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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]

Ochratoxin A (OTA), which is one of the most important mycotoxins in terms of human health, can be found in cereal products such as bread, “bazlama” (traditional flatbread), and pita bread, as well as cereals such as wheat, barley, and corn. This study aimed to determine the effect of different fermentation times, baking, and storage for various periods on the presence of OTA in sourdough bazlama. Bazlama flour was contaminated with OTA concentrations of 5 and 10 μg/kg. After two different fermentation times (1.5 and 3 h), baking at 300 ± 5°C, and storage at room temperature (25 ± 2°C) for 0, 5, and 10 days, the change in OTA levels of bazlama samples was determined by the high‐performance liquid chromatography with fluorescence detector (HPLC‐FLD) method. The effect of different storage periods on the presence of OTA is insignificant. Although a general decrease in OTA level has been determined, it has been found that long‐term fermentation (at least 3 h) was more effective, especially in flours with a high concentration (10 μg/kg) of OTA contamination. It has been determined that bazlama made from contaminated flours with OTA levels of 5 and 10 μg/kg contained OTA levels exceeding 3 μg/kg when long‐term fermentation was not used. This is the maximum permitted limit set by the Turkish Food Codex and the European Commission, indicating that it is not suitable for consumption in this position. [ABSTRACT FROM AUTHOR]

Aflatoxins (AFs) are secondary metabolites mainly produced by Aspergillus flavus and A. parasiticus and their contamination of red peppers can cause hepatocellular carcinoma, growth retardation in children, immune suppression, and death. In addition, their presence in the red peppers can affect international trade and cause significant economic burdens. Thus, the objective of this study was to assess the level of AFs contamination in packed powder (from supermarkets) and raw red pepper samples commercially available in the towns of Fiche and Mukaturi. Furthermore, this study aimed to determine the potential health and cancer risks associated with the consumption of red pepper contaminated with AFs. Red pepper samples (raw and packed powder) were collected randomly from the Fiche and Mukaturi open markets. Then AFs in the samples were extracted using methanol: water (80:20, v/v). These extract samples were then cleaned up using an immunoaffinity column (IAC) and determined with a high-performance liquid chromatography-fluorescence detector (HPLC-FLD). The finding showed that the amount of AFB1, AFB2, and AFG1 in raw red pepper was found to be 3.19 ± 0.01, 0.19 ± 0.001, and 4.07 ± 0.01 μg kg−1, respectively. The raw red pepper samples had a total of 7.66 ± 0.01 μg kg−1 of AFs. On the other hand, the amount of AFB1, AFB2, and AFG1 in Afiya-packed red pepper was found to be 7.04 ± 0.03, 2.15 ± 0.06, and 0.50 ± 0.01 μg kg−1, while Mudayi packed red pepper contained 31.60 ± 0.22, 24.40 ± 0.17, 3.37 ± 0.02 and 2.48 ± 0.004 μg kg−1 of aflatoxins, respectively. Afiya and Mudayi packed powder peppers had a total AFs content of 10.4 ± 0.07 and 61.90 ± 0.28 μg kg−1, respectively. The total AFs concentrations in packed pepper powder samples were higher than maximum toleratable limits (MTLs) set by the European Commission Regulation (EU) 2023/915 (5.00 μg kg−1 for AFB1 and 10 μg kg−1 for total AFs). AFB1 (31.60 ± 0.22 μg kg−1) had the highest level of contamination, followed by AFB2 (24.40 ± 0.17 μg kg−1) in packed pepper powder. In the adult population, the estimated daily intake (EDI) of AFB1, AFB2, AFG1, and AFG2 ranged from 0.80 to 7.90, 0.04 to 6.10, 0.02 to 1.02, and 0.05 to 0.62 μ g kg−1 body weight (bw) per day, respectively. However, the Margins of Exposure (MOE) values and combined Margin of Exposure (MoET) for these chemicals were significantly lower than the safe margin (<10 000). Therefore, this study highlights the potential health risks associated with consuming AFs-contaminated red peppers and the need for stricter regulations and monitoring to ensure food safety. [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]

Ergot alkaloids (EAs) are toxic substances naturally produced by Claviceps fungi. These fungi infest a wide range of cereals and grasses. When domestic animals are exposed to EAs through contaminated feeds, it is detrimental to them and leads to significant economic losses. For that reason, it is important to monitor feed for the presence of EAs, especially with methods enabling their determination in processed materials.

