Validation of a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of sulfonamides,trimethoprim and dapsone in muscle,egg, milk and honey

A quantitative multi-residue method that includes 13 sulfonamides, trimethoprim and dapsone was developed and validated according to Commission Decision 2002/657/EC for muscle, milk egg and honey samples. For all matrices, the same extraction procedure was used. Samples were extracted with an acetone/dichloromethane mixture and cleaned up on aromatic sulfonic acid (SO₃H) SPE cartridges. After elution and concentration steps, analytes were identified and quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Data were acquired according to the multiple reaction-monitoring approach (MRM) and analytes were quantified both by the isotope dilution and the matrix-matched approaches calculating the response factors for the scanned product ions. The developed method shows good linearity, specificity, precision (repeatability and within-laboratory reproducibility), and trueness. Estimated CCβ for sulfonamides ranged between 5.6 and 8.2 µg kg^?1 for eggs, between 11.1 and 69.9 µg kg^?1 for milk, between 64.7 and 87.9 µg kg^?1 for muscle, and between 2.7 and 5.3 µg kg^?1 for honey. CCβ values for dapsone were 3.1, 0.6, 0.7 and 1.5 µg kg^?1 and for trimethoprim were 3.1, 6.7, 81.7 and 3.0 µg kg^?1 calculated for eggs, milk, muscle and honey, respectively. Recovery for all matrices was in the range from 89.1% and 109.7%. In matrix effect testing, no significant deviations were found between different samples of muscle and milk; however, a matrix effect was observed when testing different types of honey. The validation results demonstrate that the method is suitable for routine veterinary drug analysis and confirmation of suspect samples.     

Occurrence of glyphosate and AMPA residues in soy-based infant formula sold in Brazil

Glyphosate is an herbicide widely used in the world, being applied in several crops, among them soybeans. Recently, glyphosate and its metabolite aminomethylphosphonic acid (AMPA) have been identified as possible contributors to the emergence of various diseases such as autism, Parkinson’s and Alzheimer’s diseases, as well as cancer. The child population-consuming cereal-based foods is the most exposed to the effects of pesticides because of their developmental phase and they have a higher food intake per kilogram of body weight than adults. The presence of glyphosate and AMPA residues in soy-based infant formulas was evaluated during the years 2012–2017, totalising 105 analyses carried out on 10 commercial brands from different batches. Glyphosate and AMPA were determined by liquid chromatography with fluorescence detection after derivatisation reaction. The method was validated and showed accuracy and precision with a limit of quantification (LOQ) of 0.02 mg kg^?1. Among those samples that contained levels above the LOQ, the variation of glyphosate residues was from 0.03 mg kg^?1 to 1.08 mg kg^?1 and for AMPA residues was from 0.02 mg kg^?1 to 0.17 mg kg^?1. This is the first scientific communication about glyphosate and AMPA contamination in soy-based infant formula in Brazil, The study was conducted under good laboratory practice (GLP) and supported by good scientific practice.     

Simultaneous Determination of Multiclass Pesticides and Antibiotics in Honey Samples Based on Ultra-High Performance Liquid Chromatography-Tandem Mass Spectrometry

A simple, fast, and efficient method was developed for simultaneous determination of 79 pesticides and 13 antibiotics compounds of different chemical classes of pesticides and antibiotics in honey samples by ultra-high performance liquid chromatography with tandem mass spectrometry (UHPLC-MS/MS). The sample preparation procedure includes homogenization with McIlvaine buffer 0.1 mol L^?1 (pH 4), followed by extraction with acetonitrile and cleanup with florisil, using dispersive solid phase extraction (d-SPE). The proposed method was validated with good results, such as linearity (r ²?>?0.9901), normality, and independence of the evaluated data, as well as recoveries between 70 and 120 % with relative standard deviation (RSD) <20 % for most of the compounds spiked from 0.1 to 200 μg kg^?1. The experimental method limits of detection and quantification were from 0.03 to 1.51 μg kg^?1 and from 0.1 to 5 μg kg^?1, respectively, for the pesticides. For the antibiotics, the decision limits (0.1 to 2 μg g^?1) and the detection capacity (0.12 to 2.81 μg g^?1) were below the maximum residue limits (MRLs) established for honey by the Brazilian and European legislation. The method was successfully applied to real samples from different botanical and geographic origins. From them, 44 % presented residues from 0.12 to 10 μg kg^?1 of one or more analytes. The proposed method combines the advantages of a quick sample preparation step with the selectivity and sensitivity of the UHPLC-MS/MS and proved to be suitable for routine analyses.     

