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The current study aims to develop isolation protocols for several contaminants of emerging concern (i.e., microplastics (MPs), polycyclic aromatic hydrocarbons (PAHs), and heavy metals (HMs)) from different commercial brands and raw milk samples and also to quantify and characterize the risks of these contaminants pose to human health. The quantification, shape, color, and chemical composition of MPs were achieved using optical microscopy, micro-Fourier transform infrared spectroscopy, and scanning electron microscopy coupled with energy-dispersive spectroscopy. Based on the MP dimensions highlighted by the aforementioned techniques, it can be stated that their length ranges between tens of micrometers and a few centimeters; plus, the thickness in some cases reaches more than 15 µm, and the structure of the MPs can be mostly described as a fibriform with a glossy/matte aspect. The polymeric structures identified were polyamides, poly(methyl methacrylate), polyurethane, polyester, and polyethylene. Chemical investigations (PAHs and HMs concentrations) were performed by high-performance liquid chromatography with fluorescence detection and inductively coupled plasma mass spectrometry, respectively. The pollution load index (1.091–7.676) and daily intake of MPs for adults (0.021–1.061 n·kg−1·d−1) and children (0.089–4.420 n·kg−1·d−1) were calculated. It can be concluded that the presence of MPs in milk supports the hypothesis that microplastics can act as carriers for other contaminants (HMs and PAHs), thus increasing the threat to health. [ABSTRACT FROM AUTHOR]

Exercise reduces the risk of inflammatory diseases by modulating different tissue and cell types, including those within the gastrointestinal tract. Obtaining a more comprehensive understanding of pathophysiology requires monitoring of dynamic changes in cometabolites. This study aimed to develop a method for determining gut microbiota–host cometabolites and indoleamines in human urine. Four key gut microbiota–host cometabolites were chromatographically separated by isocratic elution, with a running time of 10 min. The linearity of this method was confirmed over different concentration ranges: 1.0–400 ng/mL for melatonin (MEL), indole-3-propionic acid (3-IPA), indole (IND), and skatole (SKT). This method was extremely sensitive and stable and hence could be successfully applied to characterize the changes in gut microbiota–host cometabolites in human before- and after-running urine. The concentrations of MEL, 3-IPA, IND, and SKT in after-running urine were 84.0 ± 9.69, 25.9 ± 3.39, 343.7 ± 36.8, and 14.6 ± 1.36 ng/mL, respectively. Moreover, the concentrations in before-running urine were 54.2 ± 5.10, 14.4 ± 1.30, 250.8 ± 14.1, and 9.43 ± 1.07 ng/mL, respectively, which showed significantly less difference in concentrations (p < 0.05) in before- than after-running urine. Overall, the established method could simultaneously monitor gut microbiota–host cometabolites and hence can be further applied to clinical and comprehensive pathophysiological studies. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin commonly found in various food products, which poses potential health risks to humans and animals. Recently, more attention has been directed towards its potential neurodegenerative effects. However, there are currently no fully validated HPLC analytical methods established for its quantification in mice, the primary animal model in this field, that include pivotal tissues in this area of research, such as the intestine and brain. To address this gap, we developed and validated a highly sensitive, rapid, and simple method using HPLC-FLD for OTA determination in mice tissues (kidney, liver, brain, and intestine) as well as plasma samples. The method was rigorously validated for selectivity, linearity, accuracy, precision, recovery, dilution integrity, carry-over effect, stability, and robustness, meeting the validation criteria outlined by FDA and EMA guidelines. Furthermore, the described method enables the quantification of OTA in each individual sample using minimal tissue mass while maintaining excellent recovery values. The applicability of the method was demonstrated in a repeated low-dose OTA study in Balb/c mice, which, together with the inclusion of relevant and less common tissues in the validation process, underscore its suitability for neurodegeneration-related research. [ABSTRACT FROM AUTHOR]

Magnetic molecularly imprinted polymers (MMIPs) have fused molecular imprinting technology with magnetic separation technology, emerging as an innovative material capable of recognizing specific molecules and efficiently separating target substances. Their application to the extraction and purification of mycotoxins has great potential, due to the toxicity and economic impact of these contaminants. In this work, MMIP has been proposed as a sample treatment for the determination of main four aflatoxins (B1, B2, G1 and G2) in pig feed. The MMIP was formed through the integration of magnetic material (Fe3O4) with commercial molecularly imprinted polymers, avoiding the synthesis step and, therefore, simplifying the process. The analyses were carried out by high-performance liquid chromatography with fluorescence detection and the method was validated and limits of quantification (LOQs) between 0.09 and 0.47 ng/g were obtained, below the allowed or recommended levels by the European Union. Repeatability and intermediate precision showed relative standard deviations lower than 10% in all cases and trueness ranged from 92 to 111%. Finally, the proposed method was applied to 31 real pig feed samples, detecting aflatoxins with concentrations between 0.2 and 3.2 ng/g. [ABSTRACT FROM AUTHOR]

A method utilizing high-performance liquid chromatography-fluorescence detection (HPLC–FLD) has been developed and refined for the simultaneous detection of florfenicol (FF) and its metabolite florfenicol amine (FFA) along with three fluoroquinolone (ciprofloxacin (CIP), enrofloxacin (ENR), and sarafloxacin (SAR)) residues in different parts of eggs (whole egg, egg yolk, and egg albumen). The QuEChERS ("Quick, easy, cheap, effective, rugged, and safe") procedure utilized 0.1 M disodium EDTA solution, water, and acetonitrile as extractants; sodium sulfate, sodium chloride, and trisodium citrate as dehydrating salts; and N-propylethylenediamine and C18 as adsorbents. A dual-channel FLD method was utilized to analyze the target compounds using an XBridge BEH C18 chromatographic column (4.6 mm × 150 mm, 5 μm). The mobile phase was employed isocratically using a solution of 0.01 M sodium dihydrogen phosphate, 0.005 M sodium dodecyl sulfate, and 0.1% triethylamine (pH 4.8) in combination with acetonitrile at a ratio of 65:35 (V/V). The limits of detection (LOD) and quantification (LOQ) of the analytes ranged from 0.03 to 1.5 µg/kg and from 0.1 to 5.0 µg/kg, respectively. The recoveries of the analytes in the blank egg samples ranged from 71.9% to 94.8% when reference standard concentrations of the LOQ, half of the maximum residual limit (MRL), MRL, and twice the MRL were added. The parameters of the presented protocol were validated and subsequently applied to the analysis of real samples, demonstrating the applicability and reliability of the method. [ABSTRACT FROM AUTHOR]