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.

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]

A zero-order emission system was applied to address the variation in emission wavelengths between analytes. Also, it assists to compensate for the drop in emission intensity resulted from the changes in analyte excitation wavelength. So, a high performance liquid chromatography-fluorescence detector (HPLC-FLD) with zero order emission mode was used for the simultaneous estimation of rhodamine B (RhB) and eosin Y (EsY) based on their native fluorescence. The presented method is considered the first fluorescence method for simultaneous determination of RhB and EsY. The efficiency of the developed method was tested by determining the two dyes in a high matrix sample, chili sauce. Under the optimum condition, a linear calibration graphs for RhB and EsY in range of 0.5 -300 ng/mL with r = 0.9994 and 0.9991, respectively, were acquired. The limits of detection for RhB and EsY were 0.344 ng/mL (0.355 ng/g) and 0.215 ng/mL (0.222 ng/g), respectively. The recovery percentage and relative standard deviation of two analytes were obtained in the range of 98.38-101.86% and 0.83-1.66%, respectively. RhB was detected only in one chili sauce sample at a concentration of 0.388 ng/mL while EsY was not found in any sample. [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.

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.

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]

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.

Measurement uncertainty is a metrological concept that characterizes distributions/variations that can make the measurement results logical and is linked to the result of the measurement. The measurement uncertainty of the method was estimated using two basic approaches (top-down and bottom-up approach). The top-down approach covers in-house validation data (trueness, repeatability and intra-lab reproducibility), while the bottom-up approach involves individual contributions to all uncertainty at each stage of the analysis/process. We estimated measurement uncertainty of BPA analysis by HPLC-FLD test according to JCGM GUM and EURACHEM-CITAC guidelines. The relative expanded uncertainties at the BPA concentration by the bottom-up approach and top-down approaches were ±3.2% and ±4.8% respectively (95% confidence interval, k=2). Thus, although it is seen that the results of the two approaches are different in chromatographic BPA analysis, it is concluded that the measurement uncertainty related to BPA analysis, especially in food analysis laboratories, can be determined by a simpler topdown approach. [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]

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.

Nuts are an important food group that contributes to maintaining health; however, they can be a source of exposure to aflatoxins. This study was conducted from 2017 to 2021 to assess the incidence of aflatoxins in nuts and nut products imported to the UAE from 57 countries. Associations between container type and processing technique and aflatoxin levels were also analyzed. A total of 5401 samples of pistachios, peanuts, peanut butter, and mixed nuts were examined using HPLC-FLD analysis in conjunction with immunoaffinity cleanup. In nuts, non-conformity was detected in samples imported from 32 different countries. Mean aflatoxin values for the non-compliant samples ranged from 81.0 to 92.7 μg/kg in pistachios, peanuts, and mixed nuts. A significant difference (p

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]

Hydrogen peroxide (H2O2) is a food additive for bleaching and sterilization, owing to its strong oxidizing effect. The current study aimed to develop analytical methods to detect trace amounts of residual H2O2 in diverse foods using high-performance liquid chromatography (HPLC) equipped with diode array detector (DAD) or fluorescence detector (FLD). The vanadium(V)-peroxo complex, derived from the reaction of H2O2 with ammonium metavanadate, was used for the detection of H2O2 with DAD. H2O2 was indirectly analyzed using FLD via the detection of 7-hydroxycoumarin, derived by Fenton reaction, followed by verification using liquid chromatography with tandem mass spectrometry analysis. Both the detection methods showed good linearity with R2 > 0.997. Limit of detection and limit of quantification were 0.30 and 0.91 mg/L (8.82 and 26.76 μM) with HPLC–DAD and 0.001 and 0.003 mg/L (0.03 and 0.09 μM) with HPLC–FLD, respectively. Applicability of both the methods was successively tested through sample analysis. [ABSTRACT FROM AUTHOR]

