<1	0.00378	<1	0.00504	<1	0.00756	<1
HS-17	-	-	-	-	-	-	-	-	0.00037	<1	0.00055	<1	0.00074	<1	0.00111	<1
HS-18	-	-	-	-	-	-	-	-	0.00201	<1	0.00301	<1	0.00402	<1	0.00603	<1
HS-19	-	-	-	-	-	-	-	-	0.00389	<1	0.00584	<1	0.00778	<1	0.01167	<1
HS-20	-	-	-	-	-	-	-	-	0.00442	<1	0.00663	<1	0.00884	<1	0.01327	<1
HS-21	-	-	-	-	-	-	-	-	0.00262	<1	0.00392	<1	0.00523	<1	0.00785	<1
HS-22	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-	-
HS-24	-	-	-	-	-	-	-	-	0.00232	<1	0.00348	<1	0.00696	<1	0.00696	<1
HS-25	0.00038	<1	0.00056	<1	0.00075	<1	0.00113	<1	0.00075	<1	0.00075	<1	0.00100	<1	0.00149	<1
HS-26	0.00238	<1	0.00358	<1	0.00477	<1	0.00715	<1	0.00465	<1	0.00698	<1	0.00931	<1	0.01396	<1
HS-28	0.00113	<1	0.00113	<1	0.00151	<1	0.00226	<1	0.00148	<1	0.00222	<1	0.00296	<1	0.00444	<1
HS-30	-	-	-	-	-	-	-	-	0.00045	<1	0.00067	<1	0.00090	<1	0.00135	<1
HS-31	0.00027	<1	0.00041	<1	0.00055	<1	0.00082	<1	-	-	-	-	-	-	-	-
HS-36	0.00025	<1	0.00037	<1	0.00050	<1	0.00075	<1	0.00671	<1	0.01007	<1	0.01343	<1	0.02014	<1
Sample Code	BPA
	Algeria Adults				Algerian Children				European Adults				European Children
	EDI		HQ		EDI		HQ		EDI		HQ		EDI		HQ
HS-1	73.33		366.66		110.00		550.00		146.67		733.33		220.00		1100.0
HS-7	48.57		242.88		72.86		364.33		97.16		485.77		145.73		728.6
HS-10	128.57		642.88		192.86		964.33		257.16		1285.77		385.73		1928.66
HS-11	71.15		355.77		106.73		533.66		142.31		711.55		213.46		1067.33
HS-12	96.53		482.66		144.80		724.00		193.07		965.33		289.60		1448.00
HS-13	169.73		848.66		254.60		1273.00		339.47		1697.33		509.20		2546.00
HS-15	43.77		218.88		65.66		328.33		87.56		437.77		131.33		656.66
HS-23	108.84		544.22		163.26		816.33		217.69		1088.44		326.53		1632.66
HS-27	107.06		535.33		160.60		803.00		214.13		1070.66		321.20		1606.00
HS-29	35.82		179.11		53.733		268.66		71.64		358.22		107.46		537.33
HS-32	83.33		416.66		125.00		625.00		166.67		833.33		250.00		1250.00
HS-33	-		-		-		-		-		-		-		-
HS-34	-		-		-		-		-		-		-		-
HS-35	-		-		-		-		-		-		-		-
HS-2	72.66		363.33		109.00		545.00		145.33		726.66		218.00		1090.00
HS-3	37.15		185.78		55.73		278.66		74.31		371.55		111.46		557.33
HS-4	-		-		-		-		-		-		-		-
HS-5	-		-		-		-		-		-		-		-
HS-6	-		-		-		-		-		-		-		-
HS-8	40.08		200.44		60.13		300.66		80.18		400.88		120.26		601.33
HS-9	35.20		176.00		52.80		264.00		70.40		352.00		105.60		528.00
HS-14	-		-		-		-		-		-		-		-
HS-16	-		-		-		-		-		-		-		-
HS-17	115.28		576.44		172.93		864.66		230.58		1152.88		345.86		1729.33
HS-18	-		-		-		-		-		-		-		-
HS-19	68.13		340.67		102.20		511.00		136.27		681.33		204.40		1022.00
HS-20	-		-		-		-		-		-		-		-
HS-21	-		-		-		-		-		-		-		-
HS-22	70.44		352.22		105.66		528.33		140.89		704.44		211.33		1056.66
HS-24	-		-		-		-		-		-		-		-
HS-25	65.73		328.66		98.60		493.00		131.47		657.33		197.20		986.00
HS-26	85.24		426.22		127.86		639.33		170.49		852.44		255.73		1278.66
HS-28	79.68		398.44		119.53		597.66		159.38		796.88		239.06		1195.33
HS-30	-		-		-		-		-		-		-		-
HS-31	70.62		353.11		105.93		529.66		141.24		706.22		211.86		1059.33
HS-36	90.80		454.00		136.20		681.00		181.60		908.00		272.40		1362.00