Homotaurine (HOM) is considered a promising drug for the treatment of Alzheimer's and other neurodegenerative diseases. In the present work, a new high-performance liquid chromatography with fluorescence detection (HPLC–FLD) (λex. = 340 nm and λem. = 455 nm) method was developed and validated for the study of substance permeability in the central nervous system (CNS). Analysis was performed on a RP-C18 column with a binary gradient elution system consisting of methanol–potassium phosphate buffer solution (pH = 7.0, 0.02 M) as mobile phase. Samples of homotaurine and histidine (internal standard) were initially derivatized with ortho-phthalaldehyde (OPA) (0.01 M), N-acetylcysteine (0.01 M) and borate buffer (pH = 10.5; 0.05 M). To ensure the stability and efficiency of the reaction, the presence of different nucleophilic reagents, namely (a) 2-mercaptoethanol (2-ME), (b) N-acetylcysteine (NAC), (c) tiopronin (Thiola), (d) 3-mercaptopropionic acid (3-MPA) and (e) captopril, was investigated. The method was validated (R2 = 0.9999, intra-day repeatability %RSD < 3.22%, inter-day precision %RSD = 1.83%, limits of detection 5.75 ng/mL and limits of quantification 17.43 ng/mL, recovery of five different concentrations 99.75–101.58%) and successfully applied to investigate the in vitro permeability of homotaurine using Franz diffusion cells. The apparent permeability (Papp) of HOM was compared with that of memantine, which is considered a potential therapeutic drug for various CNSs. Our study demonstrates that homotaurine exhibits superior permeability through the simulated blood–brain barrier compared to memantine, offering promising insights for enhanced drug delivery strategies targeting neurological conditions. [ABSTRACT FROM AUTHOR]

Dispersive liquid–liquid microextraction (DLLME) was optimized for the simultaneous extraction of aflatoxins (AFB1, AFB2, AFG1, and AFG2) from powdered senna leaves and pods. Detection was performed using high-performance liquid chromatography with fluorescence detection (HPLC-FLD) and pre-column derivatization. The parameters affecting the DLLME extraction efficiency were evaluated. Chloroform (200 µL) was used as an extraction solvent, 500 µL of distilled water was used as a dispersive solvent, and the extraction was performed at pH 5.6 with no salt added. The optimized method was validated using leaves and pods according to the European Commission guidelines. The linear range for all aflatoxins was 2–50 µg/kg, with values for regression coefficients of determination exceeding 0.995. The recoveries of spiked senna leaves and pods were in the ranges of 91.77–108.71% and 83.50–102.73%, respectively. The RSD values for intra-day and inter-day precisions were in the ranges of 2.30–7.93% and 3.13–10.59%, respectively. The limits of detection and quantification varied in the ranges of 0.70–1.27 µg/kg and 2.13–3.84 µg/kg, respectively. The validated method was successfully applied for the quantification of aflatoxins in 60 real samples of dried senna leaves and pods. [ABSTRACT FROM AUTHOR]

Most cans intended to come into contact with food require the application of an internal coating made from synthetic polymers. The most widely used type of coating are epoxy resins based on bisphenol A diglycidyl ether (BADGE). It has been found that some components like cyclo-di-BADGE (CdB) and other BADGE derivatives can migrate into food. In this study, the polymeric coating of forty-eight cans was assessed using an infrared spectrometer with attenuated total reflectance. The food samples were extracted, and a targeted analysis was carried out to quantify CdB using liquid chromatography with fluorescence detection (HPLC-FLD). The first estimation of the exposure of the adult population was estimated by combining the concentration of CdB in the samples and the Spanish consumption data. In addition, a non-targeted screening by LC-MS was performed in the food sample extracts for the identification of other BADGE derivatives. Twenty samples were positive for the presence of CdB reaching concentrations of 2623 µg/kg. However, the tolerable daily intake of 1.5 µg/kg bw/day recommended for chemical compounds with high toxicological risk was not exceeded. A total of 18 epoxy oligomers could be tentatively identified in the food extracts being BADGE.H2O.BuEtOH the derivative with the highest incidence. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a non-steroidal estrogenic mycotoxin produced by the fungi of the Fusarium genera, and is a contaminant of cereals and plant products. ZEN and its metabolites are considered endocrine disruptors, and could have various toxic effects on animals and humans. In recent years, there has been a significant demographic increase in wild boar (Sus scrofa) in many mountainous and hilly areas of Italy, including the Campania region, mainly due to global climate change. The wild boar can be defined as a generalist and omnivorous species capable of varying its diet; therefore, it can play a role as an environmental bioindicator towards contaminants such as mycotoxins. This study was conducted to evaluate, for the first time, the concentrations of ZEN and its metabolites in the liver, kidney, and muscle of 82 wild boars shot in their habitat by hunters with hunting permits in different localities of Avellino province (Campania region, Southern Italy) from 2021 to 2022. The samples were collected and analyzed with an SPE clean-up and high-pressure liquid chromatography method with fluorescence detection. The results indicated that ZEN and α-Zearalenol were present in most of the samples, suggesting that a plan to monitor these mycoestrogens is essential to achieve the goals of "One Health". [ABSTRACT FROM AUTHOR]

In order to meet the analytical requirements of the European Medicines Agency (EMA), a new HPLC-FLD method was successfully developed using dansyl chloride for the derivatization and determination of the genotoxic impurities N-Nitrosodimethylamine (NDMA) and N-Nitrosodiethylamine (NDEA) in Lisinopril API and its final product. Samples' pretreatment includes liquid–liquid microextraction, denitrosation, and derivatization steps. To optimize the process, the parameters contributing to high sensitivity and yielding reliable results were thoroughly studied and optimized using one-factor-at-a-time and experimental design approaches. The analytes were pre-column derivatized with Dansyl-Cl and analyzed by HPLC-fluorescence (λem/λem = 340/530) using a C18 column and a mixture of phosphate buffer (pH = 2.8; 20 mM)/acetonitrile 55:45 v/v as the mobile phase. The six-level concentration calibration was shown to be linear, with R equal to 0.9995 for both analytes. The limit of detection (LOD) was satisfactory and equal to 4.7 and 0.04 ng/mL for NDMA and NDEA, respectively. Precision was less than 13.4% in all cases, and the average recoveries were equal to 109.2 and 98.1% for NDMA and NDEA, respectively. The proposed procedure is relatively easy, rapid, and suitable for the determination of the two nitrosamines in routine analysis tests. [ABSTRACT FROM AUTHOR]