A high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD) was evaluated for the monitoring of aflatoxin B1 (AFB1) production by Aspergillus section Flavi isolates, which were found in the commodities of Slovak origin (barley and tomatoes). Rapid validation of the method was performed and the method was applied for the determination of the mycotoxin. Based on the calculated validation characteristics, the method showed high sensitivity and reproducibility. The LOD and LOQ determined were 0.03 and 0.10 μg.mL-1 which is fully acceptable for the determination of the AFB1 in isolates from plant commodities. The calibration graph was linear in the concentration range of 0.01-10.0 μg.mL-1. The optimized method was used for the analysis of the mycotoxin in three Aspergillus section Flavi isolates. The submerse cultivation was performed at 25±1°C in yeast extract sucrose liquid medium and material was collected on the 7th, 14th and 21st day of cultivation. Simple sample extraction was carried out with use of ethyl acetate, and measurement was done with use of the mobile phase consisting of water and methanol by HPLC-FLD. Production of the AFB1 by isolates was ranging from

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]

Pufferfish is one of the most poisonous marine organisms, responsible for numerous poisoning incidents and some human fatalities due to its capability to accumulate potent neurotoxins such as tetrodotoxins (TTXs) and paralytic shellfish toxins (PSTs). In this study, tissue extracts (muscle, skin, liver, intestinal tract and gonads) obtained from sixteen pufferfish specimens of the Lagocephalus lagocephalus and Sphoeroides pachygaster species, collected along the Spanish Mediterranean coast, were analysed for the presence of voltage-gated sodium channel (also known as Na v channel) blockers using cell-based assay (CBA) and automated patch clamp (APC). No toxicity was observed in any of the S. pachygaster specimens, but toxicity was detected in the liver of most L. lagocephalus specimens. Instrumental analysis of these specimens, as well as in one Lagocephalus sceleratus specimen, by high-performance liquid chromatography coupled to fluorescence detection (HPLC-FLD) was performed, which confirmed the presence of PSTs only in L. lagocephalus specimens. This analysis reported the presence of saxitoxin (STX) and decarbamoylsaxitoxin (dcSTX) in all positive samples, being dcSTX the major analogue. These results demonstrate the ability of this species to accumulate PSTs, being the first report of the presence of PSTs in Mediterranean L. lagocephalus specimens. Furthermore, the presence of high PSTs contents in all five tested tissues of one L. lagocephalus specimen pointed the risk that the presence of this toxic fish in the Mediterranean Sea may represent for seafood safety and human health in case of accidental consumption. [Display omitted] • CBA and APC were used to assess the presence of PSTs in different pufferfish species. • No toxicity was observed in S. pachygaster. • Toxicity was detected in the liver of most L. lagocephalus. • HPLC-FLD analysis confirmed the presence of PSTs in L. lagocephalus. • STX and dcSTX were detected in all positive samples, being dcSTX the major analogue. [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.

Milk and dairy products are the most important nutritional foods among all age groups. Aflatoxin M1 (AFM1) contaminates milk and makes its consumption potentially dangerous. Infants are mostly at risk because they are typically fed as many as six and more times per day, which is indeed a disquieting concern. This study aimed at evaluating AFM1 levels especially above international (European Food Safety Authority, EFSA) (0.05 µg/kg) and local (Ghana Standards Authority, GSA) (0.5 µg/kg) standards and cancer risks associated with the ingestion of raw cow milk (n = 120) sampled from Southern Ghana (Greater Accra, Volta, Western and Eastern Regions). AFM1 were measured with High-Performance Liquid Chromatography with a Fluorescence Detector (HPLC-FLD). Risk assessments were also conducted using models prescribed by the Joint FAO/WHO Expert Committee on Additives (JECFA). Out of the 120 samples analyzed for AFM1, 67 (55.8%) tested positive, 63 (52.5%) exceeded the limits of EFSA and were between the range 0.06 ± 0.001–3.52 ± 0.5 µg/kg whereas 50(41.7%) within the range of 0.50 ± 0.03–3.52.01 ± 0.5 µg/kg exceeded GSA limits. Risk assessments of AFM1 for infants, toddlers, children, adolescents, and adults ranged between 0.06 and 2.03 ng/kg bw/day, 197.04–6666.67, 0–0.0323 ng aflatoxins/kg bw/day and 1.94 × 10-3- 0.07 cases/100,000 person/yr respectively for Estimated Daily Intake (EDI), Margin of Exposure (MOE), Average Potency, and Cancer Risks. It was concluded that the consumption of raw milk posed adverse health effects on all age categories studied for the regions investigated. The use of raw cow milk may cause some problems and endanger the health of people of different age groups due to noncompliance with prescribed regulatory limits.