.

With regard to the sum of plasticizers (DBP + DEHP), EDI values consistently followed this pattern: children > adults, and Europe > Algeria. However, the Hazard Quotient (HQ) was below 1 in almost all analyzed samples, indicating no significant risk according to the established toxicological reference values. The only exception was represented by the E. globulus honeys from Annaba (HS-10) eventually consumed by European children, since the HQ slightly exceeded the unity (HQ = 1.142), thus, suggesting a potential health risk due to the presence of relatively elevated concentrations of DBP and DEHP. Overall, exposure to plasticizers through honey consumption appears limited and only rarely approaches the toxicological level of concern.

In contrast, the scenario related to BPA highlighted a more critical situation. Although EDI values followed the same pattern of plasticizers (children > adults and Europe > Algeria), they were several orders of magnitude higher than those observed for plasticizers. However, unlike plasticizers, the HQ values associated with BPA largely exceed unity in all considered scenarios, with particularly high values observed in European children. This systematic exceedance of the safety threshold indicates a potentially significant health risk associated with BPA exposure through honey consumption. In conclusion, while exposure to plasticizers through the consumption of investigated honeys appeared generally within the acceptable safety limits, BPA emerged as a toxicological contaminant of particular concern, requiring specific attention in terms of risk monitoring and management throughout the Algerian honey production chain.

4. Conclusions

Findings of this study proved that honey from northern Algeria was contaminated from plasticizers (i.e., DMP, DEP, DBP, DEHP, DEHT and DEA) and BPA. Interestingly, the sole presence of BPA may raise compliance concerns under the evolving EU framework, which implies strict FCM restrictions and a very low TDI for this compound. This was confirmed also by the assessment of dietary exposure and toxicological risk derived from the consumption of these honeys. In fact, while exposure to plasticizers was generally within the acceptable safety limits, the exposure to BPA raised particular concern from a toxicological point of view.

Overall, a higher degree of contamination was observed in products from coastal areas than inland areas. An in-depth analysis of experimental data and the main sources of pollution in Algeria showed that the honeys most contaminated by PAEs came from areas with intense urban, economic and industrial activities (i.e., Annaba and Skikda), as well as poor waste management (i.e., Tizi Ouzou), while safer alternative plasticizers, such as DEHT and DEA, were more abundant in honeys from Mostanagem and Djelfa. The coastal areas of Annaba and Skikda and the non-coastal city of Chlef were demonstrated to also have contamination inputs of BPA.

This contamination may be linked to the several anthropogenic activities present in Algeria, which accelerate the economic development of the country while exacerbating environmental pollution, as well as improper practices of honey production/processing that rely on still-poor ecofriendly beekeeping standards. In this scenario, it is crucial to establish clear and harmonized national regulations for plasticizers and BPs in FCMs, implement a systematic monitoring of contaminants in beekeeping activities and products, and, not least, encourage the adoption of good beekeeping practices, including the use of safer materials in the supply chain. Such measures would protect public health, guarantee a safe, sustainable and commercially competitive Algerian honey on the global market, and safeguard beekeeping in an increasingly compromised environmental context.