In September and November 2016, eight marine sampling sites along the coast of the southeastern Gulf of Mexico were monitored for the presence of lipophilic and hydrophilic toxins. Water temperature, salinity, hydrogen potential, dissolved oxygen saturation, inorganic nutrients and phytoplankton abundance were also determined. Two samples filtered through glass fiber filters were used for the extraction and analysis of paralytic shellfish toxins (PSTs) by lateral flow immunochromatography (IFL), HPLC with post-column oxidation and fluorescent detection (FLD) and UHPLC coupled to tandem mass spectrometry (UHPLC-MS/MS). Elevated nutrient contents were associated with the sites of rainwater discharge or those near anthropogenic activities. A predominance of the dinoflagellate Pyrodinium bahamense was found with abundances of up to 104 cells L−1. Identification of the dinoflagellate was corroborated by light and scanning electron microscopy. Samples for toxins were positive by IFL, and the analogs NeoSTX and STX were identified and quantified by HPLC-FLD and UHPLC-MS/MS, with a total PST concentration of 6.5 pg cell−1. This study is the first report that confirms the presence of PSTs in P. bahamense in Mexican waters of the Gulf of Mexico. [ABSTRACT FROM AUTHOR]

Bisphenols (BPs) are a class of endocrine disruptors widely existing in the environment. They have a great impact on human health owing to their environmental endocrine disrupting effects, chronic toxicity, neurotoxicity, cytotoxicity and genetic toxicity. In this paper, an on-line packed fiber solid phase extraction (PFSPE) coupling with column-switching HPLC-FLD determination method was developed for the determination of eight BPs in drinking water. The poly (dibenzo-18-crown-6-ether)/polystyrene composite nanofibers (PDB18C6/PS) were prepared by electrospinning and used as an adsorbent for the on-line PFSPE column. The on-line PFSPE-HPLC equipment contained a dual ternary pump and a switching valve to enable enrichment, purification, and analysis directly in the system. The results showed that the proposed on-line PFSPE-HPLC-FLD method realized the simultaneous separation and detection of eight BPs: BPF, BPE, BPA, BPB, BPAF, BPAP, BPC and BPZ. The curves of the target analytes were prepared with good correlation coefficient values (r2 > 0.998) in the range of 50–1000 pg/mL. The limit of detection (S/N = 3) was 20 pg/mL, the limit of quantitation (S/N = 10) is 50 pg/mL. The recoveries of eight BPs were 94.8–127.3%, and the intra-day precisions (RSD) were less than 10%. The PFSPE column made of the PDB18C6/PS composite nanofibers has stable properties and can be reused at least 200 times. In the detection of drinking water samples, BPZ was detected in nearly 80% of drinking water samples, and BPA, BPAP, BPF and BPAF were also detected in some water samples. This high level of integration and automation was achieved in pretreatment of eight BPs from water samples. The proposed simple, rapid, and practical method has been successfully applied to the detection of eight BPs in drinking water, which can provide powerful technical support for drinking water quality and safety monitoring. [ABSTRACT FROM AUTHOR]

Mycotoxins are bioaccumulative contaminants impacting animals and humans. The simultaneous detection of frequent active exposures and accumulated mycotoxin level (s) in exposed organisms would be the most ideal to enable appropriate actions. However, few methods are available for the purpose, and there is a demand for dedicated, sensitive, reliable, and practical assays. To demonstrate the issue, mice were exposed to a relevant agent Ochratoxin A (OTA), and accumulated OTA was measured by fine-tuned commercial assays. Quantitative high-performance liquid chromatography with fluorescence detection, enzyme-linked immunosorbent assay, and flow cytometry assays have been developed/modified using reagents available as commercial products when appropriate. Assays were performed on excised samples, and results were compared. Accumulated OTA could be detected and quantified; positive correlations (between applied doses of exposure and accumulated OTA levels and the results from assays) were found. Dedicated assays could be developed, which provided comparable results. The presence and accumulation of OTA following even a short exposure could be quantitatively detected. The assays performed similarly, but HPLC had the greatest sensitivity. Blood contained higher levels of OTA than liver and kidney. We demonstrate that specific but flexible and practical assays should be used for specific/local purposes, to measure the exposure itself and accumulation in blood or organs. [ABSTRACT FROM AUTHOR]

Paralytic Shellfish Toxins (PSTs) are marine biotoxins, primarily produced by dinoflagellates of the genera Gymnodinium spp., Alexandrium spp. They can accumulate in shellfish and, through the food chain, be assimilated by humans, giving rise to Paralytic Shellfish Poisoning. The maximum permitted level for PSTs in bivalves is 800 μg STX·2HCl eqv/kg (Reg. EC N° 853/2004). Until recently, the reference analytical method was the Mouse Bioassay, but Reg. EU N° 1709/2021 entered into force on 13 October 2021 and identified in the Standard EN14526:2017 or in any other internationally recognized validated method not entailing the use of live animals as official methods. Then the official control laboratories had urgently to fulfill the new requests, face out the Mouse Bioassay and implement instrumental analytical methods. The "EURLMB SOP for the analysis of PSTs by pre-column HPLC-FLD according to OMA AOAC 2005.06" also introduced a simplified semiquantitative approach to discriminate samples above and below the regulatory limit. The aim of the present paper is to present a new presence/absence test with a cut-off at 600 μg STX·2HCl eqv/kg enabling the fast discrimination of samples with very low PSTs levels from those to be submitted to the full quantitative confirmatory EN14526:2017 method. The method was implemented, avoiding the use of a large number of certified reference standards and long quantification procedures, resulting in an efficient, economical screening instrument available for official control laboratories. The protocol was fully validated, obtaining good performances in terms of repeatability (<11%) and recovery (53–106%) and accredited according to ISO/IEC 17025. The method was applied to mollusks collected from March 2021 to February 2022 along the Marche region in the frame of marine toxins official control. [ABSTRACT FROM AUTHOR]