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]

The contamination of food commodities with mycotoxins could be a serious health threat to humans and animals. Therefore, identification, quantification and reduction of mycotoxins in food commodities, particularly of aflatoxins (AFs) and ochratoxin A (OTA) in grain foods, is essentially required to guarantee safe food. This study determined the levels of AFs and OTA in 135 maize grains samples belonging to eight salient maize varieties cultivated in Pakistan, and evaluated the usefulness of radiations and adsorbents to reduce their levels. High performance liquid chromatography (HPLC)-based method was validated for the determination of AFs and OTA in maize grains. The results showed that 69 and 61% samples were positive for AFs and OTA, respectively and 54 and 22% of the respective samples had AFs and OTA above the permissible limits set by Pakistan Standards and Quality Control Authority. The concentration of AFs, AFB1and OTA in grains ranged from 14.5 to 92.4, 1.02 to 2.46 and 1.41 to 53.9 μg kg−1, respectively. Among the varieties, Pearl had the highest level of total AFs and OTA, whereas YH-5427 had the highest AFB1 level. The lowest concentration of AFs and OTA was found in Malaka and 30Y87, respectively. The use of 15 kGy gamma irradiation for 24 h, sunlight-drying for 20 h and UV irradiation for 12 h almost completely degraded the mycotoxins. The microwave heating for 120 s resulted in 9–33% degradation of mycotoxins. Moreover, the treatment of grains' extract with activated charcoal (5% w/w) removed > 96% of total AFs and AFB1, and up to 43% of OTA. The use of bentonite at the same rate removed OTA, total AFs and AFB1 by 93, 73 and 92%, respectively. Thus, it is concluded that contamination of maize grains with mycotoxins was fairly high in the collected maize grain samples in Pakistan, and treatment with radiations and adsorbents can effectively reduce mycotoxins contamination level in maize grains. [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]

Histamine is a security issue in food safety as an indicator of the freshness of consumed fish product; that is why we aimed in this study to determine the histamine content in 7 fish species (Scomber japonicus; Sarda sarda; Sardinella aurita; S. maderensis; Ethmalosa fimbriata; Pomatomus saltatrix and Trachurus trachurus). Hundred and eight frozen and fresh pelagic of scombroid and non-scombroid species were collected from the Nouakchott fish market (Mauritanian Atlantic coast) in different period of 2020 and 2021. Histamine was quantified by the technique of high-performance liquid chromatography with a fluorescence detector (HPLC-FLD). Histamine was detected in all analyzed fresh and frozen species and varied from 4.21 to 201.60 ppm. All evaluated fresh fish species (S. aurita; S. maderensis; E. fimbriata; P. saltatrix and T. trachurus) were conform to the FAO/WHO and the European commission regulations. Regarding investigated frozen fish species (S. aurita; S. japonicus and S. sarda); the scombroid species (S. japonicus and S. sarda) were all under the set limits; however, even its more likely for scombroid fish to develop histamine, our results showed that histamine exceed the limit in one sample of non-scombroid fish (S. aurita). No significant variation in histamine levels between scombroid and non-scombroid fish species was obtained; thereby, the study showed that fish product commercialized at the Nouakchott fish market have a good quality and safe for human consumption. [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]