Determination and surveillance of hydrocortisone and progesterone in livestock products by liquid chromatography-tandem mass spectrometry

A simple analytical method for the determination of hydrocortisone and progesterone in bovine, swine, and chicken muscle and eggs was developed. Hydrocortisone and progesterone were extracted with acetonitrile and subsequently cleaned-up using an Oasis^® HLB mini-cartridge. The method was validated in accordance with Japanese guidelines and exhibited trueness from 86.6% to 104.3% and precision (relative standard deviations (RSDs) of repeatability and within reproducibility were under 8.7% and 11.7%, respectively). The method was applied to 103 bovine muscle, 137 swine muscle, 69 chicken muscle and 52 egg samples that were commercially available in Tokyo, Japan. The hydrocortisone concentration was 0.9–41.2 µg kg^?1 in all bovine muscle samples, with an average of 7.7 µg kg^?1 and a median of 6.2 µg kg^?1. The progesterone concentration in 50 samples exceeded the limit of quantification (LOQ) and reached a maximum of 95.4 µg kg^?1. Hydrocortisone was also detected in all swine muscle samples at concentrations of 2.0–56.0 µg kg^?1. Its average and median concentrations amounted to 13.1 and 11.3 µg kg^?1, respectively. Twenty-three samples contained progesterone levels surpassing the LOQ, with a maximum concentration of 107.0 µg kg^?1. No chicken muscle samples contained any of the analytes. The progesterone concentration was 15.5–200.0 µg kg^?1 in all egg samples, with an average of 95.4 µg kg^?1 and a median of 90.5 µg kg^?1.     

Gas Purge Microsyringe Extraction Coupled with Dispersive Liquid-Liquid Microextraction for the Determination of Acidic Compounds in Food Packaging Materials

A green, simple and sensitive method was developed for the analysis of volatile carboxylic acids (VFAs) and perfluorocarboxylic acids (PFCAs) in food packaging materials. The acidic compounds in food packaging materials were first extracted by gas purge microsyringe extraction (GP–MSE) with 1.0 mL 0.1 mol·L^?1 NaOH solution, then the analytes were dispersive liquid-liquid microextracted (DLLME) by 50 μL chloroform as extraction solvent and 200 μL acetonitrile as dispersive solvent. The 2-(5-Benzoacridine) ethyl-p-toluenesulfonate (BAETS) with excellent fluorescence property was applied to enhance the high performance liquid chromatography (HPLC) sensitivity. The obtained recoveries for the VFAs ranged from 92.0 to 101 %. The method LODs calculated at a signal-to-noise ratio (S/N) of 3 were in the range of 0.80–3.40 μg·kg^?1, while the LOQs calculated at S/N of 10 were in the range of 2.5–10.2 μg·kg^?1. All compounds were in good linearity with concentration coefficients of higher than 0.997. Perfluorooctanoic acid (PFOA) was found in all of the 15 kinds of samples analyzed with concentrations ranging from 4.86–7.56 μg·kg^?1. Acetic acid, butyric acid, and caprylic acid were found in half of the samples analyzed. The other analytes were also found in more than 30 % samples with concentrations varied between 3.96 and 293 μg·kg^?1.     

Determination and surveillance of nine acaricides and one metabolite in honey by liquid chromatography-tandem mass spectrometry

A simple and accurate analytical method for the determination of acaricides in honey was developed and validated in accordance with Japanese validation guidelines. Analytes – amitraz, N-2,4-dimethylphenyl-N-methylformamidine (DMPF), etoxazole, fenpyroximate, fipronil, hexythiazox, propargite, pyridaben and spirodiclofen – were extracted with ethyl acetate under basic conditions and subsequently cleaned up using an InertSep^® MA-1 polymer-based anion-exchange column. The method was validated by fortified recovery tests at three different concentrations (1, 5 and 10 µg kg^?1) performed with three samples daily on five different days. The method exhibited recoveries of 77–116% and precision (relative standard deviations – RSDs) of repeatability and within-laboratory reproducibility ranged from 2% to 22% and from 3% to 23%, respectively. The sample solution was successfully cleaned up to enable quantification using external solvent calibration curves. The limits of quantification (LOQs) were estimated to be 1 µg kg^?1 for all analytes. The method was applied to honey samples commercially available in Tokyo, Japan. Analysis of 250 honey samples indicated that amitraz was present in 127 samples, and that its residual concentration was less than 20 µg kg^?1. Propargite was detected in 23 samples at concentrations less than 1 µg kg^?1.     