Paprika is a widely consumed spice in the world and its authentication has gained interest considering the increase in adulteration cases in recent years. In this study, second-order fingerprints acquired by liquid chromatography with fluorescence detection (HPLC-FLD) were first used to detect and quantify adulteration levels of Chinese paprika samples. Six different adulteration cases, involving paprika production region, cultivar, or both, were investigated by pairs. Two strategies were employed to reduce the data matrices: (1) chromatographic fingerprints collected at specific wavelengths and (2) fusion of the mean data profiles in both spectral and time dimensions. Afterward, the fingerprint data with different data orders were analyzed using partial least squares (PLS) and n-way partial least squares (N-PLS) regression models, respectively. For most adulteration cases, N-PLS based on second-order fingerprints provided the overall best quantitation results with cross-validation and prediction errors lower than 2.27% and 20.28%, respectively, for external validation sets with 15–85% adulteration levels. To conclude, second-order HPLC-FLD fingerprints coupled with chemometrics can be a promising screening technique to assess paprika quality and authenticity in the control and prevention of food frauds. [ABSTRACT FROM AUTHOR]

Polycyclic aromatic hydrocarbons (PAHs) are produced during incomplete combustion of organic matter. Many of them are likely to be carcinogenic and cause mutations. In this study, the PAH4 (benzo[a]pyrene (BaP), benz[a]anthracene (BaA), chrysene (CHR), benzo[b]fluoranthene (BbF)) content in deep-fat fried pork was evaluated according to temperature and time, and a risk assessment was conducted. The high performance liquid chromatography-fluorescence detection (HPLC-FLD) method for PAH4 analysis was validated by determining linearity (R2), recovery, limit of detection (LOD), and limit of quantitation (LOQ). The linearity was R2 ≥ 0.99. The PAH4 level was dependent on the temperature, time, and nature of the edible oil. Before heat treatment, the PAH4 content of pork was 0.38 μg/kg. The PAH4 content of deep-fat fried pork ranged from 0.86 to 6.86 μg/kg according to temperature (160, 180, 200 °C) and time (3, 6, 9 min). Exposure to PAH4 via the consumption of deep-fat fried pork for different age groups among the Korean population was 0.01–0.89 μg-TEQBaP/kg/day, with the margin of exposure calculated as 7.88 × 104–5.22 × 106. The PAH4 content and risk of exposure increased proportionally with the heat treatment temperature and time. The survey provided important information in terms of evaluating the health risks that PAH compounds can cause in people's diets due to the heat treatment of pork. [ABSTRACT FROM AUTHOR]

Polysaccharides are abundant in natural resources and perform numerous physiological functions. Polysaccharide structures often lack chromophore groups; thus, current analytical methods cannot distinguish polysaccharide metabolites in the body or polysaccharide prototypes in biological samples. Thus, the measurement of polysaccharides in blood, bodily fluid, and cell-culture medium is difficult. Our early-stage research resulted in the isolation of two homogeneous polysaccharides from Pseudostellaria heterophylla, PHP0.5MSC-F and PHPH-1-2, which have anti-hyperglycemia and insulin resistance improvement effects for type 2 diabetes. In this study, the reducing terminal sugars of PHP0.5MSC-F and PHPH-1-2 were labeled with 2-aminobenzamide (2-AB) to prepare novel fluorescent probes for HPLC-coupled fluorescence detection (HPLC-FLD). Quantitative analysis was performed in reference to T40, and the detection limit for PHP0.5MSC-F was found to be 8.84 μg/mL with a linear range of 29.45–683.28 μg/mL. In reference to T70, the detection limit for PHPH-1-2 was found to be 13.89 μg/mL with a linear range of 46.29–462.76 μg/mL. This method was used to measure the bidirectional transport of polysaccharides across caco-2 cells from apical to basolateral (AP→BL) or from basolateral to apical (BL→AP) directions and to evaluate the polysaccharide bioavailability. The drug absorption capacity was determined based on the apparent permeability coefficient (Papp), and the Papp values for the two polysaccharides were found to be greater than 1 × 10−6 cm/s, which suggests easy absorption. Regarding bidirectional transport, the AP→BL Papp values were greater than the BL→AP values; thus, PHP0.5MSC-F and PHPH-1-2 mainly underwent passive transference. The two membrane permeable polysaccharides were not P-gp efflux transporter substrates. The absorption mechanism of PHP0.5MSC-F complies with passive diffusion under a concentration gradient, whereas PHPH-1-2 mainly utilizes a clathrin-mediated endocytic pathway to enter caco-2 cells. This innovative HPLC-FLD method can help to track polysaccharide internalization in vitro and in vivo to facilitate cellular uptake and biodistribution exploration. [ABSTRACT FROM AUTHOR]

Flavanols are a class of polyphenols that have become the focus of much research because of their medicinal properties. For this paper, a new method to determine polyphenols in nutraceutical samples was developed, based on solid-liquid extraction and further analysis by liquid chromatography. Analytes were recovered by ultrasound-assisted extraction, using a mixture of methanol/water/hydrochloric acid (70:29:1, v:v:v) as the solvent. Reversed-phase high-performance liquid chromatography with ultraviolet/visible (HPLC-UV/Vis) and fluorescence detection (HPLC-FLD) were developed to determine the content of the principal flavanols. The HPLC-FLD method showed better selectivity and sensitivity, so it was selected for further studies. The analytical parameters established with FLD were satisfactory in terms of linearity (R2 ≥ 0.993), repeatability (RSD% ≥ 2.2), and limits of detection of 0.1 mg L−1 or below. The nutraceutical samples were analyzed by HPLC-FLD to determine the compositional profiles of flavanols, then they were subsequently treated using chemometric methods, such as principal component analysis (PCA). Results showed that the study of compositional profiles from the selected flavanols was sufficient for description and discrimination purposes. The cranberry samples were the richest in flavanols, procyanidin A2 being the principal flavanol, and artichoke samples the least rich. [ABSTRACT FROM AUTHOR]