HPLC methods are among the most studied and validated techniques for the quantification of histamine in fish and fishery products; nonetheless, most of these methods require laborious sample extraction and purification. For this purpose, in the present study, an HPLC-FLD method without any sample clean-up procedure was developed, validated and applied for histamine quantification in various fish samples. Histamine was extracted from fish and fishery products samples with perchloric acid; then a simple precolumn derivatization with o-phthalaldehyde was performed in the presence of the reducing agent 2-mercaptoethanol. Once the derivatization was carried out, the separation was performed with a reverse phase HPLC system. In terms of validation, good linearity and specificity over a concentration range from 12.5 to 200 mg/kg were achieved. Also, convenable precision with relative standard deviations smaller than 5.92%, accuracy ranged from 0.40 to 1.84%, and satisfactory recoveries ranged from 100.44 to 101.84%, were obtained. The detection and the quantification limits were calculated at 2.8 and 4.3 mg/kg, respectively. In addition, the effectiveness of the method was evaluated by the analysis of reference materials, proficiency tests and various type of samples. Moreover, the developed method was found to be simple, rapid, and reliable. [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]

A different, reliable, and cost-effective strategy for the analysis of aflatoxins, ochratoxin A, and zearalenone in corn-based foods was proposed, including one multi-toxin immunoaffinity column (IAC) sample preparation and three different high-performance liquid chromatography fluorescence detection methods. The analytical procedures were tested and verified, keeping in mind their occurrence at trace levels in corn-based foods. With the validation of the proposed multi-toxin IAC methodology and comparison of the performance characteristics with methods using a single-toxin IAC, we confirmed the reliability of the multi-toxin IAC procedure versus the single-toxin IAC. The methods were validated by revealing satisfactory performance characteristics; for example, the obtained values of limit of detection were significantly lower than the maximum limits for all mycotoxins of concern. In addition, the recovery values were between 70.9 % and 106.1 % for all mycotoxins of interest, with precision values lower than 10.5 %.

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.

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]

The aim of this study was to assess the risk of exposure to polycyclic aromatic hydrocarbons (PAH) from yerba mate infusions in Uruguay using the margin of exposure approach (MOE) and a probabilistic method (Monte Carlo simulation). Servings/day, portion size, weekly frequency of mate consumption and body weight were the factors considered. The amount in infusions of benz[a]pyrene (B[a]P), PAH2 (sum of chrysene and B[a]P), and PAH4 (sum of benz[a]anthracene, chrysene, benz[b]fluoranthene and B[a]P) were used as markers of PAH exposure. Total content of PAH in infusions had large inter-brand variability (48–54 %) with significant differences among brands. PAH content in infusions prepared as habitually consumed was about 40 % of total content. The probability of occurrence of MOE

A different, reliable, and cost-effective strategy for the analysis of aflatoxins, ochratoxin A, and zearalenone in corn-based foods was proposed, including one multi-toxin immunoaffinity column (IAC) sample preparation and three different high-performance liquid chromatography fluorescence detection methods. The analytical procedures were tested and verified, keeping in mind their occurrence at trace levels in corn-based foods. With the validation of the proposed multi-toxin IAC methodology and comparison of the performance characteristics with methods using a single-toxin IAC, we confirmed the reliability of the multi-toxin IAC procedure versus the single-toxin IAC. The methods were validated by revealing satisfactory performance characteristics; for example, the obtained values of limit of detection were significantly lower than the maximum limits for all mycotoxins of concern. In addition, the recovery values were between 70.9 % and 106.1 % for all mycotoxins of interest, with precision values lower than 10.5 %. The proposed methodology is an excellent candidate for future standardization due to its trustfulness and efficiency for mycotoxin analysis. [ABSTRACT FROM AUTHOR]