Development of an ELISA and Immunochromatographic Assay for Tetracycline,Oxytetracycline, and Chlortetracycline Residues in Milk and Honey Based on the Class-Specific Monoclonal Antibody

A sensitive class-specific monoclonal antibody against tetracyclines (TCs) was generated and used to develop an enzyme-linked immunosorbent assay (ELISA) and an immunochromatographic assay for TC, oxytetracycline (OTC), and chlortetracycline (CTC) detection in milk and honey samples. The dynamic range of detection for TC in ELISA was 0.26–2.00 μg L^?1 with an IC₅₀ of 0.72 μg L^?1. The IC₅₀ value of OTC and CTC was 3.2 and 6.4 μg L^?1, respectively. The recovery of TC, OTC, and CTC in milk samples was 82–102, 91–105, and 90–101 %, respectively, and 88–101, 89–101, and 89–95 % in honey samples, respectively. In the immunochromatographic assay, the cutoff values for TC, OTC, and CTC were 15, 15, and 50 μg L^?1 in milk, respectively, and 40, 40, and 40 μg L^?1 in honey, respectively. The results revealed that ELISA and the immunochromatographic assay can be applied for the rapid and sensitive detection of TC, OTC, and CTC in milk and honey samples.     

Ochratoxin A in red pepper flakes commercialised in Turkey

The aim of this study was to investigate the presence of Ochratoxin A (OTA) in red pepper flakes commercialised in Turkey. A total of 75 samples were collected from different supermarkets and traditional bazaars in Istanbul during 2012–2013. OTA analysis was performed by high-performance liquid chromatography equipped with fluorescence detection after immunoaffinity column clean-up. The method was linear in the range 0.05–40 μg kg^?1 (r² = 0.9997). Twenty-seven out of 31 (87.1%) packed red pepper flake samples contained OTA at concentrations ranging from 0.6 to 1.0 μg kg^?1, whereas 100% of the unpacked red pepper flake samples contained OTA, in the range 1.1–31.7 μg kg^?1. Overall, only 4 unpacked samples (5.3%) contained OTA, with a mean value of 23.4 μg kg^?1, which is higher than the European Union limit. It is suggested that OTA content should be carefully considered in red pepper flake samples especially marketed in unpacked form.     

Determination of Ochratoxin A in Cereals and Feeds by Ultra-performance Liquid Chromatography Coupled to Tandem Mass Spectrometry with Immunoaffinity Column Clean-up

This paper describes the preparation of reusable immunoaffinity columns and the development of an ultra-performance liquid chromatography tandem mass spectrometry method combined with immunoaffinity column clean-up (IAC-UPLC-MS/MS) for the determination of ochratoxin A (OTA) in cereals and feeds. The monoclonal antibody (mAb) was produced from a stable hybridoma cell line (4H10), which belongs to the immunoglobulin G1 (κ-light chain) isotype. A competitive indirect enzyme-linked immunosorbent assay was used to characterize the mAb. The concentrations causing 50 % inhibition of binding of mAb to OTA-ovalbumin by free OTA, ochratoxin B, and ochratoxin C were 1.29, 4.78, and 0.94 ng mL^?1, respectively. The IAC-UPLC-MS/MS method offers a limit of quantification (LOQ, S/N >10) ranging from 0.5 to 1.0 μg kg^?1 and a limit of detection (LOD, S/N >3) ranging from 0.2 to 0.3 μg kg^?1 in cereal and feed samples. The IAC-UPLC-MS/MS method offers a good LOQ and LOD for OTA in cereal and feed samples. The accuracy and precision at this level fall within the EU regulatory limit. This methodology has been validated in four different matrices (millet, maize, soybean, and swine finisher diet) with highly satisfactory results and applied to the analysis of samples collected from the markets.     