Antibacterial substances such as sulfonamides are widely used in veterinary medicine to treat many bacterial diseases. After their administration to animals, up to 90% of the initial dose of the antibiotic is excreted in the feces and/or urine, which can be applied to farmland as natural or organic fertilizers. In this work, an analytical method was developed with the use of HPLC-FLD for the detection and quantification of five sulfonamides (sulfaguanidine, sulfadiazine, sulfamerazine, sulamethazine and sulfamethoxazol) in poultry and pig feces, slurry and digestates. The method was validated according to EU requirements (Commission Decision 2002/657/EC and VICH GL49). Linearity, decision limit, detection capability, detection and quantification limits, recovery, precision, and selectivity were determined, and adequate results were obtained. Using the HPLC-FLD method for all analyzed matrices, recoveries were satisfactory (77.00–121.16%), with repeatability and reproducibility in the range of 4.36–17.34% to 7.94–18.55%, respectively. Decision limit (CCα) and detection capability (CCβ) were 33.87–67.63 and 53.36–92.00 µg/kg, respectively, and limit of detection (LOD) and limit of quantification (LOQ) were 13.53–23.30 and 26.02–40.38 µg/kg, respectively, depending on the analyte. The forty-four samples of natural and organic fertilizers were analyzed, and four samples showed sulfamethoxazole in the amount from range 158 to 11,070 µg/kg. The application of antibiotics including sulfonamides for farming animals is widespread and may lead to the development of antibiotic resistance and other environmental effects. [ABSTRACT FROM AUTHOR]

Aflatoxin M1 (AFM1) is a well-known mycotoxin that can be found in the milk of animals that have ingested feed contaminated with aflatoxin B1 (AFB1). In Italy, the development of donkey farms is mainly due to growing request of donkey milk, which is considered an incomparable substitute for human mother's milk for its chemical composition and organoleptic characteristics. The aim of this study was to assess the occurrence of AFM1 in donkey milk produced in a farm in Northern Italy, also in view of the few data available about the presence of this mycotoxin in this type of milk. Therefore, 63 milk samples were collected and analyzed using a fast and sensitive HPLC and fluorescence detection (FLD) method previously optimized and validated. None of the milk samples collected were found to be contaminated at a level above the limit of quantification (LOQ) (0.0125 ng/mL), while only one sample showed traces of the mycotoxin at a concentration between the limit of detection (LOD) and LOQ (0.0044 ng/mL), well below the legal limit established for infant milk and follow-on milk (0.025 ng/mL). These results are in line with those of the few similar surveys carried out on donkey milk and seem to indicate a low risk of AFM1 contamination for this food. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin that may be present in various food and feed of plant and animal origin, including chicken meat. In Italy, backyard poultry farming is rather widespread. Animals are raised for meat, eggs and for ornamental purpose, and they are often fed with home-made diets not subject to official controls. The purpose of this study was to evaluate exposure of ornamental and backyard chickens to OTA using biliary ochratoxin A as a biomarker. Therefore, bile samples, in addition to kidney, liver and muscle, were collected from 102 chickens reared in 16 farms located in 6 Italian regions. High-performance liquid chromatography method and fluorimetric detection (HPLC-FLD) analysis were carried out firstly on bile from all animals, and OTA was detected in 12 chickens (concentration range 3.83-170.42 μg/L). Subsequently, the kidneys of these chickens were also analysed, and the mycotoxin was not detected. The analytical detection limits (LODs) of OTA in bile and kidney were 2.1 μg/L and 0.1 μg/kg, respectively. In conclusion, these animals were exposed to OTA but their meat can be considered safe, given that this mycotoxin, if present, concentrates highest in kidneys. Biliary ochratoxin A confirms its use as a valid biomarker to assess exposure of poultry to OTA. [ABSTRACT FROM AUTHOR]

Gastrodia elata Blume (G. elata), a traditional Chinese medicine, is widely used for treatment of various neuro dysfunctions. However, its quality control is still limited to the determination of gastrodin. In the present study, two novel strategies based on quantitative evaluation of total gastrodin and gastrodigenin with base hydrolysis and total gastrodigenin with base-enzymatic hydrolysis followed by HPLC-FLD were put forward and successfully applied to evaluate the quality of 47 batches of G. elata from eight localities. Meanwhile, a systematic comparison of the novel strategy with the multiple markers and the Pharmacopeia method was performed. The results showed that the parishins category could be completely hydrolyzed to gastrodin by sodium hydroxide solution, and gastrodin could further utterly hydrolyze to gastrodigenin with -D-glucosidase buffer solution. The contents of total gastrodin and gastrodigenin ranged from 1.311% to 2.034%, and total gastrodigenin from 0.748% to 1.120% at the eight localities. From the comparison, we can conclude that the two novel strategies can comprehensively reveal the characteristics of overall active ingredients in G. elata for quality control. The present study provides a feasible and credible strategy for the quality control of G. elata, suggesting a revision of the latest Chinese Pharmacopoeia or European Pharmacopoeia methods for the modernization of G. elata use. [ABSTRACT FROM AUTHOR]

Ochratoxin A (OTA) is a mycotoxin produced by different Aspergillus and Penicillium species. Since its mechanism of action is not fully understood yet, it is important to gain further insight into different interactions of OTA at the molecular level. OTA is found worldwide in many foods and drinks. Moreover, it can also be detected in human and animal tissues and body fluids, as well. Therefore, the development of highly sensitive quantitative methods for the determination of OTA is of utmost importance. OTA most likely forms complexes with divalent cations, both in cells and body fluids. In the present study, the OTA-zinc interaction was investigated and compared to OTA-magnesium complex formation using fluorescence spectroscopy and molecular modeling. Our results show that zinc(II) ion forms a two-fold higher stable complex with OTA than magnesium(II) ion. In addition, based on the enhanced fluorescence emission of OTA in its magnesium-bound form, a novel RP-HPLC-fluorescence detector (FLD) method was also established. Our results highlight that the application of magnesium chloride in alkaline eluents results in an approximately two-fold increase in sensitivity using the HPLC-FLD technique. [ABSTRACT FROM AUTHOR]

The neurotoxin β-N-methylamino-L-alanine (BMAA) is suspected to play a role in the neurological diseases amyotrophic lateral sclerosis, Alzheimer's disease, and Parkinson's disease. BMAA production by cyanobacteria has been reported and contact with cyanobacteria infested waters or consumption of aquatic organisms are possible pathways to human exposure. However, there is little consensus regarding whether BMAA is present in cyanobacteria or not, and if so, at what concentrations. The aim of this review is to indicate the current state of knowledge on the presence of BMAA in aquatic ecosystems. Some studies have convincingly shown that BMAA can be present in aquatic samples at the μg/g dry weight level, which is around the detection limit of some equally credible studies in which no BMAA was detected. However, for the majority of the reviewed articles, it was unclear whether BMAA was correctly identified, either because inadequate analytical methods were used, or because poor reporting of analyses made it impossible to verify the results. Poor analysis, reporting and prolific errors have shaken the foundations of BMAA research. First steps towards estimation of human BMAA exposure are to develop and use selective, inter-laboratory validated methods and to correctly report the analytical work. [ABSTRACT FROM AUTHOR]