Abstract Background Aspergillus and their linked metabolites such as aflatoxins (AFs) are one of the extremely significant contaminants affecting food production around the world. The contamination of AFs has been identified in various food commodities, which have been recognized as carcinogenic, mutagenic, teratogenic and immunosuppressive. The present study was undertaken to assess the AFs contamination in betel nuts (Areca catechu L.) being imported to Pakistan from South Asian countries during 2018–2019. Methods A total of 143 betel nuts consignments (India = 80, Indonesia = 39 and Sri Lanka = 24) were obtained and analyzed for the AF contamination using immunoaffinity column (IAC) clean-up procedure subsequent by liquid chromatography with fluorescence detection. Results: In Indian-origin betel nuts, about 96.3% samples were contaminated with AFs, ranging from 1.18‒331.57 µg/kg with mean contamination of 76.11 ± 1.12 µg/kg; whereas, in Indonesian and Sri Lankan shipments, 100% samples of betel nuts were found infected with AFs, ranging between 1.88‒378.94 and 4.74‒106.58 µg/kg with an average level of 123.76 ± 1.25 and 47.95 ± 0.98 µg/kg, respectively. Conclusions In conclusion, the AFs levels present an acute toxicity to human health and also hazard factors for the economy since contaminated foodstuffs do not fulfill the requirements of export/import. Therefore, instant actions should be engaged and re-evaluate agricultural procedures and regular monitoring of AFs level in food stuffs to minimize the chances of various diseases such as oral pre-cancerous oral wounds, submucous fibrosis and squamous cell carcinoma (cancer).

In sports, curcumin, a substance derived from the rhizome of Curcuma longa (turmeric) plant with antioxidant effect 8 times greater than vitamin E, has attracted the attention of scientists because of its potent antioxidant action, since in athletes subjected to intense exercise the—endogenous mechanisms of neutralization of reactive species are saturated. However, the pharmacokinetic characteristics of curcumin do not favor its medicinal use due to its low absorption, accelerated metabolism and rapid systemic elimination. Thus, the determination of plasma levels in supplemented patients is a crucial step in their pharmacodynamic evaluation. Therefore, the objective of this work was to develop and validate an analytical method by HPLC-FLD for curcumin evaluation in plasma of supplemented athletes. Luna column (C18; 150 × 4 mm; 3 µm), acetonitrile: acetic acid pH 3.2 (45:55 to 60:40) as mobile phase, flow rate of 1 mL min−1, excitation at 429/285 nm and emission at 529 nm and injection of 10 µL were the chromatographic conditions used. Plasma samples were extracted using ethylacetate and methanol (95: 5, 500 µL) and estradiol (30 µg mL−1) as internal standard, with subsequent stirring (3 min) and centrifugation (8 min) (triple extraction). The organic fraction was evaporated under N2 (20 min) and the dried residue reconstituted in acetonitrile. The method was linear between 44 and 261 ng mL−1, showing intra-day (2.05.6%) and inter-day (4.0–5.1%) precision with accuracy and selectiveness (curcumin tR = 8.7 min and internal standard tR = 13.9 min with relative recovery of 83.2%). So, it can be successfully used for curcumin evaluation in plasma samples from supplemented athletes, as well as being an alternative and advantageous method to UV–Vis and MS/MS in bioavailability studies.

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”.

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.

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.

Sialic acids are postulated to improve cognitive abilities. This study aimed to evaluate the effects of sialic acid on behavior when administered in a free form as N-acetylneuraminic acid (Neu5Ac) to pregnant mothers or rat pups. The experiment involved 40 male 21-day-old rat pups and 20 15-day-pregnant rats that were randomized into four Neu5Ac treated groups: 0 (control), or 10, 20, and 40 mg/kg. Morris water maze test and shuttle box test were performed on the rat pups and maternal Neu5Ac-supplemented offspring on day 100 to evaluate their cognitive performance. The Neu5Ac levels in the cerebral cortex and hippocampus were tested with high-performance liquid chromatography-fluorescence detection (HPLC-FLD). We found that the maternal Neu5Ac-supplemented offspring showed better cognitive performance, less escape latency in the Morris water maze test, and less electric shock time shuttle box test, compared with the untreated control. In the meantime, the Neu5Ac level in the cerebral cortex and hippocampus of the offspring was higher in the Neu5Ac treatment group than that in the untreated control group. However, no significant differences were observed between rat pups in the treated and the untreated control groups in terms of cognitive performance and Neu5Ac content in the cerebral cortex and hippocampus. Maternal Neu5Ac supplementation during pregnancy could effectively promote the brain Neu5Ac content of the offspring and enhance their cognitive performance, but Neu5Ac had no such effect on rat pups while directly supplemented with Neu5Ac.

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.

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.