Analysis of trichothecenes in laboratory rat feed by gas chromatography-tandem mass spectrometry

A method for the determination of seven trichothecenes, neosolaniol (NEO), diacetoxyscirpenol (DAS), deoxynivalenol (DON), nivalenol (NIV), fusarenon-X (FUS-X), 3-acetyldeoxynivalenol (3-ADON) and 15-acetyldeoxynivalenol (15-ADON), in laboratory rat feed by GC-MS/MS was developed. Sample extraction and purification was performed by an acidified mixture of acetonitrile/water (80–20% v/v). Limits of quantitation (LOQs) were between 1 and 10 μg kg^–1 for all studied trichothecenes. Eight concentration levels between the LOQ and 100 × LOQ were used for the calibration curves. Matrix-matched calibration was used for quantitation purposes to compensate the detector signal enhancement obtained for all the analytes. The method accuracy was evaluated by recovery assays at three concentration levels, 25, 50 and 100 μg kg^–1 (n = 9). Recoveries ranged from 62% to 97% and precision, expressed as intra- and inter-day relative standard deviations, was evaluated for all compounds. The validated method was successfully applied to the analysis of 35 laboratory rat feed samples showing mycotoxin contamination in 66% of the samples. DON was the most prevalent trichothecene followed by 15-ADON, NIV and 3-ADON. The maximum DON concentration reached in real samples was 2156 ± 4.3 μg kg^–1, while NEO, DAS and FUS-X were not detected in any sample. Multi-contamination by at least two mycotoxins was observed in 17% of the analysed feed samples.     

Co-occurrence of aflatoxins,ochratoxin A and citrinin in “egusi” melon (Colocynthis citrullus L.) seeds consumed in Ireland and the United Kingdom

The natural co-occurrence of aflatoxins (AFs), ochratoxin A (OTA) and citrinin (CIT) in melon seed samples obtained from retailers and households in Ireland and the United Kingdom (UK) was evaluated. AFs and OTA were determined by HPLC with fluorescence detection while CIT was analysed by HPLC-MS/MS. AFB₁ was detected in all (100%) samples (mean = 9.7 μg kg^?1; range = 0.2–66.5 μg kg^?1). Mean total AFs was 12.0 μg kg^?1 (range = 0.3–82 μg kg^?1). Commercially retailed samples showed a significantly higher AFB₁ contamination (p < 0.05) than the household samples. OTA occurred in 3 (13.6%) samples, while 4 (18.2%) were contaminated with CIT at very low levels. In this study, 68% of the melon seed samples were contaminated above the 2 μg kg^?1 EU limit for AFB₁ in oilseeds. These results highlight the need for the development of strategies to reduce AF contamination in “egusi” for human consumption.     

Determination of seven pyrethroid pesticide residues in vegetables by gas chromatography using carboxylated multi-walled carbon nanotubes as dispersion solid phase extraction sorbent

ABSTRACT

A novel carboxylated multi-walled carbon nanotubes (MWCNTs-COOH) dispersive solid phase extraction (d-SPE) method combined with gas chromatography (GC) with an electron capture detector (ECD) was developed for the determination of seven pyrethroid pesticides in cucumber, spinach, eggplant, tomato and carrot. We optimised d-SPE conditions including the type and volume of the extractant, the type and amount of the sorbent, and shaking time. Under the optimal conditions, the linear range was from 2.0 to 2000 μg kg^?1. The recoveries were from 88.5% to 108.2%, with the corresponding RSDs <6%, correlation coefficients 0.9987–0.9998, LOD 0.5–2.9 μg kg^?1 and LOQ 1.5–9.7 μg kg^?1. The proposed method is simple, fast, and safe, with high recovery and sensitivity, and is applicable to analysis of pyrethroid pesticides in vegetable samples.     