In 1998, during a toxicological surveillance of various marine fouling organisms in Hiroshima Bay, Japan, specimens of the ribbon worm, Cephalothrix simula (Nemertea: Palaeonemertea) were found. These ribbon worms contained toxins with extremely strong paralytic activity. The maximum toxicity in terms of tetrodotoxin (TTX) was 25,590 mouse units (MU) per gram for the whole worm throughout the monitoring period. The main toxic component was isolated and recrystallized from an acidified methanolic solution. The crystalline with a specific toxicity of 3520 MU/mg was obtained and identified as TTX by high performance liquid chromatography (HPLC)-fluorescent detection (FLD) (HPLC-FLD), electrospray ionization-mass spectrometry (ESI-MS), infrared (IR), nuclear magnetic resonance (NMR) and gas chromatography-mass spectrometry (GC-MS). The highest toxicity of C. simula exceeded the human lethal dose per a single worm. A toxicological surveillance of C. simula from 1998 to 2005 indicated approximately 80% of the individuals were ranked as "strongly toxic" (⩾1000 MU/g). Forty-eight percent of the specimens possessed toxicity scores of more than 2000 MU/g. Seasonal variations were observed in the lethal potency of C. simula. Specimens collected on January 13, 2000 to December 26, 2000 showed mean toxicities of 665-5300 MU/g (n = 10). These data prompted a toxicological surveillance of ribbon worms from other localities with different habitats in Japan, including Akkeshi Bay (Hokkaido) under stones on rocky intertidal beaches, as well as Otsuchi (Iwate) among calcareous tubes of serpulid polychaetes on rocky shores. Within twelve species of ribbon worms examined, only C. simula possessed extremely high toxicity. Therefore, C. simula appears to show generally high toxicity irrespective of their locality and habitat. [ABSTRACT FROM AUTHOR]

Spices are a popular ingredient in cuisine worldwide but can pose a health risk as they are prone to fungal infestation and mycotoxin contamination. The purpose of this study was to evaluate ochratoxin A (OTA) in 54 single-kind traditional and less traditional spices, each of which was purchased in six samples of different batches (324 samples in total) at the Czech market during 2019–2020. The HPLC-FLD method with pre-treatment by immunoaffinity columns was employed to determine OTA. The limits of detection and quantification were 0.03 ng g−1 and 0.10 ng g−1, respectively. A total of 101 (31%) samples of 19 spice kinds were positive at concentrations ranging from 0.11–38.46 ng g−1. Only turmeric was contaminated with an OTA level exceeding the European Union limits. However, most spices have no regulation, thus further extensive monitoring of various mycotoxins in various kinds of spices is necessary. Chilli and black pepper are the most studied spices for OTA contamination, however, many other kinds of spice can also be highly contaminated, but studies on them are less common, rare, or have not yet been performed. The uniqueness of this study lies in the wide range of spice types studied for the presence of OTA on the Czech market. [ABSTRACT FROM AUTHOR]

Tea is a widely consumed drink in the world which is susceptible to undergoing adulterations to reduce manufacturing costs and rise financial benefits. The development of simple analytical methodologies to assess tea authenticity, as well as to detect and quantify frauds, is an important matter considering the rise of adulteration issues in recent years. In the present study, untargeted HPLC-UV and HPLC-FLD fingerprinting methods were employed to characterize, classify, and authenticate tea extracts belonging to different varieties (red, green, black, oolong, and white teas) by partial least squares-discriminant analysis (PLS-DA), as well as to detect and quantify adulteration frauds when chicory was used as the adulterant by partial least squares (PLS) regression, to ensure the authenticity and integrity of foodstuffs. Overall, PLS-DA showed a good classification and grouping of the tea samples according to the tea variety and, except for some white tea extracts, perfectly discriminated from the chicory ones. One hundred percent classification rates for the PLS-DA calibration models were achieved, except for green and oolong tea when HPLC-FLD fingerprints were employed, which showed classification rates of 96.43% and 95.45%, respectively. Good predictions were also accomplished, also showing, in almost all the cases, a 100% classification rate for prediction, with the exception of white tea and oolong tea when HPLC-UV fingerprints were employed that exhibited a classification rate of 77.78% and 88.89%, respectively. Good PLS results for chicory adulteration detection and quantitation were also accomplished, with calibration, cross-validation, and external validation errors beneath 1.4%, 6.4%, and 3.7%, respectively. Acceptable prediction errors (below 21.7%) were also observed, except for white tea extracts that showed higher errors which were attributed to the low sample variability available. [ABSTRACT FROM AUTHOR]

Analytical methods using the fluorescence properties of bisphenols (BPA, BPF and BPS) and their complexes with β-cyclodextrin and methyl-β-cyclodextrin were developed. The methods were applied for the analysis of thermal paper and canned food. Their performance was compared with a standard HPLC approach with a diode array and fluorescence detections. For comparison purposes, basic validation parameters (linear range, limit of detection, sensitivity, precision) were evaluated. It was concluded the developed methods facilitate fast and cost-effective determination of three bisphenol species in liquid samples, similar to the HPLC performance. They are also environmentally friendly. BPA, BPF and BPS can be routinely determined with the presented approach. [ABSTRACT FROM AUTHOR]

Cytostatics are toxic pharmaceuticals, whose presence in surfaces puts healthcare workers at risk. These drugs might also end up in hospital effluents (HWW), potentially damaging aquatic ecosystems. Bicalutamide is a cytostatic extensively consumed worldwide, but few analytical methods exist for its quantification and most of them require advanced techniques, such as liquid chromatography mass spectrometry (LC-MS), which are very complex and expensive for large monitoring studies. Therefore, a simple but reliable multi-matrix high performance liquid chromatographic method, with fluorescence detection, was developed and validated to rapidly screen abnormal concentrations of bicalutamide in HWW and relevant contamination levels of bicalutamide in indoor surfaces (>100 pg/cm2), prior to confirmation by LC-MS. The method presents good linearity and relatively low method detection limits (HWW: 0.14 ng/mL; surfaces: 0.28 pg/cm2). Global uncertainty was below 20% for concentrations higher than 25 ng/mL (HWW) and 50 pg/cm2 (surfaces); global uncertainty was little affected by the matrix. Therefore, a multi-matrix assessment could be achieved with this method, thus contributing to a holistic quantification of bicalutamide along the cytostatic circuit. Bicalutamide was not detected in any of the grab samples from a Portuguese hospital, but an enlarged sampling is required to conclude about its occurrence and exposure risks. [ABSTRACT FROM AUTHOR]

Hawthorn leaves are a rich source of phenolic compounds that possess beneficial activities for human health. Ultrasonic-assisted extraction (UAE) is an extraction technique frequently used for the isolation of phenolic compounds in plants. Thus, in this study, a Box–Behnken design was used to optimize UAE conditions such as the percentage of acetone, the extraction time and solvent-to-solid ratio (v/w) in order to obtain the maximum content of total compounds by Folin–Ciocalteu and the maximum in vitro antioxidant activity by DPPH, ABTS and FRAP assays in Crataegus monogyna leaves. The optimum conditions to obtain the highest total phenolic content and antioxidant activities were 50% acetone, 55 min and 1/1000 (w/v). A total of 30 phenolic compounds were identified and quantified in C. monogyna leaf extract obtained at these optimum UAE conditions. HPLC-MS allows the identification and quantification of 19 phenolic compounds and NP-HPLC-FLD analyses showed the presence of 11 proanthocyanidins. According to the results, the most concentrated phenolic compounds in C. monogyna leaf extract obtained at optimum UAE conditions were phenolic acid derivatives such as protocatechuic acid-glucoside, dihydroxy benzoic acid pentoside and chlorogenic acid, flavones such as 2″-O-rhamnosyl-C-hexosyl-apigenin, flavonols such as hyperoside and isoquercetin and proanthocyanidins such as monomer and dimer. As a result, the optimized UAE conditions could be used to obtain an extract of C. monogyna leaves enriched with phenolic compounds. [ABSTRACT FROM AUTHOR]

Coffee, one of the most popular drinks around the world, is also one of the beverages most susceptible of being adulterated. Untargeted high-performance liquid chromatography with ultraviolet and fluorescence detection (HPLC-UV-FLD) fingerprinting strategies in combination with chemometrics were employed for the authenticity assessment and fraud quantitation of adulterated coffees involving three different and common adulterants: chicory, barley, and flours. The methodologies were applied after a solid–liquid extraction procedure with a methanol:water 50:50 (v/v) solution as extracting solvent. Chromatographic fingerprints were obtained using a Kinetex® C18 reversed-phase column under gradient elution conditions using 0.1% formic acid aqueous solution and methanol as mobile phase components. The obtained coffee and adulterants extract HPLC-UV-FLD fingerprints were evaluated by partial least squares regression-discriminants analysis (PLS-DA) resulting to be excellent chemical descriptors for sample discrimination. One hundred percent classification rates for both PLS-DA calibration and prediction models were obtained. In addition, Arabica and Robusta coffee samples were adulterated with chicory, barley, and flours, and the obtained HPLC-UV-FLD fingerprints subjected to partial least squares (PLS) regression, demonstrating the feasibility of the proposed methodologies to assess coffee authenticity and to quantify adulteration levels (down to 15%), showing both calibration and prediction errors below 1.3% and 2.4%, respectively. [ABSTRACT FROM AUTHOR]

Bisphenol A (BPA) is one of the chemicals used to produce both polycarbonate plastics and epoxy resin coatings. Research has shown that small amounts of BPA can migrate into the foods and beverages enclosed in these types of containers. In this research, an analytical method based on high-performance liquid chromatography with fluorescence detection (HPLC-FLD) was developed and validated for the determination of BPA in canned vegetables. The results were confirmed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) was performed, to identify the coating material of each tin can. Nineteen cans of vegetables were taken as study samples (eleven samples from the Spanish market, and eight samples from the Portuguese market). Excellent linear correlation (r2 = 0.9999) was observed over the range of 0.01 to 0.25 mg/L. Limit of detection (LOD) and limit of quantification (LOQ) values were calculated to be 0.005 mg/kg and 0.01 mg/kg, respectively. Good recoveries, between 72% and 90% were obtained at three different levels of concentration (RSD% = 4.6). BPA was not detected in the samples. The proposed HPLC-FLD was found to be suitable for the determination of BPA in canned vegetables. [ABSTRACT FROM AUTHOR]

A magnetic solid phase extraction technique followed by liquid chromatography with a fluorescence detector for naproxen analysis in human urine samples was developed. The method includes the extraction of naproxen with a magnetic solid synthetized with magnetite and poly 4-vinylpriridine, followed by the magnetic separation of the solid phase and desorption of the analyte with methanol. Under optimal conditions, the linear range of the calibration curve was 0.05–0.60 μg L−1, with a limit of detection of 0.02 μg L−1. In all cases values of repeatability were lower than 5.0% with recoveries of 99.4 ± 1.3%. Precision and accuracy values are adequate for naproxen (Npx) analysis in urine samples. [ABSTRACT FROM AUTHOR]

The introduction of antimicrobial residues in the food chain has a significant impact on human health. An innovative solution to avoid their presence in meat is the adaptation of current control methods for use with in vivo matrixes. Thus, the aim was to obtain paired blood and muscle samples from pigs treated with some of the main antimicrobials currently used in veterinary medicine (oxytetracycline, sulfamethoxypyridazine, enrofloxacin, amoxicillin), and to compare their rate of depletion in both matrixes. Antimicrobial concentrations in paired samples of blood and muscle were determined by liquid chromatography with tandem mass spectrometry (LC–MS/MS) or high performance liquid chromatography with fluorescence detection (HPLC-FLD). A comparison between values obtained in muscle and blood showed a similar distribution in both matrixes for oxytetracycline; for sulfamethoxypyridazine, a similar decrease rate but a concentration three times higher in blood compared to muscle was found; for enrofloxacin, we found significant differences in the rate of depletion, with similar antimicrobial concentrations in both matrixes with values close to the maximum residue limit (MRL) and higher amounts in muscle for values that lay considerably over the MRL. Conversely, amoxicillin depletion was so rapid that its appearance in carcasses does not seem to pose a risk. Therefore, blood would be a feasible matrix for the development of new in vivo tests. [ABSTRACT FROM AUTHOR]