Occurrence of deoxynivalenol in wheat flour,instant noodle and biscuits commercialised in Brazil

A total of 134 samples, consisting of 58 wheat flour, 40 instant noodle and 36 biscuits, were analysed for the presence of deoxynivalenol (DON). The samples were obtained from retail markets of the city of São Paulo during the period 2010–2014. DON was determined by high performance liquid chromatography with ultraviolet detection and immunoaffinity sample clean-up. Method validation followed international guidelines. The LOD and LOQ were 60 and 200 µg kg^?1, respectively, considering the three different types of samples analysed. The lowest recovery found in this study was 91.8% with RSD 4.5% for instant noodles. DON was detected in 91.4%, 97.5% and 97.2% of samples wheat flour, instant noodles and biscuits, respectively, resulting in a total of 94.8% with levels ranging from LOD to 1720.0 µg kg^?1.     

Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

A Highly Sensitive Method for the Determination of Thiophanate Methyl,Cyromazine, and Their Metabolites in Edible Fungi by Ultra-Performance Liquid Chromatography Using Accelerated Solvent Extraction and Cleanup with Solid-Phase Extraction

A highly sensitive ultra-performance liquid chromatographic method with diode-array detection was developed for residue determination of thiophanate methyl (TM), cyromazine (CYR), and their metabolites, carbendazim (MBC) and melamine (MEL). Edible fungi samples were treated using accelerated solvent extraction (ASE) followed by cleanup with solid-phase extraction (SPE). Under optimized conditions, good linearity was achieved with a correlation coefficient (r ²) of ≥0.9998. The limit of quantification was 0.36, 0.24, 0.4, and 0.5 μg kg^?1 for MEL, CYR, MBC, and TM, respectively. The intra- and interday precisions (in terms of the relative standard deviation (RSD)) of the four analytes were in the range of 2.3–4.5 and 3.1–6.3 %, respectively. The recoveries for TM, MBC, CYR, and MEL in four edible fungi samples at three spiked levels of 0.6, 6, and 20 μg kg^?1 for TM and MBC and 0.4, 4, and 20 μg kg^?1 for CYR and MEL were in the range of 82–105 % with RSDs below 5.6 %. The proposed method can be used for the routine determination of CYR and MEL in edible fungi with high sensitivity and accuracy as well as low consumption of reagents.     

Simultaneous Determination of Cyadox and Its Metabolites in Chicken Tissues by LC-MS/MS

A specific and sensitive LC-MS/MS method was firstly established for the simultaneous extraction and determination of cyadox and its three main metabolites—1,4-bisdesoxycyadox, 4-desoxycyadox, and quinoxaline-2-carboxylic acid—in chicken muscle, liver, kidney, and fat tissues. Samples were subjected to extraction using ethyl acetate and followed by acetonitrile–chloroform (1:4, v/v) and further purified by Oasis mixed mode anion exchange SPE cartridge. Analysis was performed on a C₁₈ column by detection with MS in multiple-reaction monitoring mode. A gradient elution program with 0.1 % formic acid solution, acetonitrile, and 1 % formic acid (adjusted to pH 8 with ammonia) was performed at a flow rate of 0.2 mL min^?1. The correlation coefficients (r) for each calibration curve are higher than 0.99 within the experimental concentration range. The recoveries of the four target analytes at three spiking levels of 2.5, 25 and 250 μg kg^?1 were between 74.5 and 93.8 %, with relative standard deviations less than 12 %. The decision limits (CCαs) of the four analytes in chicken edible tissues ranged from 0.3 to 1.5 μg kg^?1, and the detection capabilities (CCβs) were below 2.3 μg kg^?1. The developed method demonstrated a satisfactory applicability in incurred chicken tissue samples.     

Determination of Hormones Residues in Milk by Gas Chromatography-Mass Spectrometry

The article presents the use of gas chromatography-mass spectrometry (GC-MS) technique in a method for the determination of 18 anabolic hormones from synthetic stilbenes, steroids and resorcylic acid lactones (RALs) groups in raw milk and milk powder. Sample preparation consisted of liquid-liquid extraction with diethyl ether and purification by solid phase extraction (SPE). Prior to instrumental analysis, the reaction of derivatisation with the heptafluorobutyric anhydride or N-methyl-N-trimethylsilyltrifluoroacetamide was performed. Method validation was carried out according to the required performance criteria of the Commission Decision 2002/657/EC. The apparent recovery of all analytes at 1 μg L^?1 (kg^?1) level was ranged between 70.4 and 119.4 % with the coefficients of variation values less than 30 %. The decision limits (CCα) and the detection capabilities (CCβ) were in the range of from 0.11 to 0.44 μg L^?1 (kg^?1) and from 0.19 to 0.75 μg L^?1 (kg^?1), respectively. The procedure has been accredited and successfully applied as a screening method for the presence of hormone residues in the study of commercial samples of milk.     