Fatty acids (FAs) are basic components in plants. The pharmacological significance of FAs has attracted attentions of nutritionists and pharmaceutists. Sensitive and accurate detection of FAs is of great importance. In the present study, a pre-column derivatization and online mass spectrometry-based qualitative and quantitative analysis of FAs was developed. Nineteen main FAs were derivatized by 2-(7-methyl-1H-pyrazolo-[3,4-b]quinoline-1-yl)ethyl-4-methyl benzenesulfonate (NMP) and separated on reversed-phase Hypersil BDS C8 column with gradient elution. All FAs showed excellent linear responses with correlation coefficients more than 0.9996. The method obtained LOQs between 0.93 ng/mL and 5.64 ng/mL. FA derivatives were identified by both retention time and protonated molecular ion corresponding to m/z [M + H]+. A comparative study based on FA contents in peel and pulp, seeds and leaves of Nitraria tangutourum Bobr (NTB) from different geographical origins was performed with the established method. Results indicated that NTB were rich in FAs, and the types and contents of FAs varied among tissues. On the other hand, the same tissue of NTB from different geographical areas differed in the content, but not in type, of FAs. [ABSTRACT FROM AUTHOR]

Aflatoxin M 1 (AFM 1 ) is a well-known mycotoxin that can be found in the milk of animals that have ingested feed contaminated with aflatoxin B 1 (AFB 1 ). In Italy, the development of donkey farms is mainly due to growing request of donkey milk, which is considered an incomparable substitute for human mother's milk for its chemical composition and organoleptic characteristics. The aim of this study was to assess the occurrence of AFM 1 in donkey milk produced in a farm in Northern Italy, also in view of the few data available about the presence of this mycotoxin in this type of milk. Therefore, 63 milk samples were collected and analyzed using a fast and sensitive HPLC and fluorescence detection (FLD) method previously optimized and validated. None of the milk samples collected were found to be contaminated at a level above the limit of quantification (LOQ) (0.0125 ng/mL), while only one sample showed traces of the mycotoxin at a concentration between the limit of detection (LOD) and LOQ (0.0044 ng/mL), well below the legal limit established for infant milk and follow-on milk (0.025 ng/mL). These results are in line with those of the few similar surveys carried out on donkey milk and seem to indicate a low risk of AFM 1 contamination for this food.

Background: Identification and quantitative determination of cytisine, especially in biological samples and pharmaceutical formulations, is still a difficult analytical task. Cytisine is an alkaloid with a small and very polar molecule. For this reason, it is very weakly retained on reversed phase (RP) stationary phases, such as commonly used alkyl-bonded phases. The very weak retention of cytisine causes it to be eluted together with the components of biological matrices. Objective: Comparison and evaluation of various chromatographic systems for analysis of cytisine in different matrices—serum, saliva and pharmaceutical formulation—by high performance liquid chromatography (HPLC) with diode array (DAD), fluorescence (FLD) and mass spectrometry (MS) detection. Methods: The analyses were performed using HPLC in reversed phase (RP), hydrophilic interaction liquid chromatography (HILIC) and ion exchange chromatography (IEC) modes. Different sample pre-treatment methods were tested: Protein precipitation (with acetone, methanol (MeOH) or acetonitrile (ACN), and solid phase extraction (SPE) using cartridges with octadecyl (C18), hydrophilic-lipophilic balanced copolymer (HLB) or strong cation exchange sorbents (Strata X-C). Conclusion: Significant differences were observed in retention parameters with a change of the used chromatographic system. The various properties of stationary phases resulted in differences in analyte retention, peaks' shape and systems' efficiency. The weakest retention was observed using RP systems; however, the use of the Polar RP phase can be an alternative for application in green chromatography. In the strongest retention was observed using a strong cation exchange (SCX) phase. The most optimal systems were chosen for the analysis of cytisine in the pharmaceutical preparation, serum and saliva after sample pre-treatment with the new SPE procedure. Due to the sensitivity, the use of HPLC-DAD or HPLC-FLD is the most optimal for drug analysis in pharmaceutical preparations, whereas HPLC-MS is suitable for analysis of cytisine in biological samples. [ABSTRACT FROM AUTHOR]

Immunoaffinity columns (IACs) are most popularly used for mycotoxin clean-up in complex matrices prior to chromatographic analysis. But, their high cost has limited their wide application and the regeneration of IACs for multiple instances of reuse is important. This study aimed to investigate the feasibility of regeneration and reuse of IACs for purification of ochratoxin A (OTA) in spiked raw malt and dried ginger samples followed by high performance liquid chromatography-fluorescence detection. After each use, the IACs were filled with phosphate buffer saline (PBS) as the preservation solution and stored at 8 °C overnight for regeneration and reuse until the recovery rate was <70%. The results showed that matrix type, preparation procedure, and pH value of sample extraction exhibited major effects on the reuse of IACs for OTA clean-up. While, after modifying the sample preparation procedure using water as the diluent and the solution at a pH of 7 to 8, the IACs could be used eight and three times for the spiked raw malt and dried ginger samples with OTA after regeneration. Regarding the traditional procedure recommended in Chinese Pharmacopoeia (2015 edition), the IACs could be used for three and two times for the spiked raw malt and dried ginger samples with OTA, respectively. Therefore, the corresponding experimental cost could be reduced to one-eighth and one-third of the original cost. This is the first study on the regeneration and reuse of IACs for OTA clean-up in complex Chinese herbal medicines, providing a green and economical tool for a large number of samples analysis with low cost. [ABSTRACT FROM AUTHOR]

The aim of this work was to validate the method of determination of polycyclic aromatic hydrocarbons (PAHs), i.e., benzo(a)pyrene and sum of benzo(a)pyrene, benz(a)anthracene, benzo(b)fluoranthene and chrysene in different types of tea, as well as to assess the transfer of these contaminants from tea to tea infusion. The research materials were popular types of black, green, red and white tea. Quantitative and qualitative determination of PAHs was performed by High Performance Liquid Chromatography with fluorimetric detection (HPLC-FLD). The samples were prepared by QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) technique followed by cleaning-up by dispersion solid-phase extraction (d-SPE). Values of limit of detection and limit of quantification obtained in the validation of the method were lower than the respective maximum values given in Commission Regulation (EU) No. 836/2011. The level of contamination of popular teas commercially available on the Polish market with PAHs is similar to that of teas available in other countries, with a very large variation in the concentration of each of the compounds. The highest benzo(a)pyrene and Σ4PAHs contents (209 ± 42 μg/kg and 756 ± 151 μg/kg, respectively) were found for black tea leaves. The transfer of Σ4PAHs from black tea to tea infusions was 0.48%, while it was 1.55-1.72% for red, white and green teas., Competing Interests: The authors declare no conflict of interest.