Simultaneous Determination of Eight Tranquilizers in Pork by Ultra-Performance Liquid Chromatography Coupled with Electrospray Tandem Mass Spectrometry

An ultra-performance liquid chromatography/electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS) method was developed and validated for quantification of eight tranquillizers (chlorpromazine, imipramine hydrochloride, diazepam, nitrazepam, nordazepam, oxazepam, flurazepam, and haloperidol) in pork. Sample pretreatment consisted of extraction by acetonitrile, defatted by n-hexane, and further solid phase extraction by hydrophilic-lipophilic balance (HLB) extraction cartridges. The triple quadrupole mass spectrometer was operated in positive ion mode, and multiple reaction monitoring was used for drug quantification. The method was validated over the concentration ranges of 1.0 ～ 250 μg kg^?1 for the eight tranquillizers. The calibrations were performed in sample matrices, and the interference effect of sample matrices on the ionization was effectively eliminated. Good linear relationship (R ² > 0.99) was observed within the concentration range of 1.0–250 μg kg^?1. The average recoveries of the eight tranquillizers spiked at three levels ranged from 63.7 to103.2 % with the relative standard deviation below 11.8 %. The limits of detection were between 0.06 and 0.30 μg kg^?1, and the limits of quantification were between 0.2 and1.0 μg kg^?1 for all analytes in pork. This validated method has been successfully used to quantify the concentration of the eight tranquillizers in pork samples.     

Aflatoxin B1 in betel nuts (Areca catechu L.) imported to Pakistan from different regions of South Asia

Aflatoxin B₁ (AFB₁) levels were evaluated in betel nuts (Areca catechu L.) being imported to Pakistan during 2010–2011. In total, 278 betel nut samples (India = 21, Indonesia = 51, Sri-Lanka = 34 and Thailand = 172) were received from the Department of Customs and were analysed by thin layer chromatography (TLC). All Indian origin betel nuts showed AFB₁ contamination ranging from 11.7–262.0 µg kg^?1 with a mean of 92.5 µg kg^?1. Among Indonesian and Sri Lankan shipments, 80.4% and 73.5% betel nuts were contaminated with AFB₁ ranging between 3.3–39.2 and 6.5–103.4 µg kg^?1 with a mean of 11.6 and 35.0 µg kg^?1, respectively. However, only 30.2% of Thailand origin samples showed AFB₁ contamination ranging 3.3–77.0 µg kg^?1 with a mean of 6.6 µg kg^?1. The widespread occurrence of AFB₁ increases the hazard associated with betel nuts. Thus, strict control is a pre-requisite for the production and import/export of psychoactive substances as betel nuts.     

Determination of plasticisers and BPA in Sicilian and Calabrian nectar honeys by selected ion monitoring GC/MS

Twenty-six plasticisers and bisphenol A (BPA) in 39 Sicilian and Calabrian nectar honeys of different botanical south Italian origin were determined by GC-MS. Di-(2-ethylhexyl)phthalate was the most abundant plasticiser in all samples, reaching up 202.7 ± 153.1 µg kg^–1, followed by di-butylphthalate with a concentration of 40.3 ± 9.3 µg kg^–1, whereas the concentration of the other phthalates varied from not detectable to 68.2 µg kg^–1. Trace levels of adipate, sebacate and BPA were not detected in any samples. Among the different floral origin honeys, di-ethylphthalate levels were above the limit of quantification (LOQ) in orange blossom, wildflower and chestnut honey samples. Among the honeys of different geographical origin, di-methylphthalate, di-butylphthalate and di-(2-ethylhexyl)phthalate concentrations were similar. Calabrian honeys showed levels of di-ethylphthalate always lower than the LOQ, moreover di-(2-methylpropyl)phthalate levels were lower than the Sicilian ones. On the contrary, in Sicilian honeys di-(2-methylpropyl)phthalate levels were higher and di-ethylphthalate was present in 37% of